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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (12): 2035-2044.doi: 10.3724/SP.J.1006.2010.02035


Construction of SSH Library and Its Analyses of Cotton Drought Associated Genes under Drought Stress

WANG De-Long,YE Wu-Wei*,WANG Jun-Juan,SONG Li-Yan,FAN Wei-Li,CUI Yu-Peng   

  1. Cotton Research Institute, Chinese Academy of Agricultural Sciences / Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Anyang 455000, China
  • Received:2010-05-18 Revised:2010-08-01 Online:2010-12-12 Published:2010-10-09
  • Contact: YE Wu-Wei,E-mail:yeww@cricaas.com.cn;Tel:0372-2562283 E-mail:wdl_21@126.com

Abstract: A forward cDNA-SSH library was established by suppression subtractive hybridization using seedling leaf of Handan 177, a drought-tolerant cotton (Gossypium hirsuutm L.) inbred line, among which 300 positive clones were selected for sequencing. After detection by PCR for each clone, each single clone was sequenced. Totally 284 available sequences and 202 uniESTs which 28 were contigs and 174 were singlets were obtained by cluster analyses of the ESTs sequencing. The results of BlastN showed that 156 uniESTs had homologous sequences in GenBank database while the other 46 had no protein homologous. The BlastX results indicated that 116 uniESTs had significant protein homology and 40 uniESTs were unknown proteins and putative proteins. KOBAS mapped 33 ESTs of the 202 uniESTs to 55 KEGG pathways, in which there were 23 pathways at P-value<0.05. This study suggested that there were closely relationships with cotton drought tolerance among pyruvate metabolism, glyoxylate and dicarboxylate metabolism. A large group of drought stress-induced genes were found in the cDNA library, which involved in many metabolism pathways such as signal transduction, energy metabolism, protein metabolism, nucleic acid metabolism, photosynthesis, transmembran. And some genes related to drought tolerance were found, such as malate synthase genes (MS1,0001_C12; MS2, 0002_F01) and malate dehydrogenase genes(Md1, 001_C12; Md2, 002_F01), some transcription factors like NAC(001_C08), BZR1/BES1(003_G04) , zinc finger protein genes (zfp, 003_C06), and the translationally controlled tumor protein gene(TCTP,002_C04).

Key words: Drought-stress, Suppression subtractive hybridization (SSH), unigene, Drought tolerance

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