Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (15): 3087-3096.doi: 10.3864/j.issn.0578-1752.2013.15.002

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

Cloning of Choline Monooxygenase (CMO) Gene and Expression Analysis of the Key Glycine Betaine Biosynthesis-Related Genes in Tea Plant (Camellia sinensis)

 CAO  Hong-Li-12, YUE  Chuan-12, HAO  Xin-Yuan-13, WANG  Xin-Chao-1, YANG  Ya-Jun-1   

  1. 1.Tea Research Institute of the Chinese Academy of Agricultural Sciences/National Center for Tea Improvement, Hangzhou 310008
    2.Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081
    3.College of Horticulture, Northwest A & F University, Yangling 712100, Shaanxi
  • Received:2013-03-05 Online:2013-08-01 Published:2013-05-20

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Abstract: 【Objective】 Choline monooxygenase gene (CMO), one of the key enzyme genes in biosynthesize pathway of glycine betaine (GB) in plant, was cloned from tea plant (Camellia sinensis). The expression patterns of CsCMO and the other GB biosynthesis-related gene, CsBADH (betaine aldehyde dehydrogenase), under different abiotic stresses and in different cold resistant cultivars were analyzed. And the contents of GB under different abiotic stresses in tea plant were detected. 【Method】 The full-length cDNA of CsCMO was isolated using reverse transcription-PCR (RT-PCR) combined with RACE techniques from tea plant, and the bioinformatic characteristics were analyzed using online software. The expression profiles of CsCMO and CsBADH and the contents of GB were determined using quantitative real-time PCR (qRT-PCR) method and ultraviolet spectrophotometer techniques, respectively. 【Result】 The obtained cDNA of CsCMO was 1 558 bp in length with GenBank accession number JX050146, containing a 1 305 bp open reading frame (ORF) which encoded 434 amino acid residues. The CsCMO shared more than 50% amino acid sequence similarity with that of other plants. It was targeted in chloroplast and contained two consensus domains of Rieske-type [2Fe-2S] cluster and sites for mononuclear nonheme Fe. The phylogenetic tree analysis showed that CsCMO had the closest genetic relationship with Lycium barbarum. The qRT-PCR analysis indicated that the expressions of CsCMO and CsBADH were up-regulated by cold (4℃), salt (NaCl) and abscisic acid (ABA) during 96 h treatment, but they displayed different patterns under these stresses. The expression level of two genes induced by NaCl was more dramatically than other treatment. The further experiments showed that the level of CsBADH was generally higher than CsCMO in different cold tolerance cultivars, and the cultivars which had stronger cold hardiness had higher expression level of CsCMO and CsBADH than that of the cultivars which had weaker cold hardiness. The accumulation of GB was induced by abiotic stress, such as cold (4℃), salt (250 mmol•L-1 of NaCl) and ABA (100 mmol•L-1), and cold-tolerance cultivars contained higher GB levels than cold-sensitive. 【Conclusion】 CsCMO was cloned from tea plant. The expression levels of CsCMO and CsBADH were up-regulated, especially under salt stress, and the contents of GB were accumulated under cold, salt and ABA stress. The results indicated that GB had close relationship with above three stresses resistance in tea plant.

Key words: tea plant (Camellia sinensis) , glycine betaine (GB) , choline monooxygenase (CMO) , quantitative real-time PCR , abiotic stress

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