启动子WY172和WY195在暹罗炭疽菌中的活性研究  

Identification of WY172 and WY195 promoter activity in Colletotrichum siamense

作　　者：董林朋 殷金瑶 赵文渊 林春花[1] 刘文波[1] 缪卫国[1] 李潇[1] DONG Linpeng;YIN Jinyao;ZHAO Wenyuan;LIN Chunhua;LIU Wenbo;MIAO Weiguo;LI Xiao(School of Plant Protection/Ministry of Education Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests,Hainan University,Haikou,Hainan 570228,China)

机构地区：[1]海南大学植物保护学院/热带农林生物灾害绿色防控教育部重点实验室,海口570228

出　　处：《热带生物学报》2023年第5期506-513,共8页Journal of Tropical Biology

基　　金：国家自然科学基金项目(31960518)。

摘　　要：为了探究来自橡胶树白粉菌(Erysiphe quercicola)的WY172与WY195启动子在真菌内的功能,以pJNARG载体为骨架,分别构建WY172与WY195和绿色荧光蛋白(GFP)基因连接的重组表达载体(WY172-GFP和WY195-GFP载体)。通过原生质体转化法将重组载体导入侵染橡胶树的暹罗炭疽菌(Colletotrichum siamense)内。荧光显微检测结果表明WY172与WY195均可驱动GFP基因的表达,具有稳定的启动子活性。在暹罗炭疽菌侵染橡胶叶片不同进程中均可稳定驱动GFP表达;在不同环境条件下测定GFP表达量,发现长光照条件下可提高WY172启动子驱动GFP的活性,并且23℃低温处理和光处理可提高WY195的活性。本研究表明WY172与WY195在暹罗炭疽菌(Colletotrichum siamense)中具有稳定的启动子活性,并且增加光照和低温处理能诱导启动子活性的提升。Our laboratory preliminarily identified WY172 and WY195 promoter sequences from the rubber tree powdery mildew fungus(Erysiphe quercicola),which could effectively drive GUS expression in both monocotyledonous rice and dicotyledonous tobacco.In order to investigate whether WY172 and WY195 have promoter activity in E.quercicola,the pJNARG vector was used as a backbone,linking WY172-GFP and WY195-GFP to construct recombinant expression vectors(WY172-GFP and WY195-GFP vectors).The recombinant vectors were introduced into Colletotrichum siamense infecting rubber trees by protoplast transformation method.Fluorescence microscopy showed that both WY172 and WY195 could stably drive the expression of GFP gene.And in different stages of C.siamense,WY172 and WY195 could play promoter roles to stably drive the expression of GFP.In addition,we changed the environmental conditions for the growth of anthracis and measured the GFP expression under different conditions.The results showed that the long light condition enhanced the activity of WY172 to drive GFP expression,and that the activity of WY195 was improved by the long light treatment and the low temperature treatment at 23°C.This indicates that WY172 and WY195 have stable promoter activities in C.siamense,and that their activities can be induced by changing light or temperature conditions.

关 键 词：启动子 暹罗炭疽菌 原生质体转化 

分 类 号：S432.1[农业科学—植物病理学]