Source: UNIVERSITY OF FLORIDA submitted to NRP
SCALING UP THE MICROBIAL-TRIGGERED FUNCTIONAL RESPONSES TO MULTIPLE PRACTICES IN THE FORAGE SYSTEM: FROM MOLECULES TO ECOSYSTEM SERVICES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1018472
Grant No.
2019-67013-29107
Cumulative Award Amt.
$483,000.00
Proposal No.
2018-07346
Multistate No.
(N/A)
Project Start Date
Apr 1, 2019
Project End Date
Mar 31, 2024
Grant Year
2019
Program Code
[A1102]- Foundational Knowledge of Agricultural Production Systems
Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611
Performing Department
NFREC
Non Technical Summary
Dinitrogen fixing microbes are critical members of the plant microbiome regulating key aspects of plant growth, nutrient cycling and soil health. In managed pastures, the rhizosphere processes and mechanisms involved with mixed plantings are not well understood. This study will integrate cutting-edge molecular and biochemical techniques to assess soil community and functional dynamics between two forage species (bahiagrass: Paspalum notatum Flüggé; rhizoma peanut: Arachis glabrata Benth) and their nitrogen (N)-fixing microbes, as affected by plant combination, grazing, time, and environmental variables; and to determine these consequences on N uptake efficiency of pasture plants and soil N/Carbon (C) retention. Bahiagrass is distributed over 6 million acres of improved bahiagrass pasture across southeastern United States. In the southeastern states, these pastures face decreasing soil C/N. We hypothesize that specific N-microbes selected through screening grass-legume genotype combinations and integrated practices (i.e., grazing) can be used to improve pasture productivity via greater soil N utilization and C sequestration. Improving plant-soil-microbe functions will result in more resilient and environmentally sound production systems. Our aim is to provide the first ever assessment of the functional dynamics for these microbes through selection and testing of the most compatible forage genotypes, based upon microbial functions related to grass and legume N requirements, soil C sequestration and multi-practice adaptation. Our long-term goal is to improve production efficiency and persistence by harnessing plants and their microbiomes to increase pasture biodiversity, N-fixation and soil C sequestration, while reducing abiotic N inputs.
Animal Health Component
10%
Research Effort Categories
Basic
90%
Applied
10%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1024099106015%
1024099104020%
1024099107015%
1021699106010%
1021699104010%
1021699107010%
1020199200020%
Knowledge Area
102 - Soil, Plant, Water, Nutrient Relationships;

Subject Of Investigation
4099 - Microorganisms, general/other; 1699 - Pasture and forage crops, general/other; 0199 - Soil and land, general;

Field Of Science
1040 - Molecular biology; 1060 - Biology (whole systems); 1070 - Ecology; 2000 - Chemistry;
Goals / Objectives
Dinitrogen fixing microbes are critical members of the plant microbiome regulating key aspects of plant growth, nutrient cycling and soil health. In managed pastures, the rhizosphere processes and mechanisms involved with mixed plantings are not well understood. This study will integrate cutting-edge molecular and biochemical techniques to assess soil community and functional dynamics between two forage species (bahiagrass: Paspalum notatum Flüggé; rhizoma peanut: Arachis glabrata Benth) and their nitrogen (N)-fixing microbes, as affected by plant combination, grazing, time, and environmental variables; and to determine these consequences on N uptake efficiency of pasture plants and soil N/Carbon (C) retention. Here, using legumes-grass systems, we hypothesize that (a) specialized pairs of legumes-grass shift soil BNF/N activity towards legume microbial specialists that are relatively dominant and in doing so, are contributing benefit to neighboring plants; (b) the changes of rhizosphere communities due to neighboring plant effects can alter biogeochemically-related ecosystem function and services; (c) the consequences of (a) and (b) may be enhanced or compromised by grazing, time and environmental variables (i.e., location, soil type).Objective 1. Evaluate the contribution of heterospecific neighboring plants (bahiagrass-RP genotype combinations) on reshaping microbial community structures, BNF and C/N cycling. [To address hypotheses (a) and (b)]Objective 2. Evaluate the impact of environmental variables, including soil type, regional-locations, time and management (grazing v.s. non-grazing), on microbial community structures, plant N2-fixation efficiency, soil C/N storage and GHG emission [To address hypothesis (c)]
Project Methods
Field sampling and processing Plants (above and below-ground) will be sampled along with soil fertility (the composite soils from 0 to 15 cm depth). Forage will be removed from 0.25 m2 area, followed by inserting a 12-cm diameter golf hole cutting tool set to 15 cm depth to remove a whole plant sample or plug. Several lateral roots (at least 4 cm length) taken near the plant crown will be quickly excised from the plug and placed in a sterile, re-sealable plastic bag and stored on ice in a cooler. Upon return to the lab, the root samples will be processed for Raman imaging. The plug will be placed in a 1 kg, plastic pot and returned to the lab. Upon return, the plant will be separated into shoot stubble, roots, rhizomes, and nodules. The rhizosphere soil will be obtained after manually separating the root system with adhering soil by shaking and picking using forceps. The fresh root samples and rhizomes will be surface sterilized using 5% breach for 5 min, rinsed in sterile water for 1 min, wiped by clean paper towel. For legume, the nodules will be separated from roots and collected. Both root and soil will be frozen immediately in liquid nitrogen and conserved at -80C for DNA/RNA extraction.Soil chemical analysis The split set of the samples will be dried at 60 C. Dried shoot and rhizome tissue will be ground to pass through a 2-mm sieve and tissues analyzed for N, C, and plant essential nutrients, following digestion and using inductively-coupled plasma (ICP) spectroscopy. Nodules will be counted and sized. Soils will be analyzed for soil fertility: total C, N, pH, total extractable ammonium, nitrate content, % soil moisture, TKN, organic matter (Walkley Black method) and Mehlich-3 extractable P, K, S, Ca, Mg, Fe, Cu, Zn, and Mn (using AA100). The C/N ratio will be generated using dry combustion on a Vario MAX CNS Elemental Analyzer. Total extractable ammonium and nitrate concentrations will be analyzed in 2M potassium chloride extracts of each soil sample using a WestCo SmartChem 200 discrete analyzer. Potentially mineralizable N will be estimated using N mineralizable test (7-day incubation). These data will help determine how significantly environmental factors (i.e., field conditions, soil type and grazing management) may impact biological function. In addition, microplots (approximately 1.1 m2) will receive stable isotope, 15N as (NH4)2SO4 and applied as a soil drench, in February, to dormant plants.Nitrogen isotope analysis Preliminary data demonstrate that we have been able to capture the efficiency of N use using isotope tracing underlying field settings. Here we will incorporate isotope N ratio to monitor N flux and identify the fingerprint of N-fixation. The δ15N dilution technique will be applied to the soils for the contrasting genotypes of grasses and legumes. Associative N-fixation will be distinguished from other N sources by determining the δ15N for non-fixing reference plants. After establishment, plants will be sampled to determine the weight of aboveground tissue, δ15N, total N, leaf/stem ratio, specific leaf area, leaf nitrogen content, and a carbohydrate profile. Root responses will be sampled include total root length, average root radius, and root biomass, using root scanning and the WinRhizo root analysis system (Regent software, Canada); traits will be correlated with N fixation rates. Total N and δ15N will be determined using an element analyzer coupled to an isotope ratio mass spectrometer (IRMS); and the carbohydrate profile will be determined using an HPLC. Linear mixed models will be used to determine the genetic control of N fixation, predict the breeding values, and estimate the genetic correlation of N fixation with morphological and physiological traits also measured in the tests.Amplicon sequencing Total DNA will be extracted from an individual sample using CTAB approach (rhizomes, roots, nodules) and MoBio Power soil kit (soil samples). Multiplex PCR will be applied for amplicon libraries. To identify the microbial community, DNA will be extracted and quantified from four different compartments: rhizosphere soil, rhizomes, fine roots, and nodules (legume only). The 16S rDNA will be targeted to access the prokaryotic community. The NIF genes will be also applied for rhizobium in a small subset of samples to examine their capability for microbial identity. To access fungal communities, two DNA regions of ITS will be targeted using ITS1F/ITS4 primers. Fungal SSU will also be applied to target AMF community.Metatranscriptomics Soil samples will be flash frozen in liquid nitrogen at time of harvest and RNA extracted within the following week. DNA and RNA of individual soil/root samples will be co-extracted as described in Liao et al 2014. The extracted DNA/RNA will be preserved in -80C for cDNA library preparation. Two library strategies will be performed: (1) Poly-A enrichment to recover the reads that largely belong to fungi. (2) Ribo-Zero strategy from the same sample to recover the reads that mostly belongs to prokaryotes. Because rRNA and transcribed ITS regions are detected when using a poly-A enrichment strategy for cDNA libraries, we will also be able to identify the metabolically active bacterial/fungal taxa from the same RNAseq sample. One full flow run of NovaSeq 6000 S4 100-PE (3,000 Gb of data size ) will be applied for 96 bacteria and fungal cDNA respectively. About 15Gb (150M paired reads) per sample will be obtained through this approach.Soil enzyme activity analysis We will perform soil enzyme and chemical analysis for rhizosphere soils and representative bulk soil. The subset of soil samples collected from the field will be preserved at -80C for the analysis. Extracellular enzymes responsible for fast nutrient cycling (carbohydrases, phosphatases, chitinases, proteases), slow release carbon (phenol oxidases, peroxidases). The potential activities of extracellular enzymes involved in soil carbon and nutrient cycling will be analyzed using fluorometric and colorimetric procedures on a microplate reader. Those include peroxidase (the oxidases that degrade lignin); cellobiohydrolase (exocellulase), β-glucosidase (which hydrolyzes cellulose into glucose), polyphenol oxidase (which oxidizes phenols), acid phosphatase (which releases inorganic phosphate from organic matter), N-acetyl-glucosaminidase (which breaks down chitin), and leucine-aminopeptidase (which breaks down polypeptides).GHG emission analysis Static chambers will be placed on top of target plants to sample the GHG emission (N2O and CO2) of the contrasting treatments. Static chambers will be made by PVC pipe with the bottom portion (colored), inserted 7.5 cm below surface, and 5 cm above ground. Chambers will be sealed at sampling time, and deployment period will be 30 min. Gaseous samples from the headspace of the chamber will be taken at 0, 10, 20, and 30 min with a 50-mL glass syringe and transferred into 0.5-L aluminum foil gas collecting bags to store. Samples of CO2 will be taken at 0, 2.5 and 5 min. Ambient air samples (blanks) from the environment will be collected as background. At collection time, ambient temperature will be monitored and recorded. Gas samples will be analyzed using a trace gas analyzer (Elementar, Germany) using electron capture detector for N2O and TCD detector for CO2. The trace gas detector will be interfaced with an isotope ratio mass spectrometer (IRMS) to determine isotopic composition of gas samples.

Progress 04/01/23 to 03/31/24

Outputs
Target Audience:A. Train the graduate students, biologists and visiting scientists to conduct the research proposed in this study, including: Target trainees: Adesuwa Erhunmwunse (PhD student, supervisor: Liao) Luana Dantas Queiroz (PhD student, supervisor: Dubeux) Benjamin Reimer (PhD student, supervisor: Liao) Kaile Zhang (postdoc, supervisor: Liao) Pamella Sena (Visiting international graduate student: supervisor: Liao) Mario Lira Junior (Visiting professor: supervisor: Liao) Vijay Verma (Biologist, supervisor: Liao) Martin Ruiz-Moreno (Biologist, supervisor: Dubeux) George Anguelov (Biologist, supervisor: Mackowiak) José D. Pereira Neto (PhD student, supervisor: Dubeux) Daniele Loures (visiting faculty, supervisor: Dubeux) Javier Portuguez (MS student; supervisor Dubeux) Kevin R. Trumpp (MS student, supervisor Dubeux) Beatriz Bizutti Cremostin (PhD student, supervisor: Dubeux) B. The project also involves in dismission of research outcomes of this project through Extension/outreach training programs Jan 7-11, 2024; 2023: Present at AFGC Conference, AL (in 2024); Winston-Salem, NC (in 2023) Scope of AFGC: Provide options under unpredictable growing conditions, including fertilizer management schemes for grass pastures and hay fields. We presented on various topics, including grass endophyte and the management of soil nutrients for forage (focusing on warm-season grasses); Sustainable livestock systems Speakers: Liao, Invited speaker Dubeux Target audiences: Extension agents, industry personnel and producers Nov 30 2023: Present at a quarter meeting of the Florida Cattlemen's Association, Marianna FL (7-hour event). We shared the research findings focused on nutrient cycling and soil health in Florida's pastures. We also discussed the necessary steps for further investigation to address issues identified in forage production Presenters: Liao, Mackowiak, Dubeux, Reimer, Queiroz, Verma Target audience: Forage and cattle producers, stakeholders, Extension agents, government staff Nov 19 2023: Led Microbiology Class in Cub Scouts, Gadsden county FL We organized a Microbiology class and hands-on activity lasting six hours for Cub Scouts. The primary goal was to impart knowledge to the younger generation, focusing on beneficial soil microbes. We covered fundamental concepts such as the roles of decomposers and mycorrhizae play in benefiting our environment. Lead: Liao, Reimer, Zhang, Wang Target audience: 39 youth (33 males, 6 females) Nov 28-30 2023 International Symposium on Forage Legumes Scope: addressing recent advances on the use of forage legumes in grazing systems Invited speaker: Dubeux Target audience: State Faculty, industry personnel, extension agents Nov 6-9 2023 IV International Symposium on Integrated Crop-Livestock Systems Scope: Conference addressing ICL systems. Invited speaker: Dubeux Target audience: State Faculty, industry personnel, Extension agents October 29 to Nov 1 2023: ASA-CSSA-SSSA Annual Meeting Scope: Annual scientific meeting of the Tri-Societies Presenters: Queiroz, Dubeux, Trumpp, Portuguez. Target audience: State Faculty, industry personnel, Extension agents Sep 28, 2023: Pollinators, Forages, and Habitat Conservation In-service training. Presentation on the benefits of plant diversity on soil health Invited speaker: Mackowiak Target audience: State Faculty, Extension agents June 1, 2023: Led and presented at Perennial Peanut Field Day held in NFREC, Quincy. Scope: Mackowiak Coordinated this event (7 hours). Dubeux moderated producer panel on perennial peanut in grazing systems. Liao lab members presented the posters showcasing the study of microbial mediated activity in the pastures. We also shared handouts and took the audience out to our research plots and included additional updates and potential impacts from our research. Target audience: State Faculty, Extension agents, and producers May 14-17 2023: International Grassland Congress Scope: International congress on grasslands with presentation of most recent advances on scientific research. Presenter: Dubeux Target audience: State Faculty, industry personnel, Extension agents April 26 - 27 2023; March 19 2024: Conduct on-site visits lasting approximately 2 hours each with producers, during which we presented the latest research findings on soil health. Our objective is to establish a training program for collaborating producers. The program will focus on interpreting biological indices and implementing beneficial microbial-friendly practices tailored for forage production. Leads: Liao, Erhunmwunse, Zhang, Wang, Reimer Target audience: Forage producers and the ranchers actively engaged in on-farm studies within this project, including Eugene Rocks, Mack Glass and Billy Kempfer April 20, 2023: Led organizer of Grass endophyte In-Service Training: We led and delivered knowledge on fungal endophytes and the metabolites they produce in the pasture systems through lectures, posters, and round table discussions. The event drew 36 participants from 12 counties in the state of Florida. The insights gained by these Extension agents will prove valuable in enhancing their future training programs, which target a wide audience including statewide farmers, growers, forest managers, forage producers, the public, and 4-H participants. Presenter and organizer: Liao, Mackowiak, Liao lab April 13 2023: NFREC Beef & Forage Field Day Scope: Present in a field setting the most recent advances on forage systems. Presenters: All PIs Target audience: beef cattle and forage producers, industry personnel, Extension agents April 12 2023: Forage Legume Conference Scope: conference presenting most recent advances on Forage legumes in the Southeast. Presenter and organizer: Dubeux Target audience: beef cattle/forage producers, industry personnel, Extension agents March 29, 2023: Present at Forage in-service training, Marianna FL Scope: Forage-IST is an annual, day-long event to deliver the latest research and application update through presentations and field tours. This train the trainer event updates Florida county extension faculty on issues that may impact producers and to provide information to use for their own client training. We delivered several presentations and educational booths to help the audience identify the role forage plays on forage production and soil health and options for integrating the knowledge gained from this topic into their management. Presenters: All PIs Target audience: Extension agents March 17, 2023: Spring Ranchers Forum, Geneva, FL Presentation on pasture decline Invited speaker: Mackowiak Target audience: Livestock and forage producers, industry personnel, extension agents March 13, 2023: Present at Spring Forage Field Day, Osceola county. This event was led by forage Extension agent, Justesen. PIs Mackowiak and Liao co-facilitated the event by delivering presentations, posters and handouts on research update and recommendations for the topics of forage soil nutrients, forage microbes and the grass endophyte mediated compounds Target audience: beef cattle and forage producers, industry personnel, Extension agents Feb 27-28, 2023: Lead Forage Workers Tour (Marianna, FL) This training facilitates the discussion among Florida extension faculty and agents for the emerging problems of forage and to brainstorm the next step research and extension efforts for the resolution. Lead: Dubeux; Instructors: All PIs Target audience: State faculty, Extension agents, producers Feb 08, 2023: NW Florida Beef Conference (Marianna, FL) Presentation on forage fertility to improve herd nutrition Invited speaker: Mackowiak Target audience: beef cattle and forage producers, industry personnel, Extension agents Jan 11, 2023: Soil health and Cover Crop Field Day (Live Oak, FL) Presentation on the impact of land-use on soil health indicators Invited speaker: Mackowiak Target audience: Forage producers and row crop producers, industry personnel, Extension agents. Changes/Problems:No major changes to report What opportunities for training and professional development has the project provided?As indicated in the "target audience" section, the project has provided opportunities to train graduate students and student interns. Data generated in the project have been used to provide training and education outreach to extension agents and producers. Through the supervision and training activities, the graduate trainees are expected to receive the professional skills on molecular/biogeochemical technologies and bioinformatics. The professorial training for the graduate students and postdoctoral scientists includes research proposal development, experimental designs, sample collection/processing, data collection/analysis, data integration, scientific presentations, coordination for team meetings, workshop/field day/in-service training participation, and leading the manuscript preparation for his/her projects. All the trainees have been actively involved in scientific presentations and extension activities, with the evidence shown in the sections of "Product" and "Other products". (1) Adesuwa Erhunmwunse (Ph.D. student, supervisor: Liao): Adesuwa's project focused on understanding the interactive effects of multiple management (plant community, grazing, and environmental variables) on microbial-driven soil fertility and forage production. Adesuwa trained the master student (e.g., Jaime Garzon Alfonso) on the molecular work that is associated with the forage project (e.g., soil DNA extraction, qPCR analysis). Adesuwa also mentored a MS. student (Rachel Balster) to study the diversity of N-fixing bacteria living in RP nodules. Adesuwa has successfully completed her Ph.D. program in August 2023 and published 4 papers from this project (3 as first authors and one as a co-author). She is currently working as a Biologist Specialist in the Department of Biological Sciences at Idaho State University, Pocatello, Idaho. She is currently using the molecular techniques and bioinformatic skills she gained from this project to surveil wastewater generated from Idaho Municipalities for microbial pathogens. (2) Kaile Zhang (Postdoctoral Associate, supervisor: Liao): Kaile's project focuses on determining the predominant fungal taxa in pastures that bring benefits to various cropping systems, including bahiagrass and RP. The outcome of the study has resulted in the publication of one research note, a review article, and four research papers. Kaile trained a Ph.D. student (Pamella Sena) on field sampling and molecular work (e.g., soil DNA extraction, PCR, and qPCR analysis) that were related to the forage project. (3) Other students and scientists assisted on field sampling and laboratory analyses, including Vijay Verma (Biological Scientist, supervisor: Liao), Martin Ruiz-Moreno (Biological scientist, supervisor: Dubeux), George Anguelov (Biological Scientist, supervisor: Mackowiak), Luke Harlow (MS student, supervisor: Mackowiak), Kelly Thomas (MS student, supervisor: Blount), José D. Pereira Neto (sandwich PhD student, supervisor: Dubeux), Daniele Loures (visiting faculty, supervisor: Dubeux), Keving Roger Trumpp (MS student; supervisor Dubeux), Javier Portuguez (MS student, Dubeux) How have the results been disseminated to communities of interest?We have been using a combination of publications, in-person meetings and social media as our methods for disseminating information (See "Target Audience" section and "Products" section). What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? What did we do in 2023: Objective 1. We completed the second phase of the greenhouse experiment to identify free living microbes, mycorrhizae, and root associated microbes responsible for nutrient mobilization and transfer between RP and bahiagrass. For this study, we designed a rhizobox to partition RP and bahiagrass in the same pot. Laboratory and data analysis and manuscript preparation are ongoing. Experimental field site experiment (Established site at Marianna FL in 2019) For this study, we investigated changes in soil microbial communities and N cycling populations using amplicon sequencing targeting soil prokaryotes and fungi and activities of soil enzymes involved in N mineralization, and real- time qPCR to quantity the N functional groups of nitrifying, denitrifying, andnitrogen fixing bacteriain bahiagrass and RP mixtures versus their monocultures across two sampling dates (March and October).From this study, we observed that unlike prokaryotic taxa, RP influenced the dominant soil fungal taxa, such as Fusarium, Gibberella, and Humicola. Our findings showed that RP has the potential to influence N cycling process in bahiagrass system by altering the abundance of certain N cycling fungal taxa, within 2 years of RP establishment. The manuscript was published in the European Journal of Soil Biology (https://doi.org/10.1016/j.ejsobi.2023.103566) Experimental field site experiment (the long-term established sites in Quincy FL): We published the manuscript (https://doi.org/10.3390/microorganisms11123002) for this study based on the soil samples collected in 2021 across two soil depths (0-15 and 15-30 cm) from BG and RP plots that were established a decade ago (at NFREC, Quincy) to determine the long-term effects of RP and BG mixture on soil bacterial communities. We found that long-term integration of RP into bahiagrass led to responses in soil bacterial diversity and community composition. Soil bacterial community shifted from a system dominated by Proteobacteria reported in other short-term bahiagrass and rhizoma peanut systems to a system dominated by Firmicutes. We plan to publish another manuscript on these results showing the impact these systems have on soil fertility and nutrient uptake under low input management. Experimental field site experiment (Different cultivars of RP) We collected nodules from Ecoturf and Florigraze RP from NFREC, Quincy, FL in July and October to identify N fixing bacterial communities using culture dependent (selective agar media) and independent methods like amplicon sequencing. We identified the bacterial isolates, and the manuscript is under preparation to be published March 2024. From this study we found that there was a diversity of nitrogen fixing bacteria in the nodules of rhizoma peanut, including Rhizobium, Microbacterium, and Bacillus. Experimental field site experiment with arbuscular mycorrhizal fungal treatment (newly established site in 2022 at Quincy FL) This study aims to determine the N uptake efficiency and leaching alleviation of pasture as affected by the combination of forage species (bahiagrass and rhizoma peanut) and arbuscular mycorrhizal fungi (AMF). The treatments include monocultures of bahiagrass/ rhizoma peanuts and their mixers across treatments with and without AMF inoculation. We collected soil samples one month after applying N fertilizers to investigate how different forage management practices and AMF inoculation influence soil microbial dynamics. We found that ?Neither pasture management nor AMF application influenced bacterial abundance. However, they had significant and interactive impacts on fungal abundance. Surprisingly, AMF application reduced the native soil fungal population, suggesting? that external AMF addition might lead to competitive interactions with indigenous fungal communities. We also found that pasture management was the main effect on soil AMF abundance, with Bahiagrass +1/2N exhibiting greater AMF abundance than rhizoma peanut. These results were presented at the 2023 ASA-CSSA-SSSA International Conference. We plan to publish one manuscript in 2024 after collecting multiple time-point samples. Objective 2. All laboratory analysis was completed this year from the root, rhizosphere, and bulk soil samples collected from the three on-farm sites in 2020 and 2021. Two manuscripts are under preparation based on the data generated from the lab and the environment data (rainfall, soil moisture and temperature) collected.

Publications

  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Erhunmwunse A, Guerra V, Liu J-C, Mackowiak C, Blount A, Dubeux Jr. J, Liao H-L. 2023. Soil bacterial diversity responds to long-term establishment of perennial legumes in warm-season grassland at two soil depths. Microorganisms. 11(12):3002. doi.org/10.3390/microorganisms11123002
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Ehunmwunse AS, Mackowiak CL, Blount ARS. Dubeux JCB, Ogram A, Liao H-L. 2023. Short-term integration of perennial peanut into bahiagrass system influence on soil microbial-mediated nitrogen cycling activities and microbial co-occurrence networks. European Journal of Soil Biology. 119:103566. doi.org/10.1016/j.ejsobi.2023.103566
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Erhunmwunse, AS, Queiroz LMD, Zhang K, Mackowiak CL, Blount ARS, Dubeux Jr JCB, and Liao H-L. 2023. Changes in soil microbial diversity and community composition across bahiagrass and rhizome peanut pastures. Biology and Fertility of Soils. doi.org/10.1007/s00374-023-01701-z
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Garzon J, Vendramini J, Silveira M, Dubeux J, Sollenberger L, Liao H-L, Silva H, Gomes VC, Oliveira H, Moriel P. 2024. Aeschynomene overseeding and N fertilization effects in bahiagrass litter decomposition. Agronomy (manuscript ID: AJ-2023-06-0448-OA.R3, accepted). doi: 10.1002/agj2.21561
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Rusch, H., J.C.B. Dubeux, Jr., L.M.D. Queiroz, M.O. Wallau, C.H. Wilson. 2023. Species richness and functional diversity enhance winter annual forage productivity and nutritive value. Crop Science 2023; 63:3136-3150. https://doi.org/10.1002/csc2.21071
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Santos, E.R.S., J.C.B. Dubeux, Jr., C. Mackowiak, L.E. Sollenberger, G.D. Farias, B.G. Homem, D.M. Jaramillo, L. Zagato, L.M.D. Queiroz, D.L. Wright, N. DiLorenzo, M. Ruiz-Moreno. 2023. Above and belowground litter decomposition of cover crops grazed at different intensities. Grass & Forage Science, 2023, 78:376-389. https://doi.org/10.1111/gfs.12617
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Thomas, K.M.g, A.R. Blount, G.W. Knox, C.L. Mackowiak, and L.E. Sollenberger 2023. Ornamental rhizoma peanut: Perceptions and use by Florida Consumers. HortScience, 58(10) 1178-1182. https://doi.org/10.21273/HORTSCI17254-23
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Jaramillo, D.M., M. Ruiz-Moreno, J.M.B. Vendramini, L.E. Sollenberger, N. DiLorenzo, L.M.D. Queiroz, E.R.S. Santos, L. Garcia, D.S. Abreu, J.C.B. Dubeux, Jr. 2023. Methane emissions and 13C composition from beef steers consuming binary C3-C4 diets. J. Anim. Sci., Volume 101, 1-7, 2023, skad181. https://doi.org/10.1093/jas/skad181
  • Type: Journal Articles Status: Submitted Year Published: 2024 Citation: Daraz U, Erhunmwunse A, Dubeux JCB, Mackowiak C, Liao H-L, Wang XB. 2024. Soil fungal community structure and function response to rhizome perennial peanut cultivars. Applied Soil Ecology (submitted. Manuscript Number: APSOIL-D-24-00499)
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Daraz U, Erhunmwunse AS, Dubeux Jr JCB, Mackowiak C, Guerra VA, Hsu C-M, Ma J, Liao H-L, Wang X-B. 2023. Soil bacterial communities across seven rhizoma peanut cultivars (Arachis glabrata Benth.) respond to seasonal variation. Microbial Ecology. doi: 10.1007/s00248-023-02277-9.
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Guo Y, Liao H-L, Boughton EH, Martens-Habbena W, Qiu J. 2023. Interactive effects of land-use intensity, grazing and fire disturbances on soil bacterial and fungal communities in subtropical wetlands. Agriculture, Ecosystem and Environment. 345:108314. doi.org/10.1016/j.agee.2022.108314
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Garzon JE, Vendramini JMB, Silveira ML, Sollenberger LE, Dubeux JCB, Liao H-L, da Silva HMS, Gomes VC, Oliveira HMR. 2023. Overseeding aeschynomene and N fertilization effects on forage characteristics, N fixation and N2O emissions of bahiagrass pastures. Crop Science. doi.org/10.1002/csc2.20981
  • Type: Journal Articles Status: Under Review Year Published: 2024 Citation: Queiroz LMD, Dubeux Jr JCB, Sollenberger LE, Vendramini JMB, Liao H-L, Jaramillo DM, Santos ERS, Abreu DS, Lira Jr MA, Casagrande DR, Mackowiak CL, Ruiz-Moreno M. 2024. A novel methodology to track nitrogen transfer in a grass-legume mixture using enriched 15N2. Plant and Soil (In revision. PLSO-D-24-00252)
  • Type: Theses/Dissertations Status: Published Year Published: 2023 Citation: Ehunmwunse A. 2023. (Soil Microbial Diversity and Community Composition and Associated Nutrient Cycling Activities in Rhizoma Peanut-Based Bahiagrass Pastures Across Florida) Dissertation, University of Florida. ProQuest Dissertations & Theses Database. Accession No. 2023.?30631277.
  • Type: Theses/Dissertations Status: Published Year Published: 2023 Citation: Balster R. 2023. Evaluation of vegetation to improve pollinator habitat in solar farms & The characterization of bacteria in the root nodules of rhizoma peanut.
  • Type: Other Status: Published Year Published: 2024 Citation: Liao H-L, Erhunmwunse AS, Zhang K. 2024. Integrating rhizoma perennial peanut into bahiagrass pastures enhances beneficial soil microbes in the pastures. The Florida Cattleman and Livestock Journal. March-April issue.
  • Type: Other Status: Published Year Published: 2023 Citation: Liao H-L. 2023. What are the rhizosphere microbes and the roles they play in crop production and soil health. EDIS. (Peer-Reviewed: https://edis.ifas.ufl.edu/publication/SS723). doi.org/10.32473/edis-SS723-2023
  • Type: Other Status: Under Review Year Published: 2024 Citation: Liao H-L, Erhunmwunse AS 2023. Integrating rhizoma perennial peanut into bahiagrass pastures enhances beneficial soil microbes in Florida. (EDIS, Submitted in April 2023; In review, archive ID: 133946)
  • Type: Other Status: Published Year Published: 2023 Citation: Liao H-L, Chen K-H, Marcon F, Jones R, Justesen B, Walter J, Blount A, Mackowiak M, Mayo D, Wallau M. 2023. A preliminary survey of mycotoxins identified from Florida bahiagrass pastures. EDIS SL505/SS718. (Peer-Reviewed) doi.org/10.32473/edis-SS718-2023
  • Type: Other Status: Published Year Published: 2023 Citation: Liao H-L, Chen K-C, Marcon F, Jones R, Justesen B, Walter J, Blount A, Mackowiak C, Mayo D, Wallau M. 2023. Black fungal structures (black stroma) grown on the leaves of bahiagrass, limpograss and smutgrass: What they are? The Florida Cattleman and Livestock Journal. September issue.
  • Type: Other Status: Published Year Published: 2023 Citation: Liao H-L, Chen K-C, Marcon F, Justesen B, Joe Walter J, Ann Blount A, Mackowiak C. 2023. Are there beneficial fungi living in bahiagrass leaves and can occasionally produce mycotoxins? The Florida Cattleman and Livestock Journal. January issue.
  • Type: Other Status: Published Year Published: 2023 Citation: Liao H-L, Chen K-C, Marcon F, Jonse R, Justesen, B, Joe Walter J, Ann Blount A, Mackowiak C. Mayo D, Wallau M. 2023. Black fungal structures (Stroma) found on the grass leaves of your pastures. NFREC Beef/Forage Field Day Proceedings April 2023
  • Type: Other Status: Published Year Published: 2023 Citation: Mackowiak, C.L. Jun 30, 2023. Nitrogen fertilization of perennial summer grasses. Panhandle Agricultural eNews https://nwdistrict.ifas.ufl.edu/phag/2023/06/30/nitrogen-fertilization-of-perennial-summer-grasses/
  • Type: Other Status: Published Year Published: 2023 Citation: Mackowiak, C.L. 2023. Management considerations for bahiagrass late-season. The Florida Cattlemen and Livestock Journal. Aug 88(11).
  • Type: Other Status: Published Year Published: 2023 Citation: Mackowiak, C.L. 2023. Pasture potassium fertilization. The Florida Cattlemen and Livestock Journal. Apr 88(7).
  • Type: Other Status: Published Year Published: 2023 Citation: Shahid, M.A., K. Leaks, A.R. Blount, and C. Mackowiak. 2023. Perennial peanut as a potential living mulch and nitrogen source for citrus and other orchard crops in Florida. Cooperative Extension Service, IFAS, 3 pp. HS-1474.
  • Type: Other Status: Published Year Published: 2023 Citation: Vasco, C., J.C.B. Dubeux, Jr., E. Santos, C. Wickens, and L. Warren. 2023. Acceptability of rhizoma peanut forage by horses and its benefits on feeding behavior. EDIS AN390 https://doi.org/10.32473/edis-AN390-2023
  • Type: Other Status: Published Year Published: 2023 Citation: Dubeux, J.C.B., Jr., L.D. Queiroz, D. Jaramillo, L. Garcia, E.R.S. Santos, I.L. Bretas, M. Bernardini, K.R. Trumpp, J. Portuguez, H. Koury, K. Oduor, M. Ruiz-Moreno. 2023. Is a year-round grazing system feasible in North Florida? Available at Is a Year-Round Grazing System Feasible in North Florida? | Panhandle Agriculture (ufl.edu)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Zhang K, Mackowiak C, Liao H-L. 2023. Effects of pasture management and arbuscular mycorrhizal fungal inoculation on soil microbial dynamics and greenhouse gas emissions. 2023 ASA, CSSA and SSSA Annual Meeting, St. Louis poster presentation) (Abstract: https://scisoc.confex.com/scisoc/2023am/s3/papers/index.cgi?username=149811&password=944135)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Erhunmwunse AS, Reimer B, Iriarte AB, Verma VC, Mackowiak CL, Liao H-L. 2023. Identifying the microbial-mediated strategies for optimum phosphorus uptake in bahiagrass and rhizoma peanut mixture. ASA, CSSA, SSSA International Annual Meeting, St. Louis, MO. (Oral and poster) (Invitation via travel award to Erhunmwunse fully funded by SSSA). Abstract: https://scisoc.confex.com/scisoc/2023am/meetingapp.cgi/Paper/154111.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Liao H-L, Zhang K, Mendez V, Wang H, Chen K-H, Reimer B, Verma V, Justesen B, Walter J, Blount A, Mackowiak C, Wallau M, Jones R, Mayo D. January 2024. Grass endophytes and mycotoxins in Florida pastures (oral presentation). American forage and grassland council (AFGC), Mobile Alabama
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Lira Junior MA, Dantas Queiroz LM, Erhunmwunse AS, Sena PTS, Mackowiak CL, Ruiz-Moreno M, Liao H-L, Dubeux JCB Jr. 2023. Further evidence for legume-grass direct N transfer. ASA, CSSA, SSSA International Annual Meeting, St. Louis, MO. Abstract: https://scisoc.confex.com/scisoc/2023am/meetingapp.cgi/Paper/150911.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Balster R, Erhunmwunse AS, Wallau MO, Liao H-L, Mackowiak CL. 2023. Identification and characterization of bacterial communities in rhizoma peanut nodules. ASA, CSSA, SSSA International Annual Meeting, St. Louis, MO https://scisoc.confex.com/scisoc/2023am/meetingapp.cgi/Paper/151027.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Zhi R, Petticord D, Boughton E, Sparks J, Liao H-L, Qiu J. 2023. Soil fungal and arbuscular mycorrhizal communities in response to soil phosphorus gradient in managed subtropical grasslands. The Ecological Society of America (ESA).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Dubeux, J.C.B., Jr., D.M. Jaramillo, E.R.S. Santos, L. Garcia, L.M.D. Qeuiroz, K.R. Trumpp. 2023. Sustainable intensification of livestock systems using forage legumes in the Anthropocene. Proceedings of the XXV International Grassland Congress, 14-19 May 2023, Covington, Kentucky, USA. Print ISBN: 978-1-7138-8028-8. eISBN: 978-1-7138-8029-5. https://doi.org/10.52202/071171-0022 p.98-101
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Jaramillo, D.M., J.C.B. Dubeux, Jr., L.M.D. Queiroz, and L. Garcia. 2023. New insights for benefit of legume inclusion in grazing systems. Proceedings of the XXV International Grassland Congress, 14-19 May 2023, Covington, Kentucky, USA. Print ISBN: 978-1-7138-8028-8. eISBN: 978-1-7138-8029-5. https://doi.org/10.52202/071171-0152 p.637-640.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., L.E. Sollenberger, M.O. Wallau, D.R.S. Loures, M.A. Bernardini, I.L. Bretas, S.F. Novo, K.R. Trumpp, K.T. Oduor, J.D. Pereira neto, M. Ruiz-Moreno. 2023. A novel technique to label cover crop biomass using stable isotopes. Proceedings of the XXV International Grassland Congress, 14-19 May 2023, Covington, Kentucky, USA. Print ISBN: 978-1-7138-8028-8. eISBN: 978-1-7138-8029-5. https://doi.org/10.52202/071171-0263 p.1088-1091.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Dubeux, J.C.B., Jr., E.R.S. Santos, J.E. Portuguez-Acuna, L.M.D. Queiroz. 2023. Nutrient cycling and crop responses on integrated crop-livestock systems. Proceedings of the XXV International Grassland Congress, 14-19 May 2023, Covington, Kentucky, USA. Print ISBN: 978-1-7138-8028-8. eISBN: 978-1-7138-8029-5. https://doi.org/10.52202/071171-0272 p.1123-1126.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Vasco, A.C., J.C.B. Dubeux, Jr., L. Sollenberger, M. Wallau, E. Santos, J. Lance, L. Warren, C. Wickens. 2023. Diurnal variation in forage nutrient composition and metabolic parameters of horses grazing warm-season, perennial grass-legume pastures. Proceedings of the XXV International Grassland Congress, 14-19 May 2023, Covington, Kentucky, USA. Print ISBN: 978-1-7138-8028-8. eISBN: 978-1-7138-8029-5. https://doi.org/10.52202/071171-0345 p.1419-1422
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Vasco, A.C., E. Santos, J. Lance, J.C.B. Dubeux, Jr., L. Sollenberger, M. Wallau, L. Warren, C. Wickens. 2023. Herbage responses and performance of mature horses grazing warm-season perennial grass-legume mixed pastures. Proceedings of the XXV International Grassland Congress, 14-19 May 2023, Covington, Kentucky, USA. Print ISBN: 978-1-7138-8028-8. eISBN: 978-1-7138-8029-5. https://doi.org/10.52202/071171-0346 p.1423-1426.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Caram, N., L.E. Sollenberger, M.O. Wallau, J.C.B. Dubeux, Jr. 2023. Toward and optimum legume proportion in legume-grass pastures: from radiation use efficiency to animal performance. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Trumpp, K.R., J.C.B. Dubeux, Jr., H.A. Koury, J.P. Acuna, K. Oduor, I.L. Bretas, L.M.D. Queiroz, M.A. Bernardini, M. Ruiz-Moreno, L. Garcia, D.M. Jaramillo, L.E. Sollenberger, N. DiLorenzo, J.M.B. Vendramini. 2023. Belowground responses in contrasting grazing systems. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., M.A. Lira, Jr., L.E. Sollenberger, M.O. Wallau, J.M.B. Vendramini, H.L. Liao, L. Garcia-Jimenez, M. Ruiz-Moreno, K. oduor, I.L. Bretas, J.D. Pereira Neto, K.R. Trumpp, M.A. Bernardini, J.P. Acuna. 2023. Belowground responses of cool-season grass-legume mixtures using enriched 15N2 and 13CO2 as tracers. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Acuna, J.P., J.C.B. Dubeux, Jr., L.E. Sollenberger, C.L. Mackowiak, N. DiLorenzo, A. Martin-Ryals, L.M.D. Queiroz, K.R. Trumpp, K. Oduor, I.L. Bretas, L. Garcia, M. Ruiz-Moreno. 2023. Tracking nitrogen pathways on contrasting cover cropping systems utilizing 15N-labeled urea. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., L.E. Sollenberger, M.O. Wallau, K.Oduor, I.L. Bretas, J.D. Pereira neto, K.R. Trumpp, J.P. Acuna, M.A. Bernardini, M.A. Lira, Jr., B.E.B. Cremostim, F.F. Simili, M. Ruiz-Moreno. 2023. Tracking cover crop nutrient decomposition in integrated crop and livestock systems using stable isotopes. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Trumpp, K.R., J.C.B. Dubeux, Jr., J.P. Acuna, M.A. Bernardini, I.L. Bretas, L.M.D. Queiroz, K. Oduor, M. Ruiz-Moreno, L. Garcia, D.M. Jaramillo, L.E. Sollenberger, N. DiLorenzo, J.M.B. Vendramini. 2023. Ammonia volatilization from animal excreta in contrasting grazing systems. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Simili, F.F., J.C.B. Dubeux, Jr., L.M.D. Queiroz, B.E.B. Cremostim, I.L. Bretas, M.A. Lira, Jr., R.S. reis, J.P. Acuna, K.R. Trumpp, M. Ruiz-Moreno, D.M. Jaramillo, K. Oduor, M.A. Bernardini. 2023. Can sunn hemp intercropping with corn improve yield, nitrogen cycling, and nematode control? ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Queiroz, L.M.D., M.A. Lira, Jr., A.S. Erhunmwunse, P.T.S. sena, C.L. Mackowiak, M. Ruiz-Moreno, H.L. Liao, J.C.B. Dubeux, Jr. 2023. Further evidence for legume-grass direct N transfer. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Acuna, J.P., J.C.B. Dubeux, Jr., A. Martin-Ryals, L.E. Sollenberger, C.L. mackowiak, N. DiLorenzo, L. Garcia, K.R. Trumpp, I.L. Bretas, K. Oduor, M.A. Bernardini, L.M.D. Queiroz. 2023. Life cycle assessment of contrasting grazing systems within a yearling beef cattle stocker system. ASA/CSSA/SSSA International Annual Meeting Abstracts, St. Louis, MO.


Progress 04/01/19 to 03/31/24

Outputs
Target Audience:A. This project involves in training of total 32 graduate students, biologists and visiting scientists to conduct the research for this study. 11 Graduate students at the Ph.D. level, with a PI serving as the chair (Adesuwa Erhunmwunse, Benjamin Reimer, Kaile Zhang, Victor Guerra, Bruno Grossi, Carlos Garcia, Gustavo Duarte, Izabela Gomes, Jose H. Andriarimalala, José D. Pereira Neto, Beatriz Bizutti Cremostin) 8 Graduate students at the M.S. level, with a PI serving as the chair (Luana Dantas Queiroz, Jung-Chen Liu, Valerie Mendez, Luke Harlow, Kelly Thomas, Javier Portuguez, Kevin RTrumpp, Daciele Abreu) 2 International visiting M.S. and Ph.D. graduate students (Lukas Beule, Pamella Sena) 2 Postdoc (Ko-Hsuan Chen, Haihua Wang) 5 Other visiting scholars and visiting professors (Mario Lira Junior, Daniele Loures, Xiaobo Wang, Daniel Casagrande, Daniele Loures) 3 Biological scientists (Vijay Verma, Martin Ruiz-Moreno, George Anguelov) 1 Volunteer scientist (Chih-Ming Hsu) B. The project includes the dissemination of research outcomes through 48 extension/outreach training programs (listed below). Several individual training programs were coordinated by the project's PIs, with the involvement of other PIs and their respective laboratory teams actively participating in the presentations. These programs aim to reach a diverse audience, including Extension faculty, stakeholders, the general public, youth, producers, farmers, and government staff. In addition, we utilize various platforms as our primary means of dissemination, as outlined in the "Products" and "Other Products" section: These platforms include 19 social media and 28 Extension publications. The overarching goal is to enhance the knowledge of targeted audiences, motivate actions based on the acquired knowledge, and facilitate the integration of training/education materials into the management of stakeholders and farmers. Furthermore, the goal is to aid Extension agents in seamlessly incorporating these materials into their own training programs and extending outreach to environmental education targeting youth. The representative events include: a. Seven In-Service Trainings (five led by PIs) that targeted Extension professionals with approximately 180 attendances in total 2019 Ag In-Service Training (Lead organizer: Mackowiak) 2021 Soil Health In-Service Training (02/22/2021, Lead organizer:Liao) 2021 North Florida Beef and Forage In-Service Training (FBF-IST) (04/08/2021, Lead organizer: Dubeux). 2022 Beef and Forage In-Service Training, Marianna, FL (03/24/2022, Lead organizer: Dubeux) 2023 Grass endophyte In-Service Training, Quincy FL (04/20/2023, Lead organizer: Liao) 2023 Forage in-service training, Marianna FL (03/29/2023, Presentation: all PIs) 2023, Pollinators, Forages, and Habitat Conservation In-service training. Presentation on the benefits of plant diversity on soil health, FL (09/28/2023, Invited talk: Mackowiak) b. 14 Field days (seven led by PIs) that targeted producers, ranchers, and growers, approximately 1000 attendances in total 2019 Perennial Peanut field day at UF NFREC Quincy, FL (05/30/2019, Lead organizer: Blount) 2019 Beef and Forage Field Day at UF/NFREC, Marianna, FL (Lead organizer: Dubeux) 2020 Virtual Perennial Peanut Field Day at UF/NFREC Quincy, FL (06/25/2020, Lead organizer: Blount) 2020 Sod-Based Rotation Field Day (Virtual) (Presentation: Liao) 2021 Cover Crop Field Day at UF/NFREC, Suwanee Valley, FL (12/08/2021, Presentation and handout production: Mackowiak & Liao) 2022 Perennial Peanut field day at UF NFREC Quincy, FL (06/02/2022, Lead organizer: Blount) 2022 Spring Forage Field Day, Silver Spurs Ranch, Kenansville,FL. (04/06/2022, Presentation: Mackowiak) 2022 Fertilization Strategies for Pasture Management Field Day, Live Oak, FL (03/29/2022, Presentation: Mackowiak) 2022 Beef and Forage BMP Farm Tour (04/22/2022, Presentation: Dubeux) 2023, Soil health and Cover Crop Field Day, Live Oak, FL (01/11/2023, invited speaker: Mackowiak) 2023 Forage Workers Tour at UF/NFREC Marianna, FL (02/27-28/2023, Lead organizer: Dubeux) 2023 Spring Forage Field Day, Osceola county FL (03/13/2023, Presentation: Liao & Mackowiak) 2023 NFREC Beef & Forage Field Day, Marianna, FL (04/23/2023, Lead organizer: Dubeux) 2023 Perennial Peanut Field Day at UF/NFREC Quincy, FL (06/01/2023, Lead organizer: Mackowiak) c. On-farm visits and deliver research updates to the producer collaborators 2019 to 2021 (three visits per year) to the producers. The final reports were delivered to the producers (Rocks, Glass) in Spring 2023 2019 to 2024 (three visits per year) to the ranchers. The final reports were delivered to the rancher (Kempfer) in Spring 2024 d. 17 events participation for the total of 26 presentations in the research and Extension integration conferences/meetings 2019, One oral Presentations at Southern Pastures and Forage Crop Improvement Conference (Roanoke, VA, 05/23/2019) 2020, One poster presentation at Extension Professional Associations of Florida Virtual Conference (09/01/2020) 2020, Two presentations at Southern Pasture Forage Crop Improvement Conference (SPFCIC) 2021, Deliver the research and application update to Perennial Peanut Producer's Association and Florida Cattlemen's Association 2021 One poster presentation at the American Forage and Grassland Council Virtual Conference (01/12/2021). 2021, Two presentations at Perennial Peanut Producers Association meeting (PPPA)(02/10/2021) 2022, One presentation to the Georgia Wiregrass Cattlemen, Lake Park, GA(03/17/2022) 2022, One presentation at AFGC Virtual Conference(01/19/2022). 2022, One presentation at Florida Cattlemen's Association Leadership Academy (01/17/2022) 2022, One presentation at Southern Section Animal Science, Forth Worth, TX (01/24/2022) 2023, One invited presentation at NW Florida Beef Conference, Marianna, FL (02/08/2023) 2023, One invited presentation at Spring Ranchers Forum, Geneva, FL (03/17/2023) 2023, One presentation at International Grassland Congress (05/14-17/2023) 2023, One invited presentation at IV International Symposium on Integrated Crop-Livestock Systems, Brasil (11/06-09/2023) 2023, One invited presentation at International Symposium on Forage Legumes (11/28-30/2023) 2023, Four oral and three poster presentations at Florida Cattlemen's Association meeting, Marianna FL (11/30/2023) 2024, One presentation and one invited talk at AFGC Conference, AL (01/9/2024) e. Five environmental education events targeting 4-Hyouth 2019, Art, Garden and Farm Family Festival UF/NFREC, Quincy FL (10/05/2019: Booth exhibition: Liao lab; Booth visitors ~250) 2022, Booth exhibition and presentation at "STEM in the art", Tallahassee FL (11/29/2022: Booth exhibition: Liao lab, Booth visitors ~50) 2022, Tallahassee Science Festival, Tallahassee FL (10/22/2022, Booth exhibition: Liao lab, Booth visitors ~300) 2022, Summer camp, UF/IFAS Extension Gadsden County FL (07/19-21/2022, co-led by Liao lab and Extension faculty, 28 youth attendances) 2023, Lead Microbiology Class in Cub Scouts - Wallwood Outdoor Weekend Program, Gadsden County FL (11/19/2023, Station exhibition: Liao lab, 39 youth attendances) f. Three other Extension events: 2020 Presentation for the Webinar of Master Gardeners (05/20/2020, Presentation: Liao and lab team, ~35 attendance) 2020 Hosted the visit of FFA students from Gulf County visited NFREC Marianna and our research plots. (03/06/2020): (Lead organizer: Dubeux, 25 student attendances) 2022 Hosted Soil Water and Ecosystems Sciences Department (SWESD) graduate student research tour of the NFREC at Live Oak, Quincy and Marianna FL (06/17-18/2022, Lead organizer: Mackowiak) C. To targetscientific community: The program generated 61 abstract publications/research presentations and 58 peer-reviewed publications. This project was acknowledged in 28 abstracts/presentations and 17 peer-reviewed publications (Refer to "Products" section). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?As indicated in the "Target Audience" section, the project has provided opportunities to train 21 graduate students and student interns and 9 other scientists. The acquired skill set from this project has proven beneficial for numerous students and scientists, enabling them to advance in their careers, securing positions such as faculty members, biologists, and postdoctoral associates. Data generated in the project have been used to provide training and education outreach to extension agents and producers. Through the supervision and training activities, the graduate trainees are expected to receive the professional skills on molecular/biogeochemical technologies and bioinformatics. The professorial training for the graduate students and postdoctoral scientists includes research proposal development, experimental designs, sample collection/processing, data collection/analysis, data integration, scientific presentations, coordination for team meetings, workshop/field day/in-service training participation, and leading the manuscript preparation for his/her projects. All the trainees have been actively involved in scientific presentations and extension activities, with the evidence shown in the sections of "Products" and "Other products". How have the results been disseminated to communities of interest?We have been using a combination of publications, in-person meetings, Extension activities and social media as our methods for disseminating information (See "Target Audience", "Products" and "Other Products" sections). Summarized materials include: 48 Extension/outreach training programs 19 social media publications 28 Extension publications 61 abstract publications/research presentations (This project was acknowledged in 28 presentations) 58 peer-reviewed publications (This project was acknowledged in 17 publications) What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? (1)Expdesigns: We established several field experiments across different bahiagrass (BG) and rhizoma peanut (RP) cultivars. We identified mature (over 10 years) plots of RP and BG to test long-term interactions. In 2019, We identified and developed three observational field sites across regional gradients to access objective two. We also developed several mesocosm greenhouse experiments to study 15N re-allocation and P cycling across BG and RP (Referred to 2020-2024 progress reports for the details) (2) Sample and data collection: Samples: Bulk and rhizosphere soil, and roots were collected from top 15 cm layer and in some cases, samples were also collected from 15-30 cm soil depth, along with aboveground plant tissues. Time points: Samples were collected at multiple time points throughout a year and across different years for most of the experimental setups Measurements: Microbial community: DNA metabarcoding was applied to identify the relative abundance of bacteria and fungi in the soil, root, rhizome, and nodule (for RP). NGS and qPCR was integrated to estimate the absolute abundance of dominant microbial taxa. Microscopic techniques were used to estimate the abundance of some microbial groups such as AMF. Microbial activity: Root metatranscriptomics, qPCR to quantify the abundance of functional genes related to C/N/P cycling in soil (e.g., genes for N fixation, nitrification, and denitrification), and soil enzyme activities (e.g., N-acetyl-B-D-glucosamidase and leucine, aminopeptidase phosphatase) were applied. Soil physiological, chemical, and biochemical indicators: Soil reactive carbon (POXC,WSC), reactive N (TIN and PMN), total C and N, soil saturated and unsaturated hydraulic conductivity, other soil quality indicators (e.g., Fe, Mn, Zn, Cu, moisture, temperature), soil bulk density, and nitrate leaching analysis RP and BG responses: root length, root diameter, root surface area, and root C and N; plant tissue C and N Forage quality: Forage yield and aboveground and belowground plant biomass Environmental parameters: Air temperature, rainfall, soil moisture, and soil temperature. (3) Key findings: From thesis/Dissertation: Guerra VA. 2021. Belowground biomass, N conc, N fixing soil bacteria were greater in BG-RP mixture than BG monoculture with mineral fertilizer. Plant physiological and soil microbial characteristics associated with RPcould benefit the sustainability of BG pastures by replacing mineral N applications while maintaining stability in the face of drought events. Queiroz LMD. 2021: RP-BG mixtures performed differently across Florida regions, but overall the mixtures produced more biomass than grass monocultures and Ecoturf was more productive than Florigraze. We also tracked belowground transfer of N from legume to grass using stable isotopes, developing a novel methodology using H-pot technique. Liu JC. 2021: The yield gains by incorporating RP into BG depends on soil fertility distribution Ehunmwunse A. 2023: (i) Integration of RP into BG pasture supports greater soil microbial diversity and is associated with unique microbial taxa known to promote plant litter decomposition, nutrient cycling, herbicide degradation, and production of anti-microbial compounds. (ii) Soil fungal communities are easily influenced by BG-RP mixtures, while effects on soil bacterial communities are observed when RP is integrated into BG systems over time. (iii) Pasture locations and growing seasons can influence the effect of BG-RP mixtures on soil microbial communities. From paper publications BG cultivars affected soil bacterial community composition, especially the relative abundance of Nitrospira known for its roles in nitrification, whereas the soil fungal community was unaffected (Beule et al 2019) Soil bacterial taxa with N-fixing capacity differed among RP cultivars. Predicted nitrogenase genes responsible for controlling N fixation were greater in some RP cultivars such as Florigraze and Latitude than others (Wang et al 2019) Seasonal impacts on soil bacterial communities were stronger compared to RP cultivars. There was greater soil bacterial diversity in the fall with higher precipitation than the spring season (Daraz et al 2023) The relative abundance of endophytes, mycorrhizal, and saprotrophs were significantly greater among RP cultivars compared to parasites and pathogens, suggesting that incorporation of RP into grassland may promote soil fungal community shifts that may benefit plant growth and soil nutrient cycling (Daraz et al 2024, submitted; APSOIL-D-24-00499) Legumes led to a shift in soil microbial community that may affect N cycling in pastures common to the southeastern United States. For example, bacterial genera with N fixing potentials such as Bradyrhizobium spp. were greater in the pastures with legumes (Guerra et al 2022) RPhas the potential to influence N cycling process in BG system by altering the abundance of N cycling fungi within 2 years of RP establishment (Ehunmwunse et al 2023a) Long term integration of RP into BG system led to an increase in soil bacterial diversity. There was also a shift from a soil bacterial community dominated by Proteobacteria (~26%) reported in other RP studies to a soil bacterial community dominated by Firmicutes (39%). The relative abundance of the bacterial genus Crossiella, known for its antimicrobial traits, was enhanced in the presence of RP (Ehunmwunse et al 2023b) The effect of BG-RP mixtures on microbial diversity and community composition was affected by seasons and pasture locations. For example, greater bacterial diversity in BG-RP mixtures compared to BG only system was only observed in May at the Northwest farm and not at Central and Southwest farms. Location was a major factor influencing the changes in soil microbial community composition, which was mainly explained by soil pH. (Ehunmwunse et al 2023c) Mortierella elongata, a soil generalist, was identified as one of the dominant fungal taxa presenting in the BG soil (Beule et al 2019). Through greenhouse experiments, we found that M. elongata was able to promote the plant growth of multiple plant species, including BG (Zhang et al 2020) We successfully developed a protocol for subsoil DNA extraction (Guerra et al 2020) and improved DNA library construction process for microbial community studies (Chen et al 2020) in FL pastures and other soil systems. We also developed a method to track labeled nitrogen transfer from air to soil to plants underlying grass and legume systems (Queiroz et al 2024). We applied these improved protocols in this project. We investigated the influence of microbial community composition and their interactions with other organisms within alternative cropping systems incorporating BG (Zhang et al 2021). The findings include that this alternative system can increase the diversity of belowground microbial communities in both soil and roots, which fostered more complex microbial interactions, assembled more beneficial microbes, and intensified ecological connections among soil, microbes, and crops (Zhang et al 2022a; 2022b) Other preliminary findings (manuscript in preparation): Erhunmwunse et al. RP impact on phosphorus mobilizing microbes in bahiagrass system. Preliminary finding: RP has the capacity to mobilize P and micronutrients under the low soil P condition and it also promotes AMF colonization of both soil and roots. Zhang et al. AMF and pasture management shape diversity, assembly, and interactions of microbiomes in soil and roots. Preliminary finding: We found that the application of AMF and pasture management had interactive effects on microbial diversity, interactions, and assembly in the root and plant roots. Additional significant findings were detailed in (28) abstracts submitted to the scientific conferences during the project period, with acknowledgments of this project in those presentations. Refer to the "Product" section for further details.

Publications


    Progress 04/01/22 to 03/31/23

    Outputs
    Target Audience:A. This project involves in training the graduate students, biologists and visiting scientists to conduct the research. Trainees: Adesuwa Erhunmwunse(Ph.D. candidate, supervisor:Liao) Adesuwa is the key personnel for this project. She is an underrepresented scientist (an international scientist from Nigeria) Luana Dantas Queiroz(Ph.D. student, supervisor:Dubeux) Valerie Mendez(M.S. student graduated in summer 2022, supervisor:Liao) Kaile Zhang(Ph.D. graduated in summer 2022, current postdoc, supervisor:Liao) Benjamin Reimer(Ph.D student,supervisor:Liao) Vijay Verma(Biological Scientist,supervisor:Liao) Martin Ruiz-Moreno(Biological scientist,supervisor:Dubeux) George Anguelov(Biological Scientist,supervisor:Mackowiak) Luke Harlow(MS student graduated in fall 2022,supervisor:Mackowiak) Kelly Thomas(MS student graduated in summer 2022,supervisor:Blount) José D. Pereira Neto(sandwich PhD student,supervisor:Dubeux) Daniele Loures(visiting faculty,supervisor:Dubeux) Javier Portuguez(MS student;supervisor:Dubeux) KevinR. Trumpp(MS student,supervisor:Dubeux) B. The project also involves in dismission of research outcomes of this project through extension/outreach training programs 1.Feb27-28-2023:Lead Forage Workers Tour(Marianna,FL) Scope:This training facilitates the discussion amongextension faculty and agents for the emerging problems of forage and to brainstorm the next step research and extension efforts for the resolution. Lead:Dubeux; Instructors: All PIs Target audience: Academic community, Extension agents, producers 2. Nov29 2022:Booth exhibition and presentation at "STEM in the art" in Tallahassee,FL ?Scope:We demonstrated the benefits of perennial peanut as a forage and ornamental purpose in Florida. We taught the audience the importance of N fixing bacteria residing in perennial peanut nodules and practiced the art of culturing non-pathogenic bacteria on agar plates. Instructures:Adesuwa Erhunmwunse(PhD student), Benjamin Reimer(PhD student) Target audience: Leon County community residents 3. Oct22-2022:Booth exhibition and demonstrations at Tallahassee Science Festival ?Scope:Presented the role of soil microbes (mycorrhiza and rhizobia) in agriculture, using perennial peanut as an example. We shared handouts, used posters, and samples of rhizoma peanut as demonstration materials. Lead:Seven members from Liao lab, including including graduate students, postdocs, undergraduate volunteer etc. Target audience:General public from all over Florida. 4. July 19-21, 2022:Summer camp, UF/IFAS Extension Gadsden County(4-H) Scope:We taught Gadsden County Youths the importance of agriculture in connection to our different countries. We also linked our classes to our research, especially on the benefits of soil microbes in perennial peanut and grasses pastures. Lead: Six members from Liao lab including graduate students, postdocs, undergraduate volunteer etc. Target audience:5-17 years youths from Gadsden County,FL 5. June17-2022:Hosted Soil Water and Ecosystems Sciences Department (SWESD) graduate student research tour of the NFREC at Live Oak, Quincy, and Marianna(2-day event). Scope: To share what it is like to conduct research at a Research and Education Center(REC), to learn about the various disciplines represented, the research facilities and the field research. The successful inaugural event prompted the students to pursue future tours at other RECs. On the first day, Vijay Verma and Adesuwa Erhunmwunse(Ph.D. candidate) shared their research on soil microbiomes and nutrient cycling in bahiagrass/rhizoma peanut systems. On the second day, we spent the morning at NFREC-Marianna, where we have several field perennial peanut/bahiagrass trials. Dubeux spoke about the perennial peanut/bahiagrass research at Marianna Lead:Mackowiak Target Audience:SWESD Graduate Student Association 6. June02-2022:Presentation at Perennial Peanut Field Day held in North Florida Research and Education Center, Quincy. Scope:We presented posters showcasing our soil microbiome research on bahiagrass and rhizoma peanut. Adesuwa Erhunmwunse presented a poster titled"Legume cultivars effect on soil microbial communities (the hidden engineers) in bahiagrass system" and Kaile Zhang presented a poster"?How do mycorrhizal fungi improve nitrogen use efficiency in the rhizoma peanut-bahiagrass system?". We also shared handouts and took the audience out to our research plots and included additional updates and potential impacts from our research. Instructors:four members from Liao lab and Mackowiak lab Target audience:Academic community, extension agents, and producers 7. April06-2022:Present at the Spring Forage Field Day, Silver Spurs Ranch, Kenansville,FL. Presentation title:Fertilization strategies for pastures and hay fields. Scope:The field day addressed a variety of topics tailored to area producers, including cattle health, sprayer calibrations, cool-season forages and preparing for warm-season forage production. Our presentation focused on preparing bahiagrass pastures and hay fields and problems encountered when grown under poor fertilization management. Speaker:Mackowiak Target audience:County extension faculty and producers 8.Mar29-2022:Present at Fertilization Strategies for Pasture Management Field Day, Like Oak,FL Presentation title:Fertilization strategies Scope:To address cool-season and warm-season forages, fertilization and grazing management. Bahiagrass was the primary warm-season grass that was addressed. Speaker:Mackowiak Target audience:Producers and water management staff.. 9.Mar24-2022: Lead and Presentation at Beef and Forage in-service training, Marianna,FL Scope:FBF-IST is an annual, day-long event to deliver the latest research and application update through presentations and field tours. This training helps prepare Florida extension faculty and agents for client problems and to provide information for their own client training. We delivered presentation to help the audience identify the role of forage play on soil health and forage production and consider integrate the knowledge gained from this topic into their management. (e.g.,the topic delivered by Liao: "How would soil types, seasons and forage species affect soil fungi and bacteria and benefit forage production") Lead:Dubeux; Instructors: All PIs Target audience:Extension agents 10.Mar17-2022: Presentation to the Georgia Wiregrass Cattlemen, Lake Park,GA. Presentation title:How to survive the summer under crushing fertilizer prices Scope:Cattlemen from south Georgia and North Florida convened for an evening of talks to help address how to remain profitable during the 2022 difficult economic conditions. This presentation focused on grass pasture systems, with emphasis on bahiagrass pastures and hay fields Speaker:Mackowiak Target audience:Cattle/other livestock producers 11.Jan19-2022:Presentation at AFGC Virtual Conference(Zoom). Presentation title:Managing fertilizer costs in 2022. Scope:The conference was designed to provide options under unpredictable growing conditions. This included fertilizer management schemes for grass pastures and hay fields,including specifics for bahiagrass management. Speaker:Mackowiak Target audiences:County extension faculty, government staff and producers 12. Jan17-2022:Florida Cattlemen's Association Leadership Academy Presentation Title:Demonstration of field experiments using forage legumes Scope:Provide future FCA leaders examples of using forage legumes in livestock systems. Speaker:Dubeux 13.Jan24-2022:Southern Section Animal Science, Forth Worth, TX Presentation Title:Ecosystem Services Provided by Grassland Ecosystems in SoutheastUSA Scope:Address ecosystem services from grasslands, with emphasis on forage legumes. Speaker:Dubeux 14. April22-2022: 2022-Beef and Forage BMP Farm Tour Presentation Title:Integrated crop-livestock systems and use of cover crops Scope: address ICL systems and cover crops, with emphasis on forage legumes. Speaker:Dubeux Changes/Problems:No major changes to report What opportunities for training and professional development has the project provided?As indicated in the "target audience" section, the project has provided opportunities to train graduate students and student interns. Data generated in the project have been used to provide training and education outreach to extension agents and producers. Through the supervision and training activities, the graduate trainees are expected to receive the professional skills on molecular/biogeochemical technologies and bioinformatics. The professorial training for the graduate students and postdoctoral scientists includes research proposal development, experimental designs, sample collection/processing, data collection/analysis, data integration, scientific presentations, coordination for team meetings, workshop/field day/in-service training participation, and leading the manuscript preparation for his/her projects. All the trainees have been actively involved in scientific presentations and extension activities, with the evidence shown in the sections of "Product" and "Other products". (1) Adesuwa Erhunmwunse (Ph.D. student, supervisor: Liao): The project that Adesuwa conducts aims at understanding the interactive effects of multiple management (plant community, grazing, and environmental variables) on microbial-driven soil fertility and forage production. Adesuwa trained the master student (e.g., Jaime Garzon Alfonso) on the molecular work that is associated with the forage project (e.g., soil DNA extraction, qPCR analysis). Adesuwa also mentored a MS. student (Rachel Balster) to study the diversity of N-fixing bacteria living in RP nodules. (2) Kaile Zhang (Ph.D. candidate, supervisor: Liao): Kaile's project focuses on determining the predominant fungal taxa in pastures that bring benefits to various cropping systems, including bahiagrass and RP. The outcome of the study has resulted in the publication of one research note, a review article, and four research papers. (3) Valerie Mendez (M.S. student, supervisor: Liao): Valerie is involved in molecular work that are associated with forage project, including DNA extraction, 2- step PCR library construction, DNA purification quantification and Qiime 2 associated data analysis. (4) Other students and scientists assisted on field sampling and laboratory analyses, including Vijay Verma (Biological Scientist, supervisor: Liao), Martin Ruiz-Moreno (Biological scientist, supervisor: Dubeux), George Anguelov (Biological Scientist, supervisor: Mackowiak), Luke Harlow (MS student, supervisor: Mackowiak), Kelly Thomas (MS student, supervisor: Blount), José D. Pereira Neto (sandwich PhD student, supervisor: Dubeux), Daniele Loures (visiting faculty, supervisor: Dubeux), Keving Roger Trumpp (MS student; supervisor Dubeux), Javier Portuguez (MS student, Dubeux) How have the results been disseminated to communities of interest?We have been using a combination of publications, in-person meetings and social media as our methods for disseminating information (See "Target Audience" section and "Products" section). What do you plan to do during the next reporting period to accomplish the goals?We will complete the second phase of the greenhouse experiment on the study of interactions among P, P-associated microbes and forage integration. We will further complete laboratory analyses and continue with data analyses and generation of manuscripts from the data gathered from all the studies conducted under this project. We will submit the completed manuscripts to high quality journals for peer review and publication. We will also publish extension articles and engage with producers that have been involved in this project. We will receive their feedback to help define future studies. The following is a list of manuscripts that will be submitted for publication: Response of soil microbial communities and related nitrogen cycling in bahiagrass and rhizoma peanut monocultures compared to their mixtures. Long-term rhizoma peanut establishment in bahiagrass system influences soil bacterial diversity and composition across different soil depths. Microbial strategies for phosphorus and nutrient mobilization in bahiagrass and rhizoma peanut mixture. Spatial and temporal variations of soil microbial communities in bahiagrass and RP pastures. Arbuscular mycorrhizal fungi and intercropping of bahiagrass and rhizoma peanut increase nitrogen use efficiency. Water limitation impacts on bahiagrass-rhizoma peanut mixtures under low soil N fertility. Bahiagrass nutrient uptake dynamics when grown in mixture with rhizoma peanut. Root-rhizome and soil health response of rhizoma peanut incorporated into bahiagrass. Work on the manuscripts for greenhouse H-pots and on-farm trials

    Impacts
    What was accomplished under these goals? Objective 1 (1) We established and further completed a greenhouse experiment and collected belowground plant parts of BG (root+rhizome) and RP (root+rhizome+nodules) and soil samples for identification of microbial communities and genes expressed. Soil and shoot samples were also collected for nutrient analysis and we found greater bahiagrass biomass when bahiagrass was grown with RP under low P compared to bahiagrass monoculture, but this result was not consistent at the second harvest. We also observed that under low P, RP had greater shoot micronutrient concentrations, indicating that RP may be more efficient in solubilizing soil micronutrients than bahiagrass. More studies are needed to confirm the consistency of this finding. Microbial and data analysis are in progress and will be updated in the final report. A second phase of the greenhouse experiment has been initiated to identify free living microbes, mycorrhizae, and root associated microbes responsible for nutrient mobilization and transfer between RP and bahiagrass. For this study, we designed a rhizobox to partition RP and bahiagrass in the same pot. (2) Experimental field site experiment (Established site at Marianna FL in 2019) For this study, we investigated changes in soil microbial communities and N cycling populations using amplicon sequencing targeting soil prokaryotes and fungi and activities of soil enzymes involved in N mineralization, and real- time qPCR to quantity the N functional groups of nitrifying, denitrifying, andnitrogen fixing bacteriain bahiagrass and RP mixtures versus their monocultures across two sampling dates (March and October). The manuscript was submitted to Applied Soil Ecology in March 1 2023. We also collected another season of forage production and soil composition data. These data are being processed and prepared for manuscript submission on the cultivar impact on integrating RP into bahiagrass monocultures. Additionally from this study, we observed that unlike prokaryotic taxa, RP influenced the dominant soil fungal taxa, such as Fusarium, Gibberella, and Humicola. Our results demonstrated that a long-term establishment of perennial RP might help to create detectable changes in soil microbial prokaryotic communities, including those involved in N cycling activities. (3) Experimental field site experiment (the long-term established sites in Quincy FL): We have completed the manuscript for this study based on the soil samples collected in 2021 across two soil depths (0-15 and 15-30 cm) from BG and RP plots that were established a decade ago (at NFREC, Quincy) to determine the long-term effects of RP and BG mixture on soil bacterial communities. We found that long-term integration of RP into bahiagrass led to responses in soil bacterial diversity and community composition. Soil bacterial community shifted from a system dominated by Proteobacteria reported in other short-term bahiagrass and rhizoma peanut systems to a system dominated by Firmicutes. We plan to publish one manuscript on these results in 2023 and another on the impact these systems have on soil fertility and nutrient uptake under low input management. (4) Experimental field site experiment (Different cultivars of RP). We collected nodules from Ecoturf and Florigraze RP from NFREC, Quincy, FL in July and October to identify N fixing bacterial communities using culture dependent (selective agar media) and independent method like amplicon sequencing. We have stored some bacterial culture and are in the process of identification. (5) Experimental field site experiment with arbuscular mycorrhizal fungal treatment (newly established site in 2022 at Quincy FL) This study aims to determine the N uptake efficiency and leaching alleviation of pasture as affected by the combination of forage species (bahiagrass and rhizoma peanut) and Arbuscular mycorrhizal fungi (AMF). The treatments include monocultures of bahiagrass, rhizoma peanuts and their mixers across with and without AMF treatments. We collected plant root samples to evaluate the colonization efficiency of AMF after six months of trial establishment. We found that ?AMF inoculation significantly increased the density of vesicle and arbuscule in both rhizoma peanut and bahiagrass roots. R?hizoma peanut-bahiagrass integration significantly promoted vesicle density in both rhizoma peanut and bahiagrass roots under no-AMF treatments, whereas it only increased arbuscule density in bahiagrass roots with AMF inoculation. The field establishment and data were presented in the 33rd perennial peanut field day (NFREC, Quincy, FL) and the international Meeting (ASA-CSSA-SSSA) in 2022. We plan to publish one manuscript in 2023 Objective 2. A manuscript that was written based on the bulk soil samples that were collected from in May and July 2019 across the three on-farm sites (North, Central, and South FL) was recently published in the journal, Biology and Fertility of Soils. We found that the effect of bahiagrass-RP mixtures on microbial diversity and community composition was affected by season and locations. For example, greater bacterial diversity in bahiagrass-RP mixtures compared to bahiagrass only system was only observed in May at North farm and not at the other farms. Location was a major factor influencing the changes in soil microbial community composition, which was mainly explained by soil pH. Environment data (rainfall, soil moisture and temperature), root, rhizosphere, and bulk soil samples collected from the three on-farm sites in 2020 and 2021 are still under analysis. All laboratory and data analysis will be completed this year and presented in our final report.

    Publications

    • Type: Other Status: Published Year Published: 2022 Citation: Dubeux, J.C.B., L. Queiroz, L. Garcia, K. Oduor, D. Loures, J.D. Pereira Neto. 2022. Ecosystem services provided by grasslands. The Florida Cattleman and Livestock Journal, v.86, n.8, p. 38-41.
    • Type: Other Status: Published Year Published: 2022 Citation: Santos, E., J. Dubeux, C. Garcia, L. Dantas, F. Van Cleef, D. Jaramillo. 2022. Grazing cool-season cover crops in North Florida. The Florida Cattleman and Livestock Journal, January 2022, v.86, n.4, p. 34-38.
    • Type: Other Status: Published Year Published: 2022 Citation: Dubeux, J.C.B., Jr., L.D. Queiroz, J.D. Pereira Neto, D. Loures, L. Garcia, M. Ruiz-Moreno. 2022. Grow your own nitrogen fertilizer using forage legumes. Available at Grow Your Own Nitrogen Fertilizer Using Forage Legumes | Panhandle Agriculture (ufl.edu)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: 1. Santos, E.R., J.C.B. Dubeux Jr., C.L. Mackowiak, and D.L. Wright. 2022. Nitrate leaching on contrasting grazing intensities in integrated crop-livestock systems. In 2022 Annual meeting abstracts ASA/CSSA/SSSA, Baltimore, MD.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Trumpp, K., J.C.B. Dubeux Jr., D.M. Jaramillo, L. Garcia, L.M. Queiroz, E. Santos, C.L. Mackowiak, J. Pereira, and M. Ruiz-Moreno. 2022. Nitrogen cycle in contrasting grazing systems. In 2022 Annual meeting abstracts ASA/CSSA/SSSA, Baltimore, MD.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Zhang K., Erhunmwunse A, Mackowiak C, Liao H-L. 2022. Effects of Rhizoma Peanut-Bahiagrass Integration and Arbuscular Mycorrhizal Fungal Inoculation on Mycorrhizal colonization. 2022 ASA, CSSA and SSSA Annual Meeting, Baltimore (MD)-Nov. 6-9
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Zhang K., Maltais-Landry G., James M., Mendez V., Wright D., George S., Liao H-L. 2022 Absolute microbiome profiling highlights linkages between stability of bacterial and fungal communities and nutrient cycling under long-term sod-based rotation. 2022 MSA Annual Meeting, Gainesville (FL)-Jul. 10-13
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Cruz, P.J.R., D.R. Casagrande, J.C.B. Dubeux, Jr., D.V. Silva, A.J. Paiva. 2022. Stubble height of tropical mixed pasture influences forage mass, animal behavior and intake rate. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Rusch, H.L., L.M.D. Queiroz, J.C.B. Dubeux, Jr., M.O. Wallau, C.H. Wilson. 2022. Forage accumulation and tissue nitrogen of species rich and functionally diverse cool-season forages. 2022. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Naumann, H.D., J.C.B. Dubeux, Jr. 2022. Above and belowground nitrogen dynamics in tall fescue pasture interseeded with sunn hemp. 2022. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Apolin�rio, V.X.O., J.C.B. Dubeux, Jr., M.V.F. dos Santos, A.C.L. de Mello, M.V. da Cunha, I.S. de Oliveira. Animal performance in silvopasture systems compared with signalgrass in monoculture. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Novo, S.F., J.C.B. Dubeux, Jr., M. Ruiz-Moreno, L. Garcia Jimenez, D.M Jaramillo, L.M.D. Queiroz, F. van Cleef. 2022. Carbon and nitrogen stable isotopic composition of forages, soil, gas, and animal tissues in the Southeast U.S. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Wallau, M.O., J.C.B. Dubeux, Jr., Z. Grabau. 2022. Crop rotation to mitigate plant parasitic nematode buildup on cornfields. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Garcia, L., A. Cuminale, J.C.B. Dubeux, Jr., M.A. Bernardini, J. Campbell, A. Abbate, and G. Williams. 2022. Evaluation of bee presence in multi-species forage plots in North Florida. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Pereira Neto, J.D., J.C.B. Dubeux, Jr., L.M.D. Queiroz, K. Oduor, M. Ruiz-Moreno. 2022. Nutritive value, herbage, and animal performance for nitrogen-fertilized grass and grass-legume grazing systems. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Bernardini, M.A., J.C.B. Dubeux, Jr., L.M.D. Queiroz, L. Garcia Jimenez, D.M. Jaramillo, E.R.S. Santos, L.E. Sollenberger, J.M.B. Vendramini, M. Ruiz-Moreno. 2022. Rhizoma perennial peanut strip planting: how did it work over the years? ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., L.E. Sollenberger, M.O. Wallau, H.L. Liao, L. Garcia Jimenez, M. Ruiz-Moreno, M.A. Bernardini, I.L Bretas, K. Oduor, J.D. Pereira Neto, S.F. Novo. 2022. Tracking nitrogen and carbon transfer in cool-season grass-legume mixtures using enriched 15N2 and 13CO2. ASA/CSSA/SSSA International Annual Meeting Abstracts, Baltimore, MD
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Johnson, M.D., J.C.B. Dubeux, Jr., A.J. Franzluebbers. 2022. Conducting and communicating environmental impacts of research: forage production, soil health, sustainability. Journal of Animal Science, Volume 100, Issue Supplement_1, April 2022, Page 39, https://doi.org/10.1093/jas/skac028.074
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: van Cleef, F., J.C.B. Dubeux, Jr., D.D. Henry, F.M. Ciriaco, N. DiLorenzo, J. Vendramini, H. Naumann, L. Sollenberger, L. Garcia, E. Santos, D. Jaramillo. 2022. Methane and carbon dioxide emissions from excreta of beef steers fed a tannin-rich legume. 8th International Greenhouse Gas & Animal Agriculture Conference, June 5-9 2022, Orlando, FL. Proceedings& p. 118. GGAA-2022-Program-Abstracts-Online.pdf (ufl.edu)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Dubeux, J.C.B., Jr., and L. Garcia. 2022. Ecosystem services provided by grassland ecosystems in Southeast USA. Journal of Animal Science, Volume 100, Issue Supplement_1, April 2022, Page 23, https://doi.org/10.1093/jas/skac028.044
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Garzon, J.E., J.M.B. Vendramini, M.L. Silveira, L.E. Sollenberger, J.C.B. Dubeux, Jr., H.L. Liao, H.M.S. Silva, V.C. Gomes, and H.M.R. Oliveira. 2022. Nitrous oxide emissions of overseeding Aeschynomene into bahiagrass pastures in Florida. 8th International Greenhouse Gas & Animal Agriculture Conference, June 5-9 2022, Orlando, FL. Proceedings& p. 126. GGAA-2022-Program-Abstracts-Online.pdf (ufl.edu)
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Chen, K.H.p, F. Marcon, J. Duringer, A. Blount, C.L. Mackowiak, and H.L. Liao. 2022. Leaf mycobiome and mycotoxin profile of warm-season grasses structured by plant species, geography, and apparent black-stroma fungal structure. Appl. Environ. Microbiol. 88: e00942-22. https://doi.org/10.1128/aem.00942-22
    • Type: Journal Articles Status: Published Year Published: 2023 Citation: Erhunmwunse, AS, Queiroz LMD, Zhang K, Mackowiak CL, Blount ARS, Dubeux Jr JCB, and Liao H-L. 2023. Changes in soil microbial diversity and community composition across bahiagrass and rhizome peanut pastures. Biology and Fertility of Soils. doi.org/10.1007/s00374-023-01701-z
    • Type: Journal Articles Status: Submitted Year Published: 2023 Citation: Erhunmwunse AS, Liao H-L, Mackowiak CL, Ogram A, Dubeux JCB, Blount ARS. 2023. Legume integration into bahiagrass pastures affect soil fungal and not prokaryotic related nitrogen cycling activities. Applied Soil Ecology (submitted. APSOIL-D-23-00238)
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Guerra, V.A, L. Beule, C.L. Mackowiak, J.C.B. Dubeux, A.R.S. Blount, X. Wang, D.L. Rowland, and H. Liao. 2022. Soil bacterial community response to rhizoma peanut incorporation into Florida pastures. J. Environ. Qual. 51:55-65. https://doi.org/10.1002/jeq2.20307
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Zhang, K., Maltais-Landry, G., James, M., Mendez, V., Wright, D., George, S. and Liao, H.L., 2022. Absolute microbiome profiling highlights the links among microbial stability, soil health, and crop productivity under long-term sod-based rotation. Biology and Fertility of Soils, 1-19. https://doi.org/10.1007/s00374-022-01675-4
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Zhang K, Maltais-Landry G, George S, Grabau Z, Small I, Wright D, Liao H-L. 2022. Long-term sod-based rotation promotes beneficial root microbiomes and increases crop productivity. Biology and Fertility of Soils. https://doi.org/10.1007/s00374-022-01626-z
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Jaramillo, D.M., J.C.B. Dubeux, Jr., M. Ruiz-Moreno, N. DiLorenzo, J.M.B. Vendramini, L. Sollenberger, C. Mackowiak, L.M.D. Queiroz, L. Garcia, E.R.S. Santos. 2022. Stable isotopes of C and N differ in their ability to reconstruct diets of cattle fed C3-C4 forage diets. Scientific Reports (2022) 12:17138 https://doi.org/10.1038/s41598-022-21051-4
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: van Cleef, F.O.S.; J.C.B. Dubeux, Jr., C.S. Wheeler, C.C.V. Garcia, M. Ruiz-Moreno, L.E. Sollenberger, J.M.B. Vendramini, N. DiLorenzo, H. Naumann. 2022. Stable isotopes provide evidence that condensed tannins from sericea lespedeza are degraded by ruminal microbes. Scientific Reports (2022)12:14318 https://doi.org/10.1038/s41598-022-18566-1
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: van Cleef, F.O.S.; J.C.B. Dubeux, Jr., F.M. Ciriaco, D.D. Henry, M. Ruiz-Moreno, D.M. Jaramillo, L. Garcia, E.R.S. Santos, N. DiLorenzo, J.M.B. Vendramini, H.D. Naumann, L.E. Sollenberger. 2022. Inclusion of a Tannin-Rich Legume in the Diet of Beef Steers Reduces Greenhouse Gas Emissions from their Excreta. Scientific Reports (2022) 12:14220 https://doi.org/10.1038/s41598-022-18523-y
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Kohmann, M.M., M.O. Bauer, L.E. Sollenberger, L.S.B. Moreno, L.S. da Silva, S. Saraiva, J.C.B. Dubeux, Jr. 2022. Legume proportion affects bahiagrass-rhizoma peanut mixture production and nutritive value and legume composition of cattle diets. Applied Animal Science 38(6):560-569. https://doi.org/10.15232/aas.2022-02297
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Sollenberger, L., and J.C.B. Dubeux Jr. 2022. Warm-Climate, Legume-Grass Forage Mixtures Versus Grass-Only Swards: An Ecosystem Services Comparison. Brazilian Journal of Animal Science, 51:e20210198, 2022. https://doi.org/10.37496/rbz5120210198
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Santos, E.R.S., J.C.B. Dubeux, Jr., L.E. Sollenberger, M.C.B. Siqueira, F. van Cleef, D.M. Jaramillo, L. Zagato, L.M.D. Queiroz, C.C.V. Garcia, M. Ruiz-Moreno. 2022. Composition and Decomposition of Rhizoma Peanut (Arachis glabrata Benth.) Belowground Biomass. Scientific Reports (2022) 12:9967. https://doi.org/10.1038/s41598-022-14001-7
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Shepard, E.M., L.E. Sollenberger, M.M. Kohmann, L.S. da Silva, J.F. Harling, Jr., J.C.B. Dubeux, Jr., J.M.B. Vendramini. 2022. Establishing rhizoma peanut-bahiagrass mixtures. Agrosystems, Geosciences & Environment 2022;5:e20285 https://doi.org/10.1002/agg2.20285
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Silva, L.S., L.E. Sollenberger, M.K. Mullenix, M.M. Kohmann, J.C.B. Dubeux Jr., M.L. Silveira. 2022. Soil carbon and nitrogen stocks in nitrogen-fertilized grass and legume-grass forage systems. Nutr. Cycl. Agroecosyst. 122:105-117. https://doi.org/10.1007/s10705-021-10188-9
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Vasco, A.C.C.M., J.C.B. Dubeux, Jr., A.M. Arias-Esquivel, L.K. Warren, C.L. Wickens. 2022. Feeding behavior and preference of horses fed rhizoma peanut hay. Journal of Veterinary Behavior 47:35-44. https://doi.org/10.1016/j/jveb.2021.09.011
    • Type: Journal Articles Status: Published Year Published: 2022 Citation: Ferreira, I.M., B.G.C. Homem, I.B.G. de Lima, J.C.B. Dubeux Jr., T.F. Bernardes, M.A.C. Dan�s, D.R. Casagrande. 2022. Twenty-five-centimeter pre-grazing canopy height in palisade grass and forage peanut. Sci. Agric. v.79, n.2, e20200090, 2022. http://doi.org/10.1590/1678-992X-2020-0090
    • Type: Theses/Dissertations Status: Published Year Published: 2022 Citation: Zhang K. 2022. Long-Term Impacts of Sod-Based Rotation on Belowground Communities, Biologically-Mediated Nutrient Cycling, and Crop Productivity (Doctoral dissertation, SWES, University of Florida).
    • Type: Theses/Dissertations Status: Published Year Published: 2022 Citation: Mendez V. 2022. Biotic and Abiotic Factors Affect the Phyllosphere Mycobiome and Mycotoxin Concentrations of Warm-Season Pasture Grasses (Master Thesis, SWES, University of Florida)
    • Type: Theses/Dissertations Status: Published Year Published: 2022 Citation: Thomas K. 2022. Factors Affecting the Production, Performance, and Use of Ornamental Rhizoma Peanut, Arachis glabrata Benth., in Florida (Master Thesis, Agronomy Department, University of Florida)
    • Type: Theses/Dissertations Status: Published Year Published: 2022 Citation: Harlow L. 2022. Pasture decline in the southeast United States (Technical report requirement for MS professional degree, SWES department, University of Florida)
    • Type: Other Status: Published Year Published: 2022 Citation: Mackowiak, C. June 24, 2022. Impact of limiting or eliminating fertilization of bahiagrass pastures. Panhandle Agricultural eNews. https://nwdistrict.ifas.ufl.edu/phag/2022/06/24/impact-of-limiting-or-eliminating-fertilization-of-bahiagrass-pastures/
    • Type: Other Status: Published Year Published: 2022 Citation: Harlow, L. and Mackowiak, C.L. 2022. Pasture health and decline: Producer perspective from a state-wide survey. The Florida Cattlemen and Livestock Journal. 88(3).


    Progress 04/01/21 to 03/31/22

    Outputs
    Target Audience:This project involves in training the graduate students, biologists and visiting scientists to conduct the research proposed in this study, including: A. The extension trainings led by the project PD and co-PDs and the target audience: (a) Scope: Train the students and scientists to deliver the research and application updates to Perennial Peanut Producer's Association and Florida Cattlemen's Association Target trainees: José D. Pereira Neto (sandwich PhD student); Daniele Loures (visiting faculty) Luana Dantas (MS student); Martin Ruiz-Moreno (Biological scientist), Victor Guerra (PhD student); Jung-Chen Liu (PhD student); Adesuwa Erhunmwunse (PhD student); Kelly Thomas (MS student). Target audience: Perennial Peanut Producers Association; Florida Cattlemen Association (b) Presentation and handout production (12/08/2021) at Cover Crop Field Day, NFREC, Suwanee Valley. topic: Cover crops help microbes build soil C and benefit cash crops Scope: We delivered presentation and the handout to help the audience identify the role of cover crop play on soil health and crop production and consider integrate the knowledge gained from this topic into their management for their cropping system. Target audience: Extension agents, growers and the general public. (c) Lead and conduct presentations at North Florida Beef and Forage In-service training (FBF-IST) (04/08/2021). Topics: Perennial peanut cultivars, cultivar establishment differences, soil health impacts, mycotoxins in forage grasses Scope: FBF-IST is an annual, day-long event to deliver the latest research and application update through presentations and field tours. This training helps prepare Florida extension faculty and agents for client problems and to provide information for their own client training. Target audience: Florida state and county extension faculty (d) Lead and conduct the lectures for the soil Health In-service training (IST) Date of IST: 02/22/2021 Hands-on exercises: Mar-April 2021 Round-table discussion: 09/17/2021 Title: Soil Health-Banking soil carbon to benefit crop and pasture production system Scope: The Soil Health In-service training is an annual, day-long event to disseminate through presentations, hands on training, and facilitated discussions on the latest Ag-related research updates, particularly addressing soil health indicators in pasture and other agricultural systems from a soil physical, chemical, and biological perspective. This training is to help prepare Florida extension faculty and agents for client problems and to provide information for their own client training. Target audience: Florida state and county extension faculty (e) Presentation at Perennial Peanut Producers Association meeting (PPPA) (12/10/2021) and American Forage and Grassland Council Virtual Conference, (01/12/2021) Scope: We delivered the presentation to help the audience identify the role of grass species and perennial peanut play on soil health and consider integrating the knowledge gained from this topic into their management for their pastures system. Knowledge and research outcomes regarding association of grass endophytes and mycotoxin production were also delivered. A update on the soil health and environmental benefits of mixed grass-perennial peanut for low-input turf was presented. The goal is to that promotes the profitability and sustainability of grasslands through the interaction and sharing of related research, to improve the quality and quantity of forages used as hay and in pastures. Target audience: Academic community, extension agents, producers, private industry reps, turfgrass industry reps (f) One-on-one visits with the producers in May, July, and October 2021 Scope: We had a one-on-one visit with representative producers across Florida to discuss our research updates and future plans. We plan to accompany our future visits with questionnaires to identify the practices adopted by the producers on their farms and to measure the progress and quality of our extension activities to enable us to direct our research questions to their specific problems. Target audience: Forage Producers in Florida (g) Presented research plot tour at North Florida Beef and Forage Field Day (09/30/21) Scope: An eighteen-month event (alternates spring/fall) to present data and demonstrations on livestock and forages. The field tour covered three different perennial peanut varieties along with three bahiagrass varieties grown in monoculture and as mixtures, with focus on establishment comparisons and productivity. ?Target audience: Forage producers and state stakeholders in the tri-state (Florida, Geogia, and Alabama) region B. This project involves in the training of graduate students, biologists and visiting scientists to conduct research and extension proposed in this study. Target trainees: Adesuwa Erhunmwunse (Ph.D. student, supervisor: Hui-Ling Liao) Adesuwa is the key personnel for this project. She is an underrepresented scientist (an international scientist from Nigeria) Luana Dantas Queiroz (M.S. student in this project, graduated in summer 2021, supervisor: Dubeux) Jung-Chen Liu (M.S. student in this project, graduated in summer 2021, supervisor: Mackowiak) Victor Guerra (NNF Ph.D. candidate, graduated in summer 2021, supervisor: Mackowiak) Valerie Mendez (M.S. student, supervisor: Liao) Kaile Zhang (Ph.D. candidate, supervisor: Liao) Ko-Hsuan Chen (Postdoc, supervisor: Liao) Vijay Verma (Biological Scientist, supervisor: Liao) Martin Ruiz-Moreno (Biological scientist, supervisor: Dubeux) George Anguelov (Biological Scientist, supervisor: Mackowiak) Kelly Thomas (MS student, supervisor: Blount) José D. Pereira Neto (sandwich PhD student, supervisor: Dubeux) Daniele Loures (visiting faculty, supervisor: Dubeux) Pamella Sena (Exchange student, supervisor: Liao and Mackowiak) Changes/Problems:No major changes to report What opportunities for training and professional development has the project provided?As indicated in the "target audience" section, the project has provided opportunities to train graduate students and student interns. Data generated in the project have been used to provide training and education outreach to extension agents and producers. Through the supervision and training activities, the graduate trainees are expected to receive the professional skills on molecular/biogeochemical technologies and bioinformatics. The professorial training for the graduate students and postdoctoral scientists includes research proposal development, experimental designs, sample collection/processing, data collection/analysis, data integration, scientific presentations, coordination for team meetings, workshop/field day/in-service training participation, and leading the manuscript preparation for his/her projects. All the trainees have been actively involved in scientific presentations and extension activities, with the evidence shown in the sections of "Product" and "Other products". (1) Adesuwa Erhunmwunse (Ph.D. student, supervisor: Liao): The project that Adesuwa conducts aims at understanding the interactive effects of multiple management (plant community, grazing, and environmental variables) on microbial-driven soil fertility and forage production. Adesuwa will train the master student (e.g., Jaime Garzon Alfonso) on the molecular work that is associated with the forage project (e.g., soil DNA extraction, qPCR analysis). (2) Luana D Queiroz (M.S. student, supervisor: Dubeux). Luana leads the greenhouse and on-farm studies, including sample collections, data analysis and paper publication to study the nitrogen movement as affected by forage species. Luana completed her MS program and thesis publication in summer 2021. (3) Jung-Chen Liu (M.S. student, supervisor: Mackowiak). JC leads the field experiment and on-farm studies, including sample collections, data analysis and paper publication. Her study aims at understanding the effects of BG, RP and soil chemistry on forage productivity. JC completed her MS program and thesis publication in summer 2021. (4) Victor Guerra (Ph.D. candidate, supervisor, Mackowiak): Victor's study aims at identifying the microbial community in associated with BG-RP interaction in comparison with N fertilizer. Victor also studied the potential function of arbuscular mycorrhizal fungi living in BG vs. RP. His effort has led two paper publication. Victor completed his Ph.D. program and dissertation publication in summer 2021. (5) Kaile Zhang (Ph.D. candidate, supervisor: Liao): Kaile's project focuses on identifying the dominant fungal taxa presenting in the pastures that provide the beneficial effect across different cropping systems, including bahiagrass and RP. The outcome of the training had led the publications of one short research note, one review article, and two research papers. (6) Ko-Hsuan Chen (Postdoc, supervisor: Liao): Ko-Hsuan developed the 2-step in-house PCR protocol to generate the DNA library for amplicon sequencing in this project. The tech report was published in 2021 at Agronomy Journal. (7) Valerie Mendez (M.S. student, supervisor: Liao): Valerie is involved in molecular work that are associated with forage project, including DNA extraction, 2- step PCR library construction, DNA purification quantification and Qiime 2 associated data analysis. (8) Other students and scientists assisted on field sampling and laboratory analyses, including Vijay Verma (Biological Scientist, supervisor: Liao), Martin Ruiz-Moreno (Biological scientist, supervisor: Dubeux), George Anguelov (Biological Scientist, supervisor: Mackowiak), Kelly Thomas (MS student, supervisor: Blount), José D. Pereira Neto (sandwich PhD student, supervisor: Dubeux), Daniele Loures (visiting faculty, supervisor: Dubeux), Pamella Sena (Exchange student, supervisor: Liao and Mackowiak) How have the results been disseminated to communities of interest?We have been using a combination of publications, in-person meetings and social media as our methods for disseminating information (Please find the report from previous Products section). In addition, we highlighted aspects of our projects using various platforms, including publications, webinars, twitter, facebook, and instagram. Our clientele has been receptive for that form of dissemination, and the information reaches a broader audience among stockholders and general public. As mentioned in the "Target Audience section", the highlighted activities we performed in 2021 include: (a) Deliver the research and application update to Perennial Peanut Producer's Association and Florida Cattlemen's Association (b) Presentation and handout production (12/08/2021) to extension agents, growers and the general public at Cover Crop Field Day, NFREC, Suwanee Valley. Topic: Cover crops help microbes build soil C and benefit cash crops (c) Lead and conduct presentations at North Florida Beef and Forage In-service training (FBF-IST) for Florida state and county extension faculty (04/08/2021). Topics: Perennial peanut cultivars, cultivar establishment differences, soil health impacts, mycotoxins in forage grasses (d) Lead and conduct the lectures for the soil Health In-service training (IST) that target Florida state and county extension faculty and followed by training extension faculty on hands-on exercises to identify soil health indicators and conducted round-table discussion to train the extension faculty on data interpretation of these indicators. (e) Conduct presentation at Perennial Peanut Producers Association meeting (PPPA)(02/10/2021) to help the audience identify the role of grass species and perennial peanut play on soil health and consider integrate the knowledge gained from this topic into their management for their pastures system. (f) Lead poster presentation at the American Forage and Grassland Council Virtual Conference, (01/12/2021) that target academic community, extension agents, producers, private industries, institutes and foundation in America. (g) One-on-one visits with the producers in May, July, and October 2021: We had a one-on-one visit with representative producers across Florida to discuss our research updates and plans. What do you plan to do during the next reporting period to accomplish the goals?We will finish the pending greenhouse exp and laboratory analyses and we will be able to highlight some of the results in Perennial Peanut Producers annual Field Day in 2022, an upcoming Beef and Forage field day in fall, 2022 and the Soil health In-service training 2022. Additionally, we will continue to develop educational materials and publish extension articles, along with attending scientific conferences to present research updates and preparing peer-reviewed manuscripts. For example, some of the manuscript drafts that are in preparation, include: Spatial and seasonal variations on soil microbial communities and determining the correlation between soil chemical properties and soil microbial communities across Florida. Comparing the mixture of bahiagrass and different rhizoma peanut cultivars and their monoculture on soil microbial communities and diversity, enzymes and functional genes involved in N cycling (nitrogen fixation, nitrification, and denitrification). Incorporating rhizoma peanut increases nutrient uptake in bahiagrass forage systems. Rhizoma peanut defoliation frequency impacts herbage responses and soil health indicators. Water limitation impact on bahiagrass-rhizoma peanut mixtures under low soil N fertility.

    Impacts
    What was accomplished under these goals? Objective 1 (1) Greenhouse experiment (1.1) We installed an H-pot trial to assess the neighboring effect of Argentine bahiagrass and Ecoturf rhizoma peanut. We tracked N transfer using enriched 15N gas directly injected in the root zone. Data indicated belowground N transfer using stable isotope technique. Data was presented in the National Meeting (ASA-CSSA-SSSA) in 2021. Samples (root, rhizosphere soils,bulk soils) were also collected from all pots to study the bacterial and fungal communities. (1.2) A greenhouse experiment isinitiated to identify microbial-mediated strategies for nutrient uptake, especially phosphorus, in bahiagrass (BG) and rhizoma peanut (RP) mixture. This objective will be achieved by comparing the microbial communities and gene expression in the different belowground plant parts of BG (root+rhizome) and RP (roots+ rhizome+ nodules) in BGmonoculture versus BG and RP mixture under nutrient (P) limitation and adequate nutrient. (2) Experimental field site experiment (newly established site in 2019) We collected establishment data of a BG and RP cultivar mixture trial. There are three BG cultivars grown in monoculture or as binary mixtures with three RP cultivars, and these were also grown as monocultures. Forage yield and composition data were collected in 2020 and 2021, and data are being analyzed. We also collected bulk soil samples in 2020 and 2021. A goal was to determine the effect of cultivars and seasonal variations on soil microbial communities and diversity, enzymes and functional genes involved in N cycling (N fixation, nitrification, and denitrification). Soil samples collected from Argentine BG cultivar and Florigraze and Ecoturf RP (monoculture and mixture of each entry) listed above were processed in Summer 2021. We finished the amplicon library preparation as well as sequencing for 16S and ITS analysis. The bioinformatic analysis has been completed. We also measured soil extracellular enzyme activities N-acetyl-B-D-glucosamidase and leucine aminopeptidase involved in C and N cycling. We are quantifying the N cycling genes involved in nitrification, denitrification, and N fixation for these soil samples. The manuscript for this topic is under preparation. In the preliminary data, we observed that fungal communities were more responsive to forage treatments than bacterial communities. Fungal communities in the soils of BG monoculture or its mixture with RP (Argentine BG monoculture, mixture of BG and Ecoturf RP, and mixture of BG and Florigraze RP) were shifted compared the fungal community of RP monoculture soil (Ecoturf RP, and Florigraze RP). Bradyrhizobium was the main N fixing bacterial group and Nitrospira was the nitrifying bacterial group identified in our system. The data from this study was published in the NFREC-100th year Anniversary Graduate Student Poster Competition. (3) Experimental field site experiment (the long-term established sites): We collected root and soil samples on through 2021 from the BG and RP plots that were established a decade ago (at NFREC, Quincy) to determine the long-term effects of RP and BG mixture and soil depths onsoil bulk density, root length, root diameter, root surface area, and root C and N; plant tissue C and N; the gene copies of total bacteria, Actinobacteria, Alphaproteobacteria, Bacteroidetes, Betaproteobacteria, and Firmicutes as well as soil bacterial and fungal communities. Briefly, the DNA from the soil samples collected in (2) were carried out to generate amplicon sequencing targeted bacterial16S and fungal ITS. The chemical properties and root parameters have been completed and written as a chapter in a dissertation by Guerra et al. (2021). The forage productivity, nutrient uptake and impact on some soil health indicators were assessed and written as chapters in a thesis by Liu et al. (2021). The sequences for the bacterial and fungal communities have been received. We have begun analyzing the data and writing the manuscripts for these research topics. Objective 2. (1) To identify the co-relation among microbial community soil chemistry and environmental variables, we collected bulk soil in May and July 2019 across the three on-farm sites (North, Central, and South FL) and measured soil chemicals (soil pH, total C/N, P, K, Mg, Ca, and micronutrients). We collected more samples (root, rhizosphere, and bulk soil) in July, and October 2020 and 2021 Soil moisture and temperature sensors as well as rain gauges were installed in June across the three sites in 2020 and 2021. We processed the plant and soil samples collected from fields in 2019 and are completed the microbial analysis. The DNA was extracted from the bulk soils collected in May 14 -21 and July 22-26, 2019 from North, Central, and South FL (288 samples). The amplicon libraries targeted fungal ITS and bacterial 16S were sequenced. The DNA extraction and molecular analysis of the root, rhizosphere, and bulk soil collected in 2020 and 2021 are in progress. Data analysis: OTU tables were generated from the sequence reads in Qiime2. The statistical analysis of the chemical and OTU tables were carried out using R software. The manuscript for the data collected in 2019 (correlation between soil chemical properties and soil microbial communities) is near completion. The results showed that integration of RP into BG can enhance bacterial diversity and shift microbial community. However, we didn't observe such changes across all the sites. The statistical analysis showed that the sampling location and seasons can compromise the forage treatments in terms of regulating microbial diversity and community. In addition, Bacillus and Rhodoplanes were the major bacterial groups across all site locations in Florida (North, Central, and South). The fungal groups varied across different site locations. Fusarium, Epicoccum, and Pseudothielavia were dominant in North FL. Penicillium was prevalent in Central FL and Fusarium and Trichoderma was dominant in South FL. Fusarium was enriched in the presence of RP only in North and South Florida. Soil pH was a major contributing factor to changes in soil microbial communities in Florida. These results were presented in the 2021 ASA/CSSA/SSSA International Meeting. The soil moisture and temperature data generated from the sensors collected from the three on-farm sites in 2020 were graphed. The soils in Central FL (0.12 m2/m2) were drier than the soils in North (0.16 m2/m2) and South FL (0.19 m2/m2), while the average soils temperature were in the following order (Central FL (30.2oC > South FL (29.5oC) > North FL (28oC)). The environmental data from the sensors in 2021 are currently analyzed. We wrapped up the on-farm experiments and removed all the environmental sensors from the field. By the end of this year, all laboratory and molecular analysis for the bulk soil, rhizosphere, and roots should be completed and the results will be reported in our next report. (2) For biogeochemical analysis, we collected the samples from three on-farm sites as described in "(1)" across two years (2019, 2020) from the three on-farm sites. Data collected included aboveground biomass, belowground biomass, greenhouse gas emission from soil, biological N2 fixation, nutritive value of forages. In 2021, we finalized data collection and laboratory analyzes including aboveground biomass, belowground biomass, greenhouse gas emission from soil, biological N2 fixation, nutritive value of forages. The data was published by Luana Dantas in her MS thesis (Entitled "Performance and N Dynamics in Grass-Legume Systems in FL"). The MS student graduated in Summer 2021 and is working on the manuscripts. In addition, soils were collected from these three on-farm sites, as well to assess hydraulic conductivity (saturated and unsaturated). We found one location seems to have a hydrophobic surface soil. We will investigate further and perform tests to determine the degree of water repellency.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Garcia, L., J.C.B. Dubeux, Jr., L.E. Sollenberger, J.M.B. Vendramini, N. DiLorenzo, E.R.S. SantosG, D.M. JaramilloG, M. Ruiz-Moreno. 2021. Nutrient excretion from cattle grazing nitrogen-fertilized grass or grass-legume pastures. Agronomy Journal 113:3110-3123. https://doi.org/10.1002/agj2.20675
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Santos, E.R.S., J.C.B. Dubeux, Jr., C. Mackowiak, A.R.S. Blount, L. Sollenberger, D.M. Jaramillo G, L. GarciaG, D. AbreuG, R.T. Souza, M. Ruiz-Moreno. 2021. Herbage responses and nitrogen agronomic efficiency of bermudagrass-legume mixtures. Crop Sci. 61:3815-3829. https://doi.org/10.1002/csc2.20552
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Jaramillo, D.M.G, J.C.B. Dubeux Jr., L. Sollenberger, C. Mackowiak, J.M.B. Vendramini, N. DiLorenzo, L.M.D. QueirozG, E.R.S. SantosG, L. Garcia, M. Ruiz-Moreno, E. van Santen. 2021. Litter Mass, Deposition Rate, and Decomposition in N-Fertilized or Grass-Legume Grazing Systems. Crop Science https://10.1002/csc2.20475 61:2176-2189.
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Aryal, P., L.E. Sollenberger, M.M. Khomann, L.S. da Silva, E.M. Shepard, K.D. Cooley, D.L. Rowland, and J.C.B. Dubeux, Jr. 2021. Plant Growth Habit and Nitrogen Fertilizer Effects on Rhizoma Peanut Biomass Partitioning During Establishment. Grass & Forage Sci. 76:485-493 https://doi.org/10.1111/gfs.12519
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Silva, L.S., L.E. Sollenberger, M.K. Mullenix, M.M. Kohmann, J.C.B. Dubeux Jr., M.L. Silveira. 2022. Soil carbon and nitrogen stocks in nitrogen-fertilized grass and legume-grass forage systems. Nutr. Cycl. Agroecosyst. 122:105-117. https://doi.org/10.1007/s10705-021-10188-9
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Vasco, A.C.C.M., J.C.B. Dubeux, Jr., A.M. Arias-Esquivel, L.K. Warren, C.L. Wickens. 2022. Feeding behavior and preference of horses fed rhizoma peanut hay. Journal of Veterinary Behavior 47:35-44. https://doi.org/10.1016/j/jveb.2021.09.011
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Vasco, A.C.C.M., J.M. Bobel, J.C.B. Dubeux Jr., L.K. Warren, C.L. Wickens. 2021. Digestibility and nitrogen and water balance in horses fed rhizoma peanut hay. Journal of Animal Science, 2021; skab284. https://doi.org/10.1093/jas/skab284
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Jaramillo, D., H. Sheridan, K.J. Soder, J.C.B. Dubeux, Jr. 2021. Enhancing the sustainability of temperate pasture systems through more diverse swards. Agronomy 11:1912 https://www.mdpi.com/2073-4395/11/10/1912
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Silva, L.S., L.E. Sollenberger, M.M. Kohmann, J.C.B. Dubeux Jr., P. Aryal, M.L. Silveira, J.M.B. Vendramini. 2021. Existing litter mass and N Disappearance on Year-round N-fertilized Grass and Legume-based Forage Systems. Agronomy J. 113:5170-5182 https://doi.org/10.1002/agj2.20826
    • Type: Theses/Dissertations Status: Published Year Published: 2021 Citation: Guerra VA. 2021. Soil Microbial and Ecophysiological Aspects of Rhizoma Peanut Inclusion in Bahiagrass Pastures (Doctoral dissertation, SWSD, University of Florida).
    • Type: Theses/Dissertations Status: Published Year Published: 2021 Citation: Queiroz LMD. 2021. Performance and Nitrogen Dynamics in Grass-Legume Systems in Florida. (Master Thesis, Agronomy Department, UF IFAS, University of Florida).
    • Type: Theses/Dissertations Status: Published Year Published: 2021 Citation: Liu JC. 2021. Rhizoma Peanut (Arachis Glabrata Benth.) Mixture with Bahiagrass (Paspalum Notatum Fl�gge) Impacts on Soil Health. (Master Thesis, SWSD, University of Florida).
    • Type: Other Status: Published Year Published: 2021 Citation: Dubeux, JCB Jr, Garcia L, Jaramillo D, Santos E, van Cleef F, Queiroz L, Garcia C, Abreu D, Ruiz-Moreno M. 2021. Integrating perennial peanut into grazing operations. Available at Integrating Perennial Peanut into Grazing Operations | Panhandle Agriculture (ufl.edu)
    • Type: Other Status: Published Year Published: 2021 Citation: Liao, H-L 2021. The plant-growth promoting fungus, Mortierella elongata, its biology, ecological distribution and growth promoting activities. EDIS. https://edis.ifas.ufl.edu/ss679
    • Type: Other Status: Published Year Published: 2021 Citation: Mackowiak, C.L. 2021. Managing pastures and hay fields around high fertilizer prices. The Florida Cattlemen and Livestock Journal. Oct 856(1).
    • Type: Other Status: Published Year Published: 2021 Citation: Mackowiak, C.L., A.R. Blount, H.L. Liao, and JCB Dubeux. 2021. Lessons learned: Perennial peanut x bahiagrass variety mixes. 30 Sep, Beef Forage Field Day, Marianna, FL 6 pp.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Liu JC, Mackowiak CL, Dubeux JCB, Liao H-L, Blount AR. 2021. Yield gains by incorporating rhizome peanut into bahiagrass and its impact on soil fertility distribution. AFGC (American Forage & Grassland Council) 2021 Conference, Savannah, GA (Abstract and Poster Presentation, Virtual)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Harlow L, Mackowiak CL, Blount AR, Paret M, Liao H-L. 2021. Addressing the status of pasture health through producer needs assessment. AFGC (American Forage & Grassland Council) 2021 Conference, Savannah, GA (Abstract and Poster Presentation, Virtual)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Erhunmwunse AS, Mackowiak C, Blount ARS, Dubeux JCB, Liao H-L. 2021. Effects of forage species combination, temporal, and regional spatial variations on soil microbial community. ASA, CSSA, and SSSA International Annual Meeting, Salt Lake City, UT, USA (Abstract, Oral and poster concurrent presentation)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Zhang K, Maltais-Landry G, Grabau Z, George S, Wright D, Small I, Liao H-L. 2021. Long-term sod-based rotation promotes crop yield by manipulating root microbiomes. ASA, CSSA, and SSSA International Annual Meeting, Salt Lake City, UT, USA. (Abstract, Oral and poster concurrent presentation) (Abstract: https://scisoc.confex.com/crops/2021am/s3/papers/viewonly.cgi?password=964023&username=133744)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Mendez V, Chen K-H, Blount A, Mackowiak CL, Duringer J, Liao H-L. 2021. The Foliar Mycobiome and Associated Mycotoxins of Warm Season Forages. ASA, CSSA, and SSSA International Annual Meeting, Salt Lake City, UT, USA (Poster presentation) (Abstract: https://scisoc.confex.com/scisoc/2021am/meetingapp.cgi/Paper/137720)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Erhunmwunse AS, Quieroz LD, Mackowiak C, Dubeux Jr JCB, Blount ARS, Liao H-L. 2021. Effects of defoliation and perennial peanut on phosphorus and micronutrients in bahiagrass system. American Forage & Grassland Council (AFGC) 2021 Conference, Savannah, GA (Abstract and Virtual attended, Poster Presentation)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., L.E. Sollenberger, J.M.B. Vendramini, H.L. Liao, D.R. Casagrande, D.S. Abreu, D.M Jaramillo, E.R.S. Santos, C.C. Garcia, M. Ruiz-Moreno. 2021. Tracking nitrogen transfer in grass-legume system using enriched 15N2 during the warm season. ASA/CSSA/SSSA International Annual Meeting Abstracts, Salt Lake City, UT.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., L.E. Sollenberger, J.M.B. Vendramini, H.L. Liao, D.M. Jaramillo, E.R.S. Santos, H.M.S. Silva, L. Garcia, D.R.S Loures, M. Ruiz-Moreno. 2021.Belowground responses of rhizoma peanut-bahiagrass mixtures across three on-farm locations. ASA/CSSA/SSSA International Annual Meeting Abstracts, Salt Lake City, UT.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Liu JC, Mackowiak CL, Blount A, Dubeux, Jr JCB, Liao H-L. 2021. Rhizoma peanut defoliation frequency impacts on nutrient partitioning and soil health. ASA/CSSA/SSSA International Annual Meeting Abstracts, Salt Lake City, UT.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: DiLorenzo, N., J.C.B. Dubeux Jr, L. Garcia, R.D. Guevara, S. Lagrange, J. MacAdam, and J.J. Villalba. 2021. Legumes as a Strategy for Reducing Greenhouse Gas Emissions of Forage-livestock Systems. XXIV International Grassland Congress/XI International Rangeland Congress, Nairobi, Kenya, 2021. Available at XXIV International Grassland Congress /
      XI International Rangeland Congress | International Grassland Congress Proceedings (uky.edu)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Garcia, L., D.M. Jaramillo, J.C.B. Dubeux Jr., L.E. Sollenberger, J.M.B. Vendramini, N. DiLorenzo, E.R.S. Santos, M. Ruiz-Moreno, L.M.D. Queiroz. 2021. Nutrient return from plant litter and cattle excretion grazing on N-fertilized grass or grass-legume pastures in North Florida. XXIV International Grassland Congress/XI International Rangeland Congress, Nairobi, Kenya, 2021. Available at XXIV International Grassland Congress /
      XI International Rangeland Congress | International Grassland Congress Proceedings (uky.edu)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Assis, GML, J.C.B. Dubeux Jr. 2021. Warm-season legumes  Challenges and constraints to adapting warm-season legumes to transition zone climates with examples from Arachis. XXIV International Grassland Congress/XI International Rangeland Congress, Nairobi, Kenya, 2021. Available at XXIV International Grassland Congress /
      XI International Rangeland Congress | International Grassland Congress Proceedings (uky.edu)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Dubeux, J.C.B., Jr. and L.E. Sollenberger. 2021. Sustainable Intensification of Livestock Systems Using Forage Legumes. International Grassland Congress/XI International Rangeland Congress, Nairobi, Kenya, 2021. Available at XXIV International Grassland Congress /
      XI International Rangeland Congress | International Grassland Congress Proceedings (uky.edu)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Jaramillo, D.M., J.C.B. Dubeux, Jr., L.M.D. Queiroz, L. Garcia, E.R.S. Santos. 2021. Herbage and livestock responses for N-fertilized and grass-legume grazing systems. XXIV International Grassland Congress/XI International Rangeland Congress, Nairobi, Kenya, 2021. Available at XXIV International Grassland Congress /
      XI International Rangeland Congress | International Grassland Congress Proceedings (uky.edu)
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Guerra V, Beule L, Mackowiak C, Dubeux Jr JCB, Blount A, Rowland D, Wang X-B, Rowland DL, Liao H-L. 2021. Soil Bacterial Community Response to Rhizoma Peanut Incorporation into Florida Pastures. J Environmental Qual. doi.org/10.1002/jeq2.20307
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Zhang K, Schumacher L, Maltais-Landry G, Grabau Z, George S, Wright D, Small I, Liao H-L. 2021. Integrating perennial bahiagrass into the conventional rotation of cotton and peanut enhances interactions between microbial and nematode communities. Applied Soil Ecology. 170:104254. https://doi.org/10.1016/j.apsoil.2021.104254
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Zhang K, Maltais-Landry G, Liao H-L. 2021. How biota regulates C cycling underlying crop rotations. Soil Biology and Biochemistry. 156:108219. doi.org/10.1016/j.soilbio.2021.108219
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Chen K-H, Longley R, Bonito G, Liao H-L. 2021. A Two-Step PCR Protocol Enabling Flexible Primer Choice and High Sequencing Yield for Illumina MiSeq Meta-Barcoding. Agronomy. 11:1274. https://doi.org/10.3390/agronomy11071274
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Zhang K, Maltais-Landry G, George S, Grabau Z, Small I, Wright D, Liao H-L. 2022. Long-term sod-based rotation promotes beneficial root microbiomes and increases crop productivity. Biology and Fertility of Soils. https://doi.org/10.1007/s00374-022-01626-z
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Jaramillo DM, Dubeux Jr JCB, Sollenberger LE, Vendramini JMB, Mackowiak CL, DiLorenzo N, Garcia L, Queiroz LMD, Santos ERS, Homem BGC, van Cleef F, Ruiz-Moreno M. 2021. Water Footprint, Herbage, and Livestock Responses for N-Fertilized Grass and Grass-Legume Grazing Systems. Crop Sci. 61:3844-3858. http://doi.org/10.1002/csc2.20568


    Progress 04/01/20 to 03/31/21

    Outputs
    Target Audience:A. The extension trainings led by the project PD and co-PDs and the target audience: (a) Lead the soil Health In-service training (02/22/2021) Title: Soil Health-Banking soil carbon to benefit crop and pasture production system Scope: The Soil Health In-service training is an annual, day-long event to disseminate through presentations, hands on training, and facilitated discussions on the latest Ag-related research updates, particularly addressing soil health indicators in pasture and other agricultural systems from a soil physical, chemical, and biological perspective. This training is to help prepare Florida extension faculty and agents for client problems and to provide information for their own client training. Target audience: Florida state and county extension faculty (b) Lead poster presentation at the American Forage and Grassland Council Virtual Conference, (01/12/2021) Scope: An annual event that promotes the profitability and sustainability of forage through the interaction and sharing of research carried out on forage and grasslands to improve the quality and quantity of hays, pastures, and ornamental landscapes. We participated in the conference by sharing our research findings through a poster. Target audience: Academic community, extension agents, producers, private industry, institutes and foundation in America. (c) Presentation at Extension Professional Associations of Florida Virtual Conference (09/01/2020) Scope: The annual conference creates a platform for extension agents across Florida and faculty members to share the innovative activities they have carried out statewide to improve extension activities. We submitted an abstract and presented the research and extension activities for our forage project. Target audience: Florida state and county extension agents and academic communities. (d) Lead and presentation for Virtual Perennial Peanut Field Day (NFREC), (06/25/2020) Scope: Annual day-long event to promote interaction and sharing of research with membership and to promote the application of different perennial peanut species in hay, pasture, and ornamental landscapes. We presented our research projects to the Perennial Peanut Producers and the general public through a video presentation. Here is the link https://www.youtube.com/watch?v=H7a7dS2FcR4&t=37s Target audience: Perennial Peanut Producers Association (PPPA) members and the general public. (e) One-on-one visits with the producers in May, July, and October 2020 Scope: We had a one-on-one visit with representative producers across Florida to discuss our research updates and future plans. We plan to accompany our future visits with questionnaires to identify the practices adopted by the producers on their farms and to measure the progress and quality of our extension activities to enable us to direct our research questions to their specific problems. Target audience: Forage Producers in Florida (f) Presentation for the Webinar of Master Gardeners (05/20/2020) Scope: We educated the master gardeners on the important roles of microorganisms in soil and plant health. Target audience: Master Gardeners in Florida. (g) Presentation at Sod-Based Rotation Field day (Virtual). Scope: We provided the video presentation to help the audience identify the role of cover crop (mainly grass) play on soil health and consider integrate the knowledge gained from this topic into their management for rotation system. (The recorded talk: https://www.youtube.com/channel/UCsx3KN2Iu435bykZotUTUcA) Target audience: Extension agents, growers and the general public. (h) Presentation at Southern Pasture Forage Crop Improvement Conference (SPFCIC) and extension article publication. Target audience/group: SPFCIC members, Perennial Peanut Producer's Association, Florida Department of Agriculture and Consumer Services (FDACS), USDA-NRCS, County Extension Agents, regional forage producers and ranchers, and general public. (i) Host the visit of 25 FFA students from Gulf County visited NFREC Marianna and our research plots. (03/06/2020) Scope: We discussed our research plots at NFREC and discussed with them the importance of grasslands and livestock in Florida, demonstrating environmental and productive aspects. Target audience: FFA students B. This project involves in the training of graduate students, biologists and visiting scientists to conduct research and extension proposed in this study. Target trainees: Adesuwa Erhunmwunse (Ph.D. student, supervisor: Hui-Ling Liao) Adesuwa is the key personnel for this project. She is an under representative scientist (an international scientist from Nigeria); Luana Dantas Queiroz (M.S. student in this project, supervisor: Jose Dubeux); Jung-Chen Liu (M.S. student in this project, supervisor: Cheryl Mackowiak); Victor Guerra (NNF Ph.D. candidate, supervisor: Cheryl Mackowiak); Valerie Mendez (M.S. student, supervisor: Hui-Ling Liao); Kaile Zhang (Ph.D. candidate, supervisor: Hui-Ling Liao); Chih-Ming Hsu (volunteer scientist, supervisor: Hui-Ling Liao); Ko-Hsuan Chen (Postdoc, supervisor: Hui-Ling Liao); Daciele Abreu (sandwich MS student, supervisor: Jose Dubeux) Carlos Garcia (sandwich Ph.D. student, supervisor: Jose Dubeux); Martin Ruiz-Moreno (Biological scientist, supervisor Jose Dubeux); George Anguelov (Biological Scientist, supervisor Cheryl Mackowiak); Kelly Thomas (MS student, supervisor: Ann Blount) Changes/Problems:COVID-19 prevented us from organizing in-person meetings in 2020. We produced several videos and we believe we overcame most COVID-19 related setbacks. We expect the situation will improve in 2021 and that we will be able to have more in-person extension and field day meetings. What opportunities for training and professional development has the project provided?As mentioned in the "target audience" section, the project has provided opportunities to train graduate students and student interns. Data generated in the project have been used to provide training and education outreach to extension agents and producers. Through the supervision and training activities, the graduate trainees are expected to receive the professional skills on molecular/biogeochemical technologies and bioinformatics. The professorial training for the graduate students and postdoctoral scientists includes research proposal development, experimental designs, sample collection/processing, data collection/analysis, data integration, scientific presentations, coordination for team meetings, workshop/field day/in-service training participation, and leading the manuscript preparation for his/her projects. All of the trainees have been actively involved in scientific presentations and extension activities, with the evidence shown in the sections of "Product" and "Other products". (1) Adesuwa Erhunmwunse (Ph.D. student, supervisor: Liao): The project that Adesuwa conducts aims at understanding the interactive effects of multiple management (plant community, grazing, and environmental variables) on microbial-driven soil fertility and forage production. Adesuwa will train the master student (e.g., Valerie Mendez) on the molecular work that is associated with the forage project (e.g., soil and root DNA extraction). (2) Luana D Queiroz (M.S. student, supervisor: Dubeux). Luana leads the greenhouse and on-farm studies, including sample collections, data analysis and paper publication to study the nitrogen movement as affected by forage species. (3) Jung-Chen Liu (M.S. student, supervisor: Mackowiak). JC leads the field experiment and on-farm studies, including sample collections, data analysis and paper publication. Her study aims at understanding the effects of BG, RP and soil chemistry on forage productivity. (4) Victor Guerra (Ph.D. candidate, supervisor, Mackowiak): Victor's study aims at identifying the microbial community in associated with BG-RP interaction in comparison with N fertilizer. Victor also studied the potential function of arbuscular mycorrhizal fungi living in BG vs. RP. His effort has led one method paper publication and one research paper in review. (5) Kaile Zhang (Ph.D. candidate, supervisor: Liao): Kaile's project focuses on identifying the dominant fungal taxa presenting in the pastures that provide the beneficial effect across different cropping systems, including bahiagrass and RP. The outcome of the training had led a short research note published at Agronomy. Kaile also led the review article addressed the effect of rotation system (including bahiagrass-based rotation) on carbon dynamics. The manuscript has been accepted by Journal of Soil Biology and Biochemistry. (6) Chih-Ming Hsu (Volunteer Scientist, supervisor: Liao): Chih-Ming is involved in field sampling, sampling processing and molecular work for amplicon and RNA-based sequencing for this project. Mr. Hsu also trained the visiting scientists and graduate students on molecular techniques applied for this project (the trainees include, Erhunmwunse, Zhang and Mendez). The outcomes of this training led the contributions to several research papers for this project (Wang et al 2019; Beule et al 2019; Zhang et al, 2020) (7) Ko-Hsuan Chen (Postdoc, supervisor: Liao): Ko-Hsuan developed the 2-step in-house PCR protocol to generate the DNA library for amplicon sequencing in this project. The tech report is now in review (Agronomy). (8) Valerie Mendez (M.S. student, supervisor: Liao): Valerie is involved in molecular work that are associated with forage project, including DNA extraction, 2- step PCR library construction, DNA purification quantification and Qiime 2 associated data analysis. (9) Carlos Garcia (sandwich PhD student, supervisor: Dubeux); assisting on the field sampling and laboratory analyses (10) Daciele Abreu (sandwich MS student, supervisor: Dubeux). assisting on the field sampling and laboratory analyses (11) Martin Ruiz-Moreno (Biological scientist, supervisor Jose Dubeux); (12) George Anguelov (Biological Scientist, supervisor Cheryl Mackowiak); (13) Kelly Thomas (MS student, supervisor: Ann Blount) How have the results been disseminated to communities of interest?We have been using social media as our mainstream of dissemination (please see "Other Product" section for the list of social media we generated in 2020. We highlighted our projects in different platforms, including Twitter, Facebook, and Instagram. Our clientele has been receptive for that form of dissemination, and the information reaches a broader audience among producers. As mentioned in the "Target Audience section", the highlighted activities we performed in 2020 include: (a) Lead the Soil Health In-service training (02/22/2021) Title: Soil Health-Banking soil carbon to benefit crop and pasture production system Scope: The Soil Health In-service training disseminated the knowledge through presentations, hands on training, and facilitated discussions on the latest Ag-related research updates, particularly addressing soil health indicators in pasture and other agricultural systems. This training is to help prepare Florida extension faculty and agents for client problems and to provide information for their own client training. (b) Lead poster presentation at the American Forage and Grassland Council Virtual Conference, (01/12/2021). We participated in the conference with aiming at sharing of research carried out on forage to improve the quality and quantity of pastures. (c) Presentation at Extension Professional Associations of Florida Virtual Conference (09/01/2020) We presented the extension activities we performed and our future plan on extension for our forage project (d) Lead and presentation for Virtual Perennial Peanut Field Day (NFREC), (06/25/2020). We presented our research projects to the Perennial Peanut Producers and the general public through a video presentation. (e) One-on-one visits with the producers in May, July, and October 2020: We had a one-on-one visit with representative producers across Florida to discuss our research updates and future plans. (f) Presentation for the Webinar of Master Gardeners (05/20/2020): We educated the master gardeners on the important roles of microorganisms in soil and plant health. (g) Presentation at Sod-Based Rotation Field day (Virtual): We provided the video presentation to help the audience identify the role of cover crop (mainly grass) play on soil health and consider integrate the knowledge gained from this topic into their management for rotation system. (h) Presentation at Southern Pasture Forage Crop Improvement Conference (SPFCIC) and extension article publication. (i) Host the visit of 25 FFA students from Gulf County visited NFREC Marianna and our research plots. (03/06/2020): We discussed our research plots at NFREC and discussed with them the importance of grasslands and livestock in Florida, demonstrating environmental and productive aspects. What do you plan to do during the next reporting period to accomplish the goals?We will finish the pending laboratory analyses and we will be able to highlight some of the results in an extension in-service training in April 2021, Perennial Peanut Producers annual Field Day in May 2021, an upcoming Beef and Forage field day in October 2021 and the Soil health In-service training 2021.Additionally, we will continue to develop educational materials and publish extension articles, along with preparing peer-reviewed manuscripts.

    Impacts
    What was accomplished under these goals? For Objective 1. (1) Greenhouse experiment: We installed an H-pot trial to assess the neighboring effect of Argentine bahiagrass and Ecoturf rhizoma peanut. Briefly, H-pots built up with PVC pipes were set up to connect belowground tissues from BG and RP. The H-pot was planted with grass in one side and legume on the other side, connected by a PVC pipe so that the belowground transfer could be assessed. We tracked the N transfer using enriched 15N gas directly injected in the root zone. Data is being processed and will be presented in the National Meeting in 2021. Samples (root, rhizosphere soils, and bulk soils) were also collected from all pots in 07/28/2020 and 10/14/2020 to study the bacterial and fungal communities. (2) Experimental field site experiment (newly established site in 2019): Last year (2019) we established the research field site. Briefly, a split-plot design was used with BG main plots arranged in a randomized complete block design, and RP treatment as subplots (plot size 5 x 20 ft). The treatments include BG (Argentine, Pensacola, Tifton-9) and RP (Ecoturf, Florigraze, LU-1RP) monocultures and the mixtures of paired BG-RP across these cultivars. The research site is located at Marianna FL (30.871 N, -85.190 W). Three BG Flüeggé (bahiagrass) cultivars were established from seed in 2005 as randomized blocks. This year (year 2020), we collected forage yield and soil quality data with depth from established bahiagrass and perennial peanut monocultures and binary mixtures. Preliminary results were presented at ASA-CSA-SSSA International meetings, as a virtual poster in 2020 by Jung-Chen Liu, MS Student. She ranked 2nd place in the MS graduate student poster competition in the C-06 Division. We collected establishment data of a bahiagrass and perennial peanut cultivar mixture trial. There are three bahiagrass cultivars grown in monoculture or as binary mixtures with three perennial peanut cultivars, and these were also grown as monocultures. Forage yield and composition data were collected and data being analyzed. We also collected bulk soil samples from the Argentine bahiagrass cultivar and the Florigraze and Ecoturf rhizoma peanut cultivars on 3/9/2020 and 10/16/2020, from the site that was established in 2019, to determine the effect of cultivars and seasonal variations on soil microbial communities and diversity, enzymes and functional genes involved in N cycling (nitrogen fixation, nitrification, and denitrification). (3) Experimental field site experiment (the long-term established sites): We also collected root, rhizosphere, and bulk soil samples on 10/23/2020 from the bahiagrass and RP plots that were established a decade ago (at NFREC, Quincy) to determine the long-term effects of RP and bahiagrass mixture on other nutrients such as P, and micronutrients (Fe, Mn, Zn, and Cu) distribution, and the microbial communities in the different compartments (roots, rhizosphere, and bulk soil), particularly arbuscular mycorrhizal fungi, and phosphatase. The molecular works are in progress for the samples collected from above (1), (2) and (3). Briefly, the DNA from the soil samples collected in (2) were extracted and a two-step PCR was carried out to generate amplicon library. Bacterial16S rRNA and fungal ITS gene fragments were amplified using 341F/806R and ITS1/ITS4 primers. We will be sending out the amplicon library to the sequencing facility for next generation sequencing this month. We have also standardized the protocols for the enzyme analysis and will be measuring enzyme activities in the samples as well as quantification of the genes involved in N transformation (N fixation, nitrification, and denitrification) The molecular analysis of the samples collected from (1) and (3) are in progress. Data analysis: The data generated from the next generation sequencing, enzyme, and qPCR will be processed in Qiime2 and R software and statistical analysis (e.g., ANOVA, PERMANOVA, and PCA) will be applied. For Objective 2. (1) For biogeochemical analysis, we collected the second year of data from the three on-farm sites. Data collected included aboveground biomass, belowground biomass, greenhouse gas emission from soil, biological N2 fixation, nutritive value of forages. The poster with preliminary data was presented by Luana Dantas, the MS student, and she ranked first place in the MS graduate student poster competition in the C-06 Division of Crop Science Soc. of America. Soils were collected from these sites, as well to assess hydraulic conductivity (saturated and unsaturated). We found one location seems to have a hydrophobic surface soil. We will investigate further and perform tests to determine the degree of water repellency. (2) In order to perform molecular analysis of microbial community and to identify the co-relation among microbial community soil chemistry and environmental variables, we collected more samples (root, rhizosphere, and bulk soil) in May, July, and October 2020 across the three on-farm sites (North, Central, and South Florida). Soil moisture and temperature sensors as well as rain gauges were installed in June across the three sites. Sample analysis: We processed the plant and soil samples collected from fields in 2020 and are ready for microbial analysis. The DNA was extracted from the bulk soil, rhizosphere, roots samples (a total of 350 samples) collected from one of the on-farm sites (North Florida) in 2019 and the two-step PCR was carried out to generate amplicon library. Bacterial16S rRNA and fungal ITS gene fragments were amplified using 341F/806R and ITS1/ITS4 primers. The amplicon libraries were sent to the sequencing facility for next generation sequencing. The qPCR was applied to estimate the abundance of the bacterial and fungal taxa of the different samples, while the nutrient concentrations of the bulk soil, shoot, and root +rhizome were measured. The analysis of the two other on-farm sites (Central and South Florida) are in progress. Data analysis: OTU tables were generated from the sequence reads in Qiime2. The statistical analysis of the chemical and OTU tables were carried out using R software. The soil moisture and temperature data generated from the sensors were collected from the three on-farm sites and graphed. The preliminary results were generated. Summary: Our data shows that the incorporation of RP into bahiagrass benefitted the grass. There was not much impact of RP on bahiagrass shoot N. However, the bahiagrass shoot phosphorus concentration in the mixture of the RP, regardless of the RP cultivar, was greater than the bahiagrass shoot grown as monoculture. The average (across the bulk soil and rhizosphere soil) absolute abundance of the bacteria phyla were also greater in the bahaigrass grown in the mixture compared to the monoculture. While the bacteria phyla was indifferent to defoliation, the effect of defoliation (grazing) was pronounced on bahiagrass shoot N, as there was greater bahiagrass shoot N with higher defoliation intensity compared to a lower grazing intensity. Our preliminary finding also showed that the bacterial abundance differed in the different soil compartments.The temperature and soil moisture differed among the three on-farm sites. The soils in Central Florida (0.12 m2/m2) were drier than the soils in North (0.16 m2/m2) and South Florida (0.19 m2/m2), while the average soils temperature were in the following order (Central Florida (30.2oC > South Florida (29.5oC) > North Florida (28oC)). We are working on more data analysis from the other two farm sites and we will continue with the analysis of the samples from 2019 and 2020 to validate the findings obtained so far and to determine if these findings are consistent across different seasons and years. We plan to report the findings from other analysis such as enzyme measurement, microbial diversity, and the relationship between the chemical and microbial data in the next report.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Aryal, P., L.E. Sollenberger, M.M. Khomann, L.S. da Silva, E.M. Shepard, K.D. Cooley, D.L. Rowland, and J.C.B. Dubeux, Jr. 2020. Rhizoma peanut genotype and planting date affect biomass allocation patterns and establishment performance. Crop Science 60:1690-1701. https://doi.org/10.1002/csc2.20142
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Gomes, F.K., B.G.C. Homem, M.D.B.L. Oliveira, J.C.B. Dubeux, Jr., R.M. Boddey, T.F. Bernardes, D.R. Casagrande. 2020. Defoliation frequency affects litter responses and nitrogen excretion by heifers in palisadegrass-forage peanut pastures. Agronomy Journal 112:3089-3100. https://doi.org/10.1002/agj2.20240
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Zhang K, Bonito G, Hsu C-M, Hameed K, Vilgalys R, Liao H-L. 2020. Mortierella elongata Increases Plant Biomass among Non-Leguminous Crop Species. Agronomy. (Brief Report) 10:754. Doi:10.3390/agronomy10050754.
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Guerra, V., L. Beule, E. Lehtsaar, H.L. Liao, and P. Karlovsky. 2020. Improved protocol for DNA extraction from subsoils using phosphate lysis buffer. Microorganisms 8:532. https://doi.org/10.3390/microorganisms8040532
    • Type: Journal Articles Status: Under Review Year Published: 2020 Citation: Chen K-H, Longley R, Bonito G, Liao H-L. A two-step PCR protocol enabling flexible primer choice and high sequencing yield for Illumina MiSeq meta-barcoding. Agronomy. Tech Report (in review #956076)
    • Type: Journal Articles Status: Under Review Year Published: 2020 Citation: Guerra V, Beule L, Liao H-L, Dubeux J, Blount A, Rowland D, Wang X-B, Mackowiak C. 2020. Soil Bacterial Community Response to Rhizoma Peanut Incorporation into Florida Pastures. J Environmental Qual (submitted: JEQ-2020-12-0397-TR)
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Santos, E.R.S., J.C.B. Dubeux, Jr., D.M. Jaramillo, L. Garcia, L.M.D. Queiroz, C.S. Silva, and D.S. Abreu. 2020. Overseeding cool-season forages on rhizoma peanut fields. Crop, Forage, & Turfgrass 6:e20060 https://doi.org/10.1002/cft2.20060
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Santos, E.R.S., J.C.B. Dubeux, Jr., D.M. Jaramillo, L. Garcia, C. Mackowiak, A.R.S. Blount, J.D. Pereira Neto, L.M.D. Queiroz, M. Ruiz-Moreno. 2020. Herbage responses and nitrogen agronomic efficiency of bahiagrass-legume mixtures. Agronomy Journal 112:4057-4068. https://doi.org/10.1002/agj2.20278
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Cooley, K.D., L.E. Sollenberger, M.M. Kohmann, A.S. Blount, J.C.B. Dubeux, Jr., M.L. Silveira, L.S. da Silva, and P. Aryal. 2020. Rhizoma peanut herbage and root-rhizome responses to extended regrowth periods. Crop Sci. 60:2802-2813. https://doi.org/10.1002/csc2.20236
    • Type: Journal Articles Status: Accepted Year Published: 2021 Citation: Zhang K, Maltais-Landry G, Liao H-L. How biota regulates C cycling underlying crop rotations. 2020. Soil Biology and Biochemistry (review paper) (accepted SBB17475)
    • Type: Other Status: Published Year Published: 2020 Citation: Dubeux, J.C.B., Jr., L. Garcia, D. Jaramillo, E. Santos, F. van Cleef, L. Queiroz, C. Garcia, D. Abreu, M. Ruiz-Moreno. 2021. Integrating perennial peanut into grazing operations. Available at Integrating Perennial Peanut into Grazing Operations | Panhandle Agriculture (ufl.edu)
    • Type: Other Status: Published Year Published: 2020 Citation: Dubeux, J.C.B., Jr., L. Garcia, D. Jaramillo, E. Santos, F. van Cleef, L. Queiroz. 2020. How much nitrogen forage legumes can add to grazing systems in FL? The Florida Cattleman and Livestock Journal, July 2020, v. 84, n.10, p.8-10.
    • Type: Other Status: Published Year Published: 2020 Citation: 3. Mackowiak, C. September 04, 2020. UF/IFAS seeking livestock producer input with pasture health survey. Panhandle Agriculture eNews. https://nwdistrict.ifas.ufl.edu/phag/2020/09/04/uf-ifas-seeking-livestock-producer-input-with-pasture-health-survey/
    • Type: Other Status: Published Year Published: 2020 Citation: Mackowiak, C.L. 2020. Bahiagrass health depends on balanced nutrient management. The Florida Cattlemen and Livestock Journal. May 84(8).
    • Type: Other Status: Published Year Published: 2020 Citation: Mackowiak, C.L. and L. Harlow. 2020. Pasture health and pasture decline: get your voice heard! The Florida Cattlemen and Livestock Journal. Oct 84(1).
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Jaramillo, D.M., J.C.B. Dubeux, Jr., N. DiLorenzo, L.E. Sollenberger, J.M.B. Vendramini, C. Mackowiak, L.M.D. Queiroz, B.G. Homem, M. Ruiz-Moreno, E.R.S. Santos, F. van Cleef , L. Garcia, C.V. Garcia. 2020. Water footprint of beef production in contrasting N fertilized or grass-legume mixed grazing systems. ASA/CSSA/SSSA International Annual Meeting Abstracts, virtual meeting, AZ. Available at ASA-CSSA-SSSA International Annual Meeting (social27.com)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Liu, J.C., C.L. Mackowiak, J.C.B. Dubeux, Jr., H.L. Liao, A. Blount. 2020. Perennial grass-legume impacts on the vertical distribution of soil nutrient and quality indicators. ASA/CSSA/SSSA International Annual Meeting Abstracts, virtual meeting, AZ. Available at ASA-CSSA-SSSA International Annual Meeting (social27.com)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., L.E. Sollenberger, J.M.B. Vendramini, H.L. Liao, D.M. Jaramillo, E.R.S. Santos, C.V. Garcia, H.M.S. Silva, B.G.C. Homem, M. Ruiz-Moreno, D.S. Abreu, and F. van Cleef . 2020. Aboveground responses of rhizoma peanut entries in bahiagrass mixtures across multiple locations. ASA/CSSA/SSSA International Annual Meeting Abstracts, virtual meeting, AZ. Available at ASA-CSSA-SSSA International Annual Meeting (social27.com)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Erhunmwunse AS, Liao H-L, Queiroz LD, Mackowiak C, Blount ARS, Dubeux Jr. JCB. 2020. Forage production: On-Farm Studies Across Florida to Evaluate the Effects of Rhizoma Peanut Species, Grazing, and Environmental Factors on Bahiagrass Production. Extension Professional Associations of Florida (EPAF) Abstract Conference Proceeding pp :58, https://extadmin.ifas.ufl.edu/media/extadminifasufledu/epaf/docs/pdfs/abstracts/2020EPAFAbstractProceedings.pdf
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Liu JC, Mackowiak CL, Dubeux JCB, Liao HL, Blount AR. 2021. Yield gains by incorporating rhizome peanut into bahiagrass and its impact on soil fertility distribution. American Forage & Grassland Council (AFGC) 2021. Conference, Savannah, GA (Abstract and Poster Presentation, Virtual)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Harlow L, Mackowiak CL, Blount AR, Paret M, Liao H-L. 2021. Addressing the status of pasture health through producer needs assessment. American Forage & Grassland Council (AFGC) 2021. Conference, Savannah, GA (Abstract and Poster Presentation, Virtual)
    • Type: Other Status: Published Year Published: 2020 Citation: Liao H-L. 2021.The plant-growth promoting fungus, Mortierella elongata, its biology, ecological distribution and activities promoting plant growth. EDIS. https://edis.ifas.ufl.edu/ss679
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Queiroz, L.M.D., J.C.B. Dubeux, Jr., L.E. Sollenberger, J.M.B. Vendramini, H. Liao, M. Ruiz-Moreno, D.R. Casagrande, D.S. Abreu, C.V. Garcia, D.M. Jaramillo, E.R.S. Santos, F.O. van Cleef . 2020. Tracking nitrogen transfer in grass-legume system using enriched 15N2. ASA/CSSA/SSSA International Annual Meeting Abstracts, virtual meeting, AZ. Available at ASA-CSSA-SSSA International Annual Meeting (social27.com)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Erhunmwunse AS, Queiroz LD, Mackowiak CL, Dubeux Jr. JCB, Blount AR, Liao H-L. 2021. Effects of defoliation and perennial peanut on phosphorus and micronutrients in bahiagrass system American Forage & Grassland Council (AFGC) 2021. Conference, Savannah, GA (Abstract and Poster Presentation, Virtual) https://www.afgc.org/i4a/doclibrary/getfile.cfm?doc_id=832


    Progress 04/01/19 to 03/31/20

    Outputs
    Target Audience:(a) Ag In-service training 2019: Scope: Provide The Ag In-service training is an annual, day-long event to disseminate through presentations and some demonstrations, the latest Ag-related research updates to help prepare Florida extension faculty and agents for client problems and to provide information for their own client training. Target audience: Florida state and county extension faculty (b) Perennial Peanut field day (NFREC), 05/30/2019 Scope: Annual day-long event to promote interaction and sharing of research with membership and to promote the use of this species in the hay, pasture, and ornamental landscapes. Target audience: Perennial Peanut Producers Association (PPPA) members and the general public (c) On-farm visit and research, May, July and October 2019 Scope: Identify and report the C/N microbial activities to the pasture managers after their farms adapted to the integrated forage management systems (e.g., pasture mixture; grazing, etc), Three on-farm sites in FL: Marianna, Inverness, Wauchula Target Audience: Pasture managers (d) Southern Pastures and Forage Crop Improvement Conference (Roanoke, VA, 05/23/2019) Scope: An annual workshop that aims to share and exchange research and extension information among leading forage and pasture researchers and extension agents in the Southeast USA. Target Audience: Multi-state mix of university faculty, extension faculty, students, and commercial product representatives. (e) Environmental education and 4-H: July 2019: Conduct the lectures for Applied Soil Microbiology Training: Mycorrhizae short courses. (https://conference.ifas.ufl.edu/soilmicro/). The topic of Lecture: Using the magic microbial wood-wide web to manipulate usable plant food (instructor: Liao). January 2020: Lecture and workshop leading for 4-H Hort Program (Leon county extension office). The topic of the lecture: Beneficial microbes and plant-fungal interactions (instructors, Liao, Zhang, Hsu, and Erhunmwunse) Scope: To improve the Knowledge of the targeted audiences, to stimulate the actions of those targeted audiences based upon the gained knowledge Target audience: Extension agents, undergraduate students, graduate students, youth and parents. (f) 2019 Beef and Forage Field Day at UF March 2019: Field day displaying the beef and forage trials at NFREC, Marianna. This including the grazing trial with grass-legume mixtures. We highlighted the new project indicating the goals and expected outcomes. Scope: to increase the awareness of livestock producers on potential grass-legume belowground interactions. Target audience: livestock producers, extension agents, and graduate students. (g) This project involves training graduate students, biologists and visiting scientists to conduct research proposed in this study. The scope is to provide training and professional promotion in the area of agroecology and environmental microbiology. The trainees include 5 women and minorities. Target trainees: Adesuwa Erhunmwunse (Ph.D. student, supervisor: Hui-Ling Liao) Adesuwa is the key personnel for this project. She is an under representative scientist (an international scientist from Nigeria); Luana D Queiroz (M.S. student, supervisor: Jose Dubeux); Jung-Chen Liu (M.S. student, supervisor: Cheryl Mackowiak); Xiaobo Wang (Postdoc level visiting scholar, supervisor: Hui-Ling Liao); Lukas Beule (M.S. level visiting scholar, supervisor: Hui-Ling Liao); Kaile Zhang (Ph.D. student, supervisor: Hui-Ling Liao); Chih-Ming Hsu (volunteer scientist, supervisor: Hui-Ling Liao); Ko-Hsuan Chen (Postdoc, supervisor: Hui-Ling Liao); Daniel Casagrande (visiting scholar on sabbatical, supervisor: Jose Dubeux); Bruno Grossi (sandwich Ph.D. student, supervisor: Jose Dubeux); Carlos Garcia (sandwich Ph.D. student, supervisor: Jose Dubeux); Gustavo Duarte (sandwich Ph.D. student, supervisor: Jose Dubeux); Izabela Gomes (sandwich Ph.D. student, supervisor: Jose Dubeux); Jose H. Andriarimalala (sandwich Ph.D. student, supervisor: Jose Dubeux); Daciele Abreu (sandwich MS student, supervisor: Jose Dubeux); Victor Guerra (Ph.D. student, supervisor: Cheryl Mackowiak); Kelly Thomas (MS student, supervisor: Ann Blount) (h) Publication of extension articles Scope: Journal provides producers and managers of Florida cattle the best of regional information on livestock health, production, and pasture management. Target Audience (readers): State BMP regulators, extension agents/county agents, pasture managers, beef operators, forage agronomists, cattle ranchers (1) Mackowiak, C.L., H. Liao, and A. Blount. 2019. Healthy pastures rely on healthy soils. The Florida Cattlemen and Livestock Journal. Mar 82(6). (2) Dubeux, J.C.B., Jr. 2019. Developing sustainable livestock systems in Florida. The Florida Cattleman and Livestock Journal, July 2019, v. 83, n.10. p.42-50 .(3) Dubeux, J., C. Mackowiak, A. Blount, D. Wright, and L. Dantas. 2019. Overseeding rhizoma perennial peanut pasture and hay fields during the cool-season. Florida Cooperative Extension Service, IFAS, 5 pp. SS-AGR-426. (4) Van Cleef, F.; J.C.B. Dubeux, Jr. 2019. Condensed tannins in forage legumes. EDIS SS-AGR-440. https://edis.ifas.ufl.edu/ag440 Scope: EDIS and fact sheets are located at a UF-IFAS sponsored online information outlet to support agricultural information dissemination. Target Audience (readers): County extension and certified crop advisors, producers, researchers, students, state policymakers, and the general public (i) Publications of research articles A part of the research outcomes has been published in two scientific journals (to Peer J and Ecology and Evolution) and submitted one research note (to Biology and Fertility of Soils). The target audience (readers) includes whose interests focus on forage ecology, microbial ecology, crop production, soil microbiomes, soil nutrients, plant-microbe interactions, nutrient cycling and beyond. (1) Wang X-B, Hsu C-, Dubeux CB Jr, Mackowiak C, Blount A, Han X-G, Liao H-L. 2019. Effects of rhizome peanut cultivars (Arachis glabrata Benth.) on the soil bacterial diversity and predicted function in nitrogen fixation. Ecology and Evolution. 9(22):12676-12687. DOI: 10.1002/ece3.5735 (2) Beule L, Chen K-H, Hsu C-M, Mackowiak C, Dubeux JCB, Blount A, Liao H-L. 2019. Soil bacterial and fungal communities of six bahiagrass cultivars. PeerJ 7:e7014 http://doi.org/10.7717/peerj.7014 (3) Zhang K, Bonito G, Hameed K, Vilgalys R, Liao H-L. 2020. Evaluation of the impacts of Mortierella elongata isolates on plant growth. Biology and Fertility of Soils (research note; submitted, BFSO-D-20-00132) Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?As mentioned in the "target audience" section, this project involves training the graduate students, biologists, undergraduate interns and visiting scientists to conduct the researches proposed in this study (listed below). Through the supervision and training activities, the individuals are expected to receive the professional skills on molecular/biogeochemical technologies and bioinformatics. The professorial training for the graduate students and postdoctoral scientists include research proposal development, experimental designs, sample collection/processing, data collection/analysis, data integration, scientific presentations, coordination for team meetings, workshop/field day/in-service training participation, and leading the manuscript preparation for his/her projects. (1) Adesuwa Erhunmwunse (Ph.D. student, supervisor: Liao): The project that Adesuwa conducts aims at understanding the interactive effects of multiple management (plant community, grazing, and environmental variables) on microbial-driven soil fertility and forage production. Adesuwa will train the undergraduate interns (e.g., Valerie Mendez) on the molecular work that is associated with the forage project (e.g., soil and root DNA extraction; Qiime 2 associated NGS data analysis). (2) Luana D Queiroz (M.S. student, supervisor: Dubeux) (3) Jung-Chen Liu (M.S. student, supervisor: Mackowiak) (4) Xiaobo Wang (Postdoc level visiting scholar, supervisor: Liao): Xiaobo's research project focuses on investigating microbial diversity across different management strategies of the forage system. The outcome of the training had led a publication of a research paper for this project (Wang et al, 2019) (5) Lukas Beule (M.S. level visiting scholar, supervisor: Liao): Lukas' project focuses on studying belowground biodiversity in associated with forage cultivars. The outcome of the training had led a publication of a research paper for this project (Beule et al, 2019) (6) Kaile Zhang (Ph.D. student, supervisor: Liao): Kaile's project focuses on identifying the dominant fungal taxa presenting in the pastures that provide the beneficial effect across different cropping systems, including bahiagrass and RP. The outcome of the training had led a short research note submitted to Biology and Fertility of Soils for this project. (7) Chih-Ming Hsu (Volunteer Scientist, supervisor: Liao): Chih-Ming is involved in field sampling, sampling processing and molecular work for amplicon and RNA-based sequencing for this project. Mr. Hsu also trained the visiting scientists and graduate students on molecular techniques applied for this project (the trainees include, Erhunmwunse, Wang, Beule, Zhang and Mendez). The outcomes of this training has led the contributions to two research papers (Wang et al 2019; Beule et al 2019) (8) Ko-Hsuan Chen (Postdoc, supervisor: Liao): Ko-Hsuan developed the 2-step in-house PCR protocol to generate the DNA library for amplicon sequencing in this project. (9) Valerie Mendez (Undergraduate level intern, supervisor: Liao; Blount; Trainers for the molecular technology: C-M Hsu and A. Erhunmwunse): Valerie is involved in molecular work that are associated with forage project, including DNA extraction, 2-step PCR library construction, DNA purification quantification and Qiime 2 associated data analysis. (10) Daniel Casagrande (visiting scholar on sabbatical, supervisor: Jose Dubeux); assisting on developing the H-pot technique in the greenhouse (11) Bruno Grossi (sandwich PhD student, supervisor: Dubeux); assisting on the field sampling and laboratory analyses (12) Carlos Garcia (sandwich PhD student, supervisor: Dubeux); assisting on the field sampling and laboratory analyses (13) Gustavo Duarte (sandwich PhD student, supervisor: Dubeux); assisting on the field sampling and laboratory analyses (14) Izabela Gomes (sandwich PhD student, supervisor: Dubeux); assisting on the field sampling and laboratory analyses (15) Jose H. Andriarimalala (sandwich PhD student, supervisor: Dubeux); assisting on the field sampling and laboratory analyses (16) Daciele Abreu (sandwich MS student, supervisor: Dubeux). assisting on the the field sampling and laboratory analyses How have the results been disseminated to communities of interest?As stated in the target audience section, through extension programs conducted by the PDs, we provided solid scientific evidence about the actions of microbiome across different managements to diverse audiences, including stakeholders, extension agents, general public and youth. We thought the stakeholders' scientific evidence-based suggestions needed to sustainably improve ecosystem services. To enhance the awareness of the importance of microbes to soil health, we led the environmental education programs to teach the audiences with diverse ages and backgrounds. The details of the events conducted in 2019 were listed in the "target audience" section. Below we addressed "How have the results been disseminated to communities of interest": (1) Ag In-service training 2019:Powerpoint presentations included growth-promoting microbes in crop production and a forage update. Presentations were also video recorded for online access. (2) Perennial Peanut field day (NFREC), 05/30/2019: A combination of demonstrations and presentations, including perennial peanut cultivar differences in growth habit (demonstration) and response to water stress (oral presentation), were used. (3) On-farm visit and research, May, July and October 2019: Visits consist of touring the on-farm, experiment sites with the producers, discussing project progress and receiving input from producers on their impressions, as well as answer questions they might have. (4) Southern Pastures and Forage Crop Improvement Conference (Roanoke, VA, 05/23/2019): Information exchanges during the meetings and round-table topic discussions. (5) Environmental education and 4-H: Through slides and video presentation to transmit the scientific-based knowledge, hand-on practices in the workshop (e.g., microscope observation, culturing mycorrhizal fungi, inoculation of mycorrhizal fungi to the plant seedlings, plant bioassay, and mycorrhizal development observation) (6) 2019 Beef and Forage Field Day at UF: Field day organized at UF IFAS NFREC Marianna and discussions with producers and students. (7) Publication of extension articles: is located at a UF-IFAS sponsored online information outlet to support agricultural information dissemination. (8) Publications of research articles: Through publishing the research articles to the open access journals, the information was immediately available online for the scientific communities What do you plan to do during the next reporting period to accomplish the goals?We will repeat the sampling protocol we used for year 1. We plan to finish the laboratory analyses for samples collected in year 1. In addition, we will install the H-pots with perennial peanut and bahiagrass during the summer to assess belowground N transfer and resource sharing between bahiagrass and perennial peanut. The team who conduct this project will attend the scientific conferences to report their study outcomes. The team will also continuously involve and lead the extension activities to deliver the goal of objective 3.

    Impacts
    What was accomplished under these goals? The issue: Although legume integration is a practical option for enhancing ecosystem services in grasslands, it is not yet a well-established practice in the southeastern U.S. Primary Obj: We evaluate how multiple advanced components can be integrated to improve biological N use in promoting plant and soil health. We assess how these integrated practices can alter soil microbiomes and affect the outcomes of grassland ecosystem services. The Impact: This project has direct implications for improving forage nutrition, soil health, and environmental gas emissions. The key impact is to build greater insight into mechanisms that can increase the sustainability of U.S. agriculture and food systems. What did we do in 2019: (a) Obj 1: We established across three different bahiagrass (BG) cultivars, three cultivars of rhizoma peanut (RP). We also identified mature (over 10 years) plots of RP and BG to test long-term interactions and we established mesocosms for assessing fixed N transfer between plant species. (a.1) Field exp Site development and sampling: The research field site was established in May 2019 using the experimental plots originally grown BG cultivars. A split-plot design was used with BG main plots arranged in a randomized complete block design, and RP treatment as subplots (plot size 5 x 20 ft). The treatments include BG (Argentine, Pensacola, Tifton-9) and RP (Ecoturf, Florigraze, LU-1RP) monocultures and the mixtures of paired BG-RP across these cultivars. The research site is located at Marianna FL (30.871 N, -85.190 W). The soil is fine-loamy, kaolinitic, thermic Rhodic Kandiudults, Red Bay series. Three BG Flüeggé (bahiagrass) cultivars were established from seed in 2005 as randomized blocks. May 13-24, 2019: After soil sampling, the BG on the selected plots (RP monoculture=full plot; BG-RP mixes=4 strips per plot) were then killed. The RP sod was transplanted into assigned plots and a water tank was used four times to irrigate during establishment, as we entered the spring dry period. Plots were mowed twice (biomass removed) during the establishment season (summer/fall 2019) to minimize weed pressure and to maximize light interception by the establishing RP. Mar 09, 2020: Plots were mowed (10 cm stubble height) to stage them for the 2020 sampling season. All biomass was removed from plots. Sampling: Soils were collected for initial soil fertility analysis, composited per block (05/13/19); Soil samples were collected for microbial community analysis and soil chemical analysis (03/09/20). (a.1.2) greenhouse exp: The mesocosm was set up to study the 15N re-allocation across BG and RP (Jan 2020). Briefly, H-pots built up with PVC pipes were set up to connect belowground tissues from BG and RP. The H-pot was planted with grass in one side and legume on the other side, connected by a PVC pipe so that the belowground transfer could be assessed. The legume side will be enriched with 15N2 in the rhizosphere. BG and RP shoot will be analyzed for 15N to verify the N transfer. (a.1.3) Data collected (in progress): DNA metabarcoding was applied to identify the relative abundance of soil/root microbes. Integration of NGS and qPCR was applied to estimate the biomass of each dominant microbial taxa. Potential indicators for soil health will be measured, including soil reactive carbon (POxC and WSC) and reactive N (TIN and PMN), along with total C and N. Sampling of mature (over 10 years) plots of RP as monoculture versus mixed with BG will be conducted, in order to assess longer-term impacts of these systems to soil C and N indicators. Microplates will be used in the mature plots to assess N transport between RP and BG using (15NH4)2SO4. Soil saturated and unsaturated hydraulic conductivity will be measured during the summer, 2020 in order to model potential nitrate leaching under the different treatments. (a.1.4) Summary statistics, discussion of results: Our published data on the survey of microbial community across different cultivars of BG (Belue et al, 2019) and RP (Wang et al, 2019) indicate that diversity of soil microbiome was not significantly changed across different plant cultivars. However, the relative abundance of bioindicators (specific microbial taxa) was significantly changed in associated with different cultivars. Integrating the NGS data and soil chemical data collected from research field sites will allow us to identify the dynamics of these bioindicators as affected by the integration of BG-RP (vs. monocultures). In addition, the statistical models (e.g., RDA, SEM) will be applied to link the association between the abundance of bioindicators and soil chemical activities. We will report the results in the next annual report. In addition, some beneficial fungi (e.g., Mortierella elongata) were identified as one of the dominant fungal taxa presenting in the BG soil (Belue et al, 2019). Our bioassay showed that M. elongata was able to enhance the biomass of some bahiagrass cultivars (Zhang et al, submitted). The analysis of targeted beneficial microbes will be emphasized. (b) Obj 2: We identified and developed three observational field sites across regional gradients. (b.1) Major exp (b.1.1) Observation field site setup: The on-farm locations are: (Site 1: 30.678563 N, -85.331189W, Marianna Northwest FL; Site 2: 28.742953N, -82.348881W27, Rooks, Inverness, Central FL; and Site 3: 27.498424N, -81.822594W, Wauchula, South-Central FL). At each site, a randomized complete block design was used and a standard plot size of 9m x 14.5m for each plant species treatment including BG monoculture (Argentine), RP (Ecoturf)- BG (Argentine) mixture, and RP (Florigraze)-BG (Argentine) mixture. Cages were arranged as subplots (1.2mx1.2m per cage) within each plot representing the none-grazing treatment. The three sites are located in Marianna, Inverness, and Ona. Two RP cultivars (Florigraze and Ecoturf) were established on BG pastures in June 2014. (b.1.2) Sampling: In 2019, The bulk soils, rhizosphere soils, roots were collected for NGS, soil chemistry and enzyme activity analyses. The plant biomass and GHG emission analysis were conducted on-site while the samples were collected. Shoot samples were collected twice in 2019 (July and Oct) and root-rhizome samples collected once (Oct). Samples were collected in caged and uncaged areas to assess the effect of defoliation. Besides grazing, we mowed the uncaged sites with 4-wk interval to assure the defoliation was taking place. Forage samples were refrigerated and dried thereafter at 55C for processing. Samples will be analyzed for N, IVOMD, and 15N. The gas measurements also occurred twice in 2019, during the same sampling dates. Samples will be analyzed for greenhouse gases. The sampling seasons: Initial sampling prior cage setup (cages were set up right after initial sample collection, 05/2019); Harvest 1 (07/2019); Harvest 2 (10/2019). We plan to instill the environmental monitors to track the soil precipitation and above and belowground temp in April 2020. We plan to have two to three harvests in the year 2020, including May, July, and October. (b.1.3) Data collected (in progress): Data collection for Year 1 was completed. Forage, belowground tissues, and gas samples were collected as planned. Samples are still being analyzed in the laboratory. (B) Other accomplishments: Through our research and extension activities, the participant (scientist, trainee, or citizen) learns or becomes aware. For example: The fundamental or applied knowledge from our current studies was significant enough to be included in three publications (Belue et al, 2019; Wang et al, 2019; Zhang et al, submitted). The post-survey in Lecture and workshop leading for extensions showed that at least 95% of participants increase the awareness and over 15% modify their life choices among youth and stakeholders. Change in Action and Change in Condition will be reported in the following years.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Beule L, Chen K-H, Hsu C-M, Mackowiak C, Dubeux JCB, Blount A, Liao H-L. 2019. Soil bacterial and fungal communities of six bahiagrass cultivars. PeerJ 7:e7014 http://doi.org/10.7717/peerj.7014, Cited by 1
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Wang X-B, Hsu C-M, Bubeux CB Jr, Mackowiak C, Blount A, Han X-G, Liao H-L. 2019. Effects of rhizome peanut cultivars (Arachis glabrata Benth.) on the soil bacterial diversity and predicted function in nitrogen fixation. Ecology and Evolution. 9(22):12676-12687. DOI: 10.1002/ece3.5735
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Dubeux, J.C.B., Jr., M.V.F. Santos, A.C.L. de Mello, M.V. da Cunha, V.X.O Apolin�rio, H.B. Lima. 2019. Multiple ecosystem services of arboreal legumes. Legume Perspectives, 17:10-12.
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Santos, M.V.F. dos, M.V. da Cunha, J.C.B. Dubeux, Jr., R.L.C. Ferreira, M.A. Lira Jr., O.F. Oliveira. 2019. Native shrub-tree legumes of tropical America with potential for domestication. Legume Perspectives, 17:33-35.
    • Type: Journal Articles Status: Submitted Year Published: 2020 Citation: Zhang K, Bonito G, Hameed K, Vilgalys R, Liao H-L. 2020. Evaluation of the impacts of Mortierella elongata isolates on plant growth. Biology and Fertility of Soils (research note; submitted, BFSO-D-20-00132)
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Muir, J.P., J.C.B. Dubeux, Jr., M.V.F. dos Santos. 2019. Arboreal legumes for human consumption. Legume Perspectives, 17:39-42.
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Sollenberger, L.E. , M.M. Kohmann, J.C.B. Dubeux, Jr., M.L. Silveira. 2019. Grassland management affects delivery of regulating and supporting ecosystem services. Crop Science 59:441-459 doi: 10.2135/cropsci2018.09.0594
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Muir, J.P., J.C.B. Dubeux Jr. 2019. Arboreal Legumes for Multiple Uses. Legume Perspectives - The Journal of the International Legume Society. Issue 17, August 2019. Special Issue.
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