大白菜抗小菜蛾突变体硫甙含量及相关基因的表达分析  

Glucosinolate Content and Expression of Related Genes in Chinese Cabbage(Brassica campestris sub. pekinensis) Mutants Resistance to Diamondback Moth(Plutella xylostella L.)

作　　者：刘梦洋[1] 卢银[1] 韩文素[1] 王彦华[1] 赵建军[1] 申书兴[1] 

机构地区：[1]河北农业大学园艺学院/河北省蔬菜种质创新与利用重点实验室,保定071001

出　　处：《农业生物技术学报》2015年第3期320-328,共9页Journal of Agricultural Biotechnology

基　　金：河北省海外高层次人才百人计划(No.E2013100011);河北省杰出青年科学基金(No.C2013204118);十二五农村领域国家科技计划课题(No.2012AA100202-5);农业科研杰出人才培养计划;高等学校博士学科点专项基金(No.20121302110006)

摘　　要：硫代葡萄糖甙（glucosinolates,GS）是十字花科植物中重要的次生代谢物质.本研究以大白菜（Brssica campestris sub.pekinensis）甲基磺酸乙酯（ethyl methylsulfonate,EMS）诱变后代为研究对象,筛选抗小菜蛾的新种质,探索其抗性机理,明确大白菜硫甙含量与小菜蛾抗性之间的关系.通过网室和离体鉴定相结合的方法筛选到5份大白菜抗小菜蛾（Plutella xylostella L.）突变体,抗性级别达到1级.硫甙含量测定结果表明,抗小菜蛾突变体中脂肪族硫甙（2-羟基-3-丁烯基硫甙（progoitrin,PRO）、4-戊烯基硫甙（glucobrassicanapin,GBN））和吲哚族4-羟基-3-吲哚基甲基硫甙（4-hydroxyglucobrassicin,4-OH）高于野生型,大白菜脂肪族硫甙PRO含量的上升可能是小菜蛾抗性增强的主要原因.通过qRT-PCR计算突变体和野生型之间在硫甙相关基因上的相对表达量,结果表明,相对于野生型,转录因子28（myb family transcription factor 28,MYB28）、MYB29、依赖二酮戊二酸的双加氧酶（2-oxoglutarate-dependent dioxygenase,AOP2）和2-氧化酸双加氧酶（2-oxoacid-dependent dioxygenase,GSL-OH）表达量在5个突变体中的表达量分别增加了1.41～4.85、1.76～6.99、1.16～4.60、1.86～8.06和0.46～7.85倍.验证了硫甙相关基因MYB28、MYB29、AOP2和GSL-OH正向调控大白菜脂肪族硫甙合成途径,二磷酸尿核苷葡萄糖基转移酶（uridine diphosphate（UDP）-glucosyl transferase 74B1,UGT74B1）正向调控吲哚族硫甙合成途径.分析了上述基因的表达量与小菜蛾取食时间的关系,抗小菜蛾突变体相比于野生型,基因表达量峰值提前了16～20 h,可能是抗小菜蛾突变体抗性应激反应的表现.研究结果为进一步了解大白菜硫甙和小菜蛾抗性的调控关系,培育具有有益硫甙的抗虫大白菜新品种提供了基础资料.Glucosinolates（GS） is important secondary metabolites in cruciferous plant. Ethyl methylsulfonate （EMS） mutants of Chinese cabbage （Brassica campestris sub. pekinensis） were used as materials in this research, aiming to obtain resistance genotypes to diamondback moth （Plutella xylostella L.） and to identify relationships between resistance level and glucosinaolate （GS） contents. In this study 5 Chinese cabbage mutants with 1st grade resistance to diamondback moth were selected using two methods： In vitro and under net condition. Through analyzing content of GS in leaf, the levels of progoitrin （PRO）, glucobrassicanapin （GBN） and 4-hydroxyglucobrassicin （4-OH） in mutants were higher than that in wild type. The main reason caused the resistance to DBM could be the high level of PRO. Expression levels of GS related genes between wild type and mutants were compared with qRT-PCR. Expression of myb family transcription factor 28 （MYB28）, MYB29, 2-oxoglutarate-dependent dioxygenase （AOP2） and 2-oxoacid-dependent dioxygenase （GSL- OH） in five mutants were higher than that in wild type, increasing folds of 1.41-4.85, 1.76-6.99, 1.16-4.60, 1.86-8.06 and 0.46-7.85, respectively. This result indicated that GS related genes MYB28, MYB29, AOP2 and GSL-OH were positive regulation to synthesis pathway of aliphatic, and uridine diphosphate （UDP）-glucosyl transferase 74B1 （UGT74B1） was positive regulation to synthesis pathway of indolyl. Analyzing the expression of above genes along with feeding time, the expression peak in mutants was earlier for 16-20 h than that in wild type, which suggested to be the sensitive performance to DBM resistance. This study can be useful for researching regulatory relations of DBM resistance and GS, and developing new varieties with resistance to insect and valuable GS component in Chinese cabbage.

关 键 词：大白菜 硫代葡萄糖甙(GS) 小菜蛾 实时荧光定量PCR 

分 类 号：S565.4[农业科学—作物学]