Interactomic Study of Root Locus B in Tobacco Hairy Roots
Date Issued
2014
Date
2014
Author(s)
Yi, Sung-Hui
Abstract
Hairy roots are fast growing adventitious root tissues. They can accumulate high amount of plant secondary metabolites. Hairy roots are differentiated after Agrobacterium rhizogenes infects the wounded plant and transfers the T-DNA from its Ri plasmid randomly into the plant chromosome. However, some plants are resistant to the Agrobacterium-infection and therefore the utilization of hairy root culture in these plants is limited. Understanding the molecular mechanism during hairy root formation enables potential application of hairy root induction in medical herbs. In T-DNA, rolB has been considered to act the pivotal role during the differential process of hairy roots. In our previous studies, tobacco hairy roots deficient in rolB gene showed less lateral roots, less neoplastic growth and slower growth rate. To elucidate the protein-protein interactions of RolB protein in tobacco hairy roots, we conducted yeast two-hybrid screening. We found that RolB interacted with ORF13a, PHI-2 and NtbZIP. The potential protein-protein interactions were further studied by bimolecular fluorescence complementation (BiFC) assay in tobacco BY-2 cells. BiFC results showed that the interactions of RolB with PHI2 and NtbZIP were located in a DNA-containing organelle other than the nucleus. The localizations of RolB-YFP, ORF13a-YFP and PHI-2-YFP were found in both nucleus and cytosol, while NtbZIP-YFP was only detected in nucleus. ORF13a is encoded in the T-DNA as rolB. PHI-2 and NtbZIP are homologous to Arabidopsis ABA Responsive Element Binding Factors (ABFs) family. ABFs are known as the negative regulators in root growth upon ABA signaling. Our finding suggests that RolB interacts with ABFs family proteins and promotes hairy root growth possibly by regulating ABA signaling pathway.
Subjects
毛狀根
根毛農桿菌
rolB
蛋白質交互作用體學
ABFs/ABI5轉錄因子
Type
thesis
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