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Journal of Tea Science ›› 2009, Vol. 29 ›› Issue (1): 53-59.doi: 10.13305/j.cnki.jts.2009.1.009

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Cloning and Expression Analysis of CBF Gene in Cold Induced Tea Plant [Camellia Sinensis (L.)O.Kuntze]

CHEN Xuan1,2, FANG Wan-ping2, ZOU Zhong-wei2, WANG Yu-hua2, CHENG Hao1,2,*, LI Xing-hui2   

  1. 1. Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
    2. Tea Research Institute, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2008-09-26 Revised:2008-10-30 Online:2009-02-15 Published:2019-09-06

Abstract: The cDNA-AFLP was applied to identify genes expressed differentially in cold acclimatized tea plant. A cDNA fragment, TDF11 encoding a CBF (C-repeat binding factor) protein was isolated and identified. The cDNA fragment had 94%、84%、81% homology with CBF genes from Brassica napus, Arabidopsis thaliana, Nicotiana tabacum by BLAST on NCBI. The complete cDNA sequence was cloned by RACE (rapid amplification of cDNA ends), named Camellia sinensis CBF gene. Its full cDNA sequence was 981bp (GenBank accession number EU857638) and contained an open reading frame encoding a polypeptide of 275 amino acid residues. Analysis of the nucleotide sequence and deduced amino acid sequence showed 74%、57%、57% homology with CBF genes from Brassica napus, Nicotiana tabacum, Arabidopsis thaliana. To identify differential expression of CBF gene, RT-PCR experiments were performed in six samples including leaves cold-induced for different treatments. The results showed that tea CBF gene expressed after cold induced for 4 hours and up to a maximum when cold induced for 8 h, and then decreased. It hinted the CBF gene might play a key role during the development of cold resistant molecular mechanism in tea plant.

Key words: tea plant (Camellia sinensis), CBF gene, clone, expression analysis

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