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Analysis of Leaf Metabolites in a Soybean CPR5 Mutant

Undergraduate #152
Discipline: Biological Sciences
Subcategory: Plant Research
Session: 4
Room: Exhibit Hall A

KeAndre Leaks - Fort Valley State University
Co-Author(s): Eudald Illa Berenguer: -Center for Applied Genetics, University of Georgia, Athens, GA, USA Evan McCoy: -Institue of Plant Breeding, Genetics & Genomics, University of Georgia, Athens, GA, USA Maria Ortega: -Department of Plant Biology, University of Georgia, Athens, GA, USA Scott Harding: -Department of Genetics, University of Georgia, Athens, GA, USA -Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA, USA Baggi Nyamdari: -Department of Genetics, University of Georgia, Athens, GA, USA -Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA, USA CJ Tsai: -Department of Genetics, University of Georgia, Athens, GA, USA -Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA, USA Wayne Parrott: -Institue of Plant Breeding, Genetics & Genomics, University of Georgia, Athens, GA, USA, -Center for Applied Genetics, University of Georgia, Athens, GA, USA -Department of Crop and Soil Sciences, University of Georgia, Athens, GA, USA



Why this is important: Breeding pathogen-resistant soybean is a very worthy goal. Between 2010 and 2014, $26 billion was lost to soybean pathogens in the United States and Ontario, Canada (Allen et al. 2017). Hypothesis: We suspect that there will be a difference in metabolites between the cpr5 mutant gene and Wild type (Jack) because: Trichomes contain metabolites and are almost absent in cpr5 mutants Cpr5 mutants are smaller in size than wild type Cpr5 provides systemic acquired resistance in Arabidopsis which is associated with changes in metabolite abundance (Bowling et al. 1997) Methods: The plants were grown in a greenhouse and leaves were flash frozen at the V3 growth stage (Fehr and Caviness 1977). The leaves were freeze-dried and a methanol chloroform extraction was used on 5 mg of freeze-dried tissue. Liquid Chromatography Mass Spectrometry (LCMS) was used to analyze 5 microliters of each sample. We used KNApSAcK to match potential compounds with the same molecular weight that we found within the peaks. We also calculated the area of the peaks for thirteen compounds. Two were significantly different between cpr5 and the wild type. Conclusion and future research questions: We concluded that there were some significant metabolite differences between soybean possessing the cpr5 mutant gene and the wild type. Although most compounds tested were not different in the mutant and wild type, two compounds were affected by the cpr5 mutation. The compound with the retention time of 8.144 minutes has a mass of 270 g/mol which matches three entries in the KNApSAcK database. The entries are all isoflavonoids with similar molecular structures differing in the placement of a single hydroxyl (OH) group. The potential compounds are genistein, demethyltexasin, and 8-hydroxydaidzein. Isoflavonoids are known to be important for pathogen resistance in soybean (Zhang et al. 2017), but it is curious that this compound seems to disappear in the cpr5 mutant. It could be that the compound with the retention time of 8.144 minutes is only present in plant trichomes which are almost absent in cpr5 mutants. The compound with the retention time of 7.105 minutes has no matches in KNApSAcK, meaning there are no reports in the literature of a compound with this mass in soybean. The reason there were differences in certain compounds could be the lack of trichomes, pathogen resistance, or the reduced plant size. For future research, we will quantify the other compounds detected in the samples and identify the compounds with significant changes using purified standards.

Funder Acknowledgement(s): I was supported by the National Institute of Food and Agriculture Research and Extension Experiences for Undergraduates (REEU) Program.

Faculty Advisor: Sarwan Dhir, dhirs0@fvsu.edu

Role: I conducted the research in the labs of Dr. Wayne Parrot (My mentor) and Dr. Cj Tsai. I was helped by Dr. Parrott's Postdoc; Eudald Illa Berenguer and Ph.D Student; Evan McCoy. I was also helped by Dr. Tsai's Postdoc; Maria Ortega and other lab workers such as Scott Harding and Baggi Nyamdari. They taught me the techniques needed to complete this research and I took it from there.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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