Biotechnology Bulletin ›› 2016, Vol. 32 ›› Issue (4): 80-86.doi: 10.13560/j.cnki.biotech.bull.1985.2016.04.010

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Cloning and Bioinformatics Analysis of Gene GbHCT from Gossypium barbadense

CUI Xue1 ,JIADELA·Tuliuhan1, YU Yue-hua1, CHEN Quan-jia1 ,SHEN Li-jie1, YE Jia-li1, YANG Ju-li1 ,TAO Shun1 ,NI Zhi-yong1，2   

  1. 1. College of Agronomy，Xinjiang Agricultural University，Key Laboratory of Agricultural Biological Technology，Xinjiang Agricultural University，Urumqi 830052;
    2. Institute of Cotton Research，Chinese Academy of Agricultural Sciences，State Key Laboratory of Cotton Biology，Anyang 45500
  • Received:2015-07-18 Online:2016-04-25 Published:2016-04-26

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Abstract:

This study is to isolate full-length cDNA of a key enzyme gene GbHCT related to lignin metabolism in cotton（Gossypium barbadense L.），and analyze the expression of GbHCT in the different developing stages of cotton’s fiber. The primers were designed according to the HCT sequence in transcriptome data of sea island cotton，then the gene was cloned using RT-PCR，and the cDNA sequence and the amino acid sequence of GbHCT were analyzed by bioinformatics methods. The expressions of gene GbHCT in developing fibers of cotton were analyzed from the transcriptome data of sea island cotton. A gene encoding HCT，designated as GbHCT，was isolated from G. barbadense. The full-length of the GbHCT consisted of a 1，311 bp open reading frame（ORF）encoding 436 amino acids. The molecular weight of protein was 48.58 kD，with an isoelectric point（pI）of 6.03. The deduced amino acid sequence of the GbHCT had two conservative motifs HTLGD and DFGWG. The deduced amino acid sequence had a high homology with HCT from Gossypium hirsutum，Gossypium arboretum and Gossypium raimondii，however，it showed 55.7%-60.7% identities with HCTs of other plants. Analyzing the transcriptome data of sea island cotton revealed that gene GbHCT was expressed in all different development stages of fiber，and the peak of expression reached in 5 days post anthers（DPA）fiber cells，suggesting that this gene may be involved in fiber development.

Key words: Gossypium barbadense L., HCT, gene cloning, bioinformatics