Journal
PLANTA
Volume 236, Issue 3, Pages 931-941Publisher
SPRINGER
DOI: 10.1007/s00425-012-1706-y
Keywords
Agrobacterium rhizogenes; Hairy root; Hydrolyzable tannin; Punica granatum; Punicalagin
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Funding
- National Science Foundation [MCB1120323]
- UC Davis new faculty startup fund
- NIH Initiative for Maximizing Student Diversity/Development
- NSF Graduate Research Fellowship
- UC Davis Department of Plant Sciences Graduate Research Fellowship
- Henry A. Jastro Research Fellowship
- Direct For Biological Sciences [1120323] Funding Source: National Science Foundation
- Div Of Molecular and Cellular Bioscience [1120323] Funding Source: National Science Foundation
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In contrast to the numerous reports on the human therapeutic applications of hydrolyzable tannins (HTs), genes involved in their biosynthesis have not been identified at the molecular level from any plant species. Although we have previously identified candidate HT biosynthetic genes in pomegranate using transcriptomic and bioinformatic analyses, characterization of in planta enzyme function remains a critical step in biochemical pathway elucidation. We here report the establishment of a pomegranate (Punica granatum) hairy root culture system that produces HTs. Agrobacterium rhizogenes strains transformed with a binary vector harboring a yellow fluorescent protein (YFP) gene were used for hairy root induction, allowing visual, non-destructive, detection of transgene incorporation. It also demonstrated that the pomegranate hairy root culture system is suitable for expressing heterologous genes (YFP in this case). Expression of 26 putative UDP-glycosyltransferase (UGT) genes, obtained from a pomegranate fruit peel (a tissue highly abundant in HTs) RNA-Seq library, were verified in wild type and hairy roots. In addition, two candidate UGTs for HT biosynthesis were identified based on HPLC and differential gene expression analyses of various pomegranate tissues. Together with in vitro enzyme activity assays, the hairy root culture system holds great promise for revealing the undivulged HT biosynthetic pathway using pomegranate as a model system.
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