甘蔗类异黄酮还原酶（IRL）基因的克隆与表达分析

生物技术通报 ›› 2014, Vol. 0 ›› Issue (11): 130-137.

甘蔗类异黄酮还原酶（IRL）基因的克隆与表达分析

谢晓娜¹,张小秋¹,邵敏¹,朱惠¹,杨丽涛^1,2,李杨瑞^1,2

1. 广西大学农学院亚热带农业生物资源保护与利用国家重点实验室,南宁 530004；2.中国农业科学院甘蔗研究中心广西农业科院农业部广西甘蔗生物技术与遗传改良重点实验室广西甘蔗遗传改良重点实验室,南宁 530007

收稿日期:2014-03-27 出版日期:2014-11-07 发布日期:2014-11-07
作者简介:谢晓娜,女,博士研究生,研究方向：作物栽培及生理基础
基金资助:
国家”863”计划课题（2013AA102604）,国家自然科学基金项目（31360293）,国家国际合作项目（2013DFA31600）,广西科技合作与交流计划项目（桂科合1347004-2）,广西自然科学基金创新团队项目（2011GXNSFF018002）,广西自然科学基金项目（2012GXNSFDA053011

Cloning and Expression Analysis of Sugarcane Isoflavone Reductase-like(IRL)Gene

Xie Xiaona¹,Zhang Xiaoqiu¹,Shao Min¹,Zhu Hui¹,Yang Litao^1,2,Li Yangrui^1,2

1. College of Agriculture,Guangxi University,State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources,Nanning 530004；2. Sugarcane Research Center,Chinese Academy of Agricultural Sciences,Key Laboratory of Sugarcane Biotechnology and Genetic Improvement（Guangxi）,Ministry of Agriculture,Guangxi Academy of Agricultural Sciences,Guangxi Key Laboratory of Sugarcane Genetic Improvement,Nanning 530007

Received:2014-03-27 Published:2014-11-07 Online:2014-11-07

摘要/Abstract

摘要： 采用 RT-PCR 技术从甘蔗中克隆 SoIRL基因,用生物信息学方法对获得的氨基酸序列进行分析,利用荧光定量 PCR 技术研究SoIRL基因在甘蔗不同组织和不同胁迫条件下的表达特性。结果表明,克隆获得甘蔗 SoIRL,GenBank 登录号为 KF808324。该 cDNA 全长 1 169 bp,含有 1 个 927 bp 的完整开放阅读框（ORF）,编码 309个氨基酸。系统进化树分析显示,甘蔗 SoIRL 与玉米的 IRL 蛋白亲缘关系较近。qRT-PCR 分析表明 SoIRL 在甘蔗根、茎、叶中均有表达；在RSD病菌及低温（4℃）、聚乙二醇（PEG）、NaCl 和脱落酸（ABA）4种非生物胁迫下均被诱导表达,但表达模式不同。说明该基因可能参与甘蔗应答RSD过程,并可能在非生物胁迫中也发挥了作用。

关键词: 甘蔗, 类异黄酮还原酶, 基因克隆, 表达分析

Abstract: The SoIRL gene cDNA sequence was cloned from sugarcane variety GT11 using RT-PCR techniques. The bioinformatics methods were used to analyze the putative amino acid sequence, and Real-time PCR method was used to study the expression of SoIRL gene in different tissues and under different stresses. The results showed that the full-length cDNA of SoIRL（GenBank accession number：KF808324）in sugarcane was cloned. The sequence consists of 1 167 bp with an intact open reading frame of 927 bp, encoding a polypeptide of 303 amino acids. Phylogenetic tree analysis indicated that SoIRL was highlg closely related to IRL of Zea mays. Real-time PCR results showed that the SoIRL expressed in root, stalk and leaf. Furthermore, SoIRL transcription level was induced under the treatment of the bacterial of ratoon stuning diaease, low temperature, PEG, NaCl and ABA stresses, but the expression patterns were different. Gene SoIRL was firstly isolated and characterized from sugarcane（GT11）, which may participate in sugarcane resistance to RSD, and also play a role in the sugarcane resistance to chilling, drought and salt stress environments.

Key words: Sugarcane, SoIRL , Gene cloning, Expression analysis

谢晓娜,张小秋,邵敏,朱惠,杨丽涛,李杨瑞. 甘蔗类异黄酮还原酶（IRL）基因的克隆与表达分析[J]. 生物技术通报, 2014, 0(11): 130-137.

Xie Xiaona,Zhang Xiaoqiu,Shao Min,Zhu Hui,Yang Litao,Li Yangrui. Cloning and Expression Analysis of Sugarcane Isoflavone Reductase-like(IRL)Gene[J]. Biotechnology Bulletin, 2014, 0(11): 130-137.

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