水稻抗潜根线虫基因OsRAI1的克隆及功能分析  

Cloning,and Functional Analysis of Gene OsRAI1 Resistant to Hirschmanniella mucronate in Rice

作　　者：山草梅 叶蕾 张连虎 况卫刚 孙晓棠[1] 马建 崔汝强[1] SHAN Cao-mei;YE Lei;ZHANG Lian-hu;KUANG Wei-gang;SUN Xiao-tang;MA Jian;CUI Ru-qiang(College of Agronomy/Key Laboratory of Ministry of Education on Crop Physiology,Ecology and Genetic Breeding,Jiangxi Agricultural University,Nanchang 330045)

机构地区：[1]江西农业大学农学院,作物生理生态与遗传育种教育部重点实验室,南昌330045

出　　处：《生物技术通报》2021年第7期146-155,共10页Biotechnology Bulletin

基　　金：国家自然科学基金项目(32060607,31860494);江西省科技计划项目(20202ACBL205005);江西省研究生创新专项资金(YC2019-S176)。

摘　　要：水稻潜根线虫病是江西省水稻生产上危害严重的病害之一。前期通过对水稻细尖潜根线虫侵染抗/感病水稻品种的根部组织进行比较转录组分析,筛选出差异表达上调基因OsRAI1。本研究克隆获得OsRAI1全长,通过蛋白结构预测、亚细胞定位对其功能进行分析,通过IPTG诱导,对OsRAI1蛋白表达条件进行了优化。结果表明,OsRAI1基因ORF全长1065 bp,编码354个氨基酸,属于bHLH转录因子家族。该蛋白为亲水性蛋白,分子量37.90 kD,pI值4.86,脂肪系数68.33,总平均亲水性(GRAVY)-0.415,为非跨膜蛋白。蛋白互作预测显示该蛋白可能是通过两个蛋白间HLH区域相互结合作用。亚细胞定位结果表明该蛋白在细胞核中表达。可溶性蛋白在0.6 mmol/L IPTG 16℃诱导18 h后表达量最高。本研究结果为进一步阐明水稻抗潜根线虫分子机制奠定基础。The plant-parasitic nematode Hirschmanniella mucronata is one of the most serious rice nematodes in Jiangxi province.In the early work of the laboratory,the up-regulated OsRAI1 gene was screened by transcriptome analysis of resistant/susceptible rice root tissues infected by H.mucronata.The full length of the gene was cloned,its protein structure was predicted,and subcellular localization were analyzed.The expression conditions of OsRAI1 protein were optimized by IPTG induction.The results showed that the ORF of OsRAI1 gene was 1065 bp in length,encoding 354 amino acids,and it belonged to the bHLH transcription factor family.The protein was hydrophilic with molecular weight of 37.90 kD,pI value of 4.86,fat coefficient of 68.33,total average hydrophilicity(GRAVY)of-0.415,and nontransmembrane protein.The results of protein interaction prediction demonstrated that the protein may interact with each other through the HLH region between the two proteins.The results of subcellular localization showed that the protein was expressed in nucleus.The protein expression was the highest when IPTG concentration was 0.6 mmol/L and induced at 16℃for 18 h.The results lay a foundation for understanding the molecular mechanism of H.mucronate resistance.

关 键 词：水稻潜根线虫 抗病基因 bHLH转录因子 原核表达 亚细胞定位 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治]