Construction and analysis of a suppression subtractive hybridization library for the potato virus induced response gene

doi:10.11686/cyxb2017063

Abstract

Abstract: Potato virus accumulation is one of the important reasons for the decline of yield and quality in potato production. In order to screen and clone the potato virus induced response gene, we constructed a suppression subtractive hybridization (SSH) library using cDNAs from potato virus disease carrying plant leaf as the tester, and those cDNAs from a virus-free seedling leaf as the driver. To verify the effect of library, 98 randomly selected positive clones were identified by PCR from the library and sequenced, and 45 high quality non repeat sequences were obtained. By blast analysis in GenBank, we found that 14 of them were sequences belonging to the potato virus gene, 22 of them had high homology with known genes, and 9 of them had no homologous reference gene. Two frequently found ESTs in the library were analyzed using quantitative real time PCR, and their expression was induced by potato virus. The results therefore showed that the SSH library was constructed successfully. This work has thus laid a foundation for further screening for potato virus related pathogenicity, and induction of corresponding defense response genes. This may lead to analysis of the molecular mechanisms of interaction between the potato plant and the virus, and use of biotechnology to cultivate virus resistant potato varieties.

LI Zhong-Wang, CHEN Yu-Liang, OU Qiao-Ming, YE Chun-Lei, PEI Huai-Di, LIU Xin-Xing, WANG Hong-Mei, LUO Jun-Jie. Construction and analysis of a suppression subtractive hybridization library for the potato virus induced response gene[J]. Acta Prataculturae Sinica, 2017, 26(12): 152-159.

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