棉 花 学 报     Cotton Science   2009，21(6)：474-479

Construction and Bioinformatics Analysis of Suppression Subtractive Hybridization Library for Cotton Induced by Spodoptera Litura (Fabricius)
XU Dong, ZHANG Yong-jun^*, CAO Guang-chun, CHEN Yang, WU Kong-ming, GUO Yu-yuan
(State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)

Abstract：To explore insect resistant genes induced by Spodoptera litura (Fabricius) larvae from cotton, forward and reverse suppression subtractive hybridization(SSH) libraries were constructed. 1000 clones were selected from each library, the inserted fragment sizes ranged between 200 bp and 1000 bp, which meet the requirements of a plant cDNA library. 250 positive clones from the forward library were screened by using the Reverse Northern Dot-blotting technique, then sequenced and analyzed, and 35 expressed sequence Tags (ESTs) were obtained, in which 24 ESTs were obtained from the positive clone in reverse library. Based on BLASTx in GenBank analysis, it was found that those ESTs make important roles in signaling pathway regulation, gene regulation, anti-stress ability, defense responses, energy synthesis and metabolism, proliferation of cells, and so on. Those genes play an important role in regulating and defending against S.litura larvae directly and indirectly.
Key words：suppression subtractive hybridization (SSH)；expressed sequence Tags (ESTs)；cotton；
Spodoptera litura        [Full Text, 4189KB]