Source: IOWA STATE UNIVERSITY submitted to
TRANSGENIC APPROACHES IN MANAGING SUDDEN DEATH SYNDROME IN SOYBEAN
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
0231462
Grant No.
2013-68004-20374
Project No.
IOW05338
Proposal No.
2012-02341
Multistate No.
(N/A)
Program Code
A5122
Project Start Date
Jan 1, 2013
Project End Date
Dec 31, 2018
Grant Year
2017
Project Director
Bhattacharyya, M. K.
Recipient Organization
IOWA STATE UNIVERSITY
2229 Lincoln Way
AMES,IA 50011
Performing Department
Agronomy
Non Technical Summary
Soybean, one of the world's most valuable oil-seed crops, is also a major source of protein for animal feed and human diets. The U.S. is the world leader in soybean production. In 2010, the total U.S. soybean crop value was over $38.9 billion. It is estimated that the soybean crop would have been 14.4% higher if there were no incidence of diseases. Soybean sudden death syndrome (SDS) is a major disease; and in 2010, it caused crop loss valued $820M. This loss will be exacerbated if SDS continues to spread or if climate extremes create soil conditions conducive to the SDS pathogen, Fusarium virguliforme. There are no effective management tools to control SDS. SDS resistance of soybean lines is partial and governed by many quantitative trait loci. Genes encoding complete SDS resistance are yet to be discovered. Therefore, the need for alternative, complementary SDS resistance encoded by transgenes is crucial to protect this crop. In this project, we will evaluate the feasibility of several genetic approaches to create stable transgenic soybean lines with enhanced resistance to this major disease. These include: (i) suppress the pathogen's growth by expressing synthetic genes in soybeans; (ii) induce SDS resistance by expressing genes from Arabidopsis and maize in soybean; (iii) express synthetic plant antibodies to nullify toxins in soybean; and (iv) express a gene of the SDS pathogen in soybean to promote SDS resistance. SDS resistant lines will then be evaluated against selected SDS pathogen isolates representing five geographical regions, viz., Iowa, Illinois, Indiana, Argentina and Brazil. Selected transgene(s) for enhanced SDS resistance will be incorporated into elite soybean lines. To accomplish these objectives, relevant cutting-edge technologies including a micro-fluidic RNAi assay to identify vital F. virguliforme genes, phase display to identify synthetic plant antibody genes, HiSeq 2000 sequencing of 175 pathogen-isolates, and optical biosensors to identify the strongest toxin-interacting peptides will be applied. Outcomes/impacts: Gains in knowledge are: Our investigation is expected to identify gene(s) of F. virguliforme that are vital for infection and disease development and reveal if suppression of the expression of these genes in the pathogen is feasible by expressing relevant transgenes in soybean. If demonstrated, we would develop an innovative approach to manage SDS. In addition, sequence analyses of 175 SDS pathogen isolates will reveal the molecular complexity of the pathogen. Expected actions are: Incorporation of one or more of the selected transgenes will result in elite soybean lines with enhanced SDS resistance and consequently increased profitability for soybean growers who chose to use this transgenic technology to manage the SDS. Expected changes in conditions are: Our project is expected to result in reduced soybean yield losses from SDS; improved farm economy from growing elite transgenic soybean lines with enhanced SDS resistance; development of a new generation of teachers, educators and scientists, who will contribute towards enhanced crop production for sustainable food security.
Animal Health Component
(N/A)
Research Effort Categories
Basic
20%
Applied
65%
Developmental
15%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2121820104035%
2121820108020%
2121820116015%
2124020104010%
2124020108010%
2124020116010%
Knowledge Area
212 - Pathogens and Nematodes Affecting Plants;

Subject Of Investigation
1820 - Soybean; 4020 - Fungi;

Field Of Science
1040 - Molecular biology; 1160 - Pathology; 1080 - Genetics;
Goals / Objectives
Soybean is one of the world's most valuable crops and the U.S. is the world leader in soybean production. In 2010 the U.S. soybean crop value was over $38.9 billion. Soybean suffers yield loss from various biotic stresses, including soybean sudden death syndrome (SDS), which in 2010 caused losses valued at $0.82 billion. Our long-term goal is to release soybean cultivars resistant to SDS caused by the fungal pathogen, Fusarium virguliforme. This will be accomplished through collaborative efforts of 17 PIs from multiple institutions located in Alabama, Iowa, Illinois, Indiana, Nebraska, Texas, Argentina and Brazil. Objectives are: 1. Apply transgenic approaches to: i) suppress SDS pathogen growth through RNA interference, ii) generate antibodies against toxins that induce SDS, iii) overexpress maize carbonic anhydrase and Arabidopsis nonhost resistance genes to enhance SDS resistance, and iv) express an effector protein under regulation of a F. virguliforme-infection inducible promoter. 2. Incorporate successful transgenes and novel SDS resistance genes into elite soybean lines. 3. Determine the use of transgenes against a diverse collection of SDS pathogen isolates. 4. Evaluate economic and social impact of transgenes. 5. Provide education and research experience to 6-12 grade teachers and undergraduate minority students. 6. Educate stakeholders and youth via extension programs on the use of transgenic technology to ensure sustainable soybean yield. Expected Outputs: Planned activities are: 1. Trans-silencing of F. virguliforme gene(s) from transgenic soybeans for enhancing SDS resistance. 2. Expression of (i) a new generation of Anti-FvTox1 and Anti-cerato-platinin antibodies, (ii) maize ZmCA1, (iii) FvTox1-interacting GmCA1 domain, (iv) Arabidopsis nonhost resistance gene PSS1 and (v) an effector protein for enhanced SDS resistance. 3. Molecularly characterize a representative sample of 175 soybean SDS pathogen isolates, collected from Iowa, Illinois, Indiana, Argentina and Brazil, and evaluate transgenic soybean lines with enhanced SDS resistance against these isolates. 4. Encourage the adoption of transgenic soybean lines with enhanced SDS resistance for improved farm profitability. 5. Monitor the change in growers' attitudes towards growing SDS resistant transgenic soybean lines. Planned events are: 1. Educate youth and teachers in areas of plant genomics, biotechnology and plant pathology for generating teachers for future education in sustainable agriculture. Train undergraduate and graduate students for generating next generation of scientists for sustainable agriculture. 2. Summer internships and field days to educate youth, and soybean growers and stakeholders, respectively, on transgenic technology and transgenic soybean lines with enhanced SDS resistance. Expected to develop the following products: 1. Elite soybean lines containing transgenes for SDS resistance during the year 5 of project. 2. An electronic application to teach youth the transgenic technology.
Project Methods
Efforts: 1.1 Apply a micro-fluidic RNAi assay to identify vital F. virguliforme genes. Silence selected genes by micro-RNAs produced in transformed soybean roots, and later in stable transgenic soybean plants. 1.2 Create synthetic anti-FvTox1 plant antibodies using a phase display system. Apply optical biosensors to identify the strongest FvTox1-interacting peptides. 1.3 Overexpress anti-FvTox1 plant antibodies, ZmCA1 and PSS1 in soybean via Agrobacterium tumefaciens-based stable transformation procedures and evaluate transgenic lines for enhanced SDS resistance by conducting stem-cut and root inoculation assays. 1.4 Express Phytophthora sojae Avr1b gene under regulation of an infection-inducible promoter to kill cells at the SDS pathogen infection sites to promote SDS resistance. 2. Conduct marker-assisted backcrossing to introgress selected transgenes into elite soybean lines. Suitable SDS resistant high-yielding lines will be field tested in multi-state trials. 3. Sequence 175 isolates of the SDS pathogens, collected from IA, IL, IN, Argentina and Brazil, by HiSeq 2000 (Illumina). Group the isolates using suitable software programs. Select isolate(s) from each group to determine utility of transgenes against a diverse collection of isolates. 4.1 Potential profitability of transgenic soybean lines with enhanced SDS resistance will be determined via economics methodologies. 4.2 Through a survey, growers' attitudes and willingness to adopt transgenes will be determined. 5.1 Train science and agriculture teachers, and future teachers through workshop on plant biotechnology. 5.2 Provide lab-based research experience to in-service and pre-service teachers and minority students. 6.1 Develop application software for youth-education on use of transgenic technology to ensure food security. 6.2 Provide soybean growers the experience of SDS resistant transgenic soybeans in demonstration plots. Evaluation: 1. Determine success in silencing SDS pathogen genes through transgenes from soybean by measuring SDS disease symptoms. Determine if pathogen growth reduced in infected transgenic roots. 1.2 Evaluate effectiveness of plant antibody genes, ZmCA1, PSS1 and Avr1b via root inoculation and stem-cut assays of the transgenic lines. Extent of SDS disease symptom development will be statistically analyzed. 2. Evaluate elite soybean lines carrying selected transgenes in field trials during year 5. 3. Evaluate utility of selected transgenes against selected isolate(s) from each group generated from 175 molecularly characterized isolates. 4.1 Evaluate utility of transgenes through economic analyses of growing transgenic soybeans considering production costs and revenue in year 4 and 5. The farm level analysis will outline potential changes to farm profitability, while the national level analysis will indicate potential impacts for the entire soybean crop sector. 4.2 Evaluate the acceptance of the transgenic soybeans by the soybean growers by analyzing outcomes of surveys conducted among soybean growers in year 1 and 5. 5 & 6. Success of education and extension programs will be evaluated through both formative and summative evaluations.

Progress 01/01/13 to 12/31/18

Outputs
Target Audience:Target audiences include: (i) students, (ii) researchers of both public and private sectors, (iii) soybean growers, and (iv) soybean seed industry personnel. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Two postdoctoral fellows and two assistant scientists received training in plant molecular biology, plant pathology, genetics and plant breeding. One female black minority Ph.D. student received training in molecular analyses of a plant gene. One graduate student got training in software design to conduct pathogen genome assembly and analyses. One graduate student received training in the area of economics of transgenic soybean and transgene deregulation at the public sector. One female graduate student and one female undergraduate student received training to conduct research in the area of societal acceptance issues related to genetically modified organisms (GMOs) and transgenic technology. How have the results been disseminated to communities of interest? Bhattacharyya gave a key note presentation entitled "What have we learned about the plant adaptation to cold-stress by studying the model plant Arabidopsis thaliana?" at the International Conference on "Climate Change, Biodiversity and Sustainable Agriculture (ICCBSA-2018) hosted at the Assam Agricultural University, Jorhat-13, Assam, India on December 15, 2018. Bhattacharyya gave a key note presentation entitled "The endogenous transposable element Tgm9 is suitable for generating knockout mutants for functional analyses of soybean genes" at the Conference on Molecular Breeding and Green Agriculture in 21 Century hosted at Changchun, China on July 23, 2018. Bhattacharyya gave a key note presentation entitled "Engineering soybean for enhanced broad-spectrum disease and pest resistances" at the Conference on Molecular Breeding and Green Agriculture in 21 Century, Changchun, China on July 22, 2018. Bhattacharyya gave a key note presentation entitled "Arabidopsis Nonhost Resistance Genes for Enhancing Disease Resistance in Soybean" at the State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, China. July 20, 2018. Bhattacharyya gave a key note presentation entitled "Isolation and Utilization of Arabidopsis Nonhost Resistance Genes in Enhancing Disease Resistance In Soybean" at the 70th Annual Meeting, Indian Phytopathological Society and National symposium on 'Plant Health Management: Embracing Eco-Sustainable Paradigm' hosted by Assam Agricultural University, Jorhat-13, Assam, India during February 15-17, 2018. Das presented a poster entitled "Selection of informative genomic regions for closely related isolates and construction of their phylogeny" at the 10th International Conference on Bioinformatics and Computational Biology, Las Vegas, Nevada, USA during March 19-21, 2018. Leandro gave a presentation entitled "Biology, ecology and management of fungal diseases of soybean: improving resistance screening assays for SDS" at the Iowa Soybean Association Event on Jan 31, 2018. Lindberg presented a poster entitled "Emerging Social and Scientific Imaginaries for Gene Edited Foods: Implications for Governance" at the Annual meeting of the Rural Sociology Society, Portland, OR., USA during July 26 - 29, 2018. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Overall impact statement: Identified eight plant genes and four synthetic genes for enhancing SDS resistance in transgenic soybean plants, and 10 SDS resistant plant introduction lines. These resources will facilitate soybean breeding programs for SDS resistance. Identified 150 candidate pathogen genes involved in SDS development. Introduction of SDS resistant soybean varieties would increase production and lower prices, but improve producer welfare. Identified some of the key social, ethical and environmental concerns regarding GMOs that shape consumer attitudes and acceptance and how these concerns vary among different publics. Gained an enhanced understanding of the utility of food labels for enhancing public trust. This project provided (i) a research experience to 24 high school science teachers and 16 minority undergraduate students and (ii) an understanding of biotechnology concepts through a workshop to 48 teachers and 54 high school students, who were primarily African American and women. Several students expressed how the workshop greatly influenced them to pursue agricultural research. Research conducted on the use of the online resources developed in this project demonstrated that high school and entry level college students who used this resource gained a better understanding of the transgenic technology and were more likely to accept its application to the food system. Teachers who used the learning environment gained confidence in their understanding and ability to integrate this technology into life science courses. Finally, Google analytics reveals thousands of new users for these online environments in the two years since their development. Objective 1. Apply transgenic approaches. Three types of transgenes were identified that enhance SDS resistance in transgenic soybean lines under field conditions. In Type 1, two of 10 synthetic genes applied to silence vital F. virguliforme genes significantly enhanced SDS resistance in transgenic soybean lines. In Type 2, two of 18 synthetic genes encoding F. virguliforme toxin FvTox1-binding peptides significantly enhanced SDS resistance in transgenic soybean lines. In Type 3, five Arabidopsis nonhost resistance genes, PSS1, 6, 21, 25 and 30 and three soybean genes, GmARP1, GmDS1 and GmSAMT2, were shown to enhance SDS resistance in transgenic soybean lines. PSS25, PSS30, GmSAMT2 and GmDS1 also enhance soybean cyst nematode resistance of the transgenic lines. Objective 2. Incorporate successful transgenes and novel SDS resistance genes into elite soybean lines. Ten highly SDS resistant soybean plant introduction (PI) lines were identified from evaluation of 254 PI lines. Investigation of the genetic variation for SDS resistance among these lines revealed 13 novel QTL for SDS resistance. Objective 3. Determine the genetic architecture of F. virguliforme isolates in North America and F. tucumaniae isolates in South America. The 355 F. virguliforme and 120 F. tucumaniae isolates collected from different geographical regions of North and South America were genotyped and investigated for genetic structure. Phylogenetic analyses revealed two clusters, one for isolates of each species. Isolates within a species are highly similar. Two highly aggressive and two non-aggressive isolates were selected for de novo sequencing. We identified 150 aggressive isolate-specific genes and 500 genes nonaggressive isolate-specific genes. Objective 4. Evaluate economic and social impact of transgenes. To explore the economic impact of transgenes for soybeans; we gathered baseline data on crop losses due to SDS, examined the revenue and cost impacts for soybean producers of transgenic soybean varieties with enhanced SDS resistance, and calculated the national market and welfare impacts of those varieties with a simulation model. Our results show that SDS is devastating at the farm scale, however, the national losses are relatively small. At the producer level, the introduction of SDS resistant soybean varieties will lead to higher yields and gross revenues, with a mixed impact on costs. The simulation model showed that the introduction of SDS resistant soybean varieties would increase production and lower prices, but improve producer welfare. To enhance public acceptance of new technologies it is critical that scientists, regulators, and industry understand how the public perceives their risks, benefits, and trade-offs. To address this, we investigated key societal trends related to transgenic crops and foods among consumers, food, farm and biotech producers and retailers, and policymakers; public acceptance and trust issues; governance (eg food labels); and soybean perceptions and management of the problem of glyphosate resistance. This task was accomplished through media content analysis, 48 in-depth key stakeholder interviews, a national consumer survey, and a survey of 2,400 Midwest soybean growers. We also began to investigate emerging issues related public perception and governance of gene-edited foods. Our results identified some of the key social, ethical and environmental concerns regarding GMOs that shape consumer attitudes and acceptance and how these concerns vary among different publics, as well as an enhanced understanding of the utility of food labels for enhancing public trust. Objective 5. Provide education and summer research experience intership (SREI) to 6-12 grade teachers and undergraduate minority students.In this project, 48 teachers and 54 high school students participated a biotechnology workshop. Forty-two teachers and 44 students were African American, primarily from black belt counties in Alabama and the surrounding areas. Follow-up evaluations for all teachers indicated that the majority of teachers found the workshop activities to be highly useful and helped to increase their understanding of biotechnology and its concepts and would highly recommend the workshop to fellow teachers and students. Several students expressed how the workshop greatly influenced them to pursue agricultural research. ?In this project, 24 high school teachers and 16 undergraduate minority students participated in the SREI program at ISU.Teachers gained significant experience in the use of molecular biology techniques that has allowed them to better engage their students in current challenges facing agriculture and medicine. Based on the premise that each of these teachers has 150 students each year, the knowledge and skills gained will be transmitted to approximately 3.600 students annually. Over 90% of the 16undergraduate students who participated in the SREI program are now either in a STEM graduate program or working in a STEM field. Objective 6. Educate stakeholders and youth via extension programs on the use of transgenic technology to ensure sustainable soybean yield. Building and promoting online learning resources which target high school level STEM learners was the focus of objective 6. Journey of a Gene https://ge.unl.edu/journey-of-a-gene/ and Enviropig ( https://ge.unl.edu/enviropig/ ) tell the story of scientists who make genetic discoveries, apply genetic engineering and then breeding for soybeans or pigs to solve problems in our food system. Research conducted and published on the use of Journey of a Gene demonstrated that high school and entry level college students who used this resource gained a better understanding of the technology and were more likely to accept this technology in our food system. Teachers who used the learning environment gained confidence in their understanding and ability to integrate this teaching into life science courses. Finally, Google analytics reveals the potential impacts of the resources on informing students and teachers are increasing. Globally, there were 1,500 new users in 2018 for Journey of a Gene and 1,000 new users for Enviropig. These represented 6.5% and 77% increases in users from 2017.

Publications

  • Type: Journal Articles Status: Other Year Published: 2019 Citation: Ngaki, N.N., Sahoo, D., Wang, B., Bhattacharyya, M.K.* (2018) Overexpression of a putative receptorlike plasma membrane protein generates broad-spectrum immunity in soybean. Plant Biotechnology J. under revision
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Bhattacharyya, M.K. (2018) What have we learned about the plant adaptation to cold-stress by studying the model plant Arabidopsis thaliana? International Conference on Climate Change, Biodiversity and Sustainable Agriculture (ICCBSA-2018). Assam Agricultural University, Jorhat-13, Assam, India. December 13-16, 2018.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Bhattacharyya, M.K. (2018) The endogenous transposable element Tgm9 is suitable for generating knockout mutants for functional analyses of soybean genes. Conference on Molecular Breeding and Green Agriculture in 21 Century. Changchun, China. July 23, 2018.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Bhattacharyya, M.K. (2018) Engineering soybean for enhanced broad-spectrum disease and pest resistances. Conference on Molecular Breeding and Green Agriculture in 21 Century. Changchun, China. July 22, 2018.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Bhattacharyya, M.K. (2018) Arabidopsis Nonhost Resistance Genes for Enhancing Disease Resistance in Soybean. State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, China. July 20, 2018.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Bhattacharyya, M.K. (2018) Isolation and Utilization of Arabidopsis Nonhost Resistance Genes in Enhancing Disease Resistance In Soybean. 70th Annual Meeting, Indian Phytopathological Society and National symposium on 'Plant Health Management: Embracing Eco-Sustainable Paradigm' (Feb. 15-17, 2018) Assam Agricultural University, Jorhat-13, Assam, India.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Swaminathan, S., Das, A., Assefa, T., Knight, J.M., Silva, A.F.D., Carvalho, J.P.S., Hartman, G.L., Huang, X., Leandro, L.F., Cianzio, S.R. and Bhattacharyya, M.K. 2019. Genome wide association study identifies novel single nucleotide polymorphic loci and candidate genes involved in soybean sudden death syndrome resistance. PLoS ONE 14(2): e0212071. https://doi.org/10.1371/journal.pone.0212071
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Swaminathan, S., Abeysekara, N.S., Knight, J.M., Liu, M., Bhattacharyya, M.K. and Cianzio, S.R. 2018. Mapping of new quantitative trait loci for sudden death syndrome and soybean cyst nematode resistance in two soybean populations. Theoretical and Applied Genetics, 131:1047-1062.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Das, A. and Huang, X. 2018. Selection of informative genomic regions for closely related isolates and construction of their phylogeny. 10th International Conference on Bioinformatics and Computational Biology, March 19-21, 2018, Las Vegas, Nevada, USA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Leandro, L. 2018. Biology, ecology and management of fungal diseases of soybean: improving resistance screening assays for SDS  Iowa Soybean Association Event, Jan 31, 2018.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Lindberg, S., Bain, C., and Selfa, T. 2018. Emerging Social and Scientific Imaginaries for Gene Edited Foods: Implications for Governance. Annual meeting of the Rural Sociology Society. July 26  29, 2018, Portland, OR., USA.
  • Type: Journal Articles Status: Under Review Year Published: 2019 Citation: Bain, C., Lindberg, S., and Selfa, T. 2019. Emerging Sociotechnical Imaginaries for Gene Edited Foods in the United States: Implications for Governance. Agriculture and Human Values. Received Revise and Resubmit (2/5/2019)


Progress 01/01/17 to 12/31/17

Outputs
Target Audience:Target audiences include: (i) students, (ii) teachers, (iii) researchers of both public and private sectors, (iv) soybean growers, and (v) soybean seed industry personnel. Students and teachers include those who are racial and ethnic minorities and those who are socially, economically, or educationally disadvantaged. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Two postdoctoral fellows and two assistant scientists are being trained in plant molecular biology, plant pathology, genetics and plant breeding. One postdoctoral fellow and two research associates gained app development experience through generation of an app for providing knowledge to stakeholders including youth in the area of transgenic technology. One female and one male graduate student trained in microfluidic technology for studying fungal biology. One graduate student is trained in software design to conduct pathogen genome assembly and analyses. One female and one male Ph.D. students and two research associates are trained for phenotyping SDS using digital images and for molecular marker analyses. One male minority Ph.D. student is being trained for molecular analyses of quantitative trait loci. One minority female high-school student has been trained for soybean breeding techniques and DNA isolation. One graduate student is being trained in the area of economics of transgenic soybean and transgene deregulation at the public sector. One female graduate student and one female undergraduate student are being trained to conduct research in the area of societal acceptance issues related to genetically modified organisms (GMOs) and transgenic technology. A total of four teachers received training in the areas of plant molecular genetics, plant breeding, and sociology in 2017 through the summer research internship program. Twenty-one participants including ten middle and high school teachers and eleven high school students, mostly from black minority middle and high schools, received relevant training in autoclaving, micropipetting, gel electrophoresis, and DNA fingerprinting. They also gained knowledge of genetics and GMO through the use of the "Journey of a Gene" App in the Tuskegee University Plant Genomics and Biotechnology Workshop. Approximately 80 learners including the FFA teachers and their students participating in the Career Development Event (CDE) and students in University of NE-Lincoln courses gained knowledge in breeding and genetic engineering processes. How have the results been disseminated to communities of interest?We have disseminated the outcomes of this project to: Bain spoke at the Graduate Program in Sustainable Agriculture Colloquium, ISU, in November 2017. The GMO Debate: What role for Food Labels in Building Trust? Panel on Issues of Scientific Trust and Credibility in the GM Debate. Bain spoke at Midwest and MidContinental ChapterMedical Library Association Meeting. Des Moines, IA on Oct, 2016 about "The Fight over (non) GMO Labeling: A Sociological Perspective." Between December and April 2017, at Iowa State University, Southard, a student of Bain presented her work on the topic "Confronting the Challenge of Glyphosate Weed Resistance: How are Soybean Farmers' Perceiving and Managing the Problem" and "Midwest Soybean Farmers' Perceptions and Management of Glyphosate Resistant Weeds" to a group of students at Science with Practice (SWP). Bhattacharyya gave a key note presentation entitled "Arabidopsis nonhost resistance genesfor enhancing disease resistance in soybean" to the students and scientists attending the Indian Society of Mycology and Plant Pathology International Conference on "Plant Health for Human Welfare" held in the Department of Botany, University of Rajasthan, Jaipur, Rajasthan, from November 1-4, 2017. Bhattacharyya gave a keynote presentation entitled "Overexpression of a plasma membrane protein enhances resistance to multiple pathogen and pests in soybean" to the students and scientists attending the National Symposium on Molecular Insect Science, Assam Agricultural University, Jorhat, India, from February 6-8, 2017. Bhattacharyya gave a keynote presentation entitled "Identification and application of Arabidopsisnonhost immunity genes inenhancing disease resistance in soybean" to the students and faculty at the International Symposium, Central Plantation Crops Research Institute, Kerela, from December 10-12, 2016. Bhattacharyya gave a presentation entitled "Folic acid in plant health" to the students and scientists of the ICAR-NRC Plant Biotechnology, New Delhi, India, on December 2, 2016. Bhattacharyya gave a presentation entitled "Arabidopsis nonhost resistance genesfor enhancing disease resistance in soybean" to the students and scientists of the Horticultural Research Station, Assam Agricultural University, Kahikuchi, Guwahati, Assam, India, December 8, 2017. Three scientists and two Ph.D. students from Bhattacharyya lab presented the research findings at the Plant Biology meeting hosted in Austin from July 9 through 13, 2016. Bradley gave several oral presentations on the topic "Soybean Disease Update" including SDS to educate more than 550 farmers in Henderson county (KY), Hardin County (KY), Warren County (KY), Grayson County (KY), Mt. Vernon (IL; Illinois Double-Crop Meeting), Marshall County (KY), Muhlenberg County (KY), Vanderburgh County (IN), DuBois County (IN) and Princeton (KY, UKREC Field Day). Our functional App (Journey of a Gene) (http://passel.unl.edu/ge) developed has been used to educate stakeholders including youth and teachers on transgenic technology. The App has been used in FFA teacher training events in Nebraska, Iowa, Nevada and South Dakota this spring. Recently, we developed a new app for an animal genetic engineering story "Enviropig" (https://ge.unl.edu/enviropig/) as a companion learning environment based on teacher recognition of student interest in animal biology. Both apps include a training/learning guide for the FFA Career Development Experience (CDE) and are part of our efforts to expand this CDE to a national event. All districts in NE had competed in the Biotechnology CDE using the SDS in soybean story as training curriculum. The curriculum for the CDE is now directly related to the Journey of a Gene App. FFA advisors / Ag teachers are integrating biotechnology learning resources into their teaching and the App is targeting their students. Four new states will join Nebraska and Iowa in delivering this CDE at their state conventions next year. We partnered with the National Center for Ag Science Literacy to work with three biology teachers and develop lesson plans that feature Journey of a Gene and Enviropig.These lesson plans will be part of a program in Translating Applied STEM Research into Secondary Science (TASRs) and distributed at http://www.agclassroom.org/teacher/matrix/ as well as the Plant and Soil Sciences eLibrary ( http://passel.unl.edu ).These resources have been or will be promoted at the national level via workshops at national meetings for both Ag Science and STEM teachers. In year 5, we also have been working on to create a third web learning environment focused on the learning needs of farmers and consumer stakeholders to better understand the biological and social science of transgenic crops. We are making progress in creating the "Farmers, Consumers and GMOs" learning environment. We have also disseminated the outcomes of the project to general public through press and media. The activities are listed below. Southard, E., S. Lindberg and C. Bain. 2017. Midwestern Soybean Farmer's Perceptions and Management of Glyphosate Resistant Weeds. Iowa State University Extension and Outreach. Fisher, H. and C. Bain. 2017. International Regulations for US GE Agricultural Exports: An Overview. Iowa State University Extension and Outreach. Bain, C. Interviewed for "GMOs symbolize broader concerns." Feedstuffs. Vol 88 (9): September 5, 2016. What do you plan to do during the next reporting period to accomplish the goals?We plan to conduct the following activities to accomplish the goals of each of the six objectives. Objective 1. Transgenic Approaches to SDS. We proposed to accomplish three tasks to generate transgenic soybean lines with enhanced SDS resistance. Task i: Suppress fungal growth through trans-specific RNA interference (RNAi): Soybean lines showing enhanced SDS resistance under field conditions in 2017 will be tested in IA, KY and IL fields in 2018. Task ii: Generate synthetic peptides that interact with FvTox1: Soybean lines showing enhanced SDS resistance under field conditions in 2017 will be tested in IA, KY and IL fields in 2018. Task iii: Overexpress plant genes to enhance SDS resistance: Continue to characterize transgenic soybean plants expressing Arabidopsis PSS genes and one soybean gene, GmDS1. Expression of these plant genes has shown to enhance SDS resistance in transgenic soybean plants. Soybean lines with these genes that showed enhanced SDS resistance under field conditions in 2017 will be tested in IA, KY and IL fields in 2018. Objective 2. Identify novel SDS resistance genes from soybean germplasm and determine their utility against a diverse collection F. virguliforme isolates. Task (i) to screen Plant Introductions (PIs) of MGs II, III, & IV for SDS resistance has been completed. Tasks (ii) to study the inheritance of SDS resistance/ susceptibility and Task (iii) to generate RIL populations from the matings of (selected) PIs with the SDS-susceptible cultivar Spencer are in progress. The four highly SDS resistance lines identified in Task (i) were crossed to: (1) 'Spencer' to study inheritance of SDS resistance, and (2) MN1606 for further enhancing SDS resistance in soybean. The F4generations of the crosses were grown this summer in the Agronomy Farm, Iowa State University. F5generation will be grown in the ICIA nursery, Puerto Rico and F6plants will be phenotyped in Ames to map the novel SDS resistance loci. Objective 3. Determine the genetic architecture of F. virguliforme isolates in North America and F. tucumaniae isolates in South America. This objective has four tasks: (i) collect 500 single-spored isolates of the SDS pathogen; (ii) conduct GBS and analyze GBS data; (iii) perform de novo sequencing of 20 selected isolates based on the GBS data; (iv) improve assessment methods and search for evidence of race specificity in soybean-F. virguliforme interaction. We have completed Tasks (i) and (ii). For Task (iii), we have sequenced and assembled genome sequences of six selected F. virguliforme isolates; three highly pathogenic and three are nonpathogenic. In 2018, we will complete the genome comparison and identification of candidate pathogenicity genes. For Task (iv), a series of greenhouse experiments revealed that soil infestation with conidia prior to planting was found to be a quantitative and simple method that shows promise for phenotyping soybean lines for SDS resistance. We will conduct additional experiments in 2018 to finalize this conclusion. Objective 4 has two tasks: (a) To evaluate the potential economic impact of transgenes against SDS; and (b) Evaluating grower SDS knowledge and willingness to adopt transgenes Objective 4a, we will determine potential economic benefits of transgenics, and costs of generating and deregulating transgenes for public release of a transgenic line. The baseline data for the crop sector and production budget work has been completed. We will continue to work on the regulatory costs.We will also continue to explore the industry cost of developing transgenic crops, including the regulatory costs. In Objective 4b, we continue to analyze the data collected through surveys. Our content analysis, interview and survey data will be used to prepare articles for publication. We will continue to work on content analysis of the media and key advocacy organizations to assess how gene-edited foods are being framed in relation to governance, regulatory oversight, consumer acceptance and trust. We will continue on examining the issue of public acceptance issues related to gene-edited foods as suggested by the Advisory Board, including conducting in-depth interviews with key stakeholders. Objective 5: Education: Grades 6-12 Teacher Training and Summer Experiences for Undergraduates. The following two tasks of this objective have been completed. Task i. Train Grade 6-12 Teachers and Higher Education Faculty to Stimulate Interest in Current Issues Related to Food and Agricultural Sciences. Task ii. Summer Research Experience Internship (SREI) for In-service and Pre-service Grade 6-12 Teachers and Undergraduate Minority Students. To determine the outcomes of this objective, at both Tuskegee University (TU) and Iowa State University a survey is being conducted on teacher participants of all the five years. Data will be compiled to gather information on implementation of their internship experiences in their school curriculum. Data will be presented in final face-to-face meeting. Also we will summarize internship information of the past five years and the information will be presented in the final face-to-face meeting on May 17, 2018. Objective 6: Educate Stakeholders on the use of Transgenic Technology The Farmer, Consumer and GMO resource is progressing but at a slower pace than predicted. We have prioritized learning outcome goals with this resource to guide learners in evaluating the nature of the arguments in opposition to the acceptance of GMOs as biological science vs. social science based arguments. We will complete this resource with the extended time frame of the grant. We are currently gathering data on use of our Enviropig and Journey of a Gene resources by teachers, will report that in our final face-to-face meeting on May 17, 2018.

Impacts
What was accomplished under these goals? The outcomes in Year 5 of this integrated AFRI project include: (i) identification of seven plant genes and several synthetic genes that enhance sudden death syndrome (SDS) resistance in transgenic soybean plants; (ii) identification of four highly SDS resistant soybean lines and development of segregating populations for mapping SDS resistance loci; (iii) identification of 46 candidate pathogenicity genes; (iv) survey and analysis of societal responses to GMOs and soybean grower's responses to glyphosate resistance; (v) education of students and teachers for plant biotechnology through workshop and research internship programs; and (vi) evaluation of "Journey of a Gene" and "Enviropig" apps in educating a very large number of youth and stakeholders on transgenic technology. Details of progresses under each objective are presented below. The goal of theObjective 1is to generate novel genes that will confer SDS resistance in transgenic plants by conducting three tasks. In Task (i), we proposed to suppress the growth of the SDS pathogen,Fusarium virguliforme(Fv), by creating suitable transgenic soybean plants for 10 synthetic genes that will produce small interfering RNAs to suppress functions of 22 vitalFvgenes. This year we have field evaluated selected transgenic plants carrying six synthetic transgenes for silencing 11F. virguliformegenes. Six transgenic soybean lines showing significantly reduced SDS as compared to the nontransgenic Williams 82 line are selected for field trial in 2018. In Task (ii), we expressed FvTox1-interacting synthetic peptides that bind to FvTox1 toxinin plantato suppress its function. We have generated transgenic soybean plants for 18 synthetic genes encoding putative FvTox1-interacting peptides. This year we have evaluated selected transgenic soybean lines carrying six synthetic FvTox1-interacting peptide genes. Seeds of 12 lines showing enhanced SDS resistance as compared to the Williams 82 line are collected for field trial in 2018. In Task (iii), we analyzed transgenic soybean plants that express plant genes. This year we evaluated several transgenic soybean lines carrying ArabidopsisPSS1, 6, 21, 25and30genes and soybeanGmDS1,GmSAMT1andGmSEQ1genes.All eight genes exceptPSS25enhanced SDS resistance in transgenic soybeans. We also observed that transgenic lines carrying CaMV 35S promoter fusedPSS30gene mature approximately two weeks early. Early maturity did not influence the seed yield and seed size. The goal of theObjective 2is to identify novel SDS resistant soybean PI lines and determine their utility against a diverse collection ofFvisolates. We have identified four highly SDS resistant soybean PI lines from evaluation of 255 lines. We applied the genome-wide association study in this population and identified 20 genetic SNP loci associated with foliar SDS and root rot resistance. The four highly SDS resistance lines were crossed to: (i) 'Spencer' to study inheritance of SDS resistance, and (ii) MN1606 for further enhancing SDS resistance in soybean. The F4generations of the crosses were grown this summer in the Agronomy Farm, Iowa State University (ISU). F5generation will be grown in the ICIA nursery, Puerto Rico and F6plants will be phenotyped in Ames to map the novel SDS resistance loci. The goal of theObjective 3is to determine the genetic architecture ofFvisolates in North America andFvandF. tucumaniae(Ft) isolates in South America. Earlier, we classified 355Fvand 120Ftisolates from different geographical regions of North and South America and observed that theFvisolates are highly similar. Principal component analysis of 100Fvisolates indicated that genetic divergence is associated to some degree with aggressiveness of the isolates. We selected three highly pathogenic and three less-pathogenic isolates forde novosequencing. Analyses of the genome sequences revealed an ~ 300 kb region containing 46 genes present in all three pathogenic isolates and absent in all three less-pathogenic isolates. A series of greenhouse experiments revealed that soil infestation with conidia prior to planting was found to be a quantitative and simple method that shows promise for phenotyping soybean lines for SDS resistance. The goal of theObjective 4is to evaluate the economic and social impact of transgenes. The baseline data for the crop sector and production budget work has been completed. We have a draft of rudimentary decision tool to explore producer-level choices on SDS. We have also collected standardized production budgets, for use to compare the costs and benefits of transgenic varieties. We have gathered historical data on the adoption of GMO technology within the crop sector to guide analysis on the possible adoption of new technology for SDS prevention and mitigation. We have started to conduct a content analysis of the media and key advocacy organizations to assess how gene-edited foods are being framed in relation to governance, regulatory oversight, consumer acceptance and trust. We have conducted in-depth interviews with approximately 10 key stakeholders to understand how they perceive the challenges and risks in relation to governance, regulatory oversight, consumer acceptance and trust related to gene-edited foods. We have begun to work on examining the issue of public acceptance issues related to gene-edited foods. We are continuing to work on a draft white paper that summarizes the secondary literature to understand regulations on genetically engineered crops and foods among the major US trading partners (e.g. China), changes to these regulations and the factors driving these changes, and changing public attitudes towards GMOs, especially in relation to labeling. We have investigated how efforts to frame the socio-environmental problem of glyphosate resistant weeds shape calls for certain governance strategies and regulatory responses. The goal of theObjective 5is to provide education and research experiences to 6-12 grade teachers and undergraduate minority students. We organized the Tuskegee University Plant Genomics and Biotechnology Workshop for 10 teachers and 11 students from May 29 to June 2, 2017. The middle and high school teachers and high school students received training in autoclaving, micro-pipetting, gel electrophoresis, and DNA fingerprinting. They were exposed also to the "Journey of a Gene" app developed in Objective 6 for improving their knowledge of genetics and transformation. We conducted the Summer Research Experience Internship (SREI) for four teachers in ISU. Teachers began their seven-week summer internship on June 14, 2017 with a two-day orientation program prior to their research in two ISU laboratories. They attended weekly seminars on SDS. The teachers also participated in the ISU STEM Summer Institute, where they received pedagogy training to learn how to effectively translate the summer research experience into their science and/or agriculture classroom activities. On the last day of the program, the teachers gave oral and poster presentations on their research work. The goal of theObjective 6is to educate stakeholders on the use of transgenic technology, and establish demonstration plots to showcase SDS transgenic soybean lines. To educate stakeholder, earlier we developed the "Journey of a Gene" for plant genetics and transformation. Las year we have developed the "Enviropig" app educating stakeholder in animal genetic engineering and animal biology.EnviropigApp has been peer reviewed and integrated into both college and high school courses by Nebraska teachers. Both resources include a training/learning guide for the FFA Career Development Experience (CDE) and are part of our efforts to expand this CDE to a national event. We have collaborated with teachers to develop lesson plans that target state and national STEM standards and utilize Journey of a Gene and Enviropig Apps.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Marshall, J., Qiao, X., Baumbach, J., Xie, J., Dong, L. and Bhattacharyya, M. K. Microfluidic device enabled quantitative time-lapse micro-photography for phenotyping vegetative and reproductive phases in Fusarium virguliforme, which is pathogenic to soybean. Sci. Rep. 7, 44365 (2017). https://doi.org/10.1038/srep44365
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sahu, B. B., Baumbach, J., Singh, P., Srivastava, S. K., Yi, X. and Bhattacharyya, M. K. Investigation of the Fusarium virguliforme transcriptomes induced during infection of soybean roots suggests that enzymes with hydrolytic activities could play a major role in root necrosis. PLoS One 12, e0169963 (2017). https://doi.org/10.1371/journal.pone.0169963
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Bain, C. and Selfa, T. Non-GMO vs Organic labels: purity or process standards in a GMO contaminated landscape. Agriculture and Human Values. (IP 2.222) (2017). https://link.springer.com/article/10.1007%2Fs10460-017-9776-x
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Bain, C., Selfa, T., Dandachi, T. and Velardi, S. Superweeds or Survivors? framing the problem of glyphosate resistant weeds and genetically engineered crops. Journal of Rural Studies. 51: 211-221 (2017). https://doi.org/10.1016/j.jrurstud.2017.03.003
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Troupe, G., Golick, D. and Lee, D. Genetic engineering online lesson leads to increased knowledge and more accepting student attitudes. NACTA Journal, 60(2): 167-175 (2016). https://www.nactateachers.org/attachments/article/2417/7%20%20Troupe_NACTA%20Journal.pdf
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Troupe, G., Peterson, A., Golick, D., Turnbull, S. and Lee, D. Improving Genetic Engineering Secondary Education through a Classroom-Ready Online Resource. Journal of Agricultural & Food Information. Manuscript ID: 1353915 (JAFI-2017-0040.R1). Published online: 02 Nov 2017. Doi.org/10.1080/10496505.2017.1353915.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Wang, B., Sumit, R., Sahu B.B., Ngaki, N.N., Srivastava, S., Yang, Y., Swaminathan S. and Bhattacharyya, M.K. Arabidopsis nonhost resistance PSS1 gene encodes a novel glycine-rich plasma membrane protein that enhances disease resistance in transgenic soybean plants. Plant Physiology, 175: 1-13 (2017). DOI: https://doi.org/10.1104/pp.16.01982
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Swaminathan, S., Abeysekara, N.S., Knight, J.M., Liu, M., Bhattacharyya, M.K. and Cianzio, S.R. Novel quantitative trait loci associated with sudden death syndrome and soybean cyst nematode resistance in soybean. Theoretical and Applied Genetics, accepted for publication.
  • Type: Books Status: Published Year Published: 2017 Citation: The Soybean Genome by Nguyen, Henry T. (Edt)/ Bhattacharyya, Madan Kumar (Edt) Publisher: Springer Verlag ISBN-13: 9783319641966 ISBN: 3319641964 Publication Year: 2017
  • Type: Book Chapters Status: Published Year Published: 2017 Citation: Sandhu, D. and Bhattacharyya, M.K. (2017) Transposon-based functional characterization of soybean genes. In The Soybean Genome edited by Nguyen, Henry T. and Bhattacharyya, Madan Kumar. Publisher: Springer Verlag
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Southard, E. and Bain, C. (2017). The National Bioengineered Food Disclosure Standard: How Did We Get Here & What Does It Mean? Science with Practice. Iowa State University, Ames, IA. April 28.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Southard, E. and Bain, C. (2017). Confronting the Challenge of Glyphosate Weed Resistance: How are Soybean Farmers Perceiving and Managing the Problem. Symposium on Undergraduate Research and Creative Expression, Iowa State University, Ames, IA. April 11.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Southard, E. and Bain, C. (2017). Confronting the Challenge of Glyphosate Weed Resistance: How are Soybean Farmers Perceiving and Managing the Problem? College of Agriculture and Life Sciences Sustainability Symposium, Iowa State University, Ames, IA. April 13.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Southard, E. and Bain, C. (2017). Midwest Soybean Farmers Perceptions and Management of Glyphosate Resistant Weeds. Research in the Capitol, Des Moines, IA; March.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Southard, E. and Bain, C. (2017). Midwest Soybean Farmers Perceptions and Management of Glyphosate Resistant Weeds. Science with Practice, Iowa State University, Ames, IA; December 7.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bain, C. (2016). The Fight over (non) GMO Labeling: A Sociological Perspective. Midwest and MidContinental Chapter Medical Library Association Meeting. Des Moines, IA. Oct, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Das, A., Swaminathan, S., Bhattacharyya, M. K. and Huang, X. (2017). Assembly of two Fusarium virguliforme isolates and finding the region which generates the difference in virulence between two isolates. In: Computer Science Department Graduate Student Organization Poster Presentation, Iowa State University, Ames, Iowa, February, 23, 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bhattacharyya, M.K. (2016) Folic acid in plant health. ICAR-NRC Plant Biotechnology, New Delhi, India, December 2, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Bhattacharyya, M.K. (2017) Overexpression of a plasma membrane protein enhances resistance to multiple pathogen and pests in soybean. National Symposium on Molecular Insect Science, Assam Agricultural University, Jorhat, India, February 6-8, 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Bhattacharyya, M.K. (2017) Arabidopsis nonhost resistance genes for enhancing disease resistance in soybean. ISMPP International Conference on Plant Health for Human Welfare Department of Botany, University of Rajasthan, Jaipur, Rajasthan, November 1-4, 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bhattacharyya, M.K. (2016) Folic acid in plant health. College of Veterinary, Assam Agricultural University, Khanapara, Assam, India, December 6, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Bhattacharyya, M.K. (2017) Arabidopsis nonhost resistance genes for enhancing disease resistance in soybean. Horticultural Research Station, Assam Agricultural University, Kahikuchi, Guwahati, Assam, India, December 8, 2017.


Progress 01/01/16 to 12/31/16

Outputs
Target Audience:Target audiences include soybean researchers; soybean breeders; soybean pathologists; and science teachers, students and others interested in the study of genes and GM foods. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two postdoctoral fellows and two assistant scientists are being trained in the area of plant molecular biology, plant pathology, genetics and plant breeding. One female graduate student completed her master's degree in microfluidic technology for studying fungal biology. One graduate student is being trained in software design to conduct pathogen genome assembly and analyses. One female minority Ph.D. student and two research associates are being trained for phenotyping SDS using digital images and for molecular marker analyses. One male minority Ph.D. graduate student is being trained for molecular analyses of quantitative trait loci. One minority female high-school student is being trained on field soybean breeding techniques and on DNA isolation in the lab. One female graduate student and one female undergraduate student are being trained to conduct research in the area of societal acceptance issues related to genetically modified organisms (GMOs) and transgenic technology. One graduate student gained App development experience through generation of an app and incorporating this into undergraduate courses she teaches. A total of nine teachers and undergraduate students (four teachers and five undergraduates) received training in the areas of plant molecular genetics, plant breeding, and sociology during the 2016 SREI program. Of the nine SREI participants, four were African American minority students from Historically Black Universities. Eight of the SREI participants were female. Twenty one participants including ten middle and high school teachers and eleven high school students, mostly from black minority middle and high schools, received relevant training in autoclaving, micropipetting, gel electrophoresis, DNA fingerprinting and application of "Journey of a Gene" App in the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW). A total of approximately 600 learners including the FFA teachers and their highs school students who participated in the Career Development Event (CDE), science teachers in on-line MS courses, and undergraduate students in University of NE-Lincoln courses gained knowledge in the breeding and genetic engineering processes. How have the results been disseminated to communities of interest?We have disseminated the outcomes of this project to following communities of interest. Bain spoke at National Press Foundation, St Louis, Missouri on July 24-27, 2016 about "Understanding the GMO Labeling Debate " to students and scientists of both public and private sectors. She also spoke at the Annual meeting of the XIV World Congress of Rural Sociology. Toronto, Canada this year in August on " Non-GMO vs. Organic Labels: Purity or Process Standards in a GMO Contaminated Landscape " to students and scientists of both public and private sectors. Bhattacharyya gave a presentation at the Proteomics Workshop of the Plant & Animal Genome XXIII Conference, San Diego, CA, January 8-13, 2016 on "Identification of Defense-related Proteins in the Root Necrotic Mutant rn1 in Soybean" to students and scientists of both public and private sectors. In the Transposable Elements Workshop of the same conference he also spoke on "The Tgm9-Induced Indexed Insertional Mutant Collection to Conduct Community-Based Reverse Genetic Studies in Soybean". At the Soybean Breeders' & Pathologists' Workshop. St. Louis, MO, 22-24 February 2016, Bhattacharyya gave a presentation on "Novel biotech approaches in fighting sudden death syndrome in soybean" to students, researchers of public and private sectors. He also spoke on "Novel transgenic approaches in enhancing sudden death syndrome resistance in soybean" to the faculty and students of the Plant Pathology & Microbiology students of ISU. At the end of this year he is going to give a seminar on "Towardsidentification ofadaptation genes for generating climate resilientcrop plants" at the InternationalConference onClimateChange Adaptation and Biodiversity, Andaman Science Association, Port Blair, Andaman & Nicobar Islands, India, 8-10, December, 2016. Further he will be delivering a lecture on "Identification and application of Arabidopsisnonhost immunity genes inenhancing disease resistance in soybean" to the students and faculty at the International Symposium, Central Plantation Crops Research Institute, Kerela, 10-12, December, 2016. At the University of Minneapolis-St. Pual, Minnesota, September 26, 2016, Cianzio gave a seminar on "Troubles and Tribulations in breeding soybean for resistance to Sudden Death Syndrome" to the scientific community. Also she gave a presentation at the Annual Phytopathological Society Meeting, Tampa, Florida, July 30 - August 3, 2016 on "Unraveling the molecular and phenotypic relationship in soybean breeding for resistance to sudden death syndrome" to a group of plant pathologists, breeders of both the public and private sector. At Iowa State University in April and December, a student of Bain presented her work on "Understanding and evaluating changing societal attitudes towards genetically modified (GM) foods and crops" to a group of students at Science with Practice (SWP). Our functional App (Journey of a Gene) (http://passel.unl.edu/ge) developed last year has been used to educate stakeholders including youth and teachers on transgenic technology. The App has been used in FFA teacher training events in Nebraska, Iowa and South Dakota this summer. Nebraska State FFA Biotechnology career development experience (CDE) was delivered in April. All districts in NE had competed in the Biotechnology CDE using the SDS in soybean story as training curriculum. The curriculum for the CDE is now directly related to the Journey of a Gene App. FFA advisors / Ag teachers are integrating biotechnology learning resources into their teaching and the App is targeting their students. Recently, we have developed also an app for an animal genetic engineering story "Enviropig" (https://ge.unl.edu/enviropig/) as a companion learning environment based on teacher recognition of student interest in animal biology. Both resources include a training/learning guide for the FFA Career Development Experience (CDE) and are part of our efforts to expand this CDE to a national event. Four new states will join Nebraska and Iowa in delivering this CDE at their state conventions next year. We partnered with the National Center for Ag Science Literacy to work with three biology teachers and develop lesson plans that feature Journey of a Gene and Enviropig. These lesson plans will be part of a program in Translating Applied STEM Research into Secondary Science (TASRs) and distributed at http://www.agclassroom.org/teacher/matrix/ as well as the Plant and Soil Sciences eLibrary ( http://passel.unl.edu ). These resources have been be promoted at the national level via workshops at national meetings for both Ag Science and STEM teachers. In year 4, we also began a third web learning environment focused on the learning needs of farmers and consumer stakeholders to better understand the biological and social science of transgenic crops. We are making progress in creating the "Farmers, Consumers and GMOs" learning environment. We have also disseminated the outcomes of the project to general public through press and media. The activities are listed below. International Interviewed for "Bringing Back the Human Side to Ag" AgriBusiness Global. May 25, 2016. http://www.agribusinessglobal.com/agrichemicals/bringing-the-human-side-back-to-ag/ Interviewed for "Facts, not fears, the key to dealing with GM foods." The Straits Times. Singapore. April 1, 2016. http://www.straitstimes.com/tech/facts-not-fears-the-key-to-dealing-with-gm-foods National Interviewed for "GMOs symbolize broader concerns." Feedstuffs. Vol 88 (9): September 5, 2016. Featured in "Obama signs law mandating GMO labeling. AmericaNow. August 1, 2016. http://www.americanow.com/story/health/2016/08/01/obama-signs-law-mandating-gmo-labeling Featured in "Campbell's decision to label GMOs destroys Monsanto's main argument against labeling." Alternet. January 15, 2016. http://www.alternet.org/food/campbells-decision-label-gmos-destroys-monsantos-main-argument-against-labeling Interviewed for "Campbell's GMO move heralds change in labeling debate." Politico. January 12, 2016 https://www.politicopro.com/home Featured in "Campbell's will label GMOs--and the sky will not fall." Organic ConsumersAssociation.January12,2016.https://www.organicconsumers.org/blog/campbell%E2%80%99s-will-label-gmos%E2%80%94and-sky-will-not-fall Featured in "Campbell could stir up GMO labeling fight. Dairy Business. January 15, 2016.http://dairybusiness.com/dbc- tory.php?id=224599&title=#sthash.sld63HZd.dpbs Local/Regional Interviewed for "GMO labeling issue far from finished." The Dickinson Press. North Dakota. August 8, 2016. http://www.thedickinsonpress.com/news/agriculture/4089901-gmo-labeling-issue-far-finished Interviewed for "Food industry, which opposed GMO labeling, now embraces it." St Louis Dispatch. July 29, 2016. http://www.stltoday.com/business/columns/david-nicklaus/food-industry-which-opposed-gmo-labeling-now-embraces-it/article_072d3d5b-a166-5dc4-986b-2f92c7566c13.html Featured in "King, Collins should stand with Mainers on GMO labeling." Bangor Daily News. Maine. April 30, 2016 http://bangordailynews.com/2016/03/10/opinion/contributors/king-collins-should-stand-with-mainers-on-gmo-labeling/ Interviewed for "Big Food no longer has a united front on GMO labeling. St Louis Dispatch. January 11, 2016. http://www.stltoday.com/business/columns/david-nicklaus/nicklaus-big-food-no-longer-has-a-united-front-on/article_2bd2dab4-06e9-54c9-a6e9-cbdc78c72bbb.html Interviewed for "Societal Perceptions on GMOs" Iowa Agribusiness Radio Network. December 8, 2015. http://www.iowaagribusinessradionetwork.com/isu-sociologist-carmen-bain-on-societial-perceptions-on-gmos/ What do you plan to do during the next reporting period to accomplish the goals?In Objective1, following three tasks will be accomplished to generate transgenic soybean lines with enhanced SDS resistance. Task i: Suppress fungal growth through trans-specific rna interference (RNAi): We will conduct molecular analyses of selected transgenic soybean plants carrying one synthetic gene for suppressing growth of F. virguliforme for enhancing SDS resistance. The selected soybean lines have shown to express enhanced SDS resistance under field conditions. Task ii: Generate synthetic peptides that interact with the toxin FvTox1, which causes SDS: We will continue molecular analyses of the transgenic soybean plants expressing 18 FvTox1-interacting peptide genes for possible enhanced foliar SDS resistance. We will also study if FvTox1 causes root necrosis. Task iii: Overexpress plant genes to enhance SDS resistance: We will continue to characterize transgenic soybean plants expressing three Arabidopsis Pss genes and one soybean gene. Expression of these plant genes has shown to enhance SDS resistance in transgenic soybean plants. We will conduct field trials of selected transgenic soybean lines in three states, Indiana, Iowa and Kentucky. In Objective 2, we will conduct the following task. Task i. Molecular mapping of the SDS resistance quantitative trait loci (QTL): In 2017 winter, we will advance one more generation of the recombinant inbred lines (RILs), generated by crossing four PI lines to Spencer and MN1606 in ISU nursery located in Puerto Rico. Additional crosses of the four PI lines to IA3048 will be made in Ames, Iowa. In the summer of 2017, the RILs will be advanced to one more generation and also be genotyped and phenotyped for SDS resistance. In Objective 3, we will conduct the following two tasks. Task i. Race-specificity in the soybean-F. virguliforme pathosystem: We will continue this task after developing a robust phenotyping system. Digital imaging will be applied to standardize this objective phenotyping system. We have already started to develop such a system and work will continue into the early part of Year 5. Once the system is developed, a selected combination of soybean and F. virguliforme genotypes will be evaluated to determine if there is any race-specificity in the soybean-F. virguliforme interaction. Task ii. De novo sequencing of five selected F. virguliforme isolates: By conducting phylogenetic analysis of 355 F. virguliforme isolates, five distinct genetically diverse classes of the pathogen have been identified. The genotype-by-sequencing was conducted using the reference genome sequence of F. virguliforme Mont-1 isolate. To reveal additional genetic variation among the F. virguliforme isolates, we will conduct de novo sequencing of one isolate from each of the five clusters by applying the PacBio Sequencing system. In Objective 4, we will conduct the following two sub-objectives, Objective 4a and Objective 4b. In Objective 4a, we will determine the potential economic benefits of transgenics, and costs of generating and releasing a transgenic line at the public sector. In Objective 4b, we have been studying the social aspects of transgenes. In Objective 4a, we will continue to develop up-to-date baseline data for the economic analyses. We will use projected soybean prices to examine possible price differences between soybean varieties and will examine cost-of-production estimates to assess farm-level costs/benefits. Work will also continue on a soybean crop sector model to project national soybean supply, demand, acreage, and prices. Two scenarios will be examined with the model. The first scenario will assume no adoption of the transgenic line. This scenario establishes a baseline for the possible changes with the adoption of the transgenic line. The second scenario will assume a 50% adoption rate of the transgenic variety in the U.S. We will also continue to explore the industry cost of developing transgenic crops, including the regulatory costs. In Objective 4b, we will continue to investigate societal acceptance issues related to transgenic crops and foods among consumers, food, farm and biotech producers, manufacturers and retailers, and policymakers. We will accomplish this task through in-depth interviews and surveys. In Spring, 2016 we administered a public opinion survey and a grower survey. These surveys examine issues related to GMO food safety, labeling, herbicide resistance, and the development of new GE crops. Our task now is to analyze this survey data and, together with our interview data, continue to write manuscripts for publication and engage in public presentations of our findings. In Objective 5, we will continue: i) the Tuskegee University (TU) Plant Genomics and Biotechnology Workshop (TUPGBW) for grade 6-12 teachers and high school students in TU; and ii) summer research internships (SREI) for grade 6-12 teachers and undergraduate students in Iowa State University as in Year 4. In Objective 6, we will complete the "Farmers, Consumers and GMO's Learning Environment. This will reach a broader audience with the balanced learning targets of revealing how GMO crop varieties are different from non GMO versions of a crop, how they are not different and why this matters for the farmer and the consumer. Also we will create and complete a web based learning environment on testing hypotheses with biological data. This lesson plan will focus on how plant biologists work with data from an experiment designed to measure the role of genotype on the progression of SDS disease. This active learning environment will complement a teacher's use of "Journey of a Gene". Both of these outputs are deliverables not proposed in the original grant. They were motivated by the success of the online learning environment in meeting the needs of a range of life science teachers and the success of our team in advancing transgenic approaches to battle SDS, discovering that nature of farmer and consumer issues with the use of this technology and the opportunity to continue to work with the high school teachers in the summer program in lesson plan design. Budget reallocation makes these output possible.

Impacts
What was accomplished under these goals? Sudden death syndrome (SDS), caused by the fungal pathogen Fusarium virguliforme, continues to cause significant yield losses to this valuable crop worldwide.In Year 4, our efforts towards our long-term goal of releasing soybean cultivars resistant to SDS, and which are understood to be safe by the general public, include: (i) demonstration of seven plant genes and several synthetic genes in enhancing SDS resistance in transgenic soybean plants under growth chamber and greenhouse conditions; (ii) identification of 16 SDS resistant soybean plant introduction (PI) lines; (iii) genotyping and classification of 475 isolates that cause SDS in North and South America; (iv) learning societal responses to transgenes; (v) education of 30 students and teachers for plant biotechnology; and (vi) evaluation and application of an app in educating a very large number of youth and stakeholders on transgenic technology. Details of progresses under each objective are presented below. As a result of these efforts, our knowledge has increased and 14 teachers and 16 students have been provided information, which help them understand and trust the innocuous nature of transgenic soybean, which brings us closer to our overarching goal of releasing soybean cultivars resistant to SDS. The goal of theObjective 1is to generate novel genes that will confer sudden death syndrome (SDS) resistance in transgenic plants by conducting tree tasks. In Task (i), we planned to suppress the growth of the SDS pathogen,Fusarium virguliforme(Fv), by creating suitable transgenic soybean plants that will produce small interfering RNAs to destroy functions of vitalFvgenes. Ten synthetic genes including all 22 vitalFvgenes were cloned in a binary vector for transformation of soybeans. Transgenic soybean plants for all 10 synthetic genes were obtained and evaluated. One of these genes has shown to enhance SDS resistance in transgenic soybean plants. In Task (ii), our plan was to create FvTox1-interacting synthetic peptides that bind to FvTox1 toxinin plantafor suppressing its virulence function. We have generated 18 synthetic genes encoding putative FvTox1-interacting peptides and developed transgenic soybean plants for these genes and identified several of these genes that can confer SDS resistance. In Task (iii), by analyzing transgenic soybean plants that express fiveArabidopsis thalianaPssgenes and three soybean genes, we have demonstrated that fourPssand three soybean genes can enhance SDS resistance in transgenic soybean plants under growth chamber and field conditions. The goal of theObjective 2is to identify novel SDS resistant soybean plant introduction (PI) lines and determine their utility against a diverse collection ofFvisolates. Evaluation of 255 PI lines during Year 1-3, 16 highly SDS resistant soybean PI lines were identified. Field evaluation of these 16 SDS resistant lines revealed four highly SDS resistant lines. The four lines were then crossed to: 1) the susceptible cultivar 'Spencer' to study inheritance of SDS resistance; 2) two high-yielding lines for introgressing desirable agronomic traits; and 3) MN1606 for generating superior SDS resistant soybean lines. The goal of theObjective 3is to determine the genetic architecture ofFvisolates in North America andFvandF. tucumaniae(Ft) isolates in South America. In Year 3, we have genotyped and classified 355Fvand 120Ftisolates from different geographical regions of North and South America. In Year 4, we reanalyzed the genotypic data and showed that the isolates were highly homogenous with less genetic variation. Principal component analysis (PCA) of the 100F. virguliformeisolates that were phenotyped for aggressiveness indicated that genetic divergence is associated to some extent with the aggressiveness of the isolates. We are conductingde novosequencing of five highly diverse isolates to investigate the structural and genomic variation among the isolates. We have investigated the possible presence of race specificity in soybean -F. virguliformeinteraction. Preliminary results suggest that there is unlikely any race specificity in this plant-pathogen system. The goal of theObjective 4is to evaluate the economic and social impact of transgenes. We have conducted 49 in-depth interviews with key stakeholder groups and content analysis of newspapers and industry publications. In Spring 2016 we administered a national survey to 1,000 consumers focused on awareness, attitudes, perceptions and behavior regarding GMOs and non-GMO food purchases. We administered a survey to 2,400 Midwest soybean growers focused on how growers are addressing the problem of glyphosate resistance and how this might affect their views of other biotechnologies. The survey results are currently being analyzed and we are preparing a manuscript for submission to a journal. The goal of theObjective 5is to provide education and research experiences to 6-12 grade teachers and undergraduate minority students. We conducted two tasks. In Task (i), we organized the Tuskegee University (TU) Plant Genomics and Biotechnology Workshop for 10 teachers from June 13 - 17 and for 11 students from June 15- 17, 2016. The middle and high school teachers and high school students received relevant training in autoclaving, micropipetting, gel electrophoresis, and DNA fingerprinting. Pairing students with teachers for three days allowed teachers to teach the students the concepts and techniques that they learned in the first two days of the workshop. They were exposed also to the "Journey of a Gene" app developed in Objective 6 and application of genetics and biotechnology in crop improvement. In Task (ii), we conducted Summer Research Experience Internship (SREI) for in-service and pre-service four grade teachers and five minority undergraduate students in Iowa State University (ISU). Teachers began their summer internship on June 13, 2016 with a two-day orientation program prior to their SDS research in three ISU research laboratories. They attended weekly seminars relating to the SDS. The teachers also participated in the Iowa State University STEM Summer Institute, where they received pedagogy training to learn how to effectively translate the summer research experience into their science and/or agriculture classroom activities. On the last day of the program, July 29, 2016, the teachers gave a short oral presentation of their project and exhibited a poster outlining their research. One undergraduate minority student each from TU, North Carolina A & T University, two from Alabama A & M University, and one from ISU also participated in SREI program from June 6 to July 31, 2016 in three ISU research laboratories. The SREI attended weekly seminars designed to educate them for all aspects of the project. The four minority undergraduate students also participated in the George Washington Carver Summer Internship program, which provided seminars relating to graduate school preparation and extracurricular activities in the evenings and weekends. On the final day of the training, the students gave oral and poster presentations. The goal of theObjective 6is to educate stakeholders on the use of transgenic technology, and establish demonstration plots to showcase SDS transgenic soybean lines. To educate stakeholder, a functional App (Journey of a Gene:http://passel.unl.edu/ge) has been developed. An article based on the impact of "Journey of a Gene" on students learning will be published in the 2016 December issue of the peer reviewedNACTAjournal. In Year 4, we have developed also an app for an animal genetic engineering story "Enviropig" (https://ge.unl.edu/enviropig/) as a companion learning environment based on teacher recognition of student interest in animal biology. Both resources include a training/learning guide for the FFA Career Development Experience (CDE) and are part of our efforts to expand this CDE to a national event.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Abeysekara, N., Matthiesen, R.L., Cianzio, S., Bhattacharyya, M.K., and Robertson, A.E. (2016) Novel sources of partial resistance against Phytophthora sojae in PI 399036. Crop Sci. 56:114. doi: 10.2135/cropsci2015.09.0578
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Abeysekara, N.S., Desai, N., Guo, L., and Bhattacharyya, M.K. (2015) The Plant immunity inducer pipecolic acid accumulates in the xylem sap and leaves of soybean seedlings following Fusarium virguliforme infection. Plant Science 243:105114; doi:10.1016/j.plantsci.2015.11.008
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Bain, C. and T. Selfa. Non-GMO vs Organic Labels: Purity or Process Standards in a GMO Contaminated Landscape. Revise and Resubmit. Agriculture and Human Values.
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Bain, C., T. Dandachi, T. Selfa, S. Velardi. The Problem of Glyphosate Resistant Weeds and GE Crops. Environmental Sociology. Under Review.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Baumbach, J., Pudake R.N., Johnson, C., Ollhoff, A., Palmer, R.G., Bhattacharyya, M.K. and Sandhu, D. (2016) Transposon tagging of a male-sterility, female-sterility gene, St8, revealed that the meiotic MER3 DNA helicase activity is essential for fertility in soybean. PLoS One 11(3): e0150482. doi:10.1371
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Cianzio, S.R., Lundeen, P. Bhattacharyya, M.K., Swaminathan, S., Gebhart, G., and Rivera-Velez, N. (2016) Registration of AR11SDS soybean germplasm resistant to sudden death syndrome, soybean cyst nematode and with adequate iron deficiency chlorosis. Journal of Plant Registrations 10:177188. Online 2015: doi:10.3198/jpr2015.02.0010crg.
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Heinrich, J., Qiao, X., Baumbach, J., Xie, J., Dong, L., Bhattacharyya, M.K. (2016) Microfluidic device enabled quantitative time-lapse micro-photography of germination and sporulation in Fusarium virguliforme that causes sudden death syndrome in soybean. Scientific Reports, under review.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Huang, X., Das, A., Sahu, B.B., Srivastava, S.K., Leandro, L.F., ODonnell, K., and Bhattacharyya, M.K. (2016) Identification of hypervariable elements in an asexual pathogen. PLoS One 11(6): e0158183. doi:10.1371/journal.pone.0158183
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Liu, M., Li S., Swaminathan, S., Sahu, B.B., Leandro, L.F., Cardinal, A.J., Bhattacharyya, M.K., Song, Q., Walker, D.R., Cianzio, S.R. (2016) Identification of a soybean rust resistance gene in PI 567104B. Theor Appl Genet. 129: 863-877.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2016 Citation: Ngaki, N.N., Wang, B., Sahu, B.B., Srivastava, S., Farooqi, M.S., Kambakam, S., Swaminathan S., Bhattacharyya, M.K. (2016) Comparative transcriptomics of the soybean-Fusarium virguliforme interaction reveals that transcription of a novel ankyrin-repeat containing defense gene is suppressed by the pathogen to cause sudden death syndrome. PloS One, in press.
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Sahoo, D., Abeysekara, N., Cianzio, S., and Robertson, A.E., Bhattacharyya, M.K. (2016) A novel Phytophthora resistance gene, Rps12 mapped tightly to the Rps4/6 region in soybean. PLoS One, to be accepted following revision
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Swaminathan, S., Abeysekara, N.S., Liu, M, Cianzio, C.R. and Bhattacharyya, M.K. (2015) Quantitative trait loci underlying host responses of soybean to Fusarium virguliforme toxins that cause foliar sudden death syndrome. Theor Appl Genet. 129:495-506. doi: 10.1007/s00122-015-2643-5. Epub 2015 Dec 17.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Troupe, G., Golick, D., and Lee, D. (20165) Genetic engineering online lesson leads to increased knowledge and more accepting student attitudes. NACTA Journal., 60 (2): 167-175.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Wang, B., Swaminathan, S., and Bhattacharyya, M.K. (2015) Identification of Fusarium virguliforme FvTox1-interacting synthetic peptides for enhancing foliar sudden death syndrome resistance in soybean. PLoS One http://dx.doi.org/10.1371/journal.pone.0145156
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Xu, Z., Jiang, H., Sahu, B.B., Kambakam, S., Singh, P., Wang, X., Wang, Q., Bhattacharyya, M.K., and Dong, L (2016) Humidity assay for plant-pathogen interactions in miniature controlled discrete humidity environments with good throughput. Biomicrofluidics 10, 034108; http://dx.doi.org/10.1063/1.4950998
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Zhang, B., Wang, B., Morales, A.W., Scudder, J., Bhattacharyya, M.K., and Ye, J.Y., (2016) Study of the interactions of Fusarium virguliforme toxin FvTox1 with synthetic peptides by molecular simulations and a label-free biosensor. Analytical Chemistry, 88: 30243030
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Kambakam, S., Sahu, B.B., Singh, P., Ngaki, N.N., Sumit, R., Bhattacharyya, M.K. (2016). Folate Plays a Critical Role in Host and Nonhost Resistance. In: Annual Meeting of The American Society of Plant Biologists, Plant Biology 2016, Austin, Texas, July 9-13, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Ngaki, N.N., Sahoo, D., Swaminathan S., Bhattacharyya, M.K. (2016). Altered Expression of Glycine Max Disease Suscetibility 1 (GmDS1) Encoding a Plasma-membrane Protein Confers Broad-spectrum Resistance to Pathogen and Pests in Transgenic Soybean Plants. In: Annual Meeting of The American Society of Plant Biologists, Plant Biology 2016, Austin, Texas, July 9-13, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Wang, B., Sumit, R., Sahu, B.B., Ngaki, N.N., Yang, Y., Bhattacharyya, M.K. (2016). Expression of the Arabidopsis Nonhost Resistance Gene AtGRP1 Enhances Resistance to the Fungal Pathogen F. virguliforme in Transgenic Soybean Plants. In: Annual Meeting of The American Society of Plant Biologists, Plant Biology 2016, Austin, Texas, July 9-13, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Yang, Y., Kambakam, S., Ngaki, N.N., Wang, B., Sahu, B.B., Swaminathan S., Bhattacharyya, M.K. (2016). Mapping of Arabidopsis Nonhost Resistance Genes Pss5 and Pss19 that Confer Resistance to Soybean Pathogens, Phytophthora sojae and Fusarium virguliforme. In: Annual Meeting of The American Society of Plant Biologists, Plant Biology 2016, Austin, Texas, July 9-13, 2016.
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2016 Citation: Bhattacharyya, M.K. (2016) Identification and application of Arabidopsis nonhost immunity genes in enhancing disease resistance in soybean. International Symposium, Central Plantation Crops Research Institute, Kerela, 10-12, December, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bhattacharyya, M.K. (2016) Identification of Defense-related Proteins in the Root Necrotic Mutant rn1 in Soybean. Proteomics Workshop. Plant & Animal Genome XXIII, Town & Country Convention Center, San Diego, CA, January 8-13, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bhattacharyya, M.K. (2016) Novel biotech. approaches in fighting sudden death syndrome in soybean. 2016 Soybean Breeders & Pathologists Workshop. St. Louis, MO, 22-24 February 2016.
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2016 Citation: Bhattacharyya, M.K. (2016) Towards identification of adaptation genes for generating climate resilient crop plants. International Conference on Climate Change Adaptation and Biodiversity: Ecological Sustainability and Resource Management for Livelihood Security, Andaman Science Association, Port Blair, Andaman & Nicobar Islands, India, 8-10, December, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bhattacharyya, M.K. (2016) The Tgm9-Induced Indexed Insertional Mutant Collection to Conduct Community-Based Reverse Genetic Studies in Soybean. Transposable Elements Workshop. Plant & Animal Genome XXIII, Town & Country Convention Center, San Diego, CA, January 8-13, 2016.
  • Type: Theses/Dissertations Status: Published Year Published: 2016 Citation: Jill Marshall (2016) Investigation of germination, growth, sporulation, and host-induced RNA interference in Fusarium virguliforme. M.S. thesis submitted to Iowa State University


Progress 01/01/15 to 12/31/15

Outputs
Target Audience:Target audiences include: (i) students, (ii) teachers, (iii) researchers of both public and private sectors, (iv) soybean growers, and (v) soybean seed industry personnel. Students and teachers include those who are racial and ethnic minorities and those who are socially, economically, or educationally disadvantaged. Changes/Problems:In our original proposal we planned to evaluate the SDS resistance of transgenic soybean plants in the Year 4 and 5 against F. virguliforme isolates collected in Illinois, Indiana, and Iowa. Generation of seeds of transgenic plants is one year behind schedule. Currently, the main emphasis has been the identification of homozygous transgenic soybean lines. Seeds of those plants will be carefully evaluated for SDS resistance in greenhouse as well as under field conditions in Year 4 in Ames. Based on that evaluation we will select the most suitable lines for evaluation against multiple F. virguliforme isolates in all three states in the summer of Year 5. In Year 5, if feasible seeds will also be sent to Argentina and Brazil for evaluation against the SDS pathogen isolates of the South American countries. What opportunities for training and professional development has the project provided?Three postdoctoral fellows and two assistant scientists are being trained in the area of plant molecular biology, plant pathology, genetics and plant breeding. One female graduate student is getting training in fungal biology and two (one female and one male) in microfluidic technology. One graduate student is being trained in software design to conduct pathogen genome assembly and analyses. One female graduate student is being trained to conduct research in the area of societal responses to transgenic technology. One female graduate student gained App development experience through generation of an app for providing knowledge to stakeholders including youth in the area of transgenic technology. A total of thirteen teachers and undergraduate students (8 teachers and 5 undergraduates) received training in the areas of plant molecular genetics, plant breeding, computer science, computer engineering and sociology during the summer of 2015 SREI program. Of these, four were African American minority students from Tuskegee University, nine were female and four were male. Twenty participants including ten middle and high school teachers and ten high school students, mostly from black minority middle and high schools, received relevant training in autoclaving, micropipetting, gel electrophoresis, DNA fingerprinting and biotechnology in the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW). A total of approximately 300 learners including the FFA teachers and their students who participated in the Career Development Event (CDE) and students in University of NE-Lincoln courses gained knowledge in the breeding and genetic engineering processes. How have the results been disseminated to communities of interest?We have disseminated the outcomes of this project to following communities of interest. We have targeted student and teacher communities including minority African American community. Bhattacharyya spoke to high school students and high school and middle school teachers on "The Application of Transgenic Technology In Genetic Improvement Of Crop Plants" at the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW) on June 17, 2015. Bain spoke at National Press Association Food From Farm to Table Meeting, July 19-22, 2015, St Louis, Missouri on "The Fight over GMO Food Labeling" to students and scientists of both public and private sectors. She also spoke at the Annual meeting of the Rural Sociological Society, New Orleans, LA this year on "Raising the Stakes over Genetically Modified (GM) Foods: Social (Counter) Movements and the Fight over GM Labeling in the US" to students and scientists of both public and private sectors. Bhattacharyya spoke at the Plant Interactions with Pests and Pathogens Workshop of the Plant & Animal Genome XXIII Conference, San Diego, CA, January 10-14,2015 on "Arabidopsis Nonhost Resistance for Enhancing Disease Resistance in Soybean" to students and scientists of both public and private sectors. He also spoke at the 2nd International Conference on Frontiers in Biological Sciences (InCoFIBS-2015), Rourkela, Odisha, India, 22-24 January, 2015 on "Transgenic Approaches in Managing Diseases in Soybean" to student and faculty community. He also spoke at the International Conference onInnate Immunity,Barcelona, Spain, July 20-21, 2015, on "Identification and Application of Nonhost iImmunity Mechanisms for Creating Broad-Spectrum Disease Resistance in Crop Plants" to students and scientists of both public and private sectors. Huang spoke at the EITA Conference on New Media and Biomedical Research, Cambridge, Massachusetts, August, 2015 on "Developing and Using Bioinformatics Tools for Analysis of Big DNA Sequence Data" to students and scientists of both public and private sectors. A functional App (Journey of a Gene) (http://passel.unl.edu/ge), developed by us last year, has been used to educate stakeholders including youth and teachers on transgenic technology. The App has been used in FFA teacher training events in Nebraska, Iowa and South Dakota this summer. Nebraska State FFA Biotechnology career development experience (CDE) was delivered in April. All districts in NE had competed in the Biotechnology CDE using the SDS in soybean story as training curriculum. The curriculum for the CDE is now directly related to the Journey of a Gene App. FFA advisors / Ag teachers are integrating biotechnology learning resources into their teaching and the App is targeting their students.Applications for the Journey of a Gene App workshop at national conventions for FFA and NSTA have been accepted. The goal is to nationalize an FFA CDE in Biotechnology and use the App as a basis for educating and training students to prepare for these events. Three PIs, Bain, Bhattacharyya and Lee will speak at the Integrated Crop Management Conference on December 3, 2015, to be held in Iowa State University. This conference is attended by over 900 Agribusiness people including farmers, certified crop advisers, representatives from companies such as Monsanto, Pioneer, and Syngenta. Bain will speak on "Societal Perceptions and Attitudes Towards Transgenes"; while Bhattacharyya on "Application Of Transgenic Technology In Managing Crop Diseases" and Lee on "Using Apps To Educate The Public About Transgenic Technology." We have also disseminated the outcomes of the project to general public through press and media. The activities are listed below. International Interviewed for "Emotion, politics take over in US GMO labelling debate, science left out." ABC (Australia Broadcasting Corporation) July 30, 2015. Clint Jasper http://www.abc.net.au/news/2015-07-30/usa-gmo-labelling-debate/6659872 Featured in "GMO labeling debate: Controversy over genetically modified organism driven by politics and emotions, not science, sociologist says. International Business Times. July 30, 2015. http://www.ibtimes.com/gmo-labeling-debate-controversy-over-genetically-modified-organisms-driven-politics-2031571 National Interviewed for "Congress is having a messy food fight over GMO labeling." The Verge. August 4, 2015. http://www.theverge.com/2015/8/4/9094579/the-gmo-labeling-debate-is-scrambling-partisan-lines Featured in "GMO labeling becomes 'proxy' for consumer preferences." NPR's Harvest Public Media. July 31, 2015. http://harvestpublicmedia.org/article/gmo-labeling-becomes-%E2%80%98proxy%E2%80%99-consumer-preferences Featured in "Congress continues churning GMO labeling legislation." Press Examiner. August 4, 2015 http://www.pressexaminer.com/congress-continues-churning-gmo-labeling-legislation/22930 Local/Regional Featured in "Amid the GMO labeling fight, industry experts weigh in." Tampa Bay Times. July 23, 2015. http://www.tampabay.com/features/food/general/amid-the-gmo-labeling-fight-industry-experts-weigh-in/2238522 Featured in "GMO label debate a 'wicked' problem." Capital Press. July 24, 2015. http://www.capitalpress.com/Nation_World/Nation/20150724/gmo-label-debate-a-wicked-problem Featured in "GMO debate a 'wicked problem'" Joplin Globe. August 8, 2015 http://www.joplinglobe.com/news/local_news/gmo-debate-a-wicked-problem/article_e7b0296b-4475-5b0c-8f97-2f51f15c6cc8.html Featured in "GMO debate reaches new peak with 'voluntary labeling' bill." Investigate Midwest. August 4, 2015. http://investigatemidwest.org/2015/08/04/gmo-debate-reaches-new-peak-with-voluntary-labeling-bill/ Featured in "Com Sociologist says GMO controversy driven by emotions." CattleNetwork. July 30, 2015. http://www.cattlenetwork.com/news/industry/sociologist-says-gmo-controversy-not-driven-emotions Featured in "GE Critics Range as Skeptics." The Progressive Farmer. September 17, 2014. http://www.dtnprogressivefarmer.com/dtnag/common/link.do;jsessionid=39435D6C76941253EA2AAB8C2FEB86AC.agfreejvm1?symbolicName=/free/news/template1&product=/ag/news/bestofdtnpf&vendorReference=0cf7aea6-4215-4d9a-a056-582066285d77__1410970599329&paneContentId=3030&paneParentId=0&pagination_num=2 What do you plan to do during the next reporting period to accomplish the goals?In Objective1, following three tasks will be accomplished to generate transgenic soybean lines with enhanced SDS resistance. Task i: Suppress Fungal Growth through Trans-specific RNA Interference (RNAi): We will characterize the transgenic soybean plants carrying 10 synthetic genes for homozygosity and host-induced gene silencing (HIGS) of 23 genes in F. virguliforme for enhancing SDS resistance. Task ii: Generate Plant Antibodies against Toxins that Induce SDS: We will analyze the transgenic soybean plants expressing 18 FvTox1-interacting peptide genes for homozygosity and possible enhanced foliar SDS resistance. Task iii: Overexpress Plant Genes to Enhance SDS Resistance: We have identified five plant genes, two Arabidopsis (in this project) and three soybean genes (in an Iowa Soybean Association funded project), that confer SDS resistance in transgenic soybean plants. Transgenic plants carrying these genes will be molecularly characterized to identify homozygous lines, which will then be characterized for possible SDS resistance under growth chamber and field conditions. In Objective 2, we will conduct the following task. Task i. Generation of RILs for Mapping SDS resistance QTL and identifying and possible transgressive segregants with enhanced SDS resistance: We will make six crosses of the SDS susceptible cultivar 'Spencer' with PI407785 (MG IV), PI393535 (MG III), PI567649 (MG III), PI 417482 (MG III), PI 423936 (MG III), and PI 605839A (MG IV) this fall in Puerto Rico. We will make four crosses of currently grown SDS resistant lines Ripley and MN1606 with the two SDS resistant lines, PI407785 (MG IV) and PI393535 (MG III), for generating transgressive segregants with enhanced SDS resistance. We will follow the single-seed descent method for generating the RILs from all 10 crosses. In Objective 3, we will conduct the following three tasks. Task i. Determine the responses of soybean lines: Responses of selected eight soybean lines of the maturity group (MG) II and III will investigated to a set of 30 selected F. virguliforme isolates. The 30 F. virguliforme isolates will be selected from a set of 100 isolates based on their responses to 10 soybean lines observed in Year 2 and 3 of this project. Scoring be carried out by two different methods: (i) one by using 1-7 symptom scale and (ii) a percent foliar severity scale. Task ii. Determine if there is any race-specificity in the soybean-F. virguliforme interaction: To determine if there is any race-specificity in this plant-pathosystem, we will infect a set of soybean lines including Spencer and Williams 82 as susceptible checks and Ripley and MN1606 as resistant checks with a selected group of 6-8 F. virguliforme isolates. Task iii. Assembly and analyses of the F. virguliforme and F. tucumaniae genome sequences: We will assemble and analyze genome sequences of 20 isolates to be sequenced later this year. In our original proposal we planned to evaluate the SDS resistance of transgenic soybean plants in the Year 4 and 5 against F. virguliforme isolates collected in Illinois, Indiana, and Iowa. Generation of seeds of transgenic plants is one year behind schedule. We will complete the evaluation of the homozygous transgenic lines in greenhouse as well as under field conditions in Year 4 in Ames. Based on that evaluation we will select the most suitable lines for evaluation against multiple F. virguliforme isolates in all three states in the summer of Year 5. In Objective 4, we will conduct the following two sub-objectives, Objective 4a and Objective 4b. In Objective 4a, we will determine the potential economic benefits of transgenics, and costs of generating and releasing a transgenic line at the public sector. In Objective 4b, we have been studying the social aspects of transgenes. The following three tasks will be conducted in the Objective 4a. Task i. Establish Baseline Data for Crop Losses due to SDS: We will develop solid and up-to-date baseline data to perform meaningful economic analyses, as this baseline data will form the yardstick against which we measure impact. We will begin with 2010 estimates of $0.82 billion loss from SDS and use data from soybean commodity boards to assemble state-by-state and year-by-year estimates. Task ii. Evaluate the Gross and Net Revenue Potential for Various Soybean Lines: We will use historical average and projected soybean prices to examine possible price differences between the SDS resistant soybeans and other soybean varieties. We will take advantage of well-established cost-of-production methodology to provide historical soybean-specific data to assess farm-level costs/benefits. We will project crop revenues for a number of soybean lines, including the transgenic varieties developed here. Production cost budgets will be created for the transgenic varieties and compared to budgets for current soybean lines. Task iii. Determine National Level Impacts of Wide-scale Adoption of the Transgenic Lines: As the transgenic lines will likely have different gross and net revenues than traditional soybean lines, this will impact crop plantings not only in soybeans, but also in other crops, as farmers rebalance their cropping patterns to reflect the change in expected profitability from soybeans. We will construct a soybean crop sector model to project national soybean supply, demand, acreage, and prices and run two scenarios with the model. The first scenario will assume no adoption of the transgenic line. This scenario establishes a baseline for the possible changes with the adoption of the transgenic line. The second scenario will assume a 50% adoption rate of the transgenic variety in the U.S. We will also explore the industry cost of developing transgenic crops. In Objective 4b, we will continue to investigate societal acceptance issues related to transgenic crops and foods among consumers, food, farm and biotech producers, manufacturers and retailers, and policymakers. We will accomplish this task through in-depth interviews and surveys. We will administer a public opinion survey; a grower survey; and a survey to crop advisors. These surveys will examine issues related to GMO food safety, labeling, herbicide resistance, and the development of new GE crops. We will continue with in-depth interviews that focus on market challenges and opportunities related to non-GMO certification and labels. In Objective 5, we will continue: i) the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW) for grade 6-12 teachers and high school students; and ii) summer research internships (SREI) for grade 6-12 teachers and undergraduate students. A pre- and post-test will be developed to determine knowledge gained by SREI participants in the area of plant biotechnology. SREI participants will respond to this questionnaire at the beginning and end of the training period. Results will be compared to evaluate knowledge gained over the summer. In Objective 6, we will develop a responsive design website that is a learning environment for the extension audience. The learning environment will blend expert information from the Iowa State team in video, images and writing with interactive elements to engage this audience. The learning outcome goals for this project will target an understanding of the following issues, which are key to the development and integration of transgenic technologies in our food system: (i) Labeling of food products containing transgenic (GMO) ingredients. (ii) The scientific assessment of food and environmental safety risks for transgenic products. (iii) The costs of meeting regulatory testing expectations for transgenic products. (iv)Weighing risks and benefits for transgenic technologies. (v) Craft 'case-based' learning stories, deliver at live extension events.

Impacts
What was accomplished under these goals? We have continued to make significant progresses under each of the six objectives covering research, education and extension components of this integrated project. The outcomes in Year 3, which have impacted our knowledge and education of students, teachers and stakeholders, include: (i) demonstration of two plant genes in enhancing SDS resistance in transgenic soybean plants; (ii) identification of four SDS resistant soybean lines; (iii) genotyping and grouping of 475 isolates that cause SDS in North and South America; (iv) learning societal responses to transgenes; (v) education of 33 students and teachers for plant biotechnology; and (vi) evaluation and application of an app in educating a very large number of youth and stakeholders on transgenic technology. Details of progresses under each objective are presented below. The goal of the Objective 1 is to generate novel genes that will confer sudden death syndrome (SDS) resistance in transgenic plants by conducting three tasks. In Task (i), we plan to suppress the growth of the SDS pathogen, Fusarium virguliforme (Fv), by creating suitable transgenic soybean plants that will produce small interfering RNAs to destroy functions of vital Fv genes. We selected a list of 23 candidate vital Fv genes based on prior knowledge in other plant-pathogen interactions. Ten synthetic genes covering all 23 vital Fv genes were cloned in a binary vector for transformation of soybeans. Transgenic soybean plants for four of these synthetic genes have been obtained. In Task (ii), our plan was to create FvTox1-interacting synthetic peptides that bind to FvTox1 toxin in planta for suppressing its virulence function. We have generated 18 synthetic genes encoding putative FvTox1-interacting peptides and developed transgenic soybean plants for 17 of these genes. In Task (iii), by analyzing transgenic soybean plants carrying altered expression of two Arabidopsis thaliana genes we have demonstrated that they can enhance SDS resistance in transgenic soybean plants under field condition. The goal of the Objective 2 is to identify novel SDS resistant soybean lines and determine their utility against a diverse collection Fv isolates. Eight highly SDS resistant lines selected from Year 2 study were evaluated against 100 Fv isolates under greenhouse condition. Additional 28 lines of the maturity groups (MG) II and III were evaluated against three Fv isolates. Of the 16 most SDS resistant lines under greenhouse condition, four were found to be highly resistant to the SDS pathogen under field condition. The goal of the Objective 3 is to determine the genetic architecture of Fv isolates in North America and Fv and F. tucumaniae (Ft) isolates in South America. We have genotyped and classified 475 Fv and Ft isolates from different geographical regions of North and South America into several distinct groups. The goal of the Objective 4 is to evaluate the economic and social impact of transgenes. We have conducted 29 in-depth interviews with key stakeholder groups as well as conducting a content analysis of newspapers and industry publications. We published a journal article that focused on the GMO labeling debate and presented our findings in a meeting of the National Press Association. This article revealed that after nearly 20 years since the first introduction of genetically engineered (GE) corn, there is growing controversy and debates over this technology. Public opinion surveys revealed that while most of the general public still knows very little or nothing about the technology, when prompted it appears that public acceptance for GMOs has declined. This can partly be attributed to the public outreach work of key environmental and consumer advocacy organizations that are increasingly focusing on the topic of GMO, in particular leading efforts for the mandatory labeling of GMO foods. Importantly a number of food retailers and food processors are responding to the increasing interest and concerns around GMO food by introducing non-GMO labels for food products. We are now interviewing key stakeholder groups and preparing a grower survey that is focused on understanding key socio-political and economic factors that have contributed to the dramatic increase in glyphosate resistant weeds or 'superweeds'. The spread of these weeds may undermine key economic and environmental arguments that have been used by proponents of GE technology to garner support for the technology. The goal of the Objective 5 is to provide education and research experience to 6-12 grade teachers and undergraduate minority students. We conducted two tasks. In Task (i), we organized Tuskegee University Plant Genomics and Biotechnology Workshop for 10 teachers from June 15-19 and for 10 students from June 17-19, 2015. The middle and high school teachers and high school students received relevant training in autoclaving, micropipetting, gel electrophoresis, and DNA fingerprinting. Pairing students with teachers for three days allowed teachers to teach the students the concepts and techniques that they learned in the first two days of the workshop. In Task (ii), we conducted Summer Research Experience Internship (SREI) for in-service and pre-service 6-12 grade teachers and undergraduate minority students. Eight teachers began their summer internship on June 8, 2015, with a two-day orientation program. On June 10, the teachers began their research in pairs. One experienced teacher was paired with one novice teacher. Four teachers conducted research in the Bhattacharyya lab. A third pair conducted research in Leandro lab and the fourth pair in the Robinson lab. They conducted research on different aspects of the SDS project. They attended weekly seminars relating to the SDS. The teachers also participated in the Iowa State University STEM Summer Institute, where they received pedagogy training so that they can effectively translate the summer research experience into their science and/or agriculture classroom activities. On the last day of the program, July 23, 2015, the teachers gave a short oral presentation of their project and exhibited a poster outlining their research. Four undergraduate minority students from Tuskegee University (TU) and one from Iowa State University also participated in the SREI program from June 9 to July 31, 2015. Two TU undergraduate students conducted research in the Cianzio lab. A third TU undergraduate student conducted research in Huang lab and the fourth one in the Dong lab. The ISU undergraduate student conducted research under Bain. TU undergraduate students also participated in the George Washington Carver Summer Internship program, which provided seminars relating to graduate school preparation and extracurricular activities in the evenings and weekends. On the final day of the training, the students gave oral and poster presentations. The goal of the Objective 6 is to educate stakeholders on the use of transgenic technology, and establish demonstration plots to showcase SDS transgenic soybean lines. To educate stakeholders, a functional App (Journey of a Gene: http://passel.unl.edu/ge) has been developed. The App has been used in FFA teacher training events in Nebraska, Iowa and South Dakota this summer. Nebraska State FFA Biotechnology career development experience (CDE) was delivered in April. All districts in NE had competed in the Biotechnology CDE using the SDS in soybean story as training curriculum. The curriculum for the CDE is now directly related to the Journey of a Gene App. FFA advisors/Ag teachers are integrating biotechnology learning resources into their teaching and the App is targeting their students. Applications for Journey of a Gene App workshop at national conventions for FFA and NSTA have been accepted. The goal is to nationalize an FFA CDE in Biotechnology and use the App as a basis for educating and training students to prepare for these events.

Publications

  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bain, C. (2015). The Fight over GMO Food Labeling. National Press Association Food From Farm to Table Meeting. July 19-22, 2015. St Louis, Missouri.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bain, C., and Dandachi, T. (2014). Raising the Stakes over Genetically Modified (GM) Foods: Social (Counter)Movements and the Fight over GM Labeling in the US. Annual meeting of the Rural Sociological Society. New Orleans, LA. July/August.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bhattacharyya, M.K. (2015) Arabidopsis nonhost resistance for enhancing disease resistance in sotbean. Plant Interactions with Pests and Pathogens Workshop. Plant & Animal Genome XXIII Conference, San Diego, CA, January 10-14, 2015.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bhattacharyya, M.K. (2015) Transgenic approaches in managing diseases in soybean. 2nd International Conference on Frontiers in Biological Sciences (InCoFIBS-2015), Rourkela, Odisha, India, 22-24 January, 2015.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bhattacharyya, M.K. (2015) Identification and application of nonhost immunity mechanisms for creating broad-spectrum disease resistance in crop plants. International Conference on Innate Immunity. Barcelona, Spain, July 20-21, 2015.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Huang, X. (2015) Developing and Using Bioinformatics Tools for Analysis of Big DNA Sequence Data. EITA Conference on New Media and Biomedical Research, Cambridge, Massachusetts, August, 2015.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bain, C. (2015) Societal perceptions and attitudes towards transgenes. Integrated Crop Management Conference, Iowa State University, December 3, 2015.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Lee, D. (2015) Using apps to educate the public about transgenic technology. Integrated Crop Management Conference, Iowa State University, December 3, 2015.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bhattacharyya, M.K. (2015) Application of transgenic technology in managing crop diseases. Integrated Crop Management Conference, Iowa State University, December 3, 2015.
  • Type: Other Status: Published Year Published: 2015 Citation: Interviewed for "Emotion, politics take over in US GMO labelling debate, science left out." ABC (Australia Broadcasting Corporation) July 30, 2015. Clint Jasper http://www.abc.net.au/news/2015-07-30/usa-gmo-labelling-debate/6659872
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "GMO labeling debate: Controversy over genetically modified organism driven by politics and emotions, not science, sociologist says." International Business Times. July 30, 2015. http://www.ibtimes.com/gmo-labeling-debate-controversy-over-genetically-modified-organisms-driven-politics-2031571
  • Type: Other Status: Published Year Published: 2015 Citation: Interviewed for "Congress is having a messy food fight over GMO labeling." The Verge. August 4, 2015. http://www.theverge.com/2015/8/4/9094579/the-gmo-labeling-debate-is-scrambling-partisan-lines
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "GMO labeling becomes 'proxy' for consumer preferences." NPR's Harvest Public Media. July 31, 2015. http://harvestpublicmedia.org/article/gmo-labeling-becomes-%E2%80%98proxy%E2%80%99-consumer-preferences
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "Congress continues churning GMO labeling legislation." Press Examiner. August 4, 2015 http://www.pressexaminer.com/congress-continues-churning-gmo-labeling-legislation/22930
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "Amid the GMO labeling fight, industry experts weigh in." Tampa Bay Times. July 23, 2015. http://www.tampabay.com/features/food/general/amid-the-gmo-labeling-fight-industry-experts-weigh-in/2238522
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "GMO label debate a 'wicked' problem." Capital Press. July 24, 2015. http://www.capitalpress.com/Nation_World/Nation/20150724/gmo-label-debate-a-wicked-problem
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "GMO debate a 'wicked problem.'" Joplin Globe. August 8, 2015 http://www.joplinglobe.com/news/local_news/gmo-debate-a-wicked-problem/article_e7b0296b-4475-5b0c-8f97-2f51f15c6cc8.html
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "GMO debate reaches new peak with 'voluntary labeling' bill." Investigate Midwest. August 4, 2015. http://investigatemidwest.org/2015/08/04/gmo-debate-reaches-new-peak-with-voluntary-labeling-bill/
  • Type: Other Status: Published Year Published: 2015 Citation: Featured in "Com Sociologist says GMO controversy driven by emotions." CattleNetwork. July 30, 2015. http://www.cattlenetwork.com/news/industry/sociologist-says-gmo-controversy-not-driven-emotions
  • Type: Other Status: Published Year Published: 2014 Citation: Featured in "GE Critics Range as Skeptics." The Progressive Farmer. September 17, 2014. http://www.dtnprogressivefarmer.com/dtnag/common/link.do;jsessionid=39435D6C76941253EA2AAB8C2FEB86AC.agfreejvm1?symbolicName=/free/news/template1&product=/ag/news/bestofdtnpf&vendorReference=0cf7aea6-4215-4d9a-a056-582066285d77__1410970599329&paneContentId=3030&paneParentId=0&pagination_num=2
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bradely, C. (2015) Management of sudden death syndrome of soybean. University of Illinois Crop Management Conferences (4 locations  Champaign, Malta, Mt. Vernon, and Springfield, IL) with approximately 500 people in attendance.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Bradely, C. (2015) Management of sudden death syndrome of soybean. Illinois Fertilizer and Chemical Association Meeting (Peoria, IL). Approximately 300 people in attendance.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Robertson, A. (2015). Northern leaf blight and sudden death syndrome. CPS Grower Meeting. Feb 2015. Atlanta. Iowa (30 attendees)
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Robertson, A. (2015). Soybean Diseases: Sudden Death Syndrome and White mold. ISUEO crop Advantage Series. Jan 2015. Storm Lake, Iowa (100 attendees)
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Robertson, A. (2015). Soybean Diseases: Sudden Death Syndrome and White mold. ISUEO crop Advantage Series. Jan 2015. Waterloo, Iowa (80 attendees)
  • Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Robertson, A. (2015). Northern leaf blight and sudden death syndrome. ISUEO Crop Advantage Series. Jan 2015. Atlantic, Iowa (140 attendees)


Progress 01/01/14 to 12/31/14

Outputs
Target Audience: Target audiences include: (i) students and (ii) teachers, (iii) researchers; and (iv) soybean growers. Students and teachers include those who are racial and ethnic minorities and those who are socially, economically, or educationally disadvantaged. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Five postdoctoral scientists are getting training in the area of plant molecular biology and genetics and biosensors. One graduate student is getting training in fungal biology and two in microfluidic technology. One graduate student is being trained in software design for genome assembly and analyses. One graduate student is gaining experience in developing an app for providing knowledge to stakeholders including youth in the area of transgenic technology. Two undergraduate research assistants getting research experience in the area of molecular plant pathology. Three undergraduate students including two African American minority students from Tuskegee University received research experience in the area of plant molecular genetics and sociology in the summer of 2014. Seven K-12 female teachers, including one African American minority teacher, received training in the area plant molecular biology and plant pathology during the summer of 2014. Ten middle and high school teachers (all African American) and nine high school students (all African American) received relevant training in autoclaving, micropipetting, gel electrophoresis, DNA fingerprinting and biotechnology in the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW). Teachers participating in the TUPGBW received 4.0 continuing education credits. A total of approximately 300 learners including the FFA teachers and their students who participated in the Career Development Event (CDE) and students in University of NE-Lincoln courses gained knowledge in the breeding and genetic engineering processes. How have the results been disseminated to communities of interest? We have targeted students and teachers community including minority African American community. Bhattacharyya spoke about “Novel management approaches: managing diseases in soybean.” IIT, Guwahati, February 17, 2014 and SOYCON-2014 International Soybean Research Conference, Indore, India, 22-24 February, 2014. He also spoke about transgenic soybean for disease resistance to all African American minority teacher and student participants of the second Plant Genomics & Biotechnology Workshop for 7th - 12th grade teachers and high school students on June 18, 2014. Leandro spoke about the “Transgenic Approaches in Managing Sudden Death Syndrome in Soybean, this AFRI-USDA Project” to the participants of NCERA 137 – Soybean disease committee meeting on March 4, 2014, Pensacola, FL. Robertson spoke about the “Plant Pathology and Disease Resistance” to the teacher participants of Summer Research Experience Internship program of ISU on July 2014. We have also reached both youth and adult stakeholders through developing and delivering two outreach education products: (1) A Career Development Event (CDE); and (2) A Biotechnology APP to educate stakeholders for basic facts in plant genetic engineering. Nebraska State Future Farmers America (FFA) Biotechnology CDE was delivered in April, 2014. All districts in NE had competed in the Biotechnology CDE using the SDS in soybean story as training curriculum. Arrangements for a Biotechnology CDE in IA/MO have been made. The curriculum for the CDE is now directly related to the Journey of a Gene APP (http://passel.unl.edu/ge; Journey of a Gene App). FFA advisors/Ag teachers are integrating biotechnology learning resources including the APP into their teaching curriculum. FFA Agri-science teacher meetings will be a venue for training with this APP in NE, IA and SD. Application for the Journey of a Gene APP workshop has been conducted for FFA and National Science Teachers Association (NSTA) and distributed to the teacher participants of the ISU summer research experience internship program for use among the high school students. What do you plan to do during the next reporting period to accomplish the goals? In Objective 1, the following three tasks will be accomplished to generate transgenic soybean lines with enhanced SDS resistance. Task i: Suppress Fungal Growth through Trans-specific RNA Interference (RNAi): We have created nine synthetic genes to suppress F. virguliforme’s growth in transgenic soybean plants through host-induced silencing of 17 selected F. virguliforme genes. Task ii: Generate Plant Antibodies against Toxins that Induce SDS: Earlier we have shown that expression of an Anti-FvTox1 plant antibody can reduce foliar SDS development. The antibody gene was created from mammalian gene sequences and not acceptable in commercial cultivars. Therefore, by screening phage display libraries we have identified four synthetic peptides that bind to FvTox1. We will generate and analyze transgenic soybeans to determine if any of the 18 synthetic genes created from four peptides can enhance foliar SDS resistance. Task iii: Overexpress Plant Genes to Enhance SDS Resistance: We have shown that Pss1 encoding a glycine-rich protein, AtGRP1, and Pss30 encoding a folate transporter, AtFOLT1, confer enhanced resistance to the soybean pathogens, Phytophthora sojae and F. virguliforme in transgenic soybean plants. We will analyze transgenic soybean lines carrying five Arabidopsis nonhost resistance genes, Pss1, Pss6, Pss21, Pss25 and Pss30. In Objective 2, we will conduct two tasks. In task i, we will evaluate a collection of SDS resistant germplasm of the MG II, III, & IV against a selected set of ~20 diverse Fv isolates, identified based on their genotypes. The most suitable SDS resistant lines will be selected as the recurrent parents for introgressing of the transgenes for SDS resistance. Back-crossing will be continued in year 4. In the task ii, we will study the inheritance of SDS resistance in one or two PI lines showing broad-spectrum resistance to F. virguliforme (Fv) isolates. Based on the screening of recently identified PI lines with SDS resistance in year 1 and 2, we have selected seven PI lines for further evaluation against a diverse group of ~20 Fv isolates, classified by conducting genotyping by sequencing (GBS) under the objective 3. The PI lines showing resistance to most or all isolates will be crossed to the SDS susceptible cultivar ‘Spencer,’ and segregating F2:3 progeny populations will be scored for resistant and susceptible phenotypes following root inoculation with a selected Fv isolate and conduct mapping experiment to identify molecular markers tightly linked to SDS resistance loci. In Objective 3, we will conduct de novo sequencing of 20 isolates selected from five diverse clusters. We will also characterize the selected isolates from each cluster for virulence. The outcome of this objective will allow us to identify a set of isolates that will be used to determine the utility of transgenes generated in Objective 2. In Objective 4, we will continue to investigate societal acceptance issues related to transgenic crops and foods among consumers, food retailers and manufacturers, and policymakers. We will accomplish this task utilizing a range of methods including in-depth interviews, secondary analysis of the media and scholarly literature, and surveys. We will conduct a secondary analysis of the literature on consumer surveys that will attempt to answer more specific and detailed understandings of consumer attitudes towards transgenes as well as the literature on government regulations for GE crops and foods among the US trading partners. We will continue to conduct interviews with key stakeholders engaged in efforts related to GE labeling of foods. We will construct a survey of growers and crop advisors that will be aimed at understanding attitudes related to GE crops, especially related to recent challenges (e.g. glyphosate resistance) and moves towards new GE seeds (e.g. 2,4-D resistant). In Objective 5, we will continue: i) the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW) carrying continuing education credit for grade 6-12 teachers and high school students; and ii) summer research internships (SREI) for grade 6-12 teachers and undergraduate students. A pre- and post-test will be developed to determine knowledge gained by SREI participants in the area of plant biotechnology. SREI participants will respond to this questionnaire at the beginning and end of the training period. Results will be compared to evaluate knowledge gained over the summer. In Objective 6, we will conduct the NE CDE contest combined with our APP pilot in the training sessions given to Nebraska science teachers and to FFA teachers in Iowa. SREI participants will also receive training how to effectively use this APP. A next generation APP addressing feedbacks from APP users will be developed.

Impacts
What was accomplished under these goals? The goal of Objective 1 is to generate novel genes that will confer sudden death syndrome (SDS) resistance in transgenic plants. Progress has been made under each of the tree tasks of this objective. Task (i), we plan to suppress the growth of the SDS pathogen, Fusarium virguliforme (Fv), by creating suitable transgenic soybean plants that will produce small interfering RNAs specific to vital Fv genes. This approach is termed host-induced gene silencing (HIGS). Earlier we have shown that the RNA interference pathway is conserved in Fv. This novel approach has been shown to work in enhancing disease resistance in other crop species. Currently we have selected a list of 17 candidate Fv genes for HIGS based on prior knowledge in other plant-pathogen interactions. Thirteen synthetic genes for HIGS of all 17 Fv genes have been cloned in E. coli. Three of the 13 synthetic genes were already cloned into a binary vector and being used in transforming soybeans. In task (ii), our plan was to create synthetic antibodies or FvTox1-interacting synthetic peptides that will bind to FvTox1 toxin in planta and suppress its virulence function. Four FvTox1-interacting synthetic peptides were identified. From these peptides, 18 synthetic genes are created to express in transgenic plants. Transgenic soybean plants will be evaluated if any of the peptides can enhance SDS resistance. In task (iii), we proposed to express Arabidopsis nonhost resistance genes in transgenic soybean plants to enhance SDS resistance. Earlier we have shown that Arabidopsis Pss1 can enhance resistance against Fv in transgenic soybean plants. This year we have shown that Pss30 can also enhance SDS resistance in transgenic soybeans. The goal of Objective 2 is to identify novel SDS resistance genes from soybean germplasm and determine their utility against a diverse collection of Fv isolates. From a collection of 227 we recently identified 14 candidate SDS resistant plant introduction lines. The goal of Objective 3 is to determine the genetic architecture of Fv isolates in North America and F. tucumaniae isolates in South America. SDS is caused by Fv in North America and primarily by F. tucumaniae in South America. Last year, we collected 47 isolates from 20 counties of IL, 25 isolates from 15 counties of IN, and 34 isolates from 8 counties of IA. This year, we have collected 2, 60, 4, 15, 18, and 4 Fv isolates from AR, IL, KY, MI, MN, and NE, respectively. We have also received the genomic DNA of 3 F. virguliforme isolates from Canada and 16 F. tucumaniae isolates from Argentina. We have received DNA samples of F. tucumaniae isolates from Kerry O'Donnell (USDA-ARS). Our collaborator from Brazil will send us DNA of 10 Fv isolates. We have also received recently F. virguliforme infected root samples from SD, TN, KY, IA, and Canada. This year we genotyped an initial group of 168 F. virguliforme (Fv) isolates collected from different geographical regions of the North and South Americas at the Institute for Genomic Diversity (IGD), Cornell University. Isolates formed five groups, of which two groups include most of the isolates. The goal of Objective 4 is to evaluate the economic and social impact of transgenes. In year 2, we have extended our investigation into understanding societal attitudes towards transgenic crops. Our interviews and content analysis of newspapers revealed that after nearly 20 years since the first introduction of genetically engineered (GE) corn/soybeans societal attitudes concerning GE foods and crops are changing. There is growing controversy and debate over this technology; and public opinion surveys revealed that while most of the general public still knows very little or nothing about the technology, when prompted it appears that public acceptance for genetically modified organisms (GMOs) has declined. This can partly be attributed to the public outreach work of key environmental and consumer advocacy organizations who are increasingly focusing on the topic of GMO, in particular leading efforts for the mandatory labeling of GMO foods. For example, they are trying for state ballot initiatives across the nation, which is proving to be very expensive. Proponents and opponents of labeling collectively spent $85 million in the state of California and Washington on this initiative. In addition, a number of food retailers and food processors are responding to the increasing interest and concerns around GMO food by introducing non-GMO labels for food products. The economic component of Objective 4 is scheduled for year 5. The goal of Objective 5 is to provide education and research experience to 6-12 grade teachers and undergraduate minority students. We are focusing on two tasks: They are: i) workshops carrying continuing education credit for 6-12 grade teachers; and ii) summer research internships for 6-12 grade teachers and undergraduate students. In task (i), we organized the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW) for teachers from June 16-20, 2014, and for students from June 18-20, 2014. Ten middle and high school teachers and 9 high school students received relevant training in autoclaving, micropipetting, gel electrophoresis, DNA fingerprinting and biotechnology. Pairing students with teachers for three days allowed teachers to teach the students the concepts and techniques that they learned in the first two days of the workshop. Daily surveys allowed teachers and students to offer feedback regarding each teaching/learning session (e.g., clear definition of objectives, meeting the objectives, materials provided, time allocated, etc.). In task (ii), we conducted a Summer Research Experience Internship (SREI) for in-service and pre-service 6-12 grade teachers and undergraduate minority students. In Iowa State University, we conducted SREI for seven 6-12 grade teachers and three undergraduate students. Two of the students came from Tuskegee University, and were housed under the George Washington Carver (GWC) summer intern program. Five teachers conducted research in the Bhattacharyya lab for Objective 1. One pair of teachers conducted research in the Leandro lab. Two GWC interns conducted research for Objective 2 in the Cianzio and Bhattacharyya labs. One undergraduate SREI intern conducted research in the Bain lab for Objective 4. Students attended weekly seminars presented by investigators of this project. They prepared and presented their findings via poster and oral presentations at the end of their internship programs. The goal of the Objective 6 is to educate stakeholders on the use of transgenic technology, and establish demonstration plots to showcase SDS transgenic soybean lines. The SDS resistant transgenic lines, once developed, will be showcased in demonstration plots in year 5 of this project. To educate stakeholders we are developing and delivering two outreach education products: (1) A Career Development Event (CDE); and (2) A Biotechnology APP to educate stakeholders for basic facts in plant genetic engineering. Nebraska State Future Farmers America (FFA) Biotechnology CDE was delivered in April, 2014. All districts in NE had competed in the Biotechnology CDE using the SDS in soybean story as training curriculum. Arrangements for a Biotechnology CDE in IA/MO have been made. The curriculum for the CDE is now directly related to the Journey of a Gene APP, which is the second product. The functional Biotechnology App has been developed and is available at: http://passel.unl.edu/ge (Journey of a Gene App). FFA advisors/Ag teachers are integrating biotechnology learning resources including the APP into their teaching curriculum. FFA Agri-science teacher meetings will be a venue for training with this APP in NE, IA and SD. Application for the Journey of a Gene APP workshop has been conducted for FFA and National Science Teachers Association (NSTA).

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Abeysekara, N.S., and Bhattacharyya, M.K. (2014) Analyses of the xylem sap proteomes identified candidate Fusarium virguliforme proteinacious toxins. PLoS One. 9:e93667. doi: 10.1371/journal.pone.0093667.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Srivastava, S.K., Brar, H.K., Fakhoury, A.M., Bluhm, B.H., Huang, X., and Bhattacharyya, M.K. (2014) The genome sequence of the fungal pathogen Fusarium virguliforme that causes sudden death syndrome in soybean. PLoS One 9:e81832. doi: 10.1371/journal.pone.0081832.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Hughes, T.J., ODonnel, K., Rooney, A.P., Sink, S., Scandiani, M.M., Luque, A., Bhattacharyya, M.K., and Huang, X. (2014) Genetic architecture and evolution of the mating type locus in fusaria that cause soybean sudden death syndrome and bean root rot. Mycologia 106:686-697.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Cianzio, S.R., Bhattacharyya, M.K., Swaminathan, S, Westgate, M., Gebhart, G., Rivera-Velez, N, Lundeen, P., VanDerMolen, K. and Prusky, T. (2014) Registration of 'AR10SDS' soybean germplasm partially resistant to sudden death syndrome and resistance to soybean cyst nematode. J. Plant Regist. 8:200-210.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: M. K. Bhattacharyya , P. Singh, B. B. Sahu, R. Sumit, M. Xu, D. Sandhu (2014) Positional Cloning Of Arabidopsis Nonhost Resistance Genes Conferring Immunity To Two Soybean Pathogens, Phytophthora sojae and Fusarium virguliforme, Revealed Novel Disease Resistance Mechanisms. XVI International Congress on Molecular Plant-Microbe Interactions, Rhodes, Greece, July 6-10, 2014.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: M. K. Bhattacharyya, P. Singh, B. B. Sahu, R. Sumit, M. Xu, D. Sandhu (2014) Positional Cloning Of Arabidopsis Nonhost Resistance Genes Conferring Immunity To Two Soybean Pathogens, Phytophthora sojae and Fusarium virguliforme, Revealed Novel Disease Resistance Mechanisms. Oomycete Molecular Genetics Network Meeting, Norwich, UK, July 2-4, 2014.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: B. Zhang, B. Wang, A.W. Morales, J. M. Tamez-Vela, J. Scudder, M. K. Bhattacharyya, and J. Y. Ye, (2014) Study Interactions of FvTox1 with Synthetic Peptides Using a Label-Free Biosensor and Molecular Simulations. Annual Meeting of Biomedical Engineering Society, San Antonio, Texas, October 22-25, 2014.
  • Type: Other Status: Other Year Published: 2014 Citation: Bhattacharyya, M.K. (2014) Novel management approaches: managing diseases in soybean. IIT, Guwahati, February 17, 2014. Invited Presentation.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Bhattacharyya, M.K. (2014) Novel management approaches: managing diseases in soybean. SOYCON-2014 International Soybean Research Conference. Indore, India, 22-24 February, 2014.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: K. H. Brown, J. L. Baumbach, and M. K. Bhattacharyya. (2014) Investigation of the Possible Host-Induced Silencing in Fusarium virguliforme that Causes Sudden Death Syndrome in Soybean. Plant & Animal Genome XXII, San Diego, January 11-15, 2014. https://pag.confex.com/pag/xxii/webprogram/Paper12160.html
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Leandro, L. (2014) Transgenic Approaches in Managing Sudden Death Syndrome in Soybean, AFRI-USDA Project. NCERA 137  Soybean disease committee meeting March 4, 2014, Pensacola, FL.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Robertson, A. Plant Pathology and Disease Resistance. Research Experiences for teachers participants, ISU, July 2014.
  • Type: Websites Status: Published Year Published: 2014 Citation: TUPGBW promotional video: The Workshop video from Summer Workshop of 2013 was developed and posted on YouTube, and link to this video publicized widely and is expected to facilitate recruiting participants for the future workshops in Year 3, 4 and 5 of this project. https://www.youtube.com/watch?v=tOf1bSYyzew
  • Type: Websites Status: Published Year Published: 2014 Citation: Tuskegee University posted details of their workshop online at: http://www.tuskegee.edu/plant_genomics_and_biotechnology_workshop.aspx
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Bain, C. (2014). GMOs as a Wicked Problem: Social Movements, Political Consumerism and the Fight over GM Labeling. National Academy of Sciences, Meeting of the Committee on Genetically Engineered Crops: Past Experience and Future Prospects. Washington, DC. September 15-16, 2014.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Bain, C. and T. Dandachi (2014). Raising the Stakes over Genetically Modified (GM) Foods: Social (Counter)Movements and the Fight over GM Labeling in the US. Annual meeting of the Rural Sociological Society. New Orleans, LA. July/August.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Dandachi, T. and C. Bain (2014). The Genetically Modified (GM) Food Controversy: The Impact of Anti-Labeling Efforts on GM Labeling Movements in the U.S. Annual meeting of the Midwest Sociological Society. Omaha, NE. April.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Bain, C. (2014). Genetically Modified Foods and the Fight over GM Labeling in the US. Department of Horticulture, Iowa State University, 2014 Spring Seminar Series. March.


Progress 01/01/13 to 12/31/13

Outputs
Target Audience: Target audiences include: (i) students and (ii) teachers, (iii) researchers; and (iv) soybean growers. Students and teachers include those who are racial and ethnic minorities and those who are socially, economically, or educationally disadvantaged. Changes/Problems: There are several changes recommended by the advisory board. Changes are listed by objectives. Objective 1 has four tasks for developing transgenic soybean lines with enhanced resistance against Fusarium virguliforme that causes sudden death syndrome (SDS) in soybean. The tasks are: (i) develop host-induced gene silencing; (ii) express single-chain antibodies and random peptides that neutralize the toxin; (iii) over-express maize carbonic anhydrase and Arabidopsis non-host resistance genes to enhance SDS resistance; (iv) express an effector protein under the regulation of a F. virguliforme-infection inducible promoter. Task (iii) seems very promising, based on preliminary results (especially Pss1) and the board recommends that this objective be given top priority. We have expanded this task and characterized additional Pss genes that may confer resistance to F. virguliforme in transgenic soybean lines. Task (iv) does not appear promising, and the board does not recommend investing in this approach. Expression of avirulence effectors from pathogen-specific promoters has been explored for more than 20 years and the approach has not been proven useful. We agree with the advisory board and discontinued this task. Instead, we have intensely focused on cloning and transferring additional Pss genes to transgenic soybean plants for enhancing SDS resistance. Objective 2 has two tasks: (i) to screen SDS resistant Plant Introductions and develop mapping populations to identify resistance genes, and (ii) to introgress successful transgenes identified in objective 1 into resistant cultivars. The board recommended that we focus on task (i) and eliminate task (ii). It is highly unrealistic to breed the transgenes into elite lines in five years, which is the focus of task (ii). We agree with the board and concentrated our effort on task (i). Objective 3 has two tasks: (i) to collect and sequence a diverse selection of F. virguliforme isolates from Iowa, Illinois, Indiana, Argentina and Brazil; (ii) to screen the transgenic lines from objective 1 with a selection of the most diverse isolates. The board suggested including more isolates covering a wider geographical regions for task (i) using a marker technology. Subsequently, de novo sequencing of members from each diverse class can be sequenced to identify diverse isolates for screening the soybean germplasm with enhanced SDS resistance. We agree with the board and plan to classify ~400 isolates and then sequence only a few Objective 4 has two tasks. They are: task (i) to evaluate the potential economic impact of transgenes against SDS; and task (ii) to evaluate the grower SDS knowledge and willingness to adopt transgenes. The board made suggestions to revise these two tasks; accordingly in task (i), we will gather information and explore the potential for cross-project acquisition and dissemination of information on the costs of crop modification development and release at the public sector. In task (ii) we will study the social impacts of transgenes. We will focus on understanding industry and societal attitudes towards GMOs and how they are changing, as well as grower willingness to adopt transgenic varieties. This is to be accomplished through secondary data analysis of consumer surveys, together with in-depth interviews and a survey of key stakeholders, including industry organizations, policy makers, consumer and environmental advocacy organizations, food retailers, and growers. No major changes have been made in objectives 5 and 6. What opportunities for training and professional development has the project provided? Four postdoctoral scientists are receiving training in the area of plant molecular biology and genetics and biosensors. One graduate student is getting training in fungal biology and one in microfluidic technology. Two graduate students are being trained in software design. One graduate student is gaining experience in developing an app for providing knowledge to stakeholders including youth in the area of transgenic technology. Two undergraduate research assistants were employed (1 during the summer and one during the fall), including one who is currently hired through ISU’s “Science with Practice” program. The students were fully integrated into the research process. Six undergraduate students including five African American minority students from Tuskegee University received research experience in the area plant molecular genetics and sociology. Two K-12 teachers from African American minority school system of Tuskegee received training in the area plant molecular biology. Nine middle and high school teachers and 12 high school students received relevant training in autoclaving, micropipetting, gel electrophoresis, DNA fingerprinting and biotechnology in the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW). Teachers participating in the TUPGBW received 4.0 continuing education credits. There are two current targets for training/education. The FFA teacher and their students who are participating in the CDE and students in University of NE-Lincoln courses studied the breeding and genetic engineering processes. This would impact a total of approximately 300 learners during this reporting period. How have the results been disseminated to communities of interest? In Objective 1, investigator Dr. Liang Dong participated in the OPEN LAB NIGHT event on 11/19/2013 at ISU to present the microfluidic research for agronomy applications to K-12 and college freshmen. Sufficient results have not been gathered for peer reviewed publication or meeting presentation. Bhattacharyya spoke at the following meetings. (1) North Central Soybean Research Program and the United Soybean Board SDS Fall Meeting, November 15-16, 2012, Kansas City, MO. (2) Consortium of Plant Biotechnology Research Symposium, March 5-6, 2013, Washington, DC. (3) Plant Genomics & Biotechnology Workshop for 7th - 12th grade teachers and high school students was held from July 29 to August 2, 2013 (http://www.tuskegee.edu/plant_genomics_and_biotechnology_workshop.aspx). He plans to speak to the genetics graduate students of ISU on December 2, 2013. In Objectives 2, 3 and 4, sufficient reproducible information has not been gathered to disseminate. In Objective 5, teacher and students participating in the Tuskegee University Plant Genomics and Biotechnology Workshop received an electronic copy of the summary report. In Objective 6, investigator Lee has disseminated resources to support the CDE training through the website at http://aged.unl.edu/web/aged/alec_ag_biotechnology. A website (http://soysdsresistance.ag.iastate.edu/index.html) has been developed to showcase the activity of this project. What do you plan to do during the next reporting period to accomplish the goals? In Objective 1, for task (i) we plan to confirm that RNA interference of vital Fv genes from transgenic soybean plants will suppress Fv’s growth and enhance SDS resistance. Stable transgenic soybean lines with suitable genes for suppressing function of one or more vital Fv genes will be developed and evaluated. Synthetic anti-FvTox1 antibody genes will be generated and tested in transgenic soybean lines. Transgenic soybean lines carrying PSS1 will be evaluated for expression of possible enhanced SDS resistance. Additional Arabidopsis PSS genes will be cloned to investigate their roles in enhancing SDS resistance in soybean. In Objective 2, we will continue the screening of 227 soybean PI lines for SDS resistance to identify suitable SDS resistance soybean lines. Top five most resistant SDS PI lines will be crossed to (i) two SDS resistant cultivars for introgression of novel SDS resistance genes and (ii) one SDS susceptible soybean line to map the new SDS resistance genes. Population development will begin at the ISU research site with the objectives to i) develop mapping populations (F2­-derived lines) to identify quantitative trait loci (QTL), and recombinant inbred lines (RIL) for fine mapping. In Objective 3, Identification of new Fv isolates from IA, IL and IN will be continued. DNA of over 400 Fv and F. tucumanie isolates will be prepared and genotyped by sequencing (GBS). Based on the genotypes, isolates will be grouped to understand structure of the pathogen populations. In Objective 4, a comprehensive content analysis of key US newspapers and industry publications and their coverage of societal issues related to GM food and crops will be completed. In-depth, face-to-face interviews will be conducted with ~ 20 key informants related to food retailing and processing, agriculture and commodity organizations, state and national government agencies, consumer and environmental advocacy organizations. A survey questionnaire will be constructed and administered to crop advisors, food retailers and food processors. Interview and survey data from Year 1 and Year 2 will be analyzed. Two journal articles will be prepared for publications that discuss the major societal trends occurring that are related to consumer attitudes and societal and business acceptance of GM food crops. In Objective 5, we will continue: i) the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW) carrying continuing education credit for grade 6-12 teachers and high school students; and ii) summer research internships (SREI) for grade 6-12 teachers and undergraduate students. TUPGBW brochures and a promotional video are currently in production to assist with accomplishing the goal(s), disseminating findings and promoting the workshop. In Objective 6, we will conduct the NE CDE contest combined with our APP pilot in the training sessions given to Nebraska science teachers and to FFA teachers in Iowa.

Impacts
What was accomplished under these goals? The accomplishments are described below under each of the six objectives. The goal of Objective 1 is to generate novel genes that will confer SDS resistance in transgenic plants. There are three major tasks of this objective. They are: (i) suppress fungal growth, (ii) generate plant antibodies, and (iii) express plant genes. In task (i), we plan to suppress the growth of the SDS pathogen, Fusarium virguliforme (Fv), by creating suitable transgenic soybean plants that will produce small interfering RNAs specific to vital Fv genes. We have shown that the RNA interference pathway is conserved in Fv. This novel approach has been shown to suppress growth of the barley pathogen, Fusarium graminearum (Fg). Currently we are investigating a list of candidate genes including the cytochrome P450 lanosterol C14α-demethylase-encoding genes. In task (ii), our plan was to create synthetic antibodies that will bind to FvTox1 toxin which causes foliar SDS. We have identified FvTox1-interacting synthetic peptides. Anti-FvTox1 genes developed based on these FvTox1-interating synthetic peptides are currently being investigated to determine if any of them can enhance SDS resistance in transgenic soybean plants. In task (iii), we proposed to express Arabidopsis nonhost resistance genes to enhance SDS resistance in transgenic soybean plants. We have shown that Arabidopsis PSS1 can enhance resistance against Fv in transgenic soybean plants. We have demonstrated that there are additional PSS genes that confer immunity of Arabidopsis against Fv. The goal of Objective 2 is to identify novel SDS resistance genes from soybean germplasm and determine their utility against a diverse collection of Fv isolates. In early 2013, the Hartman lab reported to us that they identified 227 soybean plant introduction (PI) lines from screening thousands of soybean PI lines available at the USDA-ARS. We obtained the seeds of these selected 227 lines and re-screened for SDS resistance using a modified protocol that we developed recently here at ISU. This modified protocol subjects soybean genotypes to additional pressure of the SDS pathogen without damaging seed germination. This technique was found to produce results that are similar to the data collected under field conditions. Of the 227 lines, we have identified 14 that were highly resistant to Fv and are more resistant than MN1606, an early maturing highly SDS resistant cultivar. Seeds of a selected group of 59 lines with potential SDS resistance genes are being grown at the ISU research site in Puerto Rico for seed increase. The goal of Objective 3 is to determine the genetic architecture of Fv isolates in North America and F. tucumaniae isolates in South America. SDS is caused by Fv in North America and by F. tucumaniae in South America. In addition to already available isolate collections in North and South America, 10, 39, and 47 Fv isolates have been collected from the major soybean growing states, IA, IN and IL, respectively, during the 2013 field season. In IL, 47 isolates were collected from 22 counties; in IN, 39 from 21 counties and in IA 10 isolates from six counties. Scientists from other states and Canada agreed to provide us Fv isolates from their existing collections. In 2014 we plan to conduct genotyping by sequencing experiments to classify a large collection of around 400 SDS pathogen isolates including 105 collected from IA, IL and IN and 70 from our two collaborators from Argentina and Brazil. Subsequently, we will conduct sequencing of a limited number of isolates from each of the diverse groups. Towards this end, we have developed software for producing a reference-based assembly of short sequence reads. We plan to develop bioinformatics tools for SNP analysis of genomic reads from a number of Fv isolates. The goal of Objective 4 is to evaluate the economic and social impact of transgenes. In 2013, 15 in-depth, face-to-face interviews were conducted with top management and officials from farm organizations, food processors, government agencies, and advocacy organizations. A literature review was conducted that identifies the key debates, controversies, and regulatory issues related to transgenic crops and their acceptance among business, consumers, and foreign markets. Interview transcription and analysis is ongoing. The economic component of this objective is scheduled for year 5. The goal of Objective 5 is to provide education and research experience to 6-12 grade teachers and undergraduate minority students. We are focusing on two tasks of this objective. They are: i) workshops carrying continuing education credit for 6-12 grade teachers; and ii) summer research internships for 6-12 grade teachers and undergraduate students. In task (i), we organized the Tuskegee University Plant Genomics and Biotechnology Workshop (TUPGBW) from Monday, July 29th - Friday, August 2nd on the campus of Tuskegee University to educate youth and teachers in the areas of plant genomics and biotechnology. Nine middle and high school teachers and 12 high school students received relevant training in autoclaving, micropipetting, gel electrophoresis, DNA fingerprinting and biotechnology. Pairing students with teachers for two days allowed teachers to teach the students the concepts and techniques that they learned in the first three days of the workshop. Daily surveys allowed teachers and students to offer feedback regarding each teaching/learning session (e.g. clear definition of objectives, meeting the objectives, materials provided, time allocated, etc.). In task (ii), we conducted a Summer Research Experience Internship (SREI) for in-service and pre-service 6-12 grade teachers and undergraduate minority students. In Iowa State University, we conducted SREI for two 6-12 grade teachers and six undergraduate students. Five of the students came from Tuskegee University, and teachers came from the Tuskegee School System. The five undergraduate interns were housed under the George Washington Carver (GWC) summer intern program. Five students and two teachers conducted research in Bhattacharyya lab under Objective 1. One additional undergraduate SREI intern conducted research in Bain lab to investigate social impact of transgenes under Objective 4. Students attended weekly lab meetings, three seminars, and presented their findings via poster and oral presentations at the end of their internships. SREI participants significantly contributed towards advancement of the research component of this project. The goal of Objective 6 is to educate stakeholders on the use of transgenic technology, and establish demonstration plots to showcase SDS transgenic soybean lines. To educate stakeholders we are developing and delivering two outreach education products. 1) A Career Development Event (CDE) that is now a Nebraska statewide event has been created and a training curriculum to support teachers and their students to prepare for the regional and state events has been developed. Regional CDE contests are now underway in Nebraska. Current education resources available at the Plant and Soil Science eLibrary serve as the current training resources. 2) This educational event is serving as a learning forum structure for the second product, the Biotechnology APP. An APP developer (Crowd Favorite, Denver, CO) has been contracted and is designing the pilot version of this product. Pictures and videos of biotechnology scientists on the SDS team and a farmer who has dealt with SDS disease challenges in soybean have been captured and are in various editing stages for use in the APP. The SDS resistant transgenic lines, once developed, will be showcased in demonstration plots in year 5 of this project.

Publications