甘蔗ScHAK9基因克隆及表达分析
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1广西作物遗传改良生物技术重点开放实验室,南宁 530007;,1广西作物遗传改良生物技术重点开放实验室,南宁 530007;,1广西作物遗传改良生物技术重点开放实验室,南宁 530007;,2广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室,南宁 530007,1广西作物遗传改良生物技术重点开放实验室,南宁 530007;,2广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室,南宁 530007,2广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室,南宁 530007,1广西作物遗传改良生物技术重点开放实验室,南宁 530007;,1广西作物遗传改良生物技术重点开放实验室,南宁 530007;

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广西自然科学(2014GXNSFAA118081)、广西自然科学基金项目(2013GXNSFAA019077)、广西农业科学院基本科研业务专项项目(2015YT96)、广西农业科学院基本科研业务专项项目(桂农科2017YM35)


Molecular Isolation and Expression Analysis of ScHAK9 Gene from Sugarcane
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1Guangxi Crop Genetic Improvement and Biotechnology Laboratory Nanning 530007 ;,1Guangxi Crop Genetic Improvement and Biotechnology Laboratory Nanning 530007 ;,1Guangxi Crop Genetic Improvement and Biotechnology Laboratory Nanning 530007 ;,2Sugarcane Research Institute Guangxi Academy of Agricultural Sciences/Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi),Ministry of Agriculture, Nanning 530007,1Guangxi Crop Genetic Improvement and Biotechnology Laboratory Nanning 530007 ;,2Sugarcane Research Institute Guangxi Academy of Agricultural Sciences/Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi),Ministry of Agriculture, Nanning 530007,2Sugarcane Research Institute Guangxi Academy of Agricultural Sciences/Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi),Ministry of Agriculture, Nanning 530007,1Guangxi Crop Genetic Improvement and Biotechnology Laboratory Nanning 530007 ;,1Guangxi Crop Genetic Improvement and Biotechnology Laboratory Nanning 530007 ;

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    摘要:

    KUP/HAK/KT家族基因在介导细胞内钾的积累及维持植物的生长发育中起重要作用,本研究以新台糖22号(ROC22)甘蔗为研究材料,采用同源克隆技术获得钾转运ScHAK9基因的完整编码序列(CDs),并利用生物信息学软件对蛋白质结构和功能进行预测分析,同时利用qPCR技术分析基因的组织特异性表达以及在不同干旱条件下的表达情况。结果表明,ScHAK9基因的cDNA完整编码区长度为2352 bp,编码783个氨基酸,属于碱性疏水蛋白,结构中包含了12个跨膜结构域,蛋白主要定位在细胞质膜上;蛋白质二级结构主要由α螺旋结构、无规则卷曲结构和扩展长链组成;系统发育树分析显示ScHAK9与玉米ZmHAK9基因同源性较高。qPCR分析结果表明,ScHAK9基因在不同组织间的相对表达量具有明显差异,叶片中表达最高,其次是茎,根系中表达最低,具有组织特异性;干旱胁迫可以诱导ScHAK9基因表达,其表达量随着胁迫程度加重表现出升高趋势,且在复水后才有所下降。初步推测该基因可能在甘蔗发育和抵御干旱胁迫中起着重要作用,本研究为进一步阐明ScHAK9的功能及作用机制奠定了分子基础。

    Abstract:

    The KUP/HAK/KT family gene plays an important role in regulation of potassium accumulation. In this research, we isolated the complete coding sequence of ScHAK9 and investigated the transcriptional levels in ROC22. ScHAK9 gene was composed of 2352 nucleotides that encodes 783 amino acids with 12 predictable transmembrane domains. The deduced protein was predicted to be a basic dewatering protein and localized to the cytoplasmic membrane. The secondary structure consisted of alpha spiral structure, irregular curling structure and extended long chain. ScHAK9 gene showed highly sequence identity with the homologous genes as compared with ZmHAK9 gene(Zea mays L). Real-Time PCR showed that the ScHAK9 gene highly expressed in the leaves, followed by the stem and the root at last. ScHAK9 gene expression was induced under drought stress, and the higher expression decreased after re-watering. Thus, our work provided first insight about ScHAK9, which might play a role in drought stress resistance.

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罗海斌,蒋胜理,黄诚梅,等.甘蔗ScHAK9基因克隆及表达分析[J].植物遗传资源学报,2018,19(4):731-739.

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  • 收稿日期:2018-01-24
  • 最后修改日期:2018-05-22
  • 录用日期:2018-05-14
  • 在线发布日期: 2018-07-16
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