Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (14): 2953-2962.doi: 10.3864/j.issn.0578-1752.2013.14.011

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Expression Analysis of A Male Sterility-Related Gene VvMS2 from Grapes

 ZHENG  Huan-1, ZHANG  Ji-Yu-12, WANG  Xin-Wei-1, ZHANG  Zhen-1, JI  Chen-Fei-1, TAO  Jian-Min-1   

  1. 1.College of Horticulture, Nanjing Agricultural University, Nanjing 210095
    2.Institute of Botany, Jiangsu Province and the Chinese Academy of Sciences, Nanjing 210014
  • Received:2012-11-01 Online:2013-07-15 Published:2013-05-13

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Abstract: 【Objective】 This paper aims to isolate the VvMS2 gene from the Vitis vinifera L. cv. Wink and Zhongshanhong, in order to further study the molecular mechanism of grape male sterility and grape molecular breeding. 【Method】 Specific primers were designed based on the grape sequence (CBI29968) in NCBI which is highest homology with Arabidopsis male sterility gene MS2, and the cDNA and genomic DNA of VvMS2 were cloned by PCR. VvMS2 gene structure characteristics were analyzed using the bioinformatics software. Quantitative real-time PCR (qRT-PCR) was performed to determine the expression pattern of VvMS2 in different tissues of ‘Wink’ and at different development stages in bud between ‘Zhongshanhong’ and ‘Wink’.【Result】The full length ORF cDNA sequence of VvMS2 with 1 755 bp were cloned. Bioinformatics analysis showed that this gene encodes 584 amino acids, containing NAD-binding-4 structure domains and sterile structure domains. This gene contains 9 exons and 8 introns. The protein molecular weight is 64.74 kD, isoelectric point is 8.84. Compared with other sequences of MS2 in GenBank, the deduced amino acid sequences of VvMS2 shared 40%-71% in homology. VvMS2 only specially expressed in floral buds and at the stages of tetrad to dinucleate. The expression level in button at uninucleate stage in ‘Wink’ is remarkably higher than that in ‘Zhongshanhong’.【Conclusion】The abnormal gene expression is related with male sterility in ‘Zhongshanhong’.

Key words: grape , male sterility , VvMS2 , gene cloning , characterization , expression pattern

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