摘要
本研究利用生物信息学手段分析了玉米大斑病菌高亲和力cAMP磷酸二酯酶(High-affinity cAMP phosphodiesterase,H-PDE)和低亲和力cAMP磷酸二酯酶(Low-affinity cAMP phosphodiesterase,L-PDE)氨基酸序列同源性,发现StH-PDE与狂犬壳二胞菌(Ascochyta rabiei)、小麦黄斑叶枯病菌(Pyrenophora triticirepentis)、链格孢(Alternariaalternata)中同源蛋白的氨基酸序列相似性分别为69.21%、84.21%、85.94%,StL-PDE与狂犬壳二胞菌、壳球孢菌(Macrophominaphaseolina)、小麦黄斑叶枯病菌中同源蛋白的氨基酸序列相似性分别为57.90%,60.14%,71.38%;启动子预测发现,StH-PDE和StL-PDE启动子区存在SP1、AP1、p53、STAT5、AP-2、ATF/CREB以及GATA等转录因子结合位点。进一步利用实时荧光定量PCR技术检测其在不同碳源、氮源环境中的表达情况,结果表明,以果糖、乳糖、山梨醇和淀粉作为碳源对StH-PDE和StL-PDE的表达具有诱导作用,其中以山梨醇对StH-PDE表达影响最强,铵态氮和尿素作为氮源对StH-PDE和StL-PDE的表达有明显的诱导作用,而硝态氮诱导作用不明显。所以,明确PDE对碳氮源的响应为深入研究其基因功能奠定理论基础。
In this study, the amino acid sequence homology of high-affinity cAMP phosphodiesterase(StH-PDE) and low-affinity cAMP phosphodiesterase(StL-PDE) were analyzed by bioinformatics softwares in S. turcica. It was found that the amino acid sequence similarities of StH-PDE with its homologs in Ascochyta rabiei, Pyrenophora tritici-repentis, and Alternaria alternata were 69.21%,84.21%,and 85.94% respectively. The amino acid sequence similarities of StL-PDE with its homologs in A. rabiei, Macrophomina phaseolina, and P. tritici-repentis were 57.90%,60.14%,and 71.38% respectively. Promoter prediction revealed that SP1, AP1, p53, STAT5, AP-2, ATF/CREB, GATA and other transcription factor binding sites existed in the promoter regions of StH-PDE and StL-PDE. Further real-time fluorescence quantitative PCR was used to detect the expression levels of both genes in different carbon and nitrogen sources. The results showed that fructose, lactose, sorbitol and starch as carbon sources could induce the expression of StH-PDE and StL-PDE. Sorbitol had the strongest influence on the expression of StH-PDE ammonium nitrogen and urea as nitrogen sources could induce the expression of StH-PDE and StL-PDE, however, nitrate nitrogen had no obvious effect. Clarifying the response of PDE to carbon and nitrogen sources will lay a theoretical foundation for further study of its gene function and the molecular mechanism of cAMP signaling pathway regulating the nutritional response of filamentous fungi.
作者
李天聪
王擎
王建霞
张燕
申珅
LI Tiancong;WANG Qing;WANG Jianxia;ZHANG Yan;SHEN Shen(Hebei Key Laboratory of Plant Physiology and Molecular Pathology/College of Life Science,Hebei Agricultural University,Baoding 071000,China)
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2019年第5期15-20,共6页
Journal of Hebei Agricultural University
基金
国家自然科学基金项目(31601598)
河北省自然科学基金项目(C2018204120)
河北省高等学校青年拔尖人才科学技术研究项目(BJ2014349Y)
