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园艺学报 ›› 2016, Vol. 43 ›› Issue (12): 2412-2422.doi: 10.16420/j.issn.0513-353x.2016-0615

• 其它园艺植物 • 上一篇    下一篇

铁皮石斛DoCDPK基因的克隆与表达分析

盛 况,高燕会*,斯金平,朱玉球   

  1. (浙江农林大学,亚热带森林培育国家重点实验室培育基地,浙江省铁皮石斛产业技术创新战略联盟,浙江临安 311300)
  • 出版日期:2016-12-25 发布日期:2016-12-25

Cloning and Expression Analysis of DoCDPK Genes in Dendrobium officinale

SHENG Kuang,GAO Yan-hui*,SI Jin-ping,and ZHU Yu-qiu   

  1. (Zhejiang Agricultural and Forestry University,Nurturing Station for State Key Laboratory of Subtropical Silviculture,Zhejiang Provincial Strategic Alliance for Technical Innovation in Industry of Dendrobium Officinale,Lin’an,Zhejiang 311300,China)
  • Online:2016-12-25 Published:2016-12-25

摘要:

利用RACE技术和RT-PCR相结合,从铁皮石斛Dendrobium officinale Kimura et Migo)中克隆得到长度分别为1 863和1 729 bp的钙依赖蛋白激酶(calcium-dependent protein kinase)基因DoCDPK1DoCDPK2的cDNA全长序列,开放阅读框分别为1 539和1 536 bp,编码512和511个氨基酸,编码蛋白表现为亲水性,结构稳定,二级结构主要由α–螺旋和无规卷曲构成。qRT-PCR分析结果表明DoCDPK1DoCDPK2主要在铁皮石斛叶片中表达;4 ℃低温和400 mmol · L-1 NaCl处理后,DoCDPK1DoCDPK2在各个时间点上(2、4、8、12和24 h)的表达量有不同程度提高,而100 mmol · L-1 ABA处理后DoCDPK1DoCDPK2的表达量呈现不同变化趋势,推测DoCDPK1DoCDPK2参与低温等非生物胁迫的响应。

关键词: 铁皮石斛, CDPK基因, 基因克隆, 逆境胁迫, 基因表达

Abstract:

To study the structure,function and expression patterns of CDPK(calcium-dependent protein kinase)genes in Dendrobium officinale Kimura et Migo,cDNA sequences were cloned from Dendrobium officinale by RACE and RT-PCR. The full length of cDNA sequences of DoCDPK1 and DoCDPK2 geneswere 1 863 bp and 1 729 bp which the ORF(open reading frames)were 1 539 bp and 1 536 bp encoded 512 and 511 amino acids,respectively. And DoCDPK1 and DoCDPK2 proteins were hydrophilic and stable,whose secondary structures were made up of α-helix and random coil. The qRT-PCR analysis showed that DoCDPK1 and DoCDPK2 genes mainly expressed in leaves. The expression of DoCDPK1 and DoCDPK2 genes increased under low temperature and NaCl. However,the expression of DoCDPK1 and DoCDPK2 genes showed different expression patterns under the treatment of ABA. We speculated that DoCDPK genes took part in the abiotic stress like cold stress. The results will lay the foundation of further study of the specific regulation mechanism and relationship with other signaling pathways.

Key words: Dendrobium officinale, CDPK, gene cloning, abiotic stress, gene expression

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