闻真珍, 刘运权, 林首恒, 林茵, 刘伟. 金钗石斛VRN1-like基因的克隆及其表达分析[J]. 华南师范大学学报(自然科学版), 2013, 45(4).
引用本文: 闻真珍, 刘运权, 林首恒, 林茵, 刘伟. 金钗石斛VRN1-like基因的克隆及其表达分析[J]. 华南师范大学学报(自然科学版), 2013, 45(4).
CLONING AND EXPRESSION ANALYSIS OF A VRN1-LIKE GENE FROM DENDROBIUM NOBILE[J]. Journal of South China Normal University (Natural Science Edition), 2013, 45(4).
Citation: CLONING AND EXPRESSION ANALYSIS OF A VRN1-LIKE GENE FROM DENDROBIUM NOBILE[J]. Journal of South China Normal University (Natural Science Edition), 2013, 45(4).

金钗石斛VRN1-like基因的克隆及其表达分析

CLONING AND EXPRESSION ANALYSIS OF A VRN1-LIKE GENE FROM DENDROBIUM NOBILE

  • 摘要: 用20mg/L的噻重氮苯基脲(thidiazuron,TDZ)处理金钗石斛(Dendrobium nobile)成株叶片,观测其对开花的影响;以金钗石斛类原球茎为材料,通过同源克隆分离其VRN1-like同源基因,利用半定量RT-PCR分析细胞分裂素对该基因在腋芽和类原球茎中表达的影响. 结果表明,喷施TDZ可以代替低温处理诱导金钗石斛开花;通过同源克隆得到了金钗石斛VRN1-like基因DnVRNA的全长cDNA. 该cDNA编码的蛋白与高羊茅(Festuca arundinacea)和小麦(Triticum monococcum)中VRN1蛋白的氨基酸序列一致性分别为63%和62%. 半定量RT-PCR结果表明,DnVRNA在腋芽中的表达受TDZ的促进,而在类原球茎中,其表达也受到细胞分裂素和低温的诱导,暗示该基因可能参与细胞分裂素替代低温诱导金钗石斛成花的生物学过程.

     

    Abstract: Effects of thidiazuron (TDZ) on flowering of Dendrobium nobile were investigated, by spraying on leaves with a concentration of 20mg/L; and to further investigate the molecular mechanism of the process, theVRN1-like gene was cloned from protocorm-like bodies (PLBs) of Dendrobium nobile , the effects of cytokinins on its expression in axillary buds and PLBs were studied using semi-quantitative RT-PCR analysis. The results showed that flowering of D. nobile could be induced by application with TDZ instead of cold; the full-length cDNA of a VRN1-like gene named DnVRNA (GenBank accession No. EF535598) was cloned, which encodes a peptide with 63% and 62% similarity to the VRN1 protein from Festuca arundinacea and Triticum monococcum, respectively. Semi-quantitative RT-PCR analysis showed that the expression of DnVRNA in axillary buds was improved by TDZ, and those in PLBs were also up-regulated by cytokinins and cold, which suggesting that it would function in cytokinin-induced floral transition.

     

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