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2025年4月4日 星期五
  2017, Vol. 25 Issue (10): 1661-1667    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
玉米大斑病菌转录因子Flo8的基因克隆及表达规律分析
申珅1,李贞杨2,赵玉兰1,李盼1,董金皋3,韩建民3,郝志敏4
1. 河北农业大学 生命科学学院
2.
3. 河北农业大学生命科学学院
4. 河北农业大学植物分子病理学实验室
Gene Cloning and Expression Pattern Analysis of Transcription Factor Flo8 in Setosphaeria turcica
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摘要 lo8是cAMP信号途径下游的重要转录因子,与真菌的生长发育密切相关。本研究旨在获得玉米大斑病菌(Setosphaeria turcica)转录因子Flo8编码基因,并分析其在病菌侵染结构发育过程及病菌侵染寄主早期阶段的表达模式,为深入探索该转录因子在病菌发育及致病过程中的功能奠定基础。本研究以白色念珠菌(Candida albicans)Flo8氨基酸序列为探针在玉米大斑病菌数据库中进行同源搜索,利用PCR技术对玉米大斑病菌转录因子Flo8的编码基因进行克隆,通过GSDS、ProtParam、SOMPA 和SMART在线软件进行基因及其编码产物结构特征预测分析,利用WoLF PSORT分析蛋白的亚细胞定位;利用 qRT-PCR技术研究其在病菌侵染结构形成过程中不同阶段的转录水平。结果表明,玉米大斑病菌中存在1个含有LisH保守结构域的蛋白,属于典型的FLO8同源蛋白,将其编码基因命名为StFlo8。该基因DNA全长为2 384 bp,cDNA全长为2 037 bp,含有6个内含子,7个外显子,编码678个氨基酸。表达规律分析结果表明,与菌丝时期相比,StFlo8在孢子时期及芽管时期表达量显著降低,在附着胞时期和侵染丝时期表达量显著升高,达到菌丝时期的4倍以上;而在侵染感病寄主叶片过程中,StFlo8表达量呈现先下调后上调再下调的趋势,并在接种后18 h表达量达到最高,约为接种初期的2倍。以上结果表明StFlo8参与调控病菌附着胞发育及侵染丝形成,该结果为进一步研究StFlo8基因的功能及玉米大斑病菌的侵染机制提供了理论依据。
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申珅
李贞杨
赵玉兰
李盼
董金皋
韩建民
郝志敏
关键词 玉米大斑病菌, 转录因子, cAMP信号途径, qRT-PCR    
Abstract:Flo8 is a key transcription factor at the downstream of cAMP signal pathway and plays an important role in fungal development. In this experiment, in order to clarify the function of transcription factor Flo8 and cAMP singal pathway in regulating the pathogenicity of Setosphaeria turcica, the genes encoding transcription factor Flo8 was cloned and the expression pattern of the gene was analyzed during the development of infective structures and the early stage of infecting the host. The homologous search was carried out in the database of S. turcica using the Candida albicans Flo8 amino acid sequence as a probe. The gene encoding the transcription factor Flo8 was identified by PCR. The structural characteristics of the gene and protein were analyzed through GSDS, ProtParam, SOMPA and SMART software, respectively. Subcellular localization of protein was analyzed by WoLF PSORT online software. Quantitative real-time PCR (qRT-PCR) was used to analyze the gene expression patterns. The results showed that there was a protein sequence containing the conserved domain of LisH which belonged to the typical FLO8 protein, and its coding gene was named StFlo8. The total length of the gene was 2 384 bp, and that of the cDNA was 2 037 bp, this gene contained 6 introns and 7 exons, encoding 678 amino acids. The results of expression pattern analysis showed that, compared with the mycelial period, the expression level of StFlo8 was significantly decreased in the conidia and germ tube period, and was significantly higher in appressorium and invade mycelial period, reaching more than 4 times of that in the mycelia. During the interaction of S. turcica with the susceptible host, the expression of StFlo8 was down-up-down regulated, and reached the highest level at 18 h after inoculation, which was as about twice as that of the beginning of inoculation. The results indicated that StFlo8 was involved in the regulation of appressorium development and infected mycelial formation. The results provide a theoretical basis for the further study of the function of StFlo8 gene and the infection mechanism of S. turcica.
Key wordsSetosphaeria turcica, transcription factor, cAMP signal pathway, qRT-PCR
收稿日期: 2017-03-14      出版日期: 2017-10-30
基金资助:StPKA-c调控玉米大斑病菌侵染能力的分子机制; 高亲和力cAMP磷酸二酯酶调控玉米大斑病菌细胞形态建成的分子机制;环核苷酸磷酸二酯酶调控玉米大斑病菌细胞形态建成的分子机制;钙/钙调素依赖性蛋白激酶(CaMK)基因家族调控玉米大斑病菌致病性的功能解析
通讯作者: 郝志敏     E-mail: hzm_0322@163.com
引用本文:   
申珅 李贞杨 赵玉兰 李盼 董金皋 韩建民 郝志敏. 玉米大斑病菌转录因子Flo8的基因克隆及表达规律分析 [J]. , 2017, 25(10): 1661-1667.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2017/V25/I10/1661
 
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