毛果杨丝氨酸蛋白酶抑制子PtrSPI的抗虫能力分析  

Analysis of insect resistance ability of the serine protease inhibitor PtrSPI from Populus tomentosa

作　　者：遇文婧[1,2] 杨帅 黄颖[1] 刁桂萍[1] Yu Wenjing;Yang Shuai;Huang Ying;Diao Guiping(School of Forestry,Northeast Forestry University,Harbin 150040,Heilongjiang,China;Forestry Protection Institute,Heilongjiang Academy of Forestry,Harbin 150040,Heilongjiang,China)

机构地区：[1]东北林业大学林学院,黑龙江哈尔滨150040 [2]黑龙江省林业科学院森林保护研究所,黑龙江哈尔滨150040

出　　处：《北京林业大学学报》2021年第9期51-58,共8页Journal of Beijing Forestry University

基　　金：中央高校基本科研业务费专项(2572020BA08)。

摘　　要：【目的】研究毛果杨丝氨酸蛋白酶抑制子PtrSPI的抗虫功能,为开发新型林木抗虫生物农药奠定基础。【方法】本研究通过研究毛果杨丝氨酸蛋白酶抑制子基因PtrSPI的启动子及其在舞毒蛾幼虫取食胁迫下的表达模式,分析PtrSPI基因功能;利用真核重组蛋白PtrSPI饲喂舞毒蛾幼虫,观察记录幼虫的取食量、体质量和死亡率,并测定取食后的幼虫体内丝氨酸蛋白酶活性,探讨PtrSPI蛋白的抗虫能力。【结果】结果表明毛果杨丝氨酸蛋白酶抑制子基因PtrSPI的启动子区域共包含5个与植物抗病虫相关的元件;经舞毒蛾幼虫取食后,毛果杨叶片的PtrSPI基因呈先降后升的表达模式,且在取食30 h达到峰值,为空白对照的2.03倍;高质量浓度重组蛋白PtrSPI(300和500 mg/mL)对舞毒蛾幼虫的取食量和体质量有显著的抑制作用,而且分别在取食4和6 d后舞毒蛾幼虫死亡率均达到60%以上,而幼虫体内丝氨酸蛋白酶的活性在取食初期显著上升。【结论】本研究验证了毛果杨丝氨酸蛋白酶抑制子PtrSPI的抗虫能力,可为进一步研发新型无公害抗虫生物农药提供理论基础和研究材料。[Objective]To lay the foundation of the development of new insect-resistant biological pesticides in trees,the insect-resistant function of the serine protease inhibitor PtrSPI of Populus tomentosa was studied.[Method]To analyze the function of serine protease inhibitor gene PtrSPI,the promoter and the expression pattern under feeding stress of Lymantria dispar larvaes of the gene PtrSPI in P.tomentosa were studied.To discuss the anti-insect ability of PtrSPI,the food intake,the body mass,the mortality and the serine protease enzyme activities of L.dispar larvae were studied after using the eukaryotic recombinant protein PtrSPI feeding L.dispar larvaes.[Result]The results showed that 5 elements related to plant resistance to disease and insects were in the promoter region of the serine protease inhibitor gene PtrSPI of P.tomentosa;the expression pattern of the gene PtrSPI in P.tomentosa leaves was firstly decreased and then increased after being fed by L.dispar larvae,and the peak was 2.03 times of the control at 30 h;under the high concentration of recombinant protein(300 and 500 mg/mL),the food intake and the body mass of L.dispar larvaes were significantly inhibited,the mortality of L.dispar larvaes was up to more than 60%after 4 and 6 days,respectively,and the activities of the serine protease in L.dispar larvaes were significantly increased at the beginning of feeding.[Conclusion]This study verifies the insect resistance of the serine protease inhibitor PtrSPI of P.tomentosa,and it provides theoretical basis and research materials for further research and development of new non-pollution and insect-resistance biological pesticides.

关 键 词：丝氨酸蛋白酶抑制子 毛果杨 抗虫功能 生物农药 舞毒蛾 

分 类 号：S763.1[农业科学—森林保护学]