海岛棉GbRLCK10基因克隆及表达分析
发布时间：2018-12-13 04:59
【摘要】：该研究以拟南芥抗逆基因At1g67520为探针,利用海岛棉ESTs数据库,通过电子克隆获得海岛棉RLCK家族基因GbRLCK10,解析该基因组结构,并结合qRT-PCR技术分析该基因mRNA的组织表达特征以及在不同胁迫诱导下的表达模式,为揭示RLCK家族基因在海岛棉中的表达调控及作用机制提供理论依据。结果显示:(1)获得海岛棉类受体胞质激酶(RLCK)基因,其开放阅读框(ORF)为1 179bp,编码392个氨基酸,具有典型的Serine/Threonine结构域,属于RLCK家族,与GaRLCK10(XP_017604046.1)亲缘关系较近,命名为GbRLCK10(登录号2022184),且该基因由5个外显子和4个内含子组成。(2)实时荧光定量(qRT-PCR)检测显示,GbRLCK10基因在抗病品种‘新海21’和感病品种‘新海14’的根、茎、叶中均有表达;当黄萎病菌诱导后,GbRLCK10基因在抗病品种中对于病原菌的响应时间早于感病品种,且对黄萎病菌响应更强烈,推测该基因参与棉花对黄萎病的响应;盐(NaCl)、干旱(PEG-6000)处理‘新海21’后,GbRLCK10基因在NaCl处理下响应时间要早于PEG-6000处理,但对PEG-6000处理响应更强烈;分别用4种激素处理‘新海21’后,GbRLCK10均能被诱导表达,且在水杨酸(SA)处理后表现为先增加后下降再增加趋势,在乙烯(ET)处理后表达量为持续上升趋势,在茉莉酸甲酯(MeJA)处理后呈先升高然后下降的趋势,但GbRLCK10基因对赤霉素(GA3)响应不明显。研究表明,GbRLCK10基因具有RLCK基因家族典型特征,该基因随黄萎病菌、NaCl、干旱、激素处理时间推移而发生变化,推测GbRLCK10基因可能参与了棉花对黄萎病菌、NaCl、干旱、激素胁迫的应答反应,但其功能仍需进一步研究。
[Abstract]:Using Arabidopsis thaliana (Arabidopsis thaliana) stress resistance gene At1g67520 as probe and island cotton ESTs database, the genomic structure of island cotton RLCK family gene GbRLCK10, was obtained by electronic cloning. The expression characteristics of mRNA and the expression patterns under different stress were analyzed by qRT-PCR technique, which provided a theoretical basis for revealing the regulation and mechanism of the expression of RLCK family genes in island cotton. The results showed that: (1) the receptor cytoplasmic kinase (RLCK) gene was obtained. Its open reading frame (ORF) was 1179 BP, encoding 392 amino acids. It had a typical Serine/Threonine domain and belonged to the RLCK family. The gene was closely related to GaRLCK10 (XP_017604046.1) and was named GbRLCK10 (accession number 2022184), and the gene was composed of 5 exons and 4 introns. (2) Real-time fluorescence quantitative (qRT-PCR) analysis showed that the gene was highly related to GbRLCK10 (accession number 2022184), and the gene was composed of 5 exons and 4 introns. GbRLCK10 gene was expressed in the roots, stems and leaves of the resistant variety 'Xinhai 21' and the susceptible variety 'Xinhai 14'. When Verticillium wilt was induced, the response time of GbRLCK10 gene to pathogen in resistant varieties was earlier than that of susceptible variety, and the response to Verticillium wilt was stronger. It was speculated that the GbRLCK10 gene was involved in the response of cotton to Verticillium wilt. The response time of GbRLCK10 gene under NaCl treatment was earlier than that of PEG-6000 treatment after salt (NaCl), drought (PEG-6000) treatment, but the response to PEG-6000 treatment was stronger. After treated with four hormones, the expression of GbRLCK10 was induced, and the expression of GbRLCK10 increased first, then decreased and then increased after treatment with salicylic acid (SA), and the expression level increased continuously after treatment with ethylene (ET). After (MeJA) treatment with methyl jasmonate, the GbRLCK10 gene increased first and then decreased, but the response of GbRLCK10 gene to gibberellin (GA3) was not obvious. The results showed that GbRLCK10 gene had the typical characteristics of RLCK gene family, and it changed with Verticillium wilt, NaCl, drought and hormone treatment. It was suggested that GbRLCK10 gene might be involved in cotton drought to Verticillium wilt and NaCl,. Response to hormone stress, but its function still needs further study.
【作者单位】： 新疆农垦科学院生物技术研究所/作物种质创新与基因资源利用兵团重点实验室;中国农业科学院生物技术研究所;
【基金】：兵团应用基础研究计划(2016AG005) 国家重点研发计划(2016YFD0100203-6)
【分类号】：Q943.2;S562
，
本文编号：2375925
[Abstract]:Using Arabidopsis thaliana (Arabidopsis thaliana) stress resistance gene At1g67520 as probe and island cotton ESTs database, the genomic structure of island cotton RLCK family gene GbRLCK10, was obtained by electronic cloning. The expression characteristics of mRNA and the expression patterns under different stress were analyzed by qRT-PCR technique, which provided a theoretical basis for revealing the regulation and mechanism of the expression of RLCK family genes in island cotton. The results showed that: (1) the receptor cytoplasmic kinase (RLCK) gene was obtained. Its open reading frame (ORF) was 1179 BP, encoding 392 amino acids. It had a typical Serine/Threonine domain and belonged to the RLCK family. The gene was closely related to GaRLCK10 (XP_017604046.1) and was named GbRLCK10 (accession number 2022184), and the gene was composed of 5 exons and 4 introns. (2) Real-time fluorescence quantitative (qRT-PCR) analysis showed that the gene was highly related to GbRLCK10 (accession number 2022184), and the gene was composed of 5 exons and 4 introns. GbRLCK10 gene was expressed in the roots, stems and leaves of the resistant variety 'Xinhai 21' and the susceptible variety 'Xinhai 14'. When Verticillium wilt was induced, the response time of GbRLCK10 gene to pathogen in resistant varieties was earlier than that of susceptible variety, and the response to Verticillium wilt was stronger. It was speculated that the GbRLCK10 gene was involved in the response of cotton to Verticillium wilt. The response time of GbRLCK10 gene under NaCl treatment was earlier than that of PEG-6000 treatment after salt (NaCl), drought (PEG-6000) treatment, but the response to PEG-6000 treatment was stronger. After treated with four hormones, the expression of GbRLCK10 was induced, and the expression of GbRLCK10 increased first, then decreased and then increased after treatment with salicylic acid (SA), and the expression level increased continuously after treatment with ethylene (ET). After (MeJA) treatment with methyl jasmonate, the GbRLCK10 gene increased first and then decreased, but the response of GbRLCK10 gene to gibberellin (GA3) was not obvious. The results showed that GbRLCK10 gene had the typical characteristics of RLCK gene family, and it changed with Verticillium wilt, NaCl, drought and hormone treatment. It was suggested that GbRLCK10 gene might be involved in cotton drought to Verticillium wilt and NaCl,. Response to hormone stress, but its function still needs further study.
【作者单位】： 新疆农垦科学院生物技术研究所/作物种质创新与基因资源利用兵团重点实验室;中国农业科学院生物技术研究所;
【基金】：兵团应用基础研究计划(2016AG005) 国家重点研发计划(2016YFD0100203-6)
【分类号】：Q943.2;S562
，
本文编号：2375925
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