陆地棉GDSL脂肪酶增强拟南芥对黄萎病菌的抗性  被引量:6

A Cotton GDSL Lipase Increases the Resistance against Verticillium dahilae in Arabidopsis

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作  者:袁哈利 郝晓云[1] 郑学伟[1] 李鸿彬[1,2] 

机构地区:[1]石河子大学生命科学学院,石河子832003 [2]石河子大学农业生物技术重点实验室,石河子832003

出  处:《石河子大学学报(自然科学版)》2015年第5期529-534,共6页Journal of Shihezi University(Natural Science)

基  金:新疆兵团种质资源创新专项(2012BB050);新疆兵团杰出青年项目(2014CD003);石河子大学杰出青年项目(2012ZRKXJQ03)

摘  要:为了分析脂肪酶对于黄萎病菌的抗性功能,本研究采用RT-PCR技术从陆地棉中克隆得到1个GDSL脂肪酶(GDSL Lipase,GLIP)基因Gh GLIP,ORF全长1068 bp。采用滴花法将重组表达载体35S::Gh GLIP转入哥伦比亚型拟南芥,通过筛选、鉴定和纯合得到T3代转基因拟南芥。利用黄萎病菌悬浮液分别对野生型和转基因拟南芥植株叶片进行局部处理后,发现转Gh GLIP基因拟南芥植株能够抑制黄萎病菌诱发的叶片细胞死亡及病菌在叶片感染处的生长繁殖。酶活性测定结果显示,转Gh GLIP基因拟南芥植株的抗氧化系统酶和病程相关酶的表达获得了显著的增强;RT-PCR分析结果表明,黄萎病菌处理1 d后,转Gh GLIP基因拟南芥叶片的病程相关基因和乙烯信号相关基因的表达获得了显著的增加。本研究结果表明,棉花Gh GLIP基因可能通过参与乙烯信号,以及促进抗氧化系统酶和病程相关基因的表达,进一步增强拟南芥对于黄萎病菌的抗性。本研究可为棉花Gh GLIP基因增强植物抗病性分子机制解析,以及进一步的基因工程利用提供一定参考。This study aimed to analyze the resistance function of GDSL lipase(GLIP) against Verticillium dahliae.In this study,a cotton GDSL lipase gene,named Gh GLIP,which was previously cloned from Gossypium hirsutum L.with a 1068 bp open reading frame,was transformed into Col- 0 Arabidopsis by floral diping method,and as a result the T3 generation transgenic Arabidopsis plants were obtained through screening of homozygotes.Verticillium dahliae suspension was utilized to infect the leaves of wide- type(WT) and transgenic Arabidopsis plant carrying Gh GLIP gene,respectively.The leaves of transgenic plants displayed less cell death and reproduction of Verticillium dahlia compared with the wide- type plants.Enzyme assays showed that expressions of antioxidant enzymes and pathogen related enzymes in transgenic Gh GLIP Arabidopsis plants were significantly enhanced than that in the wild- type plants;RT- PCR analyses indicate that the expression levels of pathogen related genes and ethylene signal transduction related genes were obviously increased after1 d treatment of Verticillium dahliae.These results suggested that cotton Gh GLIP gene might increase Arabidopsis resistance against to Verticillium dahliae by participating in ethylene signal pathway and increasing the expression levels of antioxidant enzyme system and pathogen related genes.This study provides references forthe molecularmechanism elucidation of plant resistance to pathogen and furthergenetic improvement applications.

关 键 词:棉花GDSL脂肪酶 黄萎病 乙烯信号 抗氧化系统酶 病程相关基因 

分 类 号:S435.621.2[农业科学—农业昆虫与害虫防治]

 

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