西花蓟马被球孢白僵菌侵染后βGRPs基因的鉴定及表达分析

doi:10.16409/j.cnki.2095-039x.2017.02.010

中国生物防治学报 ›› 2017, Vol. 33 ›› Issue (2/3): 206-212.DOI: 10.16409/j.cnki.2095-039x.2017.02.010

西花蓟马被球孢白僵菌侵染后βGRPs基因的鉴定及表达分析

赵倩倩¹, 张桃², 王小奇³, 王海鸿¹

1. 中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室, 北京 100193;
2. 成都市植物检疫站, 成都 610041;
3. 沈阳农业大学植物保护学院, 沈阳 110000

收稿日期:2016-11-30 出版日期:2017-06-08 发布日期:2017-04-08
通讯作者: 王海鸿,副研究员,E-mail:wanghaihong2020@sina.com。
作者简介:赵倩倩,硕士研究生;E-mail:1078689122@qq.com
基金资助:
现代农业产业技术体系（CARS-25-B-07）

Identification and Expression Analysis on βGRPs of Frankliniella occidentalis

ZHAO Qianqian¹, ZHANG Tao², WANG Xiaoqi³, WANG Haihong¹

1. State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. Chengdu Plant Quarantine Station, Chengdu 610041, China;
3. College of Plant Protection, Shenyang Agricultural University, Shenyang 110000, China

Received:2016-11-30 Online:2017-06-08 Published:2017-04-08

摘要/Abstract

摘要： 本文旨在鉴定西花蓟马β-1，3-葡聚糖识别蛋白（FoβGRP）基因，分析其在受到球孢白僵菌侵染后的表达模式，探讨其在西花蓟马先天免疫过程中的作用。根据高通量转录组测序结果，比对非冗余核苷酸数据库，筛选E值小于10^-5的基因为目标基因，对筛选出的FoβGRPs基因编码的蛋白序列进行结构分析鉴定，通过构建系统进化树分析其与其他物种同源基因的进化关系，通过实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）检测分析不同活性球孢白僵菌孢子侵染西花蓟马成虫后FoβGRPs的表达模式。筛选出3条βGRP基因，分别命名为FoβGRP1、FoβGRP2和FoβGRP3，其编码的蛋白序列均含有βGRP蛋白家族特有的保守性结构域，即糖苷水解酶活性结构域；FoβGRP1与FoβGRP2和FoβGRP3分属两类不同的FoβGRP；在检测的60 h内，与对照相比，西花蓟马被高致病性和低致病性真菌菌株侵染后，3条FoβGRP基因的诱导表达量均在12 h时最高，而且对于同一基因来说，高活性真菌菌株对其的诱导水平均高于低活性的真菌菌株；经灭活真菌处理的西花蓟马体内3条FoβGRP基因表达量均呈不同程度的下降。3条FoβGRP基因可能参与到西花蓟马先天免疫反应中。本文为后续深入研究其在西花蓟马先天免疫反应中的作用奠定了基础。

关键词: 西花蓟马, β-1, 3-葡聚糖识别蛋白, 先天免疫反应

Abstract: The objective of this study is to identify the β-1,3-glucan recognition protein in Frankliniella occidentalis to analyse the express model when F. occidentalis infected by the fungusBeauveria bassiana, and to provide a theoretical foundation for clarifying the innate immunity in F. occidentalis. Target genes were screened from the data of high-throughout sequencing, the genes were identified through blasting against the redundancy nucleotide databases when the Evalue was less than 10^-5. Three β-1,3-glucan recognition protein identified in F. occidentalis were performed by protein structure analysis and phylogenetic analysis with homologous genes of other insects. The expression of β-1,3-glucan recognition protein genes of F. occidentalis infected by different active strains of B.bassiana was analyzed by quantitative real-time PCR. Three βGRP genes were identified and designated asFoβGRP1, FoβGRP2, FoβGRP3. The protein structure analysis showed that three sequences had the unique structure of βGRP family which contained conserved domains-glycoside hydrolase activity domain. Homology analysis showed that FoβGRP1 and FoβGRP2 /FoβGRP3 belong to two different clades of FoβGRP. The expression of all the three FoβGRPs peak at 12 h. The expression level of a certain gene is higher when thrips treated by high-activity strain than those treated by low-activity strain. In conclusion, FoβGRPs might be involved in the immune response to B. bassiana. This study provided the foundation to research the function of these genes in the innate immune response in thrips.

Key words: Frankliniella occidentalis, β-1,3-glucan recognition protein, innate immune response

中图分类号:

S476.12

赵倩倩, 张桃, 王小奇, 王海鸿. 西花蓟马被球孢白僵菌侵染后βGRPs基因的鉴定及表达分析[J]. 中国生物防治学报, 2017, 33(2/3): 206-212.

ZHAO Qianqian, ZHANG Tao, WANG Xiaoqi, WANG Haihong. Identification and Expression Analysis on βGRPs of Frankliniella occidentalis[J]. journal1, 2017, 33(2/3): 206-212.

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西花蓟马被球孢白僵菌侵染后βGRPs基因的鉴定及表达分析

Identification and Expression Analysis on βGRPs of Frankliniella occidentalis

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