首页> 中文期刊> 《西北植物学报》 >海岛棉GbRLCK10基因克隆及表达分析

海岛棉GbRLCK10基因克隆及表达分析

         

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1179 bp,编码392个氨基酸,具有典型的Ser-ine/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、N aC l、干旱、激素胁迫的应答反应,但其功能仍需进一步研究.%In this study ,the anti-stress At1g67520 gene from A rabidopsis thaliana was used as a probe ,a RLCK gene of GbRLCK10 was cloned through the in silico cloning and RT-PCR.To reveal the expres-sion ,regulation and mechanism of RLCK family genes in Gossypium barbadense ,we has analyzed the ge-nomic structure and the expression pattern of GbRLCK10 by combining with qRT-PCR technology under mRNA organization and different stress inducements .Results show that :(1) the ORF of GbRLCK10 was 1179 bp ,encoding 392 amino acids .GbRLCK10 contained only the Serine/Threonine kinase domain ,and belonged to receptor-like cytoplasmic kinase gene family .Phylogenetic analysis showed that GbRLCK10 closed to GaRLCK10 and named as GbRLCK10 (Genebank :2022184 ) .The gene was made up of 5 exons and 4 introns .(2) The GbRLCK10 expression were detected by qRT-PCR ,the results showed that GbRL-CK10 gene is expressed in root ,stem and leaf of resistant cultivar 'Xinhai 21' and susceptible cultivar'Xinhai 14' .With the duration of the treatment of V erticillium dahliae ,w hile the relative expression of GbRLCK10 was significantly lower and the response was later in the susceptible cultivar than that in resist-ant cultivar .Both NaCl and PEG6000 could induce the expression of GbRLCK10 ,the expression of GbRL-CK10 increased firstly and decreased .After resistant cultivar 'Xinhai 21' was treated by NaCl ,response time of GbRLCK10 is earlier than PEG6000 ,but the response to PEG6000 was more intense .Salicylic acid ,ethylene ,methyl jasmonate and gibberellic acid could also induce the expression of GbRLCK10 .Af-ter the treatment of salicylic acid ,the level of GbRLCK10 expression increased and decreased ,then in-creased again .The expression of GbRLCK10 increased and maintained at a high level when induced by eth-ylene .However ,the expression of GbRLCK10 firstly increased and then decreased when induced by meth-yl jasmonate .Compared with salicylic acid ,ethylene ,methyl jasmonate ,the gibberellic acid induced ex-pression levels of GbRLCK10 were significantly lower .We suggested that GbRLCK10 may be response to pathogens ,NaCl ,PEG6000 and hormones stress in Gossypium barbadense L .,but it's function still need further study .

著录项

  • 来源
    《西北植物学报》 |2017年第11期|2130-2138|共9页
  • 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,新疆石河子 832000;

    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,新疆石河子 832000;

    中国农业科学院 生物技术研究所,北京 100081;

    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,新疆石河子 832000;

    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,新疆石河子 832000;

    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,新疆石河子 832000;

    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,新疆石河子 832000;

    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,新疆石河子 832000;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 转化及克隆;基因的表达;
  • 关键词

    棉花; 类受体胞质激酶; GbRLCK10基因克隆; 表达分析;

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