Source: UNIVERSITY OF FLORIDA submitted to
TRADITIONAL AND NOVEL SOIL HEALTH INDICATORS FOR SUB-TROPICAL TREE FRUIT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
NEW
Funding Source
Reporting Frequency
Annual
Accession No.
1025040
Grant No.
2021-67019-34240
Project No.
FLA-SWF-006030
Proposal No.
2020-04271
Multistate No.
(N/A)
Program Code
A1401
Project Start Date
Feb 14, 2021
Project End Date
Feb 13, 2025
Grant Year
2021
Project Director
Strauss, S. L.
Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611
Performing Department
Soil & Water Sciences
Non Technical Summary
Sub-tropical tree cropping systems require intensive management, primarily due to the extremely coarse-textured and organic matter poor soil. Cover crops are a common method to improve soil health for annual crops, however, different implementation methods are needed for tree crops. In addition, data on the response of sub-tropical tree crop soils to cover crops is limited, particularly for the contribution of the soil microbiome. Soil health indicators can assist in determining how cover crops change soil health in these systems. However, while there are multiple suggested soil health indicators available, assessment has primarily occurred in temperate annual crops.This project will determine the impact of cover crops on soil health in sub-tropical tree crops through field trials with commercial producers. To assess these impacts, traditional and novel indicators of physical, chemical, and microbial soil health components will be utilized, and sampled across different timescales. This project will also examine the relationship between soil health and the diversity of plant-growth promoting functions of the soil microbiome, as these may not only be critical to soil health changes but could also serve as new biological soil health indicators.Our project directly advances the "scientific understanding of soil physical and biochemical processes and interactions" by examining the interaction of the soil microbiome with physico-chemical factors under changing soil health conditions. In addition, the exploration of a variety of soil health indicators will allow for the development of appropriate and effective methods for monitoring soil health changes in sub-tropical tree crops.
Animal Health Component
0%
Research Effort Categories
Basic
40%
Applied
60%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1020110106050%
2051099107050%
Goals / Objectives
Our overarching goals are twofold: (1) assess the impact of cover crops as a soil health management strategy for sub-tropical tree crops, including the influence of soil microbiome functional groups on physico-chemical soil health indicators and nutrient cycling, and (2) determine appropriate soil health indicators to quantify soil health changes across sub-tropical tree crops.Our supporting objectives (SO) are to:SO1: Quantify the influence of a common soil health promoting management practice (cover crops) on novel and traditional soil health indicators and crop production in tree crops, using citrus as a model system.SO2: Examine the relationship between soil health and the diversity and plant growth promoting (PGP) functions of the soil microbiome and their correlation with novel and traditional soil health indicators.SO3: Determine which soil health indicators are the most useful for monitoring subtropical tree crops and systems health.
Project Methods
Field trials for Objectives 1 and 2The two citrus groves we will study for this objective consist of three treatments, each replicated six times per grove: 1) cover crop mix #1 = 2 non-legume species; 2) cover crop mix #2 = mix #1 with 2 legumes; and 3) no-treatment control/grower standard. Based on our preliminary data and local grower´s recommendations from experiments carried out in South Florida, summer and winter non-legume and legume cover crops will be used in this experiment. Cover crops will be planted every 4 months in the alleys between trees, a common practice in sub-tropical and tropical climates. Seeds will be planted using a no-till drill, with 50% of monoculture rates for non-legumes (mix #1), and 50% monoculture rate for legumes and non-legumes (mix #2), respectively. Cover crops will be terminated by mowing and residue will remain on the soil surface; reseeding will occur on the same day as mowing, or the following day.Methods for objective 1Samples for short-term soil health indicators will be collected three times per year from the top 10 cm of soil immediately prior to mowing and the planting of the next cover crop. Short-term soil health indicators measured include: POXC, C and N mineralization/respiration, ACE protein content, microbial biomass P, and enzyme activities (C-cycling: β-glucosidase, N- and C-cycling: N-acetyl-β-D-glucosaminidase, and P-cycling: Phosphomonoesterase (Acid/Alkaline Phosphatase)), and resin- and hexanol-extractable P. At each sampling time, and for each treatment, soil cores will be collected from the row middles perpendicular to 10 trees per replicate and pooled before subsampling for indicators. Samples for enzyme activity will be frozen upon return to the lab and later processed using 4-methylumbelliferone (MUB) substrates and fluorescence. Another subsample will be kept at 4°C until processing for resin- and hexanol- extractable P in addition to C and N mineralization. The final subsample will be air-dried and sieved to 2 mm before being analyzed for POXC and ACE proteins.Long-term soil health indicators will be assessed once a year from three soil depths (0-10 cm, 10-30 cm, 30-60 cm) during the summer. Indicators will include: aggregate characterizations (following NRCS standards and laser diffraction methods), gravimetric water content, infiltration, cation-exchange capacity (CEC), bulk density, pH,soil inorganic N (NH4+and NO3-),total C and N,SOM (loss on ignition method, LOI), and qPCR of N- and P-cycling genes. For N cycling, this will include:nifHgene, that of nitrifiers by theamoAgene from ammonia-oxidizing bacteria (amoAAOB) and archaea (amoAAOA), and that of denitrifiers by thenirKandnosZI genes. For P cycling, this will include: thephoAandphoD genescoding for phosphatases. At each sampling time and for each treatment replicate, we will collect 10 intact soil cores (with known volumes for bulk density calculations) that will be separated and pooled by depth. Each pooled sample will be weighed at field moisture and then split into four subsamples. The first subsample will be kept at 4°C until processing for inorganic N using 2M KCl extractions followed by colorimetry for NH4+and NO3-. The second subsample will be air-dried, sieved to 2 mm, and then further subdivided for the following analyses: Mehlich III extractable nutrients; CEC and pH by an external laboratory; total C and total N by combustion; and LOI to determine if it can be used as a proxy for total C in these systems. Bulk density will be determined for the third subsample before being characterized for aggregation. The fourth subsample will be frozen at -80°C for microbial analysis.Crop yields, fruit size, leaf N concentration, canopy growth and trunk diameter will be assessed each year.The partial factor of productivity (PFP) will be calculated as the ratio of the total fruit yield in each plot (kg ha-1) to the total N applied through synthetic fertilizer.Cover crops and weed biomass will be measured in four 0.5m×0.5m squares randomly harvested from each plot before each cover crop termination. The annual C and N input will be calculated by multiplying the dry biomass in each plot by its C and N concentrations, respectively, assuming that all cover crop residues remained in the field. The cumulative C and N input will be obtained by adding up the annual inputs for each treatment.A sub-sample of soil collected for the long-term soil health indicators in Year 1 and Year 3 will be sent for a commercially available soil health test analysis. This data will be compared with the measurements made in this objective to assess the rankings provided by a commercial test with our regional and tree crop specific assessment.Methods for objective 2Soil samples will be collected from the same trees and treatments described in Objective 1.Sampling will occur in Year 1 and Year 3 at the same time as samples are collected for the long-term indicators in Objective 1.Bulk soil and rhizosphere will be collected and stored at -80 °C until use. Soil DNA will be extracted and sent to an outside facility for high throughput amplicon sequencing of the bacterial 16S rRNA gene using the Illumina MiSeq platform.Data will be analyzed in QIIME 2 and the Phyloseq R package. Potential bacterial functions in each sample will be predicted using PICRUSt2 software and those related to PGP (N2fixation, IAA production, phosphate solubilization, and ACC deaminase activity) will be determined using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The predicted PGP function with significantly greater abundance in the bulk soil and tree-rhizosphere of each cover crop treatment with respect to the control treatment will be selected as a criterion to isolate PGPB.PGPB will be isolated from 1 g of bulk and tree-rhizosphere soils by serial dilution plating on plates containing a selective media for isolation of PGPB. Bacterial genomic DNA will be extracted after growth of the bacterial isolates. Random amplified polymorphic DNA (RAPD)-polymerase chain reactions (RAPD-PCR) will be performed, and representative strains from each RAPD group will be selected and assayed for N2fixation, P solubilization, IAA, and ACC deaminase activity.Azospirillum brasilenseC16 will be used as a positive control for all the PGPR functions. Genomic DNA isolated from bacterial cells will be used for PCR amplifications of 16S rRNA gene and sequencing.Methods for objective 3Soil samples from five additional commercial sub-tropical tree fruit producers will be collected during the third year. These producers will currently be utilizing cover crops in at least part of their orchard, and samples will be collected from both cover-crop treated soils and soils without cover crops. At each location, three replicate samples per treatment will be collected and analyzed. Samples for short-term soil health indicators will be collected three times per year immediately prior to the next cover crop planting from the top 10 cm of soil, following the procedures detailed in Objective 1. For long-term soil health indicators, samples will be collected during the summer, to coincide with the period of highest metabolic activity of tree crops. These samples will be collected from three soil depths (0-10 cm, 10-30 cm, 30-60 cm) following the procedures detailed in Objective 1. We will collect data on management practices including the timing and rate of fertilization and cover crop planting date, to assist in our interpretation and recommendations of appropriate soil health indicators.Measurements of short and long-term soil health indicators will follow the same methods as described in Objectives 1 and 2. In addition, a sub-sample of the soil collected for the long-term soil health indicator will be sent for a commercially available soil health test analysis to provide comparison with the indicators and methods assessed in this study.?

Progress 02/14/23 to 02/13/24

Outputs
Target Audience: Researchers, producers, extension agents, citrus growers, and field specialists Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training and mentorship for one graduate student and a postdoctoral research associate are being provided by this project, as well as several research assistants. How have the results been disseminated to communities of interest?Preliminary data from the project were presented at the annual meeting of the Soil Science Society of America in November. In addition, preliminary data was also presented to citrus and other fruit tree growers at state and regional grower meetings in Florida, including the Florida Citrus Expo in August 2023 (the largest citrus grower meeting in the state). What do you plan to do during the next reporting period to accomplish the goals?Measurements of short-term indicators and cover crop biomass will occur again in February/March 2024. Fruit harvest will occur in Spring 2024. Analyses of soil enzyme activity from preserved samples will occur throughout 2024, through the work of the current graduate student working on the project in addition to a newly recruited graduate student. This work will consist of measuring cover crop effects on soil enzyme activity through time, in addition to a comparison of soil enzyme methods to identify which methodological approach is best adapted to subtropical tree systems. Analysis of the bacterial isolates for potential PGP activities will be conducted and DNA will be extracted from soil samples collected last summer for a final amplicon sequence analysis. This activity was delayed due to the former postdoc for this project leaving and a delay in a new postdoc joining the project. Correlations and assessments of short and long-term indicators will be made to identify those to target for additional tree fruit crops. These assessments will begin during the end of summer 2024. Presentations of preliminary results will be made a regional and national scientific conferences as well as grower extension meetings.?

Impacts
What was accomplished under these goals? During the third year of the project, we continued the field trials in two citrus orchards. Cover crops were again planted in June and November. The June mixture included sunnhemp, cowpeas, buckwheat, millet, and Egyptian wheat, and the November mixture included daikon radish, sunnhemp, black oats, Florida 401 rye, and hairy vetch. Prior to planting cover crops, the previous crops were mowed and a shallow discing of the soil was performed. The dissertation research of a PhD student is being funded by this project. SO1: In March, just before citrus harvest, soil samples were collected to assess short-term indicators and cover crop biomass was collected. The fruit was harvested during the end of March and subsamples of fruit were analyzed for fruit quality. Cover crops were then planted at the end of June. At the beginning of September, soil samples were collected for both long-term and short-term measurements from three soil depths. Leaves were also collected to assess tree nutrients. Short-term indicators were again assessed at the end of October, and cover crop biomass was collected on the same day. Cover crops were planted 6 days after the completion of the short-term indicator and cover crop biomass collections. Data analysis of the short-term and long-term indicators is ongoing as additional measurements are made each year. Cover crop biomass collected in March has been dried, weighed, ground, and sent for total C and N analysis. Cover crop biomass collected at the end of October has been dried and weighed and grinding is in progress. Germination of cover crops each season has been mixed due to the reliance on timing planting with rainfall. However, initial analysis of cover crop C:N indicates there is generally greater C:N of the cover crops compared to the weeds. Preliminary analysis of qPCR measurements of functional genes from the first two years indicates more differences in gene abundance between soil depths than between treatments. However, in the second year, there were greater differences between treatments for the abundance of the denitrification gene nosZI and archaeal ammonia oxidation for both orchards, though there was more variability in the abundances for the Young Orchard Site. Preliminary analysis of amplicon sequence data from Year 1 indicates that depth also plays a significant role in the overall microbial composition, but significant differences in the relative abundance of specific taxa are present between treatments, particularly in the fungi. Additional analyses are underway to identify taxa that may serve as potential indicators. Regarding the chemical soil health indicators measured, they were typically more sensitive to time than treatment. Significant cover crop effects in the Old Orchard Site include: 1) greater permanganate oxidizable carbon and potentially mineralizable nitrogen (only in Feb. 2023) in the legume and non-legume cover crop mix (LG+NL) compared to the grower standard control (GSC); and 2) greater in Feb. 2023 in LG+NL relative to GSC. In the Young Orchard Site, we found: 1) higher soil organic matter (Sept. 2023) and short-term C mineralization (Sept. and Oct. 2023) in LG+NL relative to GSC; and 2) greater potentially mineralizable nitrogen in LG+NL in March 2022, and Feb., Sept., and Oct. 2023. SO2: Soil samples collected at the beginning of September with the long-term indicators were used to isolate potential PGP bacteria. Over 300 isolates were collected. DNA from these isolates has been extracted and sent for Nanopore sequencing so they can be identified and functional assessments conducted. SO3: This objective will begin in 2024.?

Publications

  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Castellano-Hinojosa A, Maltais-Landry G, Martens-Habbena W, Strauss SL (2023) Depth-dependent effects of cover crops in citrus orchards on soil carbon and nitrogen cyclig, greenhouse gas emissions, and soil microbial communities. Applied Soil Ecology 192; doi: 10.1016/j.apsoil.2023.105071


Progress 02/14/22 to 02/13/23

Outputs
Target Audience:Researchers, producers, extension agents, citrus growers, and field specialists Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training and mentorship for one graduate student and a postdoctoral research associate are being provided by this project. How have the results been disseminated to communities of interest?The project plan and preliminary data from the proposal were presented at the Citrus Health Forum, a regional citrus grower meeting in North Florida in early 2022 and at a citrus grower meeting in South Florida in June of 2022. Preliminary data was presented by a PI and a graduate student at the 2022 Annual Soil Science Society of America conference in November in Baltimore, MD. What do you plan to do during the next reporting period to accomplish the goals?Cover crop plantings and soil collection will continue. Cover crops are scheduled to be planted twice a year at both locations. Soil sample collection for short-term indicators will occur three times during the year, and long-term collection will occur during the summer. The isolation and assessment of potential PGP microbes will begin. Preparations are in progress to begin assessments in additional tree fruit crops. Presentations of preliminary results will be made at regional and national scientific conferences, including the annual meetings of the Soil Science Society of America.?

Impacts
What was accomplished under these goals? During the second year of the project, we continued the field trials in two citrus orchards. Cover crops were again planted in June and November. The June mixture included sunnhemp, cowpeas, buckwheat, millet, sorghum sudangrass, and Egyptian wheat, and the November mixture included daikon radish, sunnhemp, black oats, Florida 401 rye, and hairy vetch. The dissertation research of a PhD student is being funded by this project. SO1: Soil samples for short-term indicators were collected in March just prior to the orange harvest. At this time, cover crop biomass was sampled, dried, and ground. Orange harvest occurred during the last week of March and the first week of April. Subsamples of fruit were collected and analyzed for fruit quality. The next cover crop planting occurred in mid-July and included sunnhemp, cowpea, sorghum sudangrass, Egyptian wheat, hybrid pearl millet, and buckwheat. In September, leaf samples were collected for plant nutrient analysis. Soil samples were collected for both short and long-term soil health indicators, including the collection of additional surface samples sent to two commercial labs (Ward Lab in Nebraska, Waters Lab in Georgia) to evaluate their soil health analyses. At the end of October, soil samples were again collected for short-term soil health indicator measurements, and cover crop biomass was collected. The winter cover crop mixtures were planted in the beginning of November and included black oats, Florida 401 rye, daikon radish, sunnhemp, and hairy vetch. In the summer of Year 2, a novel physical indicator estimating aggregate stability with laser diffraction (long-term indicator) was developed. Data analysis has begun on the short and long-term indicators measured in Year 1 and 2 of the trial. Cover crop biomass from both years has been dried, weighed, ground, and sent for total C and N analysis. SO2: Microbial DNA was extracted from soil samples collected for long-term soil health indicator assessment in Year 1 and Year 2. The extracted DNA from Year 1 was quantified and sent for amplicon sequencing. The sequencing data is currently being analyzed to begin identification of taxa that may serve as potential indicators and candidates for future PGP isolations. Extracted DNA from Year 2 will be used for quantification via qPCR of specific nitrogen cycle functional genes. Additional samples will be collected in spring of 2023 for PGP analysis.? SO3: Objective 3 will begin in the third year of the project.

Publications


    Progress 02/14/21 to 02/13/22

    Outputs
    Target Audience:Researchers, producers, extension agents, citrus growers, and field specialists Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training and mentorship for one graduate student and a postdoctoral research associate are being provided by this project. How have the results been disseminated to communities of interest?The project plan and preliminary data from the proposal were presented at the Citrus Expo (Ft. Myers, FL), a state-wide citrus grower meeting in August 2021. In addition, preliminary data and the project plan were presented at the Florida Citrus Mutual/CRDF Education Session held virtually in Summer 2021. Preliminary data was presented by a postdoctoral research associate at the 2021 Annual Soil Science Society of America conference in November in Salt Lake City, UT.? What do you plan to do during the next reporting period to accomplish the goals?Cover crop plantings and soil collection will continue. Cover crops are scheduled to be planted twice a year at both locations. Soil sample collection for short-term indicators will occur three times during the year, and long-term collection will occur during the summer. Presentations of preliminary results will be made at regional and national scientific conferences, including the annual meetings of the Soil Science Society of America.?

    Impacts
    What was accomplished under these goals? During the first year of the project, we began the field trials in two citrus groves. Cover crops were planted in June and November. The June mixture included sunnhemp, cowpeas, buckwheat, millet, and Egyptian wheat, and the November mixture included daikon radish, sunnhemp, oats, and cereal rye. A PhD student funded by this project began their studies. SO1: Baseline soil and leaf samples were collected in May prior to the planting of cover crop treatments. Approximately three months after the first set of cover crops were planted, soil samples were collected for short and long-term soil health indicators. These included POX, C and N mineralization, ACE protein content, microbial biomass P, enzyme activities, CEC, water content, SOM, pH, inorganic N, and total C and N. DNA has been extracted for baseline assessments of the abundance of N and P-cycling functional genes. In October, just prior to planting of the winter cover crops, soil samples were collected for the second set of short-term indicators. In addition, cover crop biomass was sampled, dried, and are being prepared for total C and N analysis. SO2: Preparations are underway to begin the assessment of the relationship between soil health and PGP microbes. Samples for these assessments will be collected during the next annual sampling for long-term soil health indicators in Summer 2022 after cover crops have been planted for a year. SO3: Objective 3 will begin in the third year of the project.

    Publications