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Journal of Tea Science ›› 2010, Vol. 30 ›› Issue (2): 129-135.doi: 10.13305/j.cnki.jts.2010.02.009

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Construction and Preliminary Analysis of the Suppression Subtractive Hybridization cDNA Libraries between Dormant and Sprouting Buds of Tea Plant (Camellia sinensis)

WANG Xin-chao1,2, YANG Ya-jun1,*, CHEN Liang1,*, MA Chun-lei1, YAO Ming-zhe1   

  1. 1. Research Center for Tea Germplasm and Improvement, Tea Research Institute of Chinese Academy of Agricultural Sciences; National Center for Tea Improvement, Hangzhou 310008, China;
    2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2009-11-04 Revised:2009-12-30 Published:2019-09-10

Abstract: Bud dormancy and its release are important biological phenomena in tea plant (Camellia sinensis). In order to understand the molecular mechanism of tea bud dormancy and its release, 2 SSH-cDNA libraries were constructed using dormant and sprouting buds. The forward library contained genes preferentially expressed in sprouting buds and the reverse library contained genes preferentially expressed in dormant buds. One hundred and ninety two positive clones were randomly selected for sequencing and bioinformatics analyzing from the two libraries. The results showed that differential express genes were enriched effectively in the forward and reverse libraries, and subtraction efficiency reached the demand. Most of the length of inserted fragments was 250~750 bp. It implied that the quality of the SSH-cDNA libraries was satisfied. The bioinformatics analysis results indicated that there were differences between forward and reverse libraries on gene expression profiles. In forward library, the percent of genes related with organelle part and transporter was higher than reverse one. And in the reverse library, the percent of genes related with antioxidant, translation regulator and response to stimulus was higher than the forward. The 2 SSH-cDNA libraries can be further used for gene isolation and provide a foundation for further investigation to explore the molecular mechanism of bud dormancy and its release in tea plant.

Key words: tea plant (Camellia sinensis), bud dormancy, expressed sequence tag (EST), suppression subtractive hybridization (SSH)

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