Source: NORTH DAKOTA STATE UNIV submitted to NRP
ALFALFA MANAGEMENT PRACTICES AND THEIR EFFECT ON ARBUSCULAR MYCORRHIZAL POPULATIONS - TOWARDS IMPROVING HEALTH, PRODUCTIVITY AND SUSTAINABILITY OF ALFALFA PRODUCTION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
OTHER GRANTS
Reporting Frequency
Annual
Accession No.
1020539
Grant No.
2019-70005-30239
Cumulative Award Amt.
$389,800.00
Proposal No.
2019-02820
Multistate No.
(N/A)
Project Start Date
Sep 1, 2019
Project End Date
Aug 31, 2023
Grant Year
2020
Program Code
[AFRP]- Alfalfa and Forage Program
Recipient Organization
NORTH DAKOTA STATE UNIV
1310 BOLLEY DR
FARGO,ND 58105-5750
Performing Department
Plant Sciences
Non Technical Summary
Alfalfa is a key crop to sustainable agriculture, contributing to the benefit of entire cropping systems, improvement of soil health, and numerous ecosystem services. In addition, alfalfa is a major livestock feed source nationwide. Alfalfa forage yield in rainfed environments is largely dependent on environmental conditions. South Dakota and North Dakota rank second and third in alfalfa acreage nationally, but rank 7th and 15th, respectively, in total national alfalfa production. The association of alfalfa with arbuscular mycorrhizal (AM) fungi plays an important role in alfalfa performance, especially when exposed to abiotic stresses. The presence of AM fungi has been shown to enhance nutrient uptake and to increase biomass yield, and improve forage nutritive value. These microbial communities are particularly important for alfalfa, and a better understanding the factors that contribute to the AM community composition may prove critical to increase the yield of alfalfa in stress environments. Our goals are to 1) identify the AM fungal species that colonize alfalfa in a variety of environments and soil types in North Dakota and South Dakota, 2) examine how current management practices affect these AM fungal communities, and 3) determine how management practices assist in establishing AM fungal communities with improved benefits for the host. We believe the forage community should use a more holistic approach to alfalfa management and take full advantage of AM fungi with the aim to increase yields in environments with limited resources, reduce input costs for producers, and improve the environmental sustainability of alfalfa production systems. ?
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2030199102060%
1020199107020%
2050199106020%
Knowledge Area
102 - Soil, Plant, Water, Nutrient Relationships; 205 - Plant Management Systems; 203 - Plant Biological Efficiency and Abiotic Stresses Affecting Plants;

Subject Of Investigation
0199 - Soil and land, general;

Field Of Science
1060 - Biology (whole systems); 1070 - Ecology; 1020 - Physiology;
Goals / Objectives
The main overall goal of this project is to try to develop novel strategies to reduce the existing yield gap between potential forage yield by top-tier farmers and average state yields by the colonization of alfalfa with arbuscular mycorrhizal (AM) fungi and by establishing more beneficial and sustainable AM communities. This strategy will make alfalfa a more profitable crop by increasing forage yield while reducing input costs. Our long-term goal is also to get non-alfalfa producers interested in integrating more alfalfa back into their crop rotations. The long-term goals of this project are: 1) to identify the diversity of AM fungal communities in alfalfa fields in North Dakota and South Dakota, 2) to determine how management practices influence the AM fungal communities in soils, and 3) to develop recommendations to increase the establishment of beneficial AM communities in alfalfa production systems. All of our objectives will integrate both research and extension activities.
Project Methods
Objective 1. To identify the existing AM fungal communities in alfalfa production systems in North Dakota and South Dakota - Experiment 1Soil and crown/root samples from at least 20 alfalfa fields in each state (SD and ND) will be taken at the beginning and at the end of the season each year of the project. The farms for sampling will be selected by state extension professionals, and will include fields under both yield-limiting and non-limiting conditions that represent the different environments in each state. At sampling, plant tissue samples and a soil sample from 0-6 inches and 6-24 inches in depth will be taken. The soil samples will be sent to the laboratory to analyze for N, P, K, organic matter, pH, EC, bulk density, and texture. In addition, information on plant height, growth stage, age of stand, variety and management (if available), and stand counts will be taken. Both soil and plant attributes and characteristics will be correlated to the AM fungal communities that colonized these plants. A sample for forage quality will be taken in each field. Samples will be dried and ground to 1-mm mesh. The forage quality of the samples from ND and SD will be determined with the NIR XDS Analyzer that is available in Dr. Berti's lab.Root materials will be collected, placed on ice and returned to the lab for further processing. At the lab, residual soil material will be washed off using tap water, and the root materials will be flash frozen in liquid nitrogen and stored at -80°C until further analysis. A minimum of 150 root segments will be used to determine the percentage of AM root colonization by the gridline intersection method (McGonigle et al., 1990). To identify the members of different AM fungal communities DNA will be extracted from the frozen root samples, checked for quality by running each sample on a 1.2% (w/v) agarose gel with TAE buffer, and quantified by using a ND-1000 spectrophotometer. Library construction and amplicon generation will be performed by using an Illumina Neoprep system for automated library preparation, followed by Illumina MySeq analysis. Sequence quality checks, BLAST analysis, and OTU generation will be performed using the program MOTHUR. The MaarjAM database will be used to align the sequences generated by the AM fungal primers (AMV4.5NF/AMDGR and NS31/AML2) and to generate the OTUs of the AM fungal communities (Öpik et al., 2010).Objective 2. Determine the influence of variety, K fertilizer rates and application dates on AM fungal colonization and proliferation - Experiment 2In spring 2019, field experiments will be established at two sites in North Dakota (sites will be determined according to soil test and clay ratios). The experimental design will be a randomized complete block design (RCBD) with a split-plot arrangement with four replicates. The main plot will be glyphosate-tolerant alfalfa varieties with different fall dormancy (FD) [FD3 (RR Presteez), FD4 (RR Stratica), and FD5 (L-450RR)]. Three K rates of 0, 150, and 300 lbs K2O/ acre will be applied before seeding or a split-application (half after the first cut in the seeding year and half in the fall, ~15 Sept.). Alfalfa will be planted in early May 2019, with an 8-row cone plot planter. In the seeding year, the first cut will be at 20-30% bloom and >24" in height for all plots, allowing the plants to establish. The second harvest will be done around Sept. 10 to stress the plants going into the winter. Plots will be 25 ft long and 5 ft wide with eight rows of alfalfa separated at 6" apart. The alfalfa seeding rate will be 10 lbs pure live seed/acre. Forage yield and quality will be analyzed from these plots. Soil and whole plants (root and shoot) will be sampled two weeks after emergence from only the three different varieties since K treatments will not be started until later. In the fall, plants from all treatments in this experiment will be sampled. All the samples will be frozen, and if this project is funded, they will be analyzed for AM colonization and AM community analysis (see above). Sampling will be repeated in 2020.Objective 3 Determine the AM fungal colonization of alfalfa plants grown in intercropping with corn compared with solely grown alfalfa - Experiment 3A corn-alfalfa intercropping system will be planted at one location in North Dakota and one in South Dakota. The experimental design will be a RCBD with four replicates and the following treatments: corn (T1), corn + alfalfa intercropping planted at the same time in the spring of 2020 (T2), and alfalfa alone (T3). In 2021, alfalfa will be established in the T1 treatment of the growing season 2020, and the other plots will continue with the alfalfa established under corn (T2), or the sole alfalfa treatment T3 established in 2020. Alfalfa plants will be harvested twice in 2020 and four times in year 2021. Soil and plant sampling for AM colonization and community analysis will be started two weeks after alfalfa emergence, one time in the middle of the growing season, and once in the fall. In the first production year, samples will be taken once at the end of the season. From the intercropping plots, samples of both alfalfa and corn roots will be taken. In addition, nine plots of this system that are already established on a farmer's field in Valley City, ND, will also be sampled. This on-farm replicated experiment has the same treatments as described above for Exp. 3 and was planted in 2018 as part of the USDA-NIFA CAP project. In this experiment, samples will be harvested by the end of May before the first cut.Objective 4. To determine if AM fungal communities and colonization varies with alfalfa variety - Experiment 4For this trial, existing alfalfa variety trials in both states will be sampled to determine the AM colonization and AM fungal community composition. Both, Berti and Bauder run variety testing trials which will be sampled to identify differences in AM colonization or community composition. Both trials include 'Vernal' as check variety. In addition, it is planned to plant a new variety trial with at least three old and three new varieties in North Dakota in 2020. The old varieties will include Vernal, Ladak, and Spredor, and the new varieties will be selected after discussion with farmers. Sampling for the AM analysis and forage yield and quality evaluations will be as described above in Objective 1.Objective 5. To determine the impact of mycorrhizal soil additives on alfalfa yield and quality - Experiment 5 Previous studies with soybean plants that were conducted in the Bücking lab, clearly demonstrated the important effect that the AM community composition plays for nutritional benefits and yield response. For these experiments, we will grow (in both states) the old alfalfa variety 'Vernal' and one of the varieties selected for Obj. 4. We will use (as described above) a RCBD and will compare the soil additives that we used for our soybean experiments. The soil additives differ in their AM fungal diversity and species composition (ranging from 4 to 9 fungal species). We will determine the yield and quality response of the alfalfa plants and will examine the plants for their AM colonization and community composition three times in Year 1 and Year 2 as described above.

Progress 09/01/19 to 08/31/23

Outputs
Target Audience: Farmers, ranchers, extension agents,crop consultants, researchers, graduate and undergraduate students, NRCS, and USDA personnel. Changes/Problems:The project was delayed due to the pandemic and the move of co-PI Buecking from SDSU to the University of Missouri. But all objectives have been completed and reported as indicated in the proposal. What opportunities for training and professional development has the project provided?Training: This project is providing training for two MS students at North Dakota State University and one MS and one PhD student at South Dakota State University/University of Missouri (the students started their degree program at South Dakota State University, and will complete their degree at the University of Missouri). Co-PI Buckingmoved her laboratory from South Dakota State University to University of Missouri, and both of her students moved to the University of Missouri to complete their MS or PhD degree. Both MS students at NDSU graduated and their thesis has been published and online at NDSU library. Peer -reviewed publications are being prepared but not yet submitted. Amy Greenberg, "Potassium fertilization and its impact on yield, nutritive value, root reserves, and winter hardiness of alfalfa", MS - North Dakota State University May 2019 -May 2021-graduated. Samuel Bibby, "Alfalfa-corn intercropping at different row spacings", MS - North Dakota State University, May 2020-August 2022 Salina Tripathi, "Effect of different management practices on the arbuscular mycorrhizal colonization of alfalfa", MS - South Dakota State University, January 2020- December 2023 from University of Missouri. Prema Mutyala, "Arbuscular mycorrhizal communities in field grown alfalfa populations", PhD - South Dakota State University, January 2020-expected graduation May 2024, from University of Missouri. In addition, Dr. Buecking trained a postdoctoral research associate and an undergraduate student (Grace Johnson) at the University of Missouri. The postdoctoral associate Dr. Dhruv Aditya Srivastava was only involved in the project at the end of the project and provided guidance for the metagenome analysis, and writing of scientific manuscripts (Das et al., 2023). Overall, he has participated in a grant writing workshop, and has improved his teaching and mentoring skills. He also wrote and won a grant from the University of Missouri Postdoc association. The postdoctoral scientist and Dr. Buecking mentored the undergraduate student Grace Johnson. She conducted a potassium stress experiment with alfalfa and learned microscopy and physiology of plants and AM fungi. She also presented part of her work as her poster during the Show Me Research Week at the University of Missouri and won the second prize for her presentation of around 100 undergraduate students that presented their work. This project also provided samples and served as field site for two other research projects in which Dr. Berti was co-PI: NAFA 10/2021-09/2022. Identification of rhizobium inoculants tailored for performance with new alfalfa varieties and diverse soil types. $74,325 PI Barney Geddes, NDSU. This project used the root samples collected in Objective 1 to evaluate the diversity of Rhizobia species in alfalfa in ND. One PhD student in Geddes lab is working on this project. USDA-NACA 09/2020-08/2022. Agronomic and molecular responses of maize and sunflower to competition with cover crops and alfalfa. $ 60,000, PI James V. Anderson USDA/ARS Fargo, ND. This project used the corn-alfalfa intercropping samples in Objective 3 to determine the differential gene expression in alfalfa and corn when intercropped. One MS student (Mckayla Neubauer) worked this project at NDSU and she is expected to graduate May 2024. How have the results been disseminated to communities of interest?This study has been showcased in research updates included in the "quarterly" update section of the Midwest Forage Association Forage Focus magazine as well as SDSU Extension online articles. In addition, Objective 4 variety trial plots have been discussed and shown at the SDSU Southeast Research Farm field day (held in July 2021), as well as the SDSU/UNL Forage Field Day (held in August 2021). Both field days were farmer friendly with audiences mainly consisting of producers and agronomists. Corn-alfalfa intercropping and potassium fertilization in alfalfa and effect on AM were featured in two field days at NDSU in 2022 and 2023. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Obj. 1 In 2021, alfalfa roots were sampled at 30 locations in ND and 22 locations in SD. Root samples were of different age (1-30 years) and differ in their management practices. The AM fungal colonization ranged from 12 to 88% and 18 to 82% with an average of 40% and 53% in the root samples from ND and SD, respectively. The average colonization rate in the ND samples was lower than in SD due to a higher number of sites with a very low colonization rate (≤ 21%). We found no significant effect of any the explanatory variables on arbuscular mycorrhizae (AM) fungal colonization in SD. In contrast, the ND samples showed a significant effect of grazing and irrigation on AM colonization at p≤0.05. Plant density also influenced AM fungi colonization at p<0.1. Livestock grazing has also previously been found to have a positive impact on AM fungi colonization. The removal of biomass in moderation can stimulate plant roots to produce more root exudates that could have a positive effect on AM fungal colonization. Obj. 2 Soil in Lisbon and Milnor, ND differ in their clay mineralogy, with a smectite to illite ratio > 3.5, in Milnor and a smectite to illite ratio <3.5 in Lisbon. Treatments were: three alfalfa varieties (Presteez, Stratica, and L-450) and 0, 168, and 336 kg K2O/ha in single- and split-application. Soil K was higher with a split-application, compared with a single-application of K at the same rate. Total seasonal forage yield was significantly lower when no K was applied. Independent of the location, K fertilization rates, and variety, plants showed a similar AM colonization rate of 50 to 60% in 2019. At both sites, the AM colonization of all varieties was high, and was not affected by the K treatment. In Lisbon in 2020, there was an increase in AM colonization in the variety L-450, and a decrease in Presteez in response to a higher K application. The AM colonization of L-450 and Stratica increased at high K rates, and in Presteez at low K rates. Obj. 3 This experiment was planted on May 2020 in Prosper and Hickson, ND. The experiment was a RCBD with four replicates and five treatments. The treatments were: 1) corn (76-cm row spacing), 2) corn (76-cm) + alfalfa intercrop, 3) alfalfa alone, 4) corn (152-cm row spacing), and 5) corn (152-cm) + alfalfa intercrop. In 2020, corn planted at 152-cm row spacing yielded 16% less than corn with 76-cm row spacing; however, there was no significant difference in corn yield with or without an alfalfa intercrop. In 2021, gravimetric water was significantly lower in treatments intercropped compared with treatments with corn alone when comparing the same corn row spacing. When comparing all two-year sequences, alfalfa intercropped with corn at 76-cm row spacing followed by established alfalfa saved an estimated $297/ha compared with seeding alfalfa in the spring after corn. Optimizing this specific cropping system for growers in the Midwest could increase profitability. Alfalfa and corn roots were separately analyzed for their AM colonization. Compared with Prosper, the AM colonization of alfalfa was consistently higher at Hickson (~55% to 60% in Hickson, and ~30% to 40% in Prosper), but the AM colonization in alfalfa alone did not differ from the alfalfa intercropped with corn. The spacing between the corn rows (76- or 152-cm) or the intercropping with alfalfa did not affect the AM colonization of the corn plants. Intercropping did not have an effect on the diversity of the AM community composition of alfalfa. We calculated the Simpson´s Diversity index and the relative abundance of each species in the community and found no significant effect of intercropping on alpha diversity. The AM fungal communities at both sites were mainly represented by species of the family Glomeraceae (GLO) (Funneliformis sp., Septoglomus sp., Glomus sp., Rhizophagus sp.) with mean relative abundances ranging from 68% to 100% irrespective of plot location and intercropping condition. Other fungal species in the samples belonged to the Claroideoglomeraceae (CLAR) (Claroideoglomus sp.) and Diversisporaceae (DIV) (Diversispora sp., Redeckera sp.). The analysis showed significant differences in AM fungal composition between the two locations in ND. However, the intercropping treatment did not contribute significantly to the difference in the AM community composition. Species belonging to the CLAR and DIV and unclassified fungi were present in higher percentages in the samples collected from Prosper than in those from Hickson. Consequently, GLO showed a significantly lower relative abundance in samples from Prosper. Obj. 4 The experiment was planted in Fargo, ND and Beresford, SD in May 2021. The experimental design was an RCBD with four replicates and eight alfalfa varieties. Vernal, Sholty (M. falcata), Aquamax and 54Q29 are conventional and Stratica, Presteez, L450 RR, and Vamoose are glyphosate-resistant. In 2021, alfalfa was harvested once in both locations due to a long-lasting drought period; no differences in yield were observed. In 2022, MycoApply, a mycorrhizal additive that is composed of 130,000 propagules per pound of four different AM species, G. intraradices, G. mosseae, G. aggregatum, and G. etunicatum was applied to half of plots after the first cut of the season. Alfalfa yield in 2022 varied among varieties but the MycoApply treatment did not have a significant effect in forage yield or nutritive value. The varieties Sholty and Vamoose were the lowest yielding in 2022 in SD. In 2022, in Fargo, Sholty had significantly lower yield than all varieties tested. Glyphosate tolerant varieties were higher yielding than conventional with the exception of 54Q29. In 2023, no differences in forage yield from treated or untreated treatments or variety effects were observed at either location. We found a statistically significant difference in the alpha diversity between the different alfalfa varieties harvested in 2022 in SD. Both, Stratica and Vernal had a higher alpha diversity than 54Q29, Aquamax, and Sholty. In ND, no difference in the alpha diversity among the different alfalfa varieties was found. Obj. 5 The alpha diversity of the AM communities was determined by an inverse Simpson index that was calculated based on a rarefied OTU (>97% similarity) abundance table. The AM fungal additive did not have a significant effect on the AM fungal alpha diversity. However, alfalfa plants grown in SD showed a higher alpha diversity of their fungal communities than the ND samples. The AM fungal colonization of alfalfa was quite high, and a lack of shift in the AM fungal community through the AM additive might be due to the high abundance of natural AM fungi before the inoculation. GLO fungi showed the highest abundance in the ND and SD samples and this dominance could also mask any difference in the AM community composition that MycoApply might have induced. Similar to our previous results, the AM communities in alfalfa roots were mainly dominated by species of the GLO family with mean relative abundance ranging from 75% to 93%. In SD, the AM communities also contained species of the following families: CLAR, Paraglomeraceae (PAR) (Paraglomus sp.), Gigasporaceae (GIG) (Gigaspora sp., Dentiscutata sp., Cetraspora sp., Racocetra, sp. Scutellospora sp.). The relative abundance percentage of some AM fungi families differed among plot locations with CLAR, PAR, and GIG present in higher percentages in samples collected from SD compared with those from ND. The ND samples had a higher relative abundance percentage of unclassified AM fungi. There was a shift in AM fungi from the CLAR family with the addition of AM inoculum in SD. This shift might be more pronounced in SD as the communities from the CLAR family were in lower percentage before inoculation. In ND, AM fungal communities from the GLO family were in higher percentages in the plants inoculated with AM fungi additive.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Bibby, S., M.T. Berti, A.F. Wick, D.P. Horvath, J.V. Anderson, A. Wittenberg, A. Greenberg, and K. Mozea. 2021. Corn-alfalfa intercropping with different row spacings. American Forage and Grassland Council Conference, Savanah, GA (virtual) 11-12 January 2021
  • Type: Theses/Dissertations Status: Published Year Published: 2021 Citation: Greenberg, A. 2021.Potassium fertilization and its impact on yield, nutritive value, root reserves, and winter hardiness of alfalfa . MS Thesis. North Dakota State University.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Berti, M.T. 2021. Why is potassium so important in alfalfa?. Alfalfa-U series. High Plains Journal 16 February 2021 (virtual). Available at http://www.hpj.com/alfalfau. Invited speaker.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Greenberg, A., M.T. Berti, D.P. Samarappuli, A. Peterson, S. Cabello, A. Wittenberg, and K. Mozea. 2021. Potassium fertilization and its impact on yield, quality, and winter hardiness of alfalfa. American Forage and Grassland Council Conference, Savanah, GA (virtual) 11-12 January 2021.
  • Type: Book Chapters Status: Published Year Published: 2023 Citation: Das D., Tripathi S., Mutyala P., Srivastava D.A., B�cking H.2023. Development and resource exchange processes in root symbioses of legumes, Book: Symbiosis in Nature, publisher IntechOpen, DOI: 10.5772/intechopen.111540
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Johnson G., Tripathi S., Srivastava D.A., Buecking H. 2023. Role of arbuscular mycorrhizal interactions in the potassium uptake of plants. Show Me Research Week, University of Missouri, April 18-20, 2023
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Tripathi S., B�cking H. 2022. The effect of management practices on AM fungi for sustainable alfalfa production. MU Plant symposium, University of Missouri.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Geddes, B., and M.T. Berti. 2023. Identification of Rhizobium inoculants tailored for performance with new alfalfa varieties and diverse soil types. Midwest Forage Association Annual Symposium, Wisconsin Dells, 21-22 February 2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Neubauer, M., M.T. Berti, S. Bibby, J.V. Anderson, and D.P. Horvath. 2021. How does intercropping of corn and alfalfa cause gene expression to vary? ASA-CSSA-SSSA International Annual Meetings, Salt Lake City, 7-10 November, 2021. (poster)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Berti, M.T. 2022. The fate of potassium fertilizer in alfalfa. Midwest Forage Association Annual Symposium, Wisconsin Dells , WI, 21-23 February.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Bibby, S., M.T. Berti, J.V. Anderson, E. Deckard, A.F. Wick, M. Neubauer, and K. Mozea. 2022. Corn-alfalfa intercropping with different row spacings. [Abstract] ASA-CSSA-SSSA International Annual Conference, Baltimore, MD, 6-9 November 2022. https//scisoc.confex.com/scsoc/2022am/meetingapp.cgi/Paper/142464
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Berti , M.T. 2022. Environmental benefits of intercropping systems in the northern Great Plains In Symposium-North to South and East to West: Cover Crop Use within Production Systems across the U.S. [Abstract] ASA-CSSA-SSSA International Annual Conference, Baltimore, MD, 6-9 November 2022. https//scisoc.confex.com/scsoc/2022am/meetingapp.cgi/Paper/141783
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Berti, M.T. 2023. Benefits of crop intercropping. South Dakota Soil Health Coalition Annual conference, Sioux Falls, SD, 24-25 January 2023
  • Type: Other Status: Published Year Published: 2022 Citation: Berti, M.T., and S. Bibby, 2021. Intercropping with different row spacings affects intercepted solar radiation. Forage Focus Magazine December 2021. Midwest Forage Association, St. Paul, MN.p. 17
  • Type: Other Status: Published Year Published: 2020 Citation: Berti, M.T. and A. Greenberg. 2020. Potassium alfalfa fertilization in seeding year did not affect stand persistence. Forage Focus December 2020 p. 8, Midwest Forage Association, St. Paul, MN
  • Type: Theses/Dissertations Status: Published Year Published: 2022 Citation: Bibby, S. 2022.Alfalfa-corn intercropping at different row spacings MS Thesis- North Dakota State University
  • Type: Theses/Dissertations Status: Under Review Year Published: 2024 Citation: Tripathi, S. Effect of different management practices on the arbuscular mycorrhizal colonization of alfalfa, MS Thesis. University of Missouri.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Berti, M.T., 2022. Managing cropping systems to enhance ecosystem services delivery. 37th Plant Sciences Graduate Student Symposium. Winnipeg, MB, Canada, 4-5 March 2022. Invited keynote speaker
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Berti, M.T., A. Cecchin, J.V. Anderson, and S. Bibby. 2022. Intercropping maize and alfalfa: A cropping system to mitigate the negative environmental impact of maize for ethanol production in the USA. A review European Biomass Conference, Marseille, France 9-12 May 2022. (virtual)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Neubauer, M., M.T. Berti, S. Bibby. J.V. Anderson, D. Horvath. 2022. Corn-alfalfa intercropping responses explained through transcriptome analysis. [Abstract] ASA-CSSA-SSSA International Annual Conference, Baltimore, MD, 6-9 November 2022. https//scisoc.confex.com/scsoc/2022am/meetingapp.cgi/Paper/141946


Progress 09/01/21 to 08/31/22

Outputs
Target Audience:Farmers, ranchers, scientists, graduate and undergraduate students, USDA-ARS and NRCS personnel, county extension agents, and crop consultants. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training: This project providedtraining for two MS students at North Dakota State University and one MS and one PhD student at South Dakota State University/University of Missouri (the students started their degree program at South Dakota State University, and will complete their degree at the University of Missouri). Co-PI Buckingmoved her laboratory from South Dakota State University to University of Missouri, and both of her students moved to the University of Missouri to complete their MS or PhD degree. A Amy Greenberg, "Potassium fertilization and its impact on yield, nutritive value, root reserves, and winter hardiness of alfalfa", MS - North Dakota State University May 2019 -May 2021-graduated. Samuel Bibby, "Alfalfa-corn intercropping at different row spacings", MS - North Dakota State University, May 2020- August 2022 Salina Tripathi, "Effect of different management practices on the arbuscular mycorrhizal colonization of alfalfa", MS - South Dakota State University, January 2020- graduation expected December 2022 from University of Missouri Prema Mutyala, "Arbuscular mycorrhizal communities in field grown alfalfa populations", PhD - South Dakota State University, January 2020-expected graduation August 2024 from University of Missouri Professional Development This study has been showcased in research updates included in the "quarterly" update section of the Midwest Forage Association Forage Focus magazine as well as SDSU Extension online articles. NDSU MS students presented their work at several conferences. In addition, objective 4 variety trial plots have been discussed and shown at the SDSU Southeast Research Farm field day, as well as the SDSU/UNL Forage Field Day. Both field days were farmer friendly with audiences mainly consisting of producers and agronomists. How have the results been disseminated to communities of interest?Resultswere presented by students at the ASA-CSSA-SSSA meeting in Salt Lake city in November 2022. Also, results were presented at the Midwest Forage Association Symposium. Extension publications have included part of the results of the project. What do you plan to do during the next reporting period to accomplish the goals?Objective 1. Complete sequencing of samples taken in 2021 across ND and SD. Sequencing to analyze AM communities will be done with18srRNA amplicon targeted sequencing. Analyze the results, interpret and publish. Objective 2. Complete sequencing of samples taken in this experiment in 2019 and 2020. Objective 3. Complete sequencing of samples taken in tis experiment in 2020. Objective 4. Agronomic, yield and forage quality data will be collected from July-October2022. Sampling for AMF colonization will be done in July 2022. Samples will be sent to Dr. Buecking for colonization analysis and posterior sequencing. The second, third and fourth cut ofthe alfalfa variety trial in ND and SD will be harvested in 2022. Samples will be ground and analyze din the NIR for forage quality. Sample roots after the second cut, send them to Dr. Buecking's lab forcolonization assesment and sequencingto determine alfalfa communities. Objective 5. Sample roots after the second harvest with and without AMF inoculant andsend them to Dr. Buecking's lab forcolonization assessment and sequencing. Publications containing the results of all objectives will be prepared and submitted by August 2023. We expect to write at least four publications.

Impacts
What was accomplished under these goals? Objective 1 To identify the diversity of existing AM fungal communities in alfalfa production systems in North Dakota and South Dakota The collection and preparation of samples has been completed as planned. The samples were assessed using the gridline intersection method for AMF colonization (McGonigle et al.1990). We studied how different environmental and soil variables such as (1) estimated age of the stand, (2) growth stage, (3) plant height in cm, (4) plant density, (5) irrigation, (6) grazing, (7) field conditions (dry, moderate, moist), (8) soil salinity, (9) presence of insects, (10) active nodules, or (11) organic production affect the AM colonization of the host plants and the composition of the AM fungal communities. Multiple linear regression analysis was performed to test the effect of these explanatory variables on the response variable (AM colonization). The AMF colonization percentage ranged from 12% to 88% (average of 40.3%) and from 18.2% to 82.2% (average of 53.4 %) in the root samples from North Dakota and South Dakota, respectively.The results of the multiple linear regression analysis of the SD samples showed no significant effect of the explanatory variables on AM fungal colonization. In order to avoid an inaccurate estimation of variance due to multicollinearity among explanatory variables, a variation factor (VIF) analysis with a cut-off value of >10 was used to exclude variables that showed multi-collinearity. By contrast, the North Dakota samples showed a significant effect of grazing and irrigation on the AM colonization at p≤0.05. Plant density also influenced the AM colonization at p< 0.1. Grazing, irrigation, and plant density were subsequently tested alone in simple regression models. Grazing had a significant positive effect on AM colonization percentage (Adj R2 = 9.2%, p< 0.1). However, the effect of plant density and irrigation effect was not found to be significant when tested alone. In general, livestock grazing has a negative effect on AM abundance. Grazing decreases the above-ground biomass, and thereby reduces the carbon supply for the AM fungus that is necessary to expand its soil mycelium. However, this effect depends on the grazing intensity. Heavy grazing has a strong effect on AM abundance, while light grazing has no significant effect (Yang et al. 2020). Depending on plant density, AM benefits for the host plants can differ. Duan et al. (2021) for example observed that high plant densities can increase the shoot biomass, grain yield and water use efficiency of wheat. Objective 2. To determine the influence of alfalfa variety, K fertilizer rates, and application dates on AM fungal colonization and proliferation The field experiment has been completed. Results have been reported at conferences. The roots were sampled in 2019 and 2020. AMF colonization assesment has been completed and previously reported.In progress,18srRNA amplicon targeted sequencing to analyze the AM communities. Objective 3. To determine the AM fungal colonization in alfalfa plants grown in intercropping systems with corn compared with alfalfa in monoculture The field experiment has been completed. Agronomic, yield data and AMF colonization are completed and reported.Only sequencing of the AM communities remains. Objective 4. To determine if AM fungal colonization and benefits varies with alfalfa variety This experiment was established in 2021, a year of extreme drought both in ND and SD. Varieties established but sampling was postponed for 2022. Only one cut was possible and forage quality was not determined. Glyphosate-tolerant varieitiesyielded more than conventional alfalfa varieties and yellow-flower alfalfa (Medicago falcata). Objective 5. To determine the impact of mycorrhizal soil additives on alfalfa yield and quality A mycorrhizal inoculant, Glomus mosseae, was applied after the first cut of alfalfa in half of the plots of the variety trial in Objective 4. Sampling for roots will be done in July 2022 after the second cut of alfalfa. Yield and forage quality of alfalfa will be recorded at the end of the season in 2022. AMF colonization analysis and sequencing will be done from August 2022 until March 2023. Analysis, interpretation of the information, and publication will follow.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Berti, M.T., A. Cecchin, J.V. Anderson, and S. Bibby. 2022. Intercropping maize and alfalfa: A cropping system to mitigate the negative environmental impact of maize for ethanol production in the USA. A review European Biomass Conference, Marseille, France 9-12 May 2022. (virtual)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Berti, M.T. 2022. Modifying cropping systems management to mitigate environmental impact and increase ecosystem services delivery. Environmental and Conservation Sciences Seminar Series. North Dakota State University. 20 January 2022. Invited speaker
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Neubauer, M., M.T. Berti, S. Bibby, J.V. Anderson, and D.P. Horvath. 2021. How does intercropping of corn and alfalfa cause gene expression to vary? ASA-CSSA-SSSA International Annual Meetings, Salt Lake City, 7-10 November, 2021. (poster)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Berti, M.T., 2022. Managing cropping systems to enhance ecosystem services delivery. 37th Plant Sciences Graduate Student Symposium. Winnipeg, MB, Canada, 4-5 March 2022. Invited keynote speaker
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Bibby, S., M.T. Berti, A.F. Wick, D.P. Horvath, J.V. Anderson, A. Greenberg, K. Mozea, and M. Neubauer. 2021. Corn alfalfa intercropping with different row spacings. ASA-CSSA-SSSA International Annual Meetings, Salt Lake City, 7-10 November, 2021.
  • Type: Other Status: Published Year Published: 2022 Citation: Berti, M.T. and S. Bibby. 2022. Establishing alfalfa while growing corn. Progressive Forage Magazine. Issue 2, February 2022. p. 28-31


Progress 09/01/20 to 08/31/21

Outputs
Target Audience:Farmers, ranchers. Changes/Problems:The Co-Pd Bucking moved from South Dakota State University to University of Missouri and both graduate students in the project moved as well. This change did not signify any delaysince many activities in the project have been delayed due to Covid19 restrictions. A new subcontratc to Univ. of Missouri is already in place. What opportunities for training and professional development has the project provided?This project is providing training for two MS students at North Dakota State University and one MS and one PhD student at South Dakota State University/University of Missouri (the students started their degree program at South Dakota State University, and will complete their degree at the University of Missouri). Co-PD Buckingmoved her laboratory from South Dakota State University to University of Missouri, and both of her students moved to the University of Missouri to complete their MS or PhD degree. All four students already have started their respective degree program and one MS student graduated in May 2021. Amy Greenberg, "Potassium fertilization and its impact on yield, nutritive value, root reserves, and winter hardiness of alfalfa", MS - North Dakota State University May 2019 -May 2021-graduated. Samuel Bibby, "Alfalfa-corn intercropping at different row spacings", MS - North Dakota State University, May 2020- expected graduation August 2022. Salina Tripathi, "Effect of different management practices on the arbuscular mycorrhizal colonization of alfalfa", MS - South Dakota State University, January 2020- graduation expected August 2022 from University of Missouri. Prema Mutyala, "Arbuscular mycorrhizal communities in field grown alfalfa populations", PhD - South Dakota State University, January 2020-expected graduation August 2024 from University of Missouri. Professional Development This study has been showcased in research updates included in the "quarterly" update section of the Midwest Forage Association Forage Focus magazine as well as SDSU Extension online articles. In addition, objective 4 variety trial plots have been discussed and shown at the SDSU Southeast Research Farm field day (held in July 2021), as well as the SDSU/UNL Forage Field Day (held in August 2021). Both field days were farmer friendly with audiences mainly consisting of producers and agronomists. How have the results been disseminated to communities of interest?Preliminary results have been presented at local and national conferences by the students in the project. What do you plan to do during the next reporting period to accomplish the goals?Objective 1. To identify the diversity of existing AM fungal communities in alfalfa production systems in North Dakota and South Dakota The roots collected in ND and SD in May of 2021 are currently being analyzed for their AM colonization, and will be prepared for metagenome analysis. This will be conducted at the University of Missouri. Objective 2. To determine the influence of alfalfa variety, K fertilizer rates, and application dates on AM fungal colonization and proliferation This study objective has been completed. At least two publications will be prepared and submitted. Objective 3. To determine the AM fungal colonization in alfalfa plants grown in intercropping systems with corn compared with alfalfa in monoculture The experiment planted in 2020 at Hickson and Prosper continued in 2021 with only alfalfa. Alfalfa yield in Year 2 compared with spring-seeded alfalfa will be compared. In addition, this experiment was repeated in 2021 at the same locations, only to record corn and alfalfa yield in Year 1, AM sampling will not be done. Root samples of both corn and alfalfa taken in 2020 will be prepared for metagenome analysis. Objective 4. To determine if AM fungal colonization and benefits varies with alfalfa variety The variety trial was established in 2021, but due to drought only one cut in ND and two cuts in SD were possible. The experiment will continue in 2022. Objective 5. To determine the impact of mycorrhizal soil additives on alfalfa yield and quality This objective was originally planned to start 2021. The variety trial was planted but the mycorrhizal soil additive was not applied due to persisting drought. We plan to apply the soil additive in the spring of 2022.

Impacts
What was accomplished under these goals? The three goals of the project were divided in five objectives. Accomplishment for each objective are indicated below: To identify the diversity of existing AM fungal communities in alfalfa production systems in North Dakota and South Dakota This objective was planned for the summer of 2020, but it was postponed to 2021 due to Covid-19 travel restrictions. In 2021, alfalfa roots were sampled at 30 locations in North Dakota and 22 locations in South Dakota between May 25-29. Samples for this experiment were taken from different sites in North Dakota (Fargo, Hickson, Milnor, Lisbon, Carrington, Sykeston, Williston, Charbon Township, Watford City, Dunn Center, Dickinson, Richardton, Mott, Hettinger, SD National Grasslands, New Leipzig, Strasburg, Linton, Napoleon, Alkaline Lake, REC Streeter) that were planted between 1988 (Napoleon) and 2020 (Hickson) and differ in their management practices. The samples in South Dakota were taken from Beresford, Tyndall, Mt. Vernon, Baltic, Madison, Wilmot, Clark, Ipswitch, Selby, Timber Lake, Buffalo, Newell, Spearfish, Oelrichs, Long Valley, Colome, Pierre, Ree Heights, and Brookings). Soil samples were taken at each site and N-NO3, P, K, organic matter, pH, and electrical conductivity were evaluated. Samples are currently being prepared for AMF colonization analysis and metagenome analysis. 2. To determine the influence of alfalfa variety, K fertilizer rates, and application dates on AM fungal colonization and proliferation Two separate sites were established for this study. One location is East of Lisbon, ND (46°26'N, -97°11'W, 325 m). The second location is in Milnor, ND (46°16'N, -97°28'W, 337 m). The two locations differ in their clay mineralogy and their smectite-to-illite ratio. With a smectite to illite ratio greater than 3.5, the soil at Milnor immobilizes potassium, while the soil at Lisbon with a smectite to illite ratio less than 3.5, allows potassium to be more mobile. The three alfalfa varieties Presteez RR, Stratica RR, and L-450 RR were used for this study. Each of the varieties has a different fall dormancy score. The experiments were conducted in Lisbon and Milnor, ND, in 2019 and 2020. Three different K treatments were applied: 0, 168, and 336 kg K2O ha-1 in single- and split-application. Half of the experimental units were stressed by harvesting mid-September, while the other half was non-stressed by harvesting in October. Soil K was higher with a split-application, compared with a single-application of K at the same rate. Total seasonal forage yield was significantly lower when no K was applied. Stressed alfalfa had lower root protein in both years and starch was lower in Milnor 2019 and Lisbon 2020 compared with the non-stressed treatment. We also analyzed the arbuscular mycorrhizal (AM) colonization of the roots. Independent of the location (Lisbon and Milnor), the K fertilization, and the variety, the plants showed a similar mycorrhizal colonization rate of 50 to 60% in 2019. Compared with 2019, the AM colonization in 2020 of the different varieties increased particularly in Lisbon. While the AM colonization at Milnor did not differ, the AM colonization of the varieties L-450RR and Stratica increased at high K fertilization rates, and the colonization rate of Presteez at low K fertilization rates and reached up to 75%. 3. To determine the AM fungal colonization in alfalfa plants grown in intercropping systems with corn compared with alfalfa in monoculture This experiment was planted on May 21, 2020 at Prosper and Hickson, ND. Each plot was laid out in a randomized complete block design consisting of four replicates and five treatments. Treatments 1-5 included corn at two row spacings: 30 and 60 inches. The specific experimental treatments are as follows: 1) corn (30 inches), 2) corn (30 inches) + alfalfa intercrop, 3) alfalfa alone, 4) corn (60 inches), 5) corn (60 inches) + alfalfa intercrop. Every treatment with corn includes two rows in the middle of the plot for harvest and one row on each side as a border. In Year 1, only treatments 1, 2, 4, and 5 of corn were harvested. Treatment 3 was harvested once in year one. In year two, treatments 1 and 4 were seeded into alfalfa and all other treatments contained established alfalfa. In 2020, established alfalfa was harvested four times weather permitting and new seedings were harvested once. The overall scale of success within these treatments is measured in corn grain yield in the first year and alfalfa establishment and forage yield in Year 2. In the year 2020, corn planted with 60-inch row spacing yielded 16% less than corn with 30-in row spacing; however, there was no significant difference in corn yield with or without an alfalfa intercrop. Data collected on each experimental unit also included measurements of water stress, weed suppression, and soil health. The data sets collected over this two-year study will allow for more specific recommendations regarding alfalfa-corn intercropping. Optimizing this specific cropping system for growers in the northern Great Plains could increase profitability as well as forage nutritive value and crop efficiency. Alfalfa and corn roots were separately analyzed for their AM colonization. Compared with Prosper, the AM colonization of alfalfa was consistently higher at Hickson (~55% to 60% in Hickson, and ~30% to 40% in Prosper), but the AM colonization in the plots in which alfalfa was planted alone (treatment 3) did not differ from the plots in which alfalfa was planted with corn (treatment 2 and 5). While the alfalfa colonization in Hickson was higher than in Prosper, the AM colonization of corn did not differ at both sites, and was with ~25% to 35% relatively low. The spacing between the corn rows (30 or 60 inches) or the intercropping with alfalfa did not affect the AM colonization of the corn plants. 4. To determine if AM fungal colonization and benefits vary with alfalfa variety This experiment was planted at Fargo, ND and Beresfrod SD in May 2021. The experimental design is an RCBD with four replicates and eight alfalfa varieties. Four varieties (Vernal, Sholty (M. falcata), Aquamax, and Pioneer 54Q29) are conventional and four varieties (Stratica, Presteez, L450 RR, and Vamoose) are glyphosate-tolerant. Alfalfa was harvested onceat both locationsdue to a long-lasting drought period not growing back well enough for a second cutting in either location. 5. To determine the impact of mycorrhizal soil additives on alfalfa yield and quality This objective needed to be postponed to the summer 2021 due to Covid19 restrictions that limited our ability to start new experiments. However, the 2021 season has been extremely dry, thus the inoculation with AM - Myco product was postponed to spring 2022.

Publications


    Progress 09/01/19 to 08/31/20

    Outputs
    Target Audience:Farmers, ranchers, extension agents, crop consultants, researchers, graduate and undergraduate students, NRCS, and USDA personnel. Changes/Problems:Objective 1 and 4 planned for 2020 have been postponed for 2021 due to Covid19 restrictions to plant new experiments. What opportunities for training and professional development has the project provided?This project is providing training for two MS students at North Dakota State University and two MS students at South Dakota State University. All four students already have started their MS program. How have the results been disseminated to communities of interest?Results from this research have been presented at national conferences and farmers workshops. What do you plan to do during the next reporting period to accomplish the goals?1. To identify the diversity of existing AM fungal communities in alfalfa production systems in North Dakota and South Dakota This objective was planned for the summer of 2020, however due to Covid19 will be postponed to 2021. At this time travel in North Dakota and South Dakota is limited only to essential research. 2. To determine the influence of alfalfa variety, K fertilizer rates, and application dates on AM fungal colonization and proliferation Analysis of root samples collected in the fall of 2019 for AMF colonization and root carbohydrates and protein content will be completed in the summer of 2020. Genotyping of AMF will start by the end of 2020.Root samples from this experiment will be taken in July 2020 and sent to South Dakota State University for analysis. This experiment will be harvested for yield and alfalfa will be analyzed for forage nutritive value in 2020. 3. To determine the AM fungal colonization in alfalfa plants grown in intercropping systems with corn compared with alfalfa in monoculture In Year 1, only corn will be harvested in treatments 1, 2, 4, and 5. Treatment 3 will be harvested once in year one. In Year 2, treatments 1 and 4 will be seeded into alfalfa and all other treatments will contain established alfalfa. In year two established alfalfa will be harvested four times weather permitting and new seeding will be harvested once. Forage yield and quality of alfalfa and biomass and grain yield of corn will be evaluated. Samples for AMF analysis will be taken at the end of July and sent to Dr. Heike Bucking for analysis. 4. To determine if AM fungal colonization and benefits varies with alfalfa variety This objective has been postponed to start summer 2021 due to Covid 19 restrictions to plant new experiments. 5. To determine the impact of mycorrhizal soil additives on alfalfa yield and quality This objective was originally planned to start 2021.

    Impacts
    What was accomplished under these goals? The three goals of the project were divided in five objectives. Accomplishment for each objective are indicated below. 1. To identify the diversity of existing AM fungal communities in alfalfa production systems in North Dakota and South Dakota This objective was planned for the summer of 2020, however due to Covid19 will be postponed to 2021. At this time travel in North Dakota and South Dakota is limited only to essential research. 2. To determine the influence of alfalfa variety, K fertilizer rates, and application dates on AM fungal colonization and proliferation Two separate sites were established for this study. One location is East of Lisbon, ND (46°26'N, -97°11'W, 325 m. The second location is in Milnor, ND (46°16'N, -97°28'W, 337 m elevation). The two locations differ in their smectite-to-illite ratio of clay mineralogy. Milnor has soil that immobilizes potassium, having a smectite to illite ratio greater than 3.5 and Lisbon has soil that allows potassium to be more mobile having a smectite to illite ratio less than 3.5.Three alfalfa varieties were used for this study. These varieties are Presteez RR, Stratica RR, and L-450 RR. Each of the varieties have a different fall dormancy score. The experiment set up is a randomized complete block design with a split-plot arrangement and four replicates. The main plot was the variety and the subplots a factorial combination of K rates and application timing. Each plot is split in half, having one-half of the plot being harvested in the middle of September, stressing the alfalfa, and having the second-half of the plot being harvest before first frost in October, non-stressing the alfalfa. Rate and timing of K is the treatment. Treaments were : T1 no K application, T2 168 kg/ha of K at seeding; T3 168 kg/ha split-application (seeding and September) , T4 336 kg K/ha at seeding , T5 336 kg K/ha split-application, same time as T3. Root materials were collected in October 2019 placed on ice and returned to the lab for further processing. At the lab, residual soil material was washed off using tap water, and the root materials were frozen in liquid nitrogen and stored at -80°C until further analysis, or prepared to determine the AM colonization. To determine the AM colonization, the roots were cleared with 10% KOH solution at 80C for 30 min, rinsed, and stained with 5% ink at 80C for 15 min. Results In the 2019 , there were no significant effects found from K fertilization rates and timings and total yield P = 0.05. The total forage yield (Mg/ha) was 7.0, 7.3, 7.0, 7.4, and 7.2 for treatments 1, 2, 3, 4, and 5, respectively. There was also no significant interaction between treatments, cuts, nor varietal differences. When analyzing K effects on forage quality, there was no effect on ADL, NDF, CP, and NDFD. There was, however, effects among K treatments and total percentage of ash and TDN. With increased K application, there was an increase in ash content. Treatment 1 had the lowest ash% of 7.5, 7.8, and 8.4% at cuts 1, 2 (stressed), and 2 (non-stressed), respectively. The highest ash content was treatment 4 with 8.1, 8.3, and 8.7% at cuts 1, 2 (stressed), and 2 (non-stressed), respectively. Increasing K application, decreased total digestible nutrients (TDN). Treatment 1 had the highest TDN at 61.0%, and treatment 4 had the lowest at 59.4%. The K treatments that were split-applied had higher TDN than K treatments that were applied entirely at seeding. There was also a significant effect between varietal fall dormancy and cutting with the total TDN. The variety with FD3 had the highest TDN across all three cuts and locations. Colonization of roots by AMF varied between 40-80% but analysis to determine differences among treatments still has not been completed. 3. To determine the AM fungal colonization in alfalfa plants grown in intercropping systems with corn compared with alfalfa in monoculture This experiment was planted on May 21, 2020 at Prosper and Hickson, ND. The planned South Dakota location for this experiment will be postponed to 2021 due to restrictions for new experiments in South Dakota due Covid19. Each plot was laid out in a randomized complete block design consisting of four replicates and five treatments. Treatments 1-5 will include corn at two row spacings 30 and 60 inches. The specific experimental treatments are as follows: 1) corn (30 inches), 2) corn (30 inches) + alfalfa intercrop, 3) alfalfa alone, 4) corn (60 inches), 5) corn (60 inches) + alfalfa intercrop. Every treatment with corn includes two rows in the middle of the plot for harvest and one row on each side as a border. 4. To determine if AM fungal colonization and benefits varies with alfalfa variety This objective has been postponed to start summer 2021 due to Covid 19 restrictions to plant new experiments. 5. To determine the impact of mycorrhizal soil additives on alfalfa yield and quality This objective was originally planned to start 2021.

    Publications

    • Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Greenberg, A., M.T. Berti, D.P. Samarappuli, A. Peterson, S. Cabello, A. Wittenberg, and K. Mozea. 2020. Impact of potassium fertilization on yield, quality, and winter hardiness of alfalfa. American Forage and Grassland Council Conference Greenville, SC, 5-8 January 2020.
    • Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Berti, M.T. 2020. Potassium Impact on yield, quality, and winter hardiness. Midwest Forage Associations Annual Conference. Wisconsin Dells, WI, 18-19 February 2020.
    • Type: Other Status: Published Year Published: 2020 Citation: Berti, M.T., and H. B�cking. 2020. Alfalfa productivity and nutrient uptake is related to interaction with soil microbiome. Forage Focus March 2020. Midwest Forage Association, St. Paul, MN p.18-19.
    • Type: Other Status: Published Year Published: 2020 Citation: Berti, M.T., and A. Greenberg. 2020. Potassium fertilization does not increase alfalfa forage yield in the seeding year. Forage Focus. March 2020. Midwest Forage Association, St. Paul, MN p. 3-4
    • Type: Other Status: Published Year Published: 2020 Citation: Holin, F. and M. Berti. 2020. Researcher works to Reduce Potassium Deficiency in Alfalfa. Clippings, Midwest Forage Association, St. Paul, MN. February 27, 2020.