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《Jiangsu Journal of Agricultural Sciences》 2008-01
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Construction and Analysis of A SSH Library of Upland Cotton Upon Infection with Verticillium dahliae

ZHANG Ying-xia1,2,YANG Yu-wen2,NI Wan-chao2,CAO Xiang-rong1,SHEN Xin-lian2 (1.College of Life Science,Nanjing Normal University,Nanjing 210097,China;2.Institute of Agro-biotechnology,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)  
To explore disease resistant-related candidate genes of Verticillium wilt from upland cotton,a high Verticillium wilt resistant cotton cultivar was used for construction of a suppression substrative hybridization(SSH)library.The cDNAs from the inoculated seedlings 48 h after disease inoculation were used as the tester and those from the control seedlings as the driver.The total RNA and mRNA were successfully isolated using a method of modified CTAB,followed by the synthesis of double strand cDNA.After two rounds of PCR,the up-regulated fragments responding for induction of Verticillium wilt were accumulated in the SSH library.The purified PCR products were ligated to the vector and transferred into E.coli.434 clones with 0.45 kb to 0.70 kb inserts were obtained.All the clones have been grouped to 20 kinds by fingerprinting and each clone of every kind was chosen to be sequenced.Sequence were blasted to NCBI database.Most of them had high similarity to genes or ESTs from the SSH libraries induced by other pathogens.Their amino acid sequences showed homology to some resistant-related genes,such as cytochrome P450 mono-oxygenase,Thaumatin-like protein,chitinase and PR proteins.Information of this study could be used to isolate the Verticillium wilt resistant-related genes and understand the molecular mechanisms of disease response in cotton.
【Fund】: 江苏省农业科学院基金项目(005036110528)
【CateGory Index】: S435.62
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