玉米大斑病菌Stgb-1基因正反义表达载体的构建与功能初探  

Construction of sense and antisense expression vectors and preliminary functional research of Stgb-1 gene in Setosphaeria turcica

作　　者：郝志敏[1] 李丽花[1] 李志勇[2] 韩建民[1] 陆晴[3] 佟文跃 董金皋[1] 

机构地区：[1]河北农业大学植物分子病理学实验室,河北保定071001 [2]河北省农林科学院谷子研究所,河北石家庄054000 [3]河北唐山市农业科学研究院,河北唐山063601

出　　处：《河北农业大学学报》2010年第6期1-6,共6页Journal of Hebei Agricultural University

基　　金：国家自然科学基金项目(30471126);河北省自然科学基金项目(C2008000335)

摘　　要：玉米大斑病菌(Setosphaeria turcica)是严重威胁玉米生产安全的重要植物病原真菌。本研究在前期克隆得到的玉米大斑病菌G蛋白β亚基编码基因Stgb-1的基础上,利用pSO-I质粒成功构建Stgb-1基因正反义表达载体,并将反义表达载体用于转化玉米大斑病菌的原生质体,获得了3个突变体菌株,分别命名为anti-GB-1、anti-GB-2和anti-GB-3。其中,anti-GB-2、anti-GB-3的菌落形态与野生型菌株差异较为明显,anti-GB-1则与野生型菌株没有表型差异。利用anti-GB-1对Stgb-1基因的功能进行分析,发现在反义表达诱导物奎宁酸溶液(15 mmol/L)中,野生型菌株的孢子萌发率为93.7%,而anti-GB-1的孢子萌发率仅为16%。推测Stgb-1基因调控了玉米大斑病菌的分生孢子萌发过程。Setosphaeria turcica,the cause of northern leaf blight of corn,is one of the major plant pathogens.Stgb-1（GenBank Acession No.EF407555）,a gene encoding heterotrimeric G protein beta subunit in S.turcica,was ligated with pSO-I plasmid which contained a quinic acid-inducible promoter.Both the sense and antisense expression vectors were obtained and the later was used to transform the protoplasm of S.turcica.As a result,three Stgb-1 anti-sense expression mutants verified by PCR were obtained and named as anti-GB-1,anti-GB-2,and anti-GB-3 respectively.Anti-GB-1 was used to investigate the function of Stgb-1 gene for its similar phenotype with the wild type strain.The results showed that the spore germination ratio in quinic acid（15 mmol/L） of anti-GB-1 was only 16%,while that of wild type strain was about 93.7%.Based on all the above results,it was speculated that Stgb-1 gene regulated the spore germination in Setosphaeria turcica.

关 键 词：玉米大斑病菌 异三聚体G蛋白 反义表达 转化 

分 类 号：S432.44[农业科学—植物病理学]