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棉铃虫在真菌感染和细菌感染期间免疫转录组的高通量分析。

High throughput profiling of the cotton bollworm Helicoverpa armigera immunotranscriptome during the fungal and bacterial infections.

作者信息

Xiong Guang-Hua, Xing Long-Sheng, Lin Zhe, Saha Tusar T, Wang Chengshu, Jiang Haobo, Zou Zhen

机构信息

State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China.

University of Chinese Academy of Sciences, Beijing, 100049, China.

出版信息

BMC Genomics. 2015 Apr 18;16(1):321. doi: 10.1186/s12864-015-1509-1.

Abstract

BACKGROUND

Innate immunity is essential in defending against invading pathogens in invertebrates. The cotton bollworm, Helicoverpa armigera (Hübner) is one of the most destructive lepidopteran pests, which causes enormous economic losses in agricultural production worldwide. The components of the immune system are largely unknown in this insect. The application of entomopathogens is considered as an alternative to the chemical insecticides for its control. However, few studies have focused on the molecular mechanisms of host-pathogen interactions between pest insects and their pathogens. Here, we investigated the immunotranscriptome of H. armigera larvae and examined gene expression changes after pathogen infections. This study provided insights into the potential immunity-related genes and pathways in H. armigera larvae.

RESULTS

Here, we adopted a high throughput RNA-seq approach to determine the immunotranscriptome of H. armigera larvae injected with buffer, fungal pathogen Beauveria bassiana, or Gram-negative bacterium Enterobacter cloacae. Based on sequence similarity to those homologs known to participate in immune responses in other insects, we identified immunity-related genes encoding pattern recognition receptors, signal modulators, immune effectors, and nearly all members of the Toll, IMD and JAK/STAT pathways. The RNA-seq data indicated that some immunity-related genes were activated in fungus- and bacterium-challenged fat body while others were suppressed in B. bassiana challenged hemocytes, including the putative IMD and JAK-STAT pathway members. Bacterial infection elevated the expression of recognition and modulator genes in the fat body and signal pathway genes in hemocytes. Although fat body and hemocytes both are important organs involved in the immune response, our transcriptome analysis revealed that more immunity-related genes were induced in the fat body than that hemocytes. Furthermore, quantitative real-time PCR analysis confirmed that, consistent with the RNA-seq data, the transcript abundances of putative PGRP-SA1, Serpin1, Toll-14, and Spz2 genes were elevated in fat body upon B. bassiana infection, while the mRNA levels of defensin, moricin1, and gloverin1 were up-regulated in hemocytes.

CONCLUSIONS

In this study, a global survey of the host defense against fungal and bacterial infection was performed on the non-model lepidopteran pest species. The comprehensive sequence resource and expression profiles of the immunity-related genes in H. armigera are acquired. This study provided valuable information for future functional investigations as well as development of specific and effective agents to control this pest.

摘要

背景

先天免疫对于无脊椎动物抵御入侵病原体至关重要。棉铃虫,即棉铃实夜蛾(Helicoverpa armigera (Hübner))是最具破坏性的鳞翅目害虫之一,在全球农业生产中造成巨大经济损失。该昆虫免疫系统的组成部分在很大程度上尚不清楚。昆虫病原体的应用被认为是控制该害虫的化学杀虫剂的一种替代品。然而,很少有研究关注害虫及其病原体之间宿主 - 病原体相互作用的分子机制。在此,我们研究了棉铃虫幼虫的免疫转录组,并检测了病原体感染后的基因表达变化。本研究为棉铃虫幼虫潜在的免疫相关基因和途径提供了见解。

结果

在此,我们采用高通量RNA测序方法来确定注射缓冲液、真菌病原体球孢白僵菌或革兰氏阴性菌阴沟肠杆菌的棉铃虫幼虫的免疫转录组。基于与已知参与其他昆虫免疫反应的同源物的序列相似性,我们鉴定出编码模式识别受体、信号调节剂、免疫效应器以及Toll、IMD和JAK/STAT途径几乎所有成员的免疫相关基因。RNA测序数据表明,一些免疫相关基因在受到真菌和细菌攻击的脂肪体中被激活,而其他一些基因在受到球孢白僵菌攻击的血细胞中被抑制,包括假定的IMD和JAK - STAT途径成员。细菌感染提高了脂肪体中识别和调节基因以及血细胞中信号通路基因的表达。尽管脂肪体和血细胞都是参与免疫反应的重要器官,但我们的转录组分析显示,脂肪体中诱导的免疫相关基因比血细胞更多。此外,定量实时PCR分析证实,与RNA测序数据一致,假定的PGRP - SA1、丝氨酸蛋白酶抑制剂1、Toll - 14和Spz2基因的转录丰度在球孢白僵菌感染后在脂肪体中升高,而防御素、蚕抗菌肽1和格洛弗菌素1的mRNA水平在血细胞中上调。

结论

在本研究中,对非模式鳞翅目害虫物种针对真菌和细菌感染的宿主防御进行了全面调查。获得了棉铃虫免疫相关基因的全面序列资源和表达谱。本研究为未来的功能研究以及开发控制这种害虫的特异性和有效药剂提供了有价值的信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/16e8/4490664/c62490260747/12864_2015_1509_Fig1_HTML.jpg

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