Jin Yongling, Cong Bin, Wang Liyan, Gao Yugang, Zhang Haiyan, Dong Hui, Lin Zhiwei
Shenyang Agriculture University, Shenyang of Liaoning province, China, 110061 (
Heilongjiang Bayi Agricultural University,Daqing of Heilongjiang province, China,, 163319 ((
J Insect Sci. 2016 May 3;16(1). doi: 10.1093/jisesa/iew014. Print 2016.
Epacromius coerulipes (Ivanov) is one of the most widely distributed locusts. To date, the main methods to kill locusts still rely on chemical controls, which can result in the selection of locusts with resistance to chemical pesticides. Butene-fipronil is a new pesticide that was discovered by the structural modification of fipronil. This pesticide has been used to control various agricultural pests and has become an important pesticide product to control pests that exhibit resistance to other pesticides, including locusts. To extend its useful half-life, studies of the initiation and progression of resistance to this pesticide are needed. Herein, two E. coerulipes strains, a pesticide-sensitive (PS) and a pesticide-resistant (PR) strain, were chosen to undergo de novo assembly by paired-end transcriptome Illumina sequencing. Overall, 63,033 unigenes were detected; the average gene length was 772 bp and the N50 was 1,589 bp. Among these unigenes, ∼ 25,132 (39.87% of the total) could be identified as known proteins in bioinformatic databases from national centers. A comparison of the PR and PS strains revealed that 2,568 genes were differentially expressed, including 1,646 and 922 genes that were up- and down-regulated, respectively. According to the Gene Ontology (GO) database, among biological processes the metabolic process group was the largest group (6,900 genes, 22.47%) and contained a high frequency of differentially expressed genes (544 genes, 27.54%). According to the Clusters of Orthologous Groups (COG) categories, 28 genes, representing 2.98% of all genes, belonged to the group of genes involved in the biosynthesis, transportation, and catabolism of secondary metabolites. The differentially expressed genes that we identified are involved in 50 metabolic pathways. Among these pathways, the metabolism pathway was the most represented. After enrichment analysis of differential gene expression pathways, six pathways--ribosome; starch, and sucrose metabolism; ascorbate and aldarate metabolism; drug metabolism-cytochrome P450; metabolism of xenobiotics by cytochrome P450; and glutathione metabolism--showed a high degree of enrichment. Among these pathways, drug metabolism-cytochrome P450, metabolism of xenobiotics by cytochrome P450, and glutathione metabolism have been associated with pesticide metabolism. Furthermore, 316 unigenes in the E. coerulipes transcriptome encode detoxifying enzymes and 76 unigenes encode target proteins of pesticides. Among these genes, 23 genes that encode detoxifying enzymes in the resistance group were found to be up-regulated. The transcriptome sequencing results of E. coerulipes established a genomics database of E. coerulipes for the first time. This study also establishes a molecular basis for gene function analysis of E. coerulipes Moreover, it provides a theoretical resource for mechanistic studies on pesticide resistance through the screening and investigation of resistance genes.
蓝斑翅蝗(Epacromius coerulipes (Ivanov))是分布最为广泛的蝗虫之一。迄今为止,杀灭蝗虫的主要方法仍依赖化学防治,这可能导致选择出对化学农药具有抗性的蝗虫。丁烯氟虫腈是通过对氟虫腈进行结构修饰而发现的一种新型农药。这种农药已被用于防治各种农业害虫,并已成为防治对其他农药(包括蝗虫)具有抗性的害虫的重要农药产品。为延长其有效半衰期,需要对这种农药抗性的起始和发展进行研究。在此,选择了两个蓝斑翅蝗品系,一个对农药敏感(PS)品系和一个对农药抗性(PR)品系,通过双末端转录组Illumina测序进行从头组装。总体而言,共检测到63,033个单基因;平均基因长度为772 bp,N50为1,589 bp。在这些单基因中,约25,132个(占总数的39.87%)可在国家中心的生物信息数据库中鉴定为已知蛋白质。PR品系和PS品系的比较显示,有2,568个基因差异表达,其中分别有1,646个和922个基因上调和下调。根据基因本体论(Gene Ontology,GO)数据库,在生物学过程中,代谢过程组是最大的组(6,900个基因,占22.47%),并且包含高频率的差异表达基因(544个基因,占27.54%)。根据直系同源簇(Clusters of Orthologous Groups,COG)类别,28个基因(占所有基因的2.98%)属于参与次生代谢物生物合成、运输和分解代谢的基因组。我们鉴定出的差异表达基因涉及50条代谢途径。在这些途径中,代谢途径的占比最大。对差异基因表达途径进行富集分析后,核糖体、淀粉和蔗糖代谢、抗坏血酸和醛糖代谢、药物代谢 - 细胞色素P450、细胞色素P450对外源生物的代谢以及谷胱甘肽代谢这六条途径显示出高度富集。在这些途径中,药物代谢 - 细胞色素P450、细胞色素P450对外源生物的代谢以及谷胱甘肽代谢与农药代谢有关。此外,蓝斑翅蝗转录组中的316个单基因编码解毒酶,76个单基因编码农药的靶标蛋白。在这些基因中,发现抗性组中23个编码解毒酶的基因上调。蓝斑翅蝗的转录组测序结果首次建立了蓝斑翅蝗的基因组数据库。本研究还为蓝斑翅蝗的基因功能分析奠定了分子基础。此外,通过抗性基因的筛选和研究,为农药抗性的机制研究提供了理论资源。