浙江农业学报 ›› 2018, Vol. 30 ›› Issue (4): 521-527.DOI: 10.3969/j.issn.1004-1524.2018.04.01

• 植物保护 • 上一篇    下一篇

二化螟两种P450基因CYP4M38和CYP4M39的时空表达分析

白琪1, 2, 鲁艳辉2, 郑许松2, 吕仲贤2, *   

  1. 1.浙江师范大学 化学与生命科学学院,浙江 金华321004;
    2.浙江省农业科学院 植物保护与微生物研究所,省部共建浙江省植物有害生物防控国家重点实验室培育基地,浙江 杭州310021
  • 收稿日期:2017-07-01 出版日期:2018-04-20 发布日期:2018-04-19
  • 通讯作者: 吕仲贤,E-mail:luzxmh@163.com
  • 作者简介:白琪(1992—),女,河南商丘人,硕士,主要从事害虫生物防治研究。E-mail:qiqi2015210683@163.com
  • 基金资助:
    浙江省科技计划项目重大科技专项(2015C02014); 国家重点研发计划(2016YFD0200800); 重点实验室培育基金(2010DS700124-ZZ1601); 国家自然科学基金(31672050)

Spatiotemporal expression profiling of two P450 genes, CYP4M38 and CYP4M39, in rice stem borer, Chilo suppressalis (Lepidoptera: Crambidae)

BAI Qi1, 2, LU Yanhui2, ZHENG Xusong2, LYU Zhongxian2, *   

  1. 1.College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China;
    2. State Key Laboratory of Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
  • Received:2017-07-01 Online:2018-04-20 Published:2018-04-19

摘要: 细胞色素P450在昆虫对外源物质代谢中发挥重要作用,是昆虫重要的解毒酶系。CYP4家族是昆虫细胞色素P450基因家族中的重要分支,与昆虫的解毒代谢密切相关。为初步明确CYP4家族基因在二化螟中的表达信息,本文测定分析了二化螟两种P450基因CsCYP4M38和CsCYP4M39的序列同源性及时空表达谱,序列同源性分析发现CsCYP4M38与烟草天蛾MsCYP4M2氨基酸序列同源性最高,序列一致性高达56.86%;CsCYP4M39与烟草天蛾MsCYP4M1氨基酸序列同源性最高,序列一致性高达59.96%,构建系统发育树显示,CsCYP4M38与CsCYP4M39属于CYP4家族,且分别与MsCYP4M2和MsCYP4M1进化距离较近;时空表达谱测定结果表明,两种基因在不同发育阶段和不同组织中的表达存在差异性,CsCYP4M38在2龄幼虫、5龄幼虫及雌成虫内表达量相对较高,在1龄幼虫中表达量最低,CsCYP4M39在幼虫期表达量相对较高,且表达量随龄期增加呈现先上升后下降的趋势,在卵、蛹及成虫中表达量相对较低。CsCYP4M38和CsCYP4M39在脂肪体、中肠及表皮中的表达量相对较高,且均在脂肪体内的表达量最高。明确基因表达谱是研究其功能的基础,CsCYP4M38和CsCYP4M39在不同阶段的表达量具有一定的变化性,说明这两种基因在二化螟不同发育历期中的功能不同,而两种基因均在脂肪体中表达量最高,这可能与脂肪体是昆虫的主要的解毒代谢场所有关。

关键词: 二化螟, 细胞色素P450, CsCYP4M38, CsCYP4M39, 基因表达

Abstract: Cytochrome P450 plays an important role in insect metabolism of exogenous substances, which is an important detoxification system of insects. The CYP4 family is an important branch of the insect cytochrome P450 gene family, which is closely related to the detoxification and metabolism of insects. In this study, the sequence homology and spatiotemporal expression profiles of CsCYP4M38 and CsCYP4M39 were investigated in order to clarify the expression information of CYP4 family gene in Chilo suppressalis. The results showed that CsCYP4M38 and CsCYP4M39 had highest sequence homology to MsCYP4M2 (56.86%) and MsCYP4M1 (59.96%), respectively. The phylogenetic tree showed that CsCYP4M38 and CsCYP4M39 belonged to the CYP4 family. The results of spatiotemporal expression profiling showed that the expression patterns of two genes in different developmental stages and different tissues were different. The CsCYP4M38 was expressed higher in the second instar larvae, 5th instar larvae and female adults, but lower in the 1st instar larvae. The CsCYP4M39 expressed higher in the larval stage, and the expression of CsCYP4M39 increased and then decreased. The expression level of CsCYP4M39 was lower in the egg, pupae and adult of C. suppressalis. The expression levels of CsCYP4M38 and CsCYP4M39 in the fat body, midgut and epidermis were relatively high, and both expressed highest in the fat body. The expression of CsCYP4M38 and CsCYP4M39 in different stages showed variability, which indicated that these two genes had different functions in different developmental stages of C. suppressalis, and both genes were expressed highest in the fat body, indicating the fat body might be the main detoxification tissue of insects.

Key words: Chilo suppressalis, cytochrome P450, CsCYP4M38, CsCYP4M39, gene expression

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