Progress 04/15/23 to 04/14/24
Outputs
Target Audience:April 15, 2024 -Annual Report Seeds For The Future: Spring 2023 through to April 15, 2024 Project title: Undergraduate training in applied plant microbiology for regenerative agriculture and habitat restoration in the Intermountain West Subaward No.: AS5339-691611 2023 Cohort A website was developed to communicate to the public and potential applicants about our Seeds For The Future program (www.gonzaga.edu/SeedsForTheFuture). To obtain the 2023 cohort we advertised our program with 20 regional colleges and universities in our region. From this recruitment we obtained 15 applicants and selected 7 undergraduate students from 4 regional universities to be our 2023 Cohort for our program. Participants 2023: seven participants on two university campuses; Students represented three states in the Inland Northwest; two males, two nonbinary, and three females; underrepresented students included one Asian student and two LBGQT student; three students were high-financial need students; and all seven students were undergraduates at local universities at the time of application. One participant from the 2022 program returned for a second year in 2023; funds from Gonzaga University helped to support this student. Research Location Program Duration Home University Name Gonzaga University 10-weeks summer Pacific Lutheran University Sara Wifall Gonzaga University 10-weeks summer Gonzaga University Allie Greenhouse Gonzaga University 10-weeks summer Gonzaga University Jon Mai Gonzaga University 10-weeks summer Gonzaga University Michael Robitaille University of Idaho 10-weeks summer Utah State University Signee Storrud University of Idaho 10-weeks summer North Idaho College Wyatt Tindall University of Idaho 10-weeks summer Gonzaga University Avery Kuehfuss Projects 2023 and Academic Year 2023-2024. In Summer 2023 our mentor to fellow ratio was 1:2, down from 1:5 in 2022 at Gonzaga University. At the University of Idaho the ratio was 1:1.5, down from 1:2 in 2022. Treasure Valley Community The lower mentor to fellow ratio were lower and allowed for a stronger mentoring and allowed us to take the students to a national science meeting, providing a transformational experience for the student fellows. During the 2023-2024 academic year, both the Gonzaga University and Treasure Valley Community College involved students in program. Gonzaga University involved 4 students in projects from January to May with a ratio of 1:4 Title: The Role of pH and Fungal Inoculation on Bottlebrush Squirreltail (Elymus elymoides) Growth. Student: Sara Wifall. Mentor: Julie Beckstead and Brian Connolly Title: Examining the Resilience of Squirreltail Grass (Elymus elymoides) Against a Zinc Gradient. Student: Connor Robitaille. Mentors: Julie Beckstead and Brian Connolly Title: Antagonistic Activity is observed from seed endophytes: A potential tool to aid seeds in establishment during restoration. Student: Alexandrite Greenhouse. Mentor: Julie Beckstead Title: A Pain in the Grass: Combining Germination Cues and Soil Freezing toLimit Cheatgrass (Bromus tectorum) Emergence. Student: Jon Mai. Mentors: Julie Beckstead and Brian Connolly Title: Thermotolerance of Bacterial Endophytes from Callitropsis bakeri. Student: Avery Kuehfuss. Mentors: George Newcombe and Abby Ferson-Mitchell Title: Acacia koa, Acacia melanoxylon, and Acacia mangium endophytes interact with Fusarium oxysporum f. sp. koae in antagonistic assays. Mentors: George Newcombe and Abby Ferson-Mitchell Title: The Effects Nematode Interactions have on the Relative Growth Rate of Hericium erinaceus. Student: Wyatt Tindall. Mentors: George Newcombe and Abby Ferson-Mitchell Changes/Problems:Project Director Newcombe is retiring from the University of Idaho at the end of June 2024. Prof. Newcombe will continue to help with the output from the project (e.g., publication of the manuscript that is being drafted - current draft included elsewhere in this report) despite his retirement. What opportunities for training and professional development has the project provided?Projects 2023 and Academic Year 2023-2024. In Summer 2023 our mentor to fellow ratio was 1:2, down from 1:5 in 2022 at Gonzaga University. At the University of Idaho the ratio was 1:1.5, down from 1:2 in 2022. Treasure Valley Community The lower mentor to fellow ratio were lower and allowed for a stronger mentoring and allowed us to take the students to a national science meeting, providing a transformational experience for the student fellows. During the 2023-2024 academic year, both the Gonzaga University and Treasure Valley Community College involved students in program. Gonzaga University involved 4 students in projects from January to May with a ratio of 1:4 Title: The Role of pH and Fungal Inoculation on Bottlebrush Squirreltail (Elymus elymoides) Growth. Student: Sara Wifall. Mentor: Julie Beckstead and Brian Connolly Title: Examining the Resilience of Squirreltail Grass (Elymus elymoides) Against a Zinc Gradient. Student: Connor Robitaille. Mentors: Julie Beckstead and Brian Connolly Title: Antagonistic Activity is observed from seed endophytes: A potential tool to aid seeds in establishment during restoration. Student: Alexandrite Greenhouse. Mentor: Julie Beckstead Title: A Pain in the Grass: Combining Germination Cues and Soil Freezing toLimit Cheatgrass (Bromus tectorum) Emergence. Student: Jon Mai. Mentors: Julie Beckstead and Brian Connolly Title: Thermotolerance of Bacterial Endophytes from Callitropsis bakeri. Student: Avery Kuehfuss. Mentors: George Newcombe and Abby Ferson-Mitchell Title: Acacia koa, Acacia melanoxylon, and Acacia mangium endophytes interact with Fusarium oxysporum f. sp. koae in antagonistic assays. Mentors: George Newcombe and Abby Ferson-Mitchell Title: The Effects Nematode Interactions have on the Relative Growth Rate of Hericium erinaceus. Student: Wyatt Tindall. Mentors: George Newcombe and Abby Ferson-Mitchell Calendar 2023 We held weekly Zoom lab meetings with all institutions. Activities consisted of discussion of primary literature, sharing of research projects, and trouble-shooting research problems that surfaced. Students were required to write a short reflection for each primary literature article read along with reflections about the field trips. Connections were made with seed producers, government agencies involved in preservation of crop plants and native plants. Field trips for the 2023 program were built around these connections and some were repeated from 2022. Date Events and Field Trips May 23 First day for 10-week summer research program May 23 Field Trip 1: Gonzaga only. Visit Clearwater Seeds Company, Spokane, WA May 25 Field Trip 2: University of Idaho - Botanical Garden Field Trip, Moscow, ID. Field Trip 3: Gonzaga - Hanford Reach National Monument to see cheatgrass invaded public lands and native plant communities. Seed collections obtained. June 21 Field Trip 4: Both Gonzaga University and University of Idaho. Visit Plant Germplasm Introduction and Testing - USDA AgriculturalResearchService lab, Pullman, WA. July 22 Field Trip 5:Traveled to Boise Idaho for Botany 2023, annual meetings of the Botanical Society of America. July 23 Field Trip 6: Both Gonzaga University and University of Idaho. Visited Boise Ridge, which is part of the Boise National Forest and Bogus Basin Recreation Area, for an all-day hike offered through the Botany Meetings. Besides the awesome views, we identified many native plants and participated in a discussion about forest management options in the face of climate change. July 24-26 Oral and poster presentations presented by the mentors and several of the students at the Botany 2023 Meetings. Students attended a variety of workshops, luncheons and meeting sessions at the Botany Meetings. July 26 Field Trip 7: Both Gonzaga University and University of Idaho. Visited Idaho Botanical Garden at Boise, Idaho. We talked with the garden manager and staff and enjoyed learning about the many plants on display. Seeds For The Future Symposium 2022. Both Gonzaga University and University of Idaho. Students each presented their research. July 26 Seeds For The Future Symposium 2023. Both Gonzaga University and University of Idaho. Students each presented their research. July 27 Traveled back to Spokane WA from the Botany 2023 annual meetings. July 29 Last day for 10-week summer research program Professional Development Workshops 2023 (total of 11 plus additional at Botany Meetings) Safety Training Workshop and Discussion Poster Presentation Workshop Data Collection and Data Analysis Workshop Ethics Workshop: Mentor-Mentee Relationship Ethics Workshop: Authorship Writing a Professional Statement Workshop Writing a Resume and CV Workshop Report Writing and Presentation Preparations Workshop Data Management and Statistical Analysis Graduate School Preparations Writing Grant Proposal Workshop Additional workshops were attended at the Botanical Society of America Meetings. Outreach 2023 Participants developed hands-on science activities associated with seeds and microbes aimed for elementary science classrooms. Participants played the previously developed science game from the 2022 participants, Glorious Germination card game. Science learning games were played with over 100 students in 2023, expanding the plant-microbe knowledge of these students. A publication in an education journal is in preparation. How have the results been disseminated to communities of interest?Outputs Supported 7 students for 10-weeks in the Seeds For The Future Program in Summer 2023. Eight presentations at the Botanical Society of America Annual Meetings in Boise Idaho in July 2023; five poster presentations, two lightening talks, and one oral presentation. One poster presentation from a 2022 undergraduate participant. Outreach: Development of two learning games about seeds and their endophytes by undergraduate participants. 1) High School game called "Glorious Germination" and 2) Grades 3rd-5th game called "Seeds' Hidden Helpers". Games are being disseminated to regional schools and publications in education journals are in the works. Games were played with over 100 undergraduate students and also with 1-6 grade students at the Salish School of Spokane. Communication with larger community: 1) Science learning game materials made available on the program's website, 2) YouTube video about the science learning game made available online, 3) news article about plant curriculum featured in the Gonzaga University Bulletin. Development of plant-related suite of courses for Gonzaga University, including modification to two existing courses (now Plant Population Ecology and Plant Community Ecology) and a newly designed Plant Propagation and Restoration course with course-based undergraduate research experiences associated with the topic of this grant. Networking, career exploration, and direct interactions with seed industry partners (i.e., private seed growers, private seed processing companies, and federal seed banks) and plant related careers (i.e., botanical gardens, US Forest Service, and Bureau of Land Management, etc.). Over 11 professional development workshops offered to summer undergraduate students in the 2023 program. Botany Meeting Presentation (*indicates Seeds For The Future Participants) Beckstead J, *D Eisenbrandt, *E Hallstrand, *A Greenhouse, *M Rowley, and *M Washington. 2023. Do Microbes Help Or Hinder Germination? A Learning Game Designed By USDA-REEU Undergraduate Students To Explore Seed Endophyte Interactions. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. Ferson-Mitchell A, M Alomran, *C Ditton, *K Wiser, *M Morse, *T Van Ostrand, and G Newcombe. 2023. Primary Bacterial Symbionts of Creosote Bush. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Greenhouse A, *E Hallstrand, J Beckstead. 2023. Antagonistic activity is observed from seed endophytes: A potential tool to aid seeds in establishment during restoration. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Kuehfuss A, A Ferson-Mitchell, and G Newcombe. 2023. Thermotolerance of Bacterial Endophytes fromCallitropsis bakeri. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Mai J, B Connolly, and J Beckstead. 2023. Frost Flash: The Effects of Abrupt Freezing Events on Bromus tectorum and Elymus elymoides. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Robitaille C, B Connolly, and J Beckstead. 2023. Examining the Resilience of Squirreltail Grass (Elymus elymoides) Against a Zinc Gradient. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Storrud S, A Ferson-Mitchell, and G Newcombe. 2023. Acacia koa,Acacia melanoxylon, andAcacia mangiumEndophytes Interact withFusarium oxysporum f. sp. koaein Antagonistic Assays. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Tindall W, A Ferson-Mitchell, and G. Newcombe. 2023. The Effects Nematode Interactions have on the Relative Growth Rate ofHericium erinaceus. July 25, 2023. Botany Conference, Boise, ID. What do you plan to do during the next reporting period to accomplish the goals?Initial draft of a manuscript for publication [to be submitted to a refereed journal by the end of 2024]: A bacterial pathogen of Bromus tectorum isolated from seeds of Larrea tridentata Abigail Ferson-Mitchell1*, Maryam Alomran2, Avery Kuefuss1, Julie Beckstead3, Brenda Schroeder 4, and George Newcombe1 1 Department of Forest, Rangeland, and Fire Sciences, College of Natural Resources, University of Idaho, Moscow ID 83843. 2 Department of Biology, College of Science, Princess Nourah bint Abdulrahman University, Riyadh 11671, Saudi Arabia. 3 Department of Biology, Gonzaga University, Spokane WA 99258. 4 Department of Entomology, Plant Pathology, and Nematology, College of Agricultural and Life Sciences, University of Idaho, Moscow ID 83843. Abstract Roughly 1/3 of the pathogens of cheatgrass in North America are biotrophs that cause obvious disease with typical signs and symptoms. Some of the remaining pathogens also cause obvious disease, but others cryptically depress germination or growth or fecundity without signs and symptoms. Among this remainder are cryptic pathogens that can be isolated from plants other than cheatgrass. To our knowledge no pathogen of another plant has ever been tested for negative effects on cheatgrass. Here, we tested strains of Bacillus bacteria that were not obvious pathogens but they had displayed negative effects on emergence of B. tectorum in preliminary research. One strain was isolated from B. tectorum but four more were from creosote bush, Larrea tridentata (family Zygophyllaceae), a common shrub in deserts of the American Southwest. In our various germination and emergence assays the only consistently negative strain was one, 'SB-4', from creosote bush. This cryptic pathogen completely suppressed cheat grass germination after just a single hour of direct contact. We also compared inoculation methods: two methods that depend on direct contact between Bacillus cells and seeds of B. tectorum, and a third method, the soil drench, that does not ensure direct contact. Only the direct-contact methods reduced emergence of B. tectorum; soil drench had a weak effect on emergence. Furthermore, 48 hours of direct contact reduced emergence significantly more than 24 hours. In fact, the strongest pathogenic strain, 'SB-4', completely suppressed emergence of B. tectorum following 48 hours of direct contact. The search for useful pathogens of invasive cheatgrass, previously confined to microbes isolated from cheatgrass, should include cryptic pathogens from unrelated plants.'SB-4' in particular merits further testing. Introduction Bromus tectorum, or cheatgrass, is a very common invader of plant communities in western North America (Mack, 1981). Native to Eurasia, the initial phase of its invasion of North America may have been spurred by enemy release, a key factor in many plant invasions (Mitchell and Power, 2003). Enemy release can gradually be eroded by further pathogen introductions, particularly if the latter are pathogens from the native range. Although pathogens of Bromus tectorum have been slowly accumulating in North America since B. tectorum invaded (Meyer et al, 2016), the plant remains highly successful. A total of 52 fungal pathogens have now been recorded on B. tectorum in the USA (Farr and Rossman, 2024). The 52 are 'obvious' pathogens in the sense that they are associated with characteristic signs and symptoms, and they belong to taxa with proven pathogenicity. Despite the total of 52, greater than that of any single in the native range of cheatgrass, individual plants in North America tend still to be free of obvious pathogens. Even stand die-offs have remained local and sporadic (Ricks et al, 2021); no single pathogen at present affects the entire, invaded range in North America. Bacterial pathogens of B. tectorum have been less studied, but there is one well known example that will be further developed below. Of the 52 fungal pathogens on B. tectorum in the USA, 17 belong to taxa entirely comprised of specialized, biotrophic parasites: 3 powdery mildews (i.e., order Erysiphales), 7 rust fungi (i.e., order Pucciniales), and 7 smut fungi (i.e., class Ustilaginomycetes). The other 35 are necrotrophic fungi that are endophytic in some plants, but pathogenic in yet others. An example of this bimodality is Fusarium culmorum, an endophyte not only in B. tectorum, but also in Populus trichocarpa; an endophyte from the latter was experimentally demonstrated to retain its pathogenicity towards wheat (Newcombe et al, 2020). An endophytic isolate of F. culmorum from poplar only caused disease in inoculated wheat, and not in inoculated P. trichocarpa. The same fungus, F. culmorum, has also been reported from B. tectorum (Sprague, 1950). However, no experimental inoculations with a cheatgrass isolate have been carried out. Inoculations with most of the other necrotrophic taxa in the National Fungus Collections database have also never been attempted. But, its function as either an endophyte or a pathogen in B. tectorum has never been investigated. In western North America, there must be many potential cheatgrass pathogens among endophytes of the thousands of co-occurring plants.This would be true not only for fungi but for bacteria as well. Interest in relationships among plants, on the one hand, and species of the genus Bacillus, on the other, has focused on deploying these bacteria in agricultural ecosystems (Saxena et al, 2020). Central to that interest is the ability of Bacillus to promote the growth and control the pathogens of a wide range of crop plants. However, the roles of Bacillus in natural plant communities are still largely unstudied. So as the need for greater agricultural yields increases with increasing human populations, the deployment of biscuits in such systems will increase commensurately. However, spillover into natural ecosystems is relatively unstudied. It has been difficult to improve the health of plants and animals by manipulating their respective microbiomes; complex causality is at the root (Anderson, 2023). But we are getting closer to our goal for plants. Seed microbes are our focus because they are highly antagonistic/combative (Fira et al, 2018; Newcombe et al, 2018). Our current focus, Bacillus species, are not only known for their combativeness but now we know that they can be sourced or isolated from seeds (Newcombe et al, 2023).
Impacts
What was accomplished under these goals?
?Outputs Supported 7 students for 10-weeks in the Seeds For The Future Program in Summer 2023. Eight presentations at the Botanical Society of America Annual Meetings in Boise Idaho in July 2023; five poster presentations, two lightening talks, and one oral presentation. One poster presentation from a 2022 undergraduate participant. Outreach: Development of two learning games about seeds and their endophytes by undergraduate participants. 1) High School game called "Glorious Germination" and 2) Grades 3rd-5th game called "Seeds' Hidden Helpers". Games are being disseminated to regional schools and publications in education journals are in the works. Games were played with over 100 undergraduate students and also with 1-6 grade students at the Salish School of Spokane. Communication with larger community: 1) Science learning game materials made available on the program's website, 2) YouTube video about the science learning game made available online, 3) news article about plant curriculum featured in the Gonzaga University Bulletin. Development of plant-related suite of courses for Gonzaga University, including modification to two existing courses (now Plant Population Ecology and Plant Community Ecology) and a newly designed Plant Propagation and Restoration course with course-based undergraduate research experiences associated with the topic of this grant. Networking, career exploration, and direct interactions with seed industry partners (i.e., private seed growers, private seed processing companies, and federal seed banks) and plant related careers (i.e., botanical gardens, US Forest Service, and Bureau of Land Management, etc.). Over 11 professional development workshops offered to summer undergraduate students in the 2023 program. Botany Meeting Presentation (*indicates Seeds For The Future Participants) Beckstead J, *D Eisenbrandt, *E Hallstrand, *A Greenhouse, *M Rowley, and *M Washington. 2023. Do Microbes Help Or Hinder Germination? A Learning Game Designed By USDA-REEU Undergraduate Students To Explore Seed Endophyte Interactions. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. Ferson-Mitchell A, M Alomran, *C Ditton, *K Wiser, *M Morse, *T Van Ostrand, and G Newcombe. 2023. Primary Bacterial Symbionts of Creosote Bush. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Greenhouse A, *E Hallstrand, J Beckstead. 2023. Antagonistic activity is observed from seed endophytes: A potential tool to aid seeds in establishment during restoration. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Kuehfuss A, A Ferson-Mitchell, and G Newcombe. 2023. Thermotolerance of Bacterial Endophytes fromCallitropsis bakeri. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Mai J, B Connolly, and J Beckstead. 2023. Frost Flash: The Effects of Abrupt Freezing Events on Bromus tectorum and Elymus elymoides. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Robitaille C, B Connolly, and J Beckstead. 2023. Examining the Resilience of Squirreltail Grass (Elymus elymoides) Against a Zinc Gradient. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Storrud S, A Ferson-Mitchell, and G Newcombe. 2023. Acacia koa,Acacia melanoxylon, andAcacia mangiumEndophytes Interact withFusarium oxysporum f. sp. koaein Antagonistic Assays. Botanical Society of America Annual Meeting, Boise, ID. July 22-26. *Tindall W, A Ferson-Mitchell, and G. Newcombe. 2023. The Effects Nematode Interactions have on the Relative Growth Rate ofHericium erinaceus. July 25, 2023. Botany Conference, Boise, ID. ?
Publications
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