大豆咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)基因克隆及结构分析  

Cloning and Bioinformatics of CCoAOMT Relating to Resistance of Soybean to Cyst Nematodes

作　　者：郭子雯 司修洋 焦莉苹 刘大伟[1] GUO Ziwen;SI Xiuyang;JIAO Liping;LIU Dawei(College of Plant Protection,Northeast Agricultural University,Harbin,Heilongjiang 150030,China;College of Agriculture and Biotechnology,Zhejiang University,Hangzhou,Zhejiang 310058,China)

机构地区：[1]东北农业大学植物保护学院,黑龙江哈尔滨150030 [2]浙江大学农业与生物技术学院,浙江杭州310058

出　　处：《福建农业学报》2023年第5期616-623,共8页Fujian Journal of Agricultural Sciences

基　　金：黑龙江省自然科学基金(LH2019C034)。

摘　　要：【目的】对大豆中咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)基因进行克隆和生物信息学分析,为研究木质素代谢途径关键基因CCoAOMT在大豆抗胞囊线虫的作用机制奠定基础。【方法】以高抗大豆胞囊线虫(Heterodera glycines Ichinohe)3号生理小种品种灰皮支黑豆根系为材料,利用RT-PCR技术进行克隆。【结果】克隆得到1条大豆CCoAOMT基因的cDNA全长序列,将其命名为GmCCoAOMT,GenBank登录号为MW480860。该基因全长为848 bp,含有1个741 bp的开放阅读框,编码246个氨基酸组成的蛋白,分子量为27.6 kDa,等电点为5.67;亚细胞定位预测显示,CCoAOMT蛋白可能位于细胞质,该蛋白含有1个保守的AdoMet_MTases结构域,属于AdoMet_MTases超级家族;在与其他豆科植物CCoAOMT编码的蛋白序列比对及系统进化树分析中,GmCCoAOMT编码的蛋白序列与其他豆科植物具有较高的相似性。【结论】获得大豆CCoAOMT基因的cDNA全长序列,通过构建进化树确定大豆与豇豆、木豆、菜豆和赤豆的CCoAOMT蛋白进化关系较近,而与白羽扇豆和鹰嘴豆进化关系较远。【Objective】Full-length cDNA of Caffeoyl-CoA-O-methyltransferase gene(CCoAOMT)in the lignin metabolism pathway of soybean was cloned,and bioinformatics analyzed to study the resistance mechanism to cyst nematode(Heterodera glycines Ichinohe)of the plant.【Method】CCoAOMT sequence from roots of a cyst nematode-resistant Huipizhiheidou soybean plant was cloned by RT-PCR.【Results】The full length cDNA of GmCCoAOMT was cloned and then submitted to GenBank with the accession number of MW480860.The length of GmCCoAOMT was 848 bp,containing an ORF of 741bp and encoding 246 amino acids with a predicted molecular weight of 27.6 kDa and a theoretical isoelectric point of 5.67.It was located in the cytoplasm with a conserved AdoMet_MTases domain and belonged to that superfamily.The multiple alignment on amino acid sequence and phylogenetic tree indicated a high similarity between the gene and those of other leguminous plants.【Conclusion】The cDNA of GmCCoAOMT was successfully cloned and shown with a close evolutionary relationship with Vigna unguiculata(Linn.)Walp.,Cajanus cajan(Linn.)Millsp.,Phaseolus vulgaris Linn.and Vigna angularis(Willd.)Ohwi et Ohashi but far from Lupinus albus and Cicer arietinum Linn.

关 键 词：大豆 大豆胞囊线虫 咖啡酰辅酶A-O-甲基转移酶 基因克隆 生物信息学分析 

分 类 号：S435[农业科学—农业昆虫与害虫防治]