水稻烯酰辅酶A水合酶的功能预测分析  

Functional Prediction Analysis of Enoyl-CoA Hydratase in Rice

作　　者：单崇蕾 山草梅 陈兰兰 况卫刚 林桠春 张连虎 崔汝强[1] SHAN Chonglei;SHAN Caomei;CHEN Lanlan;KUANG Weigang;LIN Yachun;ZHANG Lianhu;CUI Ruqiang(School of Agricultural Sciences,Jiangxi Agricultural University,Nanchang,Jiangxi 330045,China)

机构地区：[1]江西农业大学农学院,江西南昌330045

出　　处：《热带作物学报》2023年第3期468-475,共8页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金地区基金项目(No.32060607)。

摘　　要：水稻潜根线虫是一类寄生于水稻根部的迁移性内寄生线虫,广泛分布于各类水稻田中,成为严重危害水稻的寄生线虫,其中水稻细尖潜根线虫是江西省最严重的水稻潜根线虫之一。目前培育抗病品种是防治水稻潜根线虫病最为经济、有效的措施。通过转录组分析水稻细尖潜根线虫侵染的和未受水稻细尖潜根线虫的感病水稻品种‘霸王鞭’根组织基因表达情况,筛选出差异表达显著上调基因OsECH1,并对OsECH1蛋白及其同源蛋白进行系统发育分析。结果表明:OsECH1蛋白与其他同源蛋白具有相同的保守结构域PLN02874,属于ECH家族。同时发现在水稻‘日本晴’中有5个同源基因,其中OsECH1基因位于染色体1上;克隆获得OsECH1基因全长并利用同源重组技术构建重组表达载体,通过农杆菌转化获得转基因水稻;经RT-qPCR分析该基因表达水平对水稻细尖潜根线虫侵染的影响。结果表明,在水稻中过表达OsECH1基因会减少水稻细尖潜根线虫的侵染,沉默OsECH1基因会增加水稻细尖潜根线虫的侵染;OsECH1蛋白亚细胞定位于细胞核中;通过His-tag pull down分析,该蛋白可能与5种防御相关蛋白互作。5种防御相关蛋白分别属于vATP-synt_E家族蛋白、Ricin-B家族蛋白、过氧化氢酶和B_lectin家族。RT-qPCR结果表明OsECH1对这5种蛋白均具有调控作用,该结果为进一步揭示OsECH1基因的功能提供基础。上述结果不仅为了解OsECH1基因的分子机制奠定基础,也为进一步阐述水稻与潜根线虫的互作机制提供新方向。Hirschmanniella spp.are migratory endoparasitic nematodes which are parasitic in the root of rice.They are widely distributed in different types of fields and have become serious parasitic nematodes to rice.The plant-parasitic nematode Hirschmanniella mucronate is one of the most serious rice nematodes in Jiangxi,China.At present,breeding and planting other disease-resistant varieties is the most economical and effective measure to control the disease.The significant up-regulated OsECH1 gene was screened by transcriptome analysis of healthy susceptible rice‘Bawangbian’root tissues and susceptible rice‘Bawangbian’root tissues infected by H.mucronata.Phylogenetic analysis of OsECH1 and its homologous proteins showed that OsECH1 and other homologous proteins had the same structure,including the same conserved domain PLN02874.And OsECH1 belonged to the ECH protein family.Besides,five homologous genes were found in the genome of‘Nipponbare’group and OsECH1 was located in Chr1.The full length of OsECH1 was cloned and the recombinant expression vector was constructed by homologous recombination technique.Meanwhile,the transgenic rice was transformed by GV1301.RT-qPCR was used to analyze the effect of OsECH1 gene expression level on the infection of H.mucronate in rice.The results showed that overexpressing OsECH1 in rice could reduce the in-fection of H.mucronata,while silencing OsECH1 in rice could significantly increase the infection of H.mucronata.The results of subcellular localization of OsECH1 showed that the protein was expressed in nucleus.And the results of His-tag pull down demonstrated that the protein may interacted with five defence-related proteins.The five de-fense-related proteins belonged to vATP-synt_E protein family,Ricin-B protein family,catalase and B_lectin protein family respectively.Meanwhile,RT-qPCR results showed that OsECH1 had regulatory effects on the five proteins.The results would provide a basis for further understanding the function of OsECH1 gene,and for understanding the mo

关 键 词：水稻潜根线虫 烯酰辅酶A水合酶 抗病基因 

分 类 号：S511[农业科学—作物学]