摘要
为进一步明确麦二叉蚜Schizaphis graminum唾液蛋白Sg1655在调控小麦防御反应中的作用,基于麦二叉蚜唾液腺转录组数据,利用PCR技术克隆获取Sg1655开放阅读框,利用生物信息学软件对其序列进行分析;通过实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)技术检测Sg1655在麦二叉蚜不同组织及不同龄期中的表达;利用细菌III型分泌系统将Sg1655导入到小麦叶片内,检测其瞬时表达对小麦胼胝质积累的影响。结果表明,Sg1655开放阅读框全长375 bp,编码125个氨基酸残基,N端1~21位为信号肽序列,预测蛋白分子量为14.90 kD。麦二叉蚜Sg1655氨基酸序列与玉米蚜Rhopalosiphum maidis同源序列相致性最高,为89.52%;麦二叉蚜Sg1655与玉米蚜同源序列聚为一个分支,亲缘关系最近。Sg1655在麦二叉蚜各组织中均有表达,其中在唾液腺中表达量最高,Sg1655在麦二叉蚜各个龄期均有表达,且在不同龄期之间无显著差异。小麦叶片内Sg1655瞬时表达可显著抑制小麦胼胝质积累,表明该蛋白可作为潜在效应子参与抑制小麦防御反应。
To further elucidate the role of the functionally unknown salivary protein Sg1655 in regulating the defense response of wheat,this study was carried out based on transcriptomic data from the salivary glands of Schizaphis graminum.The open reading frame of Sg1655 was obtained via PCR,and sequence analysis was performed using bioinformatics software.The expression level of Sg1655 in different aphid tissues and instars was measured using real-time quantitative PCR(RT-qPCR).Sg1655 was delivered into wheat leaves using a bacterial type Ⅲ secretion system to examine the effect of its transient expression on callose accumulation in wheat.The results showed that the full-length open reading frame of Sg1655 was 375 bp,encoding 125 amino acid residues,with the N-terminal 1-21 sequence representing the signal peptide.The predicted protein molecular weight was 14.90 kD.Multiple sequence alignment revealed that the amino acid sequence of Sg1655 from S.graminum shared the highest sequence identity(89.52%)with the homologous sequence of Rhopalosiphum maidis.Sg1655 clustered into one branch with the homologous sequences of R.maidis,indicating a close genetic relationship between them.Sg1655 was found to be expressed in all aphid tissues,with the highest expression level in the salivary gland.In addition,Sg1655 expression was detected in all instars of S.graminum,with no significant difference in expression levels among various instars.Transient expression of Sg1655 significantly suppressed the accumulation of callose in wheat,indicating that the salivary protein Sg1655 may be involved in suppressing wheat defense responses as a potential effector.
作者
孙钰
张雨濛
刘晓蓓
陈巨莲
张勇
解海翠
Sun Yu;Zhang Yumeng;Liu Xiaobei;Chen Julian;Zhang Yong;Xie Haicui(Hebei Key Laboratory of Crop Stress Biology,Hebei Normal University of Science and Technology,Qinhuangdao 066600,Hebei Province,China;National Key Laboratory of Integrated Management of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China)
出处
《植物保护学报》
CAS
CSCD
北大核心
2024年第1期20-27,共8页
Journal of Plant Protection
基金
国家自然科学基金(31901881)
河北省属高校基本科研业务费专项(2020JK001)。
关键词
麦二叉蚜
唾液蛋白
Sg1655
基因克隆
序列比对
时空表达谱
功能分析
Schizaphis graminum
salivary protein
Sg1655
gene cloning
sequence alignment
spatio-temporal expression profile
functional analysis
