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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (02): 286-293.doi: 10.3724/SP.J.1006.2011.00286

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

Cloning of Cotton CBF Gene and Its Cold Tolerance Expression in Transgenic Tobacco

GUO Hui-Ming,LI Zhao-Chun,ZHANG Han,XIN Yue-Zhi,CHENG Hong-Mei*   

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing100081, China
  • Received:2010-08-02 Revised:2010-10-09 Online:2011-02-12 Published:2010-12-15

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Abstract: Low temperature is an adverse environment condition affecting the growth and productivity of crops, it is also one of limiting factors for cotton yeild and quality in China.CBF is a kind of transcription factor that can regulate expression of a number of genes related with abiotic stresses. Therefore, it is important to study the characteristics of cotton CBFs and their response to abiotic stresses. In this study, CBF gene was isolated from the genomic DNA of cotton cultivars Gh12, Gh36 and Gb7124. Cotton CBF gene encodes 184 amino acids, containing CBF-family signature “PKRRAGRKKFQETRHP” and “FADSAW”. Southern blotting result showed that CBF genes were presented as the form of gene family in the genome of cotton. Northern blotting result indicated that GbCBF1 gene was induced by low temperature, drought, salt and ABA. GbCBF1 was constructed into plant expression vector pCambia2301, in which the gene was driven by 35S and NOS promoters separately. Plant expression vectors were then transferred into tobacco NC89 using Agrobacterium-mediated transformation method. Twenty six trangentic tobacco lines were obtained after kanamycin screening and PCR detection. PCR and Reverse tanscription PCR methods were used to analyze part of T1 transgenic tobacco, the results showed that GbCBF1gene could be transcripted and inherit in offspring normally. The analytical result demonstrateted that the electrolytic leakage rate of transgenic tobacco was lower than that of wild type tobacco generally, however, free proline content and soluble sugar content of transgenic tobacco were higher than those of wild type tobacco under low temperature stress. In conclusion, GbCBF1 enhances cold tolerance in transgenic tobacco.

Key words: Cotton, Transgenic tobacco, CBF, Transcription factor, Cold tolerance

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