番茄LeRma1基因的克隆与表达
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黑龙江省教育厅面上项目(12531204,12531178);植物生物学黑龙江省高校重点实验室开放课题(ZK201205);哈尔滨师范大学青年学术骨干资助计划(KGB201218,09KXQ-01);黑龙江省普通高等学校青年学术骨干支持计划;哈尔滨师范大学硕士研究生创新科研项目(HSDSSCX2014-32)


Clone and Expression of LeRma1 from Lycopersicon esculentum Mill.
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    摘要:

    以番茄‘哈大粉801’为试材,利用RT-PCR技术,克隆得到1个E3泛素蛋白连接酶基因LeRma1(GenBank登录号XM_004243764.1)。对LeRma1基因进行序列分析,并对LeRma1基因在番茄植株的不同部位以及在非生物胁迫(干旱、盐、碱、高温、低温)下的表达和生理特性进行研究,为培育和改良番茄品种提供理论依据。结果表明:(1)序列分析显示,LeRma1基因的cDNA全长序列729 bp,编码242个氨基酸,分子量为27.05 kD,理论pI 7.97;同源分析显示,番茄LeRma1蛋白与马铃薯的一致性最高(91%)。(2)半定量PCR检测表明, LeRma1基因在番茄根、茎、叶、花、果实中均有表达,且表达差异不明显。(3)干旱胁迫下,LeRma1基因在番茄叶片中优势表达,而在整个干旱过程中根部的LeRma1基因表达量变化不明显;抗旱相关基因LEA、 DREB2A、ABI3在干旱胁迫过程中,番茄叶片中均有表达,且其表达量呈上升趋势,而在根部DREB2A、ABI3基因基本没有检测到。(4)干旱胁迫过程中,番茄植株中丙二醛(MDA)含量呈显著升高趋势,质膜系统严重损伤,体内保护酶(SOD、POD、CAT)活性上升,且根部活性总体明显高于叶片。(5)在非生物逆境(盐、碱、高温、低温)胁迫过程中,LeRma1基因在番茄叶片和根部的表达几乎都有增强的趋势,且在叶片中均是胁迫3 h后诱导起始增强表达。研究认为,LeRma1基因是一个受干旱胁迫诱导增强表达的基因,且在叶片中优势表达,说明LeRma1基因对植物耐受干旱胁迫所起的作用存在一定的组织差异性,而且LeRma1基因可能参与番茄的干旱应答及信号转导过程,在番茄抵抗其他非生物胁迫中LeRma1基因也可能具有一定的作用。

    Abstract:

    The experiment cloned an E3 ubiquitin ligase gene LeRma1(GenBank accession No XM_004243764.1)by RT-PCR method with tomato ‘Hadafen 801’ used as experimental materials.To provide a theoretical basis for the cultivation and improved varieties of tomato,we analyzed the gene sequence,while studied the expression and physiological characteristics of LeRma1 gene in different parts of the tomato plants under abiotic stress(drought,salt,alkali,high temperature,low temperature).The results showed that:(1)LeRma1 gene sequence analysis showed that the length of LeRma1 gene was 729 bp,which encoded 242 amino acids.It was predicted that the molecular mass of its protein was 27.05 kD,and pI was 7.97.Homology analysis showed that the deduced amino acid sequence exhibited high homology with Solanum tuberosum(91% similarity).(2)RT-PCR results indicating that the gene were expressed in roots,stems,leaves,flowers and fruits of tomato,and the changes in expression levels were not significantly different.(3)Expression of LeRma1 had the advantage in leaves,and the changes in expression levels were not significantly different in roots under the whole drought stress.The other drought-related genes DREB2A,LEA,ABI3 are all expression in leaves,and they were enhanced expression compared with the control,while DREB2A,ABI3 have not detected in roots under the whole drought stress.(4)Major membrane lipid peroxidation product of MDA content was increased significantly,and plasma membrane systems were severely damaged,the protective enzyme activity increased significantly,and activity in roots was significantly higher than that in leaves.(5)LeRma1 gene was enhanced expression in roots and leaves of tomato,and the gene in the leaves all starting enhanced expression after stress 3 h under abiotic stress(salt,alkali,high temperature,low temperature).The results showed that LeRma1 was the gene of enhancement expression by drought-induced and it had the advantage in leaves.It showed that there were some differences that the function of the LeRma1 genes in different tissues(organizational differences)under drought stress,and LeRma1 genes may be participate response of drought stress tolerance and the signal transduction process,it also might play an important role in the tomato under the other abiotic stresses.

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陈默,金晓霞,于丽杰,等.番茄LeRma1基因的克隆与表达[J].西北植物学报,2015,35(9):1735-1745

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  • 在线发布日期: 2015-10-01
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