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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (10): 1763-1770.doi: 10.3724/SP.J.1006.2011.01763

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Construction and Analysis of Tobacco SSH Library Induced by Phytophthora parasitica var. nicotianae

SU Zhen-Gang1,YANG Ai-Guo1,SUN Yu-He1,LUO Cheng-Gang1,LIU Guan-Shan1,ZHOU Jia2,LI Yuan-Yuan1,YANG Fan1,ZHAO Bai-Ying1,WANG Yuan-Ying1,*   

  1. 1 Tobacco Research Institute of Chinese Academy of Agricultural Sciences / Key Laboratory of Tobacco Genetic Improvement and Biotechnology, Qingdao 266101, China; 2 Yunnan Reacend Tobacco Technology (Group) Co. Ltd, Kunming 650106, China
  • Received:2011-03-15 Revised:2011-06-25 Online:2011-10-12 Published:2011-07-28
  • Contact: 王元英, E-mail: wyytob@126.com, Tel: 0532-88703857 E-mail:szhg521@163.com

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Abstract: Black shank of tobacco is one of the major disease which harmful to tobacco in the world, the incompatible interaction between tobacco and Phytophthora parasitica var. nicotianae was studied to provide more information for integrated control. A cDNA-SSH library was established by suppression subtractive hybridization using roots and stems sampled at 0.5, 1, 3, 6, 10, and 16 d after inoculation of Gexin 3, a horizontal resistant cultivar to race 0 of Phytophthora parasitica var. nicotianae, and 960 positive clones were picked out. Among them, 240 positive clones were verified by Reverse Northern dot-blot. A total of 57 differentially expressed EST sequences were screened out, and 33 high quality non-redundant ESTs were obtained by cluster analyses of the ESTs sequencing.The results of BlastN showed that the resistant genes of tobacco mainly related to disease defense, photosynthesis, signal transduction, and energy metabolism. Further analysis of gene function indicated thatPR1b protein, cysteine proteinase, EF1-α, α-tubulin, cytochrome P450, putative spermine synthase, aquaporin, peroxisomal membrane protein,glycine for the elongation factor-1α may be involved in the process of the incompatible interaction between tobacco and Phytophthora parasitica var. nicotianae.

Key words: Tobacco, Phytophthora parasitica var. nicotianae, Suppression subtractive hybridization (SSH), Reverse Northern dot-blot, Incompatible interaction

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