Abstract Detail

Genomics / Proteomics

Anguraj Vadivel, Arun Kumaran [1], Dhaubhadel, Sangeeta [2].

Regulation of Isoflavonoid Biosynthesis and GmMYB176 interactome in soybean.

Soybean seeds are rich source of proteins, oil and plant natural compounds including isoflavonoids. Isoflavonoids are legume-specific metabolite with a role in nitrogen fixation, defence mechanism in plants and also in human health and nutrition. Isoflavonoid biosynthesis in soybean is found to be regulated by a MYB transcription factor, GmMYB176, by regulating chalcone synthase 8 (CHS8) gene expressions in the pathway. We have demonstrated that GmMYB176 alone is not sufficient for CHS8 gene regulation and hypothesized that GmMYB176 acts cooperatively with other factor (s). The goal of this study is to elucidate the GmMYB176 interactome for CHS8 gene regulation and isoflavonoid biosynthesis in soybean. Two translational fusions of GmMYB176 (GmMYB176-YFP and YFP-GmMYB176) were created with yellow fluorescent protein (YFP). The fusion proteins were used as baits in separate experiments to pull down their interacting proteins by co-immunoprecipitation and identified by liquid chromatography tandem mass spectrometry. Proteins such as 14-3-3, WD40, bHLH, ATP synthases, kinases, phosphatase and uncharacterized proteins were identified. The interaction of some of the candidates with GmMYB176 was validated in planta by bimolecular fluorescence complementation assay. DNA binding activity of these candidates was also studied. Understanding the role of these candidates on CHS8 expression and isoflavonoid biosynthesis will allow genetic manipulation of isoflavonoid level in soybean and/or introduce isoflavonoid pathway in non-legumes.

1 - Western University, Department of Biology, 1151 Richmond street, London, Ontario, N6A 3K7, Canada
2 - Agriculture and Agri-Food Canada, 1391 Sandford Street, London , Ontario, N5V 4T3, Canada

Keywords:
Co-immunoprecipitation
Transcriptional regulation
protein protein interactions
Mass Spectrometry
Isoflavonoid
Biosynthetic pathway regulation.

Presentation Type: Oral Paper:Papers for Topics
Session: 48
Location: Salon 19/20/The Shaw Conference Centre
Date: Tuesday, July 28th, 2015
Time: 1:45 PM
Number: 48002
Abstract ID:866
Candidate for Awards:CSPB President's Award for Best Student Presentation