RNA Interference Analysis of Receptor-like Kinase Gene （StRLK） in Potato

doi:10.3969/j.issn.1000-7091.2012.04.004

Abstract

Abstract: Plant receptor-like protein kinases （RLKs） are the subfamily of protein kinases and participate plant signal transduction. RLKs play an important role in growth,development and disease resistance in plants. In the pre- vious study,we have obtained the potato receptor-like protein kinase gene （StRLK） from diploid potato genotype ED13 with high resistance to Ralstonia solanacearum. The specific fragment of StRLK was used to construct RNAi vector pCHF1-StRLK through the intermediate cloning vector pUCCRNAi and plants expression vector pCHF1. Then the pCHF1-StRLK was transfered into ED13 via agrobacterium-mediated transformation using the stems of ED13 as explants,and finally five transgenic regeneration lines were obtained. The five lines were tested with polymers Chains Reaction （PCR） using the specific primers of CaMV35S promoter,and the results showed that about 500bp PCR products were got in all of them,indicats that all of the five lines are transgenic lines. RT-PCR analysis of StR- LK gene expression in the five transgenic lines were also performed using the StRLK specific primers with ED13 as control. The results showed that the StRLK gene expression were inhibited in different level in the five different lines, demonstrated that pCHF1-StRLK had played interference activity and the interference effection was related to gene insertion site in the potato genome,which would provide some references in the function study of StRLK gene.

Key words: Diploid potato, StRLK gene, Genetic transformation RNA interference

CLC Number:

ZHU Yun, FAN Na-na, YU Yao-hua, ZHANG Bai-shun, YANG Yu, CHAO Xiang-jian, GUO Bao-tai, JIN Li-ping, GUO Xiao, LI Guang-eun. RNA Interference Analysis of Receptor-like Kinase Gene （StRLK） in Potato[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(4): 18-22. doi: 10.3969/j.issn.1000-7091.2012.04.004.

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