Yang Yajun, Xu Hongxing, Lu Yanhui, Wang Caiyun, Lu Zhongxian
State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.
PLoS One. 2018 Jan 23;13(1):e0191686. doi: 10.1371/journal.pone.0191686. eCollection 2018.
Cnaphalocrocis medinalis (Guenée) is one of the important insect pests in rice field. Bt agents were recommended in the C. medinalis control and Bt rice is bred as a tactic to control this insect. However, the tolerance or resistance of insect to Bt protein is a main threat to the application of Bt protein. In order to investigate the response of C. medinalis transcriptome in defending a Cry1C toxin, high-through RNA-sequencing was carried in the C. medinalis larvae treated with and without Cry1C toxin. A total of 35,586 high-quality unigenes was annotated in the transcriptome of C. medinalis midgut. The comparative analysis identified 6,966 differently expressed unigenes (DEGs) between the two treatments. GO analysis showed that these genes involved in proteolysis and extracellular region. Among these DEGs, carboxylesterase, glutathione S-transferase and P450 were differently expressed in the treated C. medinalis midgut. Furthermore, trypsin, chymotrypsin, and carboxypeptidase were identified in DEGs, and most of them up-regulated. In addition, thirteen ABC transporters were downregulated and three upregulated in Cry1C-treated C. medinalis midgut. Based on the pathway analysis, antigen processing and presentation pathway, and chronic myeloid leukemia pathway were significant in C. medinalis treated with Cry1C toxin. These results indicated that serine protease, detoxification enzymes and ABC transporter, antigen processing and presentation pathway, and chronic myeloid leukemia pathway may involved in the response of C. medinalis to Cry1C toxin. This study provides a transcriptomal foundation for the identification and functional characterization of genes involved in the toxicity of Bt Cry protein against C. medinalis, and provides potential clues to the studies on the tolerance or resistance of an agriculturally important insect pest C. medinalis to Cry1C toxin.
稻纵卷叶螟(Guenée)是稻田中的重要害虫之一。苏云金芽孢杆菌制剂被推荐用于防治稻纵卷叶螟,并且培育出了转Bt基因水稻作为防治这种害虫的一种策略。然而,昆虫对Bt蛋白的耐受性或抗性是Bt蛋白应用的主要威胁。为了研究稻纵卷叶螟转录组在抵御Cry1C毒素方面的反应,对经Cry1C毒素处理和未处理的稻纵卷叶螟幼虫进行了高通量RNA测序。在稻纵卷叶螟中肠转录组中总共注释了35586个高质量单基因。比较分析确定了两种处理之间有696个差异表达的单基因(DEGs)。基因本体(GO)分析表明,这些基因涉及蛋白水解和细胞外区域。在这些差异表达基因中,羧酸酯酶、谷胱甘肽S-转移酶和细胞色素P450在处理后的稻纵卷叶螟中肠中差异表达。此外,在差异表达基因中鉴定出了胰蛋白酶、胰凝乳蛋白酶和羧肽酶,其中大多数上调。另外,在经Cry1C处理的稻纵卷叶螟中肠中,有13个ABC转运蛋白下调,3个上调。基于通路分析,抗原加工和呈递通路以及慢性髓性白血病通路在经Cry1C毒素处理的稻纵卷叶螟中有显著变化。这些结果表明,丝氨酸蛋白酶、解毒酶和ABC转运蛋白、抗原加工和呈递通路以及慢性髓性白血病通路可能参与了稻纵卷叶螟对CryC毒素的反应。本研究为鉴定和功能表征参与Bt Cry蛋白对稻纵卷叶螟毒性作用的基因提供了转录组学基础,并为研究重要农业害虫稻纵卷叶螟对Cry1C毒素的耐受性或抗性提供了潜在线索。