引用本文: | 许金港, 任欣欣, 王海冰, 王白雪, 荆胜利, 刘清松.水稻OCPI2基因的克隆与表达分析[J].广西植物,2024,44(12):2187-2196.[点击复制] |
XU Jingang, REN Xinxin, WANG Haibing, WANG Baixue,
JING Shengli, LIU Qingsong.Cloning and expression analysis of OCPI2 gene in rice[J].Guihaia,2024,44(12):2187-2196.[点击复制] |
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水稻OCPI2基因的克隆与表达分析 |
许金港1,2, 任欣欣2, 王海冰2, 王白雪2, 荆胜利2, 刘清松2,3*
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1. 西南林业大学 林学院, 昆明 650224;2. 信阳师范大学 生命科学学院, 河南 信阳 464000;3. 河南大学 生命科学学院/棉花生物育种与综合利用全国重点实验室, 河南 开封 475004
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摘要: |
为探究蛋白酶抑制剂基因在水稻抵御植食性昆虫为害中的功能,该研究以水稻品种‘中花11'为材料,克隆了水稻丝氨酸蛋白酶抑制剂基因OCPI2编码区序列,并通过生物信息学软件对其序列特征进行了分析和系统发育树的构建,同时采用实时荧光定量PCR技术探究了该基因在植食性昆虫取食和植物激素诱导下的表达特征。结果表明:(1)水稻OCPI2基因编码区序列全长219 bp,编码72个氨基酸,预测蛋白分子量为7.72 kDa,理论等电点为5.21,不含信号肽,无跨膜结构。(2)OCPI2蛋白与乌拉尔图小麦(Triticum urartu, EMS61613.1)同源蛋白亲缘关系较近。(3)OCPI2基因具有1个potato_inhibit保守结构域,属于丝氨酸蛋白酶抑制剂家族。(4)二化螟和褐飞虱取食、机械损伤以及水杨酸甲酯处理均能诱导OCPI2基因的表达,茉莉酸甲酯处理则持续抑制OCPI2基因表达。以上研究结果表明OCPI2基因可能参与了水稻对植食性昆虫的诱导防御反应,为深入研究OCPI2基因在水稻抗虫防御反应中的功能提供理论依据。 |
关键词: 水稻, 蛋白酶抑制剂, OCPI2, 基因克隆, 诱导表达特征 |
DOI:10.11931/guihaia.gxzw202403028 |
分类号:Q943 |
文章编号:1000-3142(2024)12-2187-10 |
基金项目:国家自然科学基金(31901896); 河南省科技攻关项目(222102110116); 中国科协青年人才托举工程(2023QNRC001)。 |
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Cloning and expression analysis of OCPI2 gene in rice |
XU Jingang1,2, REN Xinxin2, WANG Haibing2, WANG Baixue2,
JING Shengli2, LIU Qingsong2,3*
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1. College of Forestry, Southwest Forestry University, Kunming 650224, China;2. College of Life Sciences, Xinyang Normal University,
Xinyang 464000, Henan, China;3. College of Life Sciences, State Key Laboratory of Cotton Bio-Breeding
and Integrated Utilization, Henan University, Kaifeng 475004, Henan, China
1. College of Forestry, Southwest Forestry University, Kunming 650224, China; 2. College of Life Sciences, Xinyang Normal University,
Xinyang 464000, Henan, China; 3. College of Life Sciences, State Key Laboratory of Cotton Bio-Breeding
and Integrated Utilization, Henan University, Kaifeng 475004, Henan, China
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Abstract: |
To investigate the function of protease inhibitor genes in rice defense against herbivorous insects, the coding sequence of the protease gene OCPI2 from the rice variety ‘Zhonghua 11' was cloned. The sequence characteristics of OCPI2 gene were analyzed, and a phylogenetic tree was constructed using bioinformatics software. Real-time quantitative PCR was utilized to examine the expression characteristics of OCPI2 gene under herbivorous insect feeding and plant hormone treatment. The results were as follows:(1)The coding region of OCPI2 gene was 219 bp, encoding a protein of 72 amino acids. The OCPI2 protein had a molecular weight of 7.72 kDa and a theoretical isoelectric point of 5.21, with no signal peptide and no transmembrane structure.(2)The OCPI2 protein was closely related to the homologous protein in Triticum urartu(EMS61613.1).(3)The OCPI2 gene contained a potato_inhibit conserved domain and belonged to the serine protease inhibitor family.(4)The expression of OCPI2 gene was induced by feeding from the rice striped stem borer(Chilo suppressalis)and the brown planthopper(Nilaparvata lugens), mechanical damage, and methyl salicylate treatment, whereas methyl jasmonate treatment consistently suppressed OCPI2 gene expression. Taken together, these results suggest that OCPI2 gene may be involved in the induced defense response of rice to herbivorous insects. This study provides a theoretical basis for a deeper understanding of the function of OCPI2 gene in rice defense against insect herbivores. |
Key words: rice, protease inhibitor, OCPI2, gene cloning, induced expression signature |
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