马铃薯甲虫LdATPaseE调控幼虫蜕皮的分子机制

doi:10.6048/j.issn.1001-4330.2021.06.018

新疆农业科学 ›› 2021, Vol. 58 ›› Issue (6): 1124-1137.DOI: 10.6048/j.issn.1001-4330.2021.06.018

• 园艺特产·贮藏保鲜加工·植物保护 • 上一篇下一篇

马铃薯甲虫LdATPaseE调控幼虫蜕皮的分子机制

廖兰兰^1,2, 王俊³, 付开赟^2,4, 丁新华², 何江², 吐尔逊·阿合买提², 郭文超⁵

1.塔里木大学植物科学学院,新疆阿拉尔 843300;
2.新疆农业科学院植物保护研究所/农业部西北荒漠绿洲作物有害生物综合治理重点实验室,乌鲁木齐 830091;
3.新疆农业农村厅植物保护站,乌鲁木齐 830049;
4.新疆农业科学院博士后工作站/新疆农业大学博士后流动站,乌鲁木齐 830091;
5.新疆农业科学院微生物应用研究所,乌鲁木齐 830091

收稿日期:2020-03-10 出版日期:2021-06-20 发布日期:2021-06-15
通信作者: 郭文超(1966-),男,河北人,研究员,博士,博士生导师,研究方向为农业害虫综合防治,(E-mail)gwc1966@163.com; 付开赟(1987-),浙江人,助理研究员,在站博士后,研究方向为昆虫生理生化与分子生物学,(E-mail)fukaiyun87@163.com
作者简介:廖兰兰(1993-),女,四川人,硕士研究生,研究方向为资源利用与植物保护,(E-mail)1456804924@qq.com
基金资助:
新疆维吾尔自治区自然科学基金青年科学基金(2016D01B043)

LdATPaseE in the Regulation of Larval Molting Mechanism in Leptinotarsa decemlineata

LIAO Lanlan^1,2, WANG Jun³, FU Kaiyun^2,4, DING Xinhua², HE Jiang², Tursun Ahmat², GUO Wenchao⁵,

1. College of Plant Science, Tarim University, Alar Xinjiang 843300, China;
2. Key Laboratory of Integrated Crop Pest Management in Northwestern Oasis of China,MOARA, Institute of Plant Protection,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China;
3. Plant Protection Station of Xinjang Uygur Autoncmous Regiou, Urumqi 830049;
4. Post-Doctoral Mobile Station of Xinjiang Agricultural University/Post-Doctoral Workstation of Xinjiang Academy of Agricultural Sciences,Urumqi 830091, China;
5. Institute of Applied Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China

Received:2020-03-10 Online:2021-06-20 Published:2021-06-15
Correspondence author: GUO Wenchao (1966-),male,born in Hebei,Professor,doctoral supervisor,Research direction: Integrated management of Agricultural invasive pests,( E-mail)gwc1966@163.com; FU Kaiyun (1987-),male,born in Zhejiang,Post-doctoral at Post-doctoral Mobile Station of Xinjiang Agricultural University/Postdoctoral Workstation of Xinjiang Academy of Agricultural Sciences,Professor associate,Research direction: Integrated management of Agricultural invasive pests,(E-mail)fukaiyun87@163.com

摘要/Abstract

摘要： 【目的】研究LdATPaseE通过类胰岛素信号通路影响保幼激素和蜕皮激素通路,调节马铃薯甲虫幼虫化蛹的分子机制。【方法】LdATPaseE cDNA序列通过马铃薯甲虫转录组数据分析和RT-PCR克隆获得;qPCR检测其在马铃薯甲虫各发育阶段,以及四龄幼虫不同组织的相对表达量;采用喂食幼虫dsRNA的方法,分析该基因在马铃薯甲虫幼虫的生长发育过程中的影响,并测定类胰岛素、保幼激素和蜕皮激素通路基因对该基因的表达响应机制。【结果】喂食二龄幼虫dsLdATPaseE后,成功敲低了靶标基因,阻止二龄幼虫的生长,显著增加了二龄幼虫的死亡率。三龄和四龄幼虫喂食dsLdATPaseE-1和dsLdATPaseE-2,在极低浓度下即敲低了靶标基因,阻止了幼虫生长,显著降低马铃薯甲虫幼虫的化蛹率。敲低LdATPaseE还显著升高了LdInR与Ld4EBP的表达量,抑制一个蜕皮激素合成基因的转录,降低20E滴度并降低了一个20E响应基因的表达。敲低LdATPaseE还抑制了一个JH合成酶基因的表达,降低了JH滴度,下调了一个JH早期响应基因的表达量。【结论】沉默LdATPaseE后,其可能通过抑制IIS信号途径,降低20E和JH的滴度、下调20E和JH信号从而影响幼虫生长和发育。

关键词: 马铃薯甲虫; RNA干扰; ATPaseE; 类胰岛素; 保幼激素; 蜕皮激素

Abstract: 【Objective】 Previous experiments showed that feeding different doses of LdATPaseE to Colorado Potato Beetles' larvae resulted in a gradual decline in the feeding amount, mobility, digging ability, pupation rate, and emergence rate of the larvae. This experiment attempts to explain the LdATPaseE gene expression pattern and how the insulin-like signaling pathway affects juvenile hormone and ecdysone hormone, thereby regulating the pupa and emergence of Coloradopotato beetle larvae.【Methods】 The LdATPaseEcDNA was obtained by transcriptome data and RT-PCR cloning from Colorado Potato Beetles. Sequence alignment and phylogenetic analysis were used to determine the integrity and phylogeny of the gene's cDNA. The expression of larvae in different tissues of Colorado Potato Beetles at different developmental stages was detected by qPCR. The effect of the gene on the growth and development of Colorado Potato Beetle larvae was observed by feeding the larvae dsRNA, and the effects of juvenile hormone and ecdysone on the expression of the gene were determined.【Results】 dsLdATPaseE-1 and dsLdATPaseE-2 were fed to second-instar larvae, leading to successfully knocking down of the target gene's expression level, and prevented the growth of second-instar larvae and significantly increased the mortality of second-instar larvae. Feeding dsLdATPaseE-1 and dsLdATPaseE-2 at the third and fourth instar stage of larvae, the target gene was knocked down at even very low concentrations, preventing larval growth and significantly reducing the pupation rate of potato beetle larvae. In addition, knockdown of LdATPaseE also significantly increased the expression of LdInR and Ld4EBP, inhibited the transcription of an ecdysone synthesis gene, and decreased the 20E titer and the expression of a 20E response gene. Silence of LdATPaseE also inhibited the expression of a JH synthase gene, reduced the JH titer, and down-regulated the expression of a JH early response gene.【Conclusion】 Upon silencing LdATPaseE, it may affect larval growth and development by inhibiting its suppressioninsulin/insulin-like pathways, thereby reducing the titer of 20E and JH, down-regulating 20E and JH signals.

Key words: Leptinotarsa decemlineata; RNA interference; ATPaseE; insulin/insulin-like; juvenile hormone; molting hormone

中图分类号:

S188
S435.32

廖兰兰, 王俊, 付开赟, 丁新华, 何江, 吐尔逊·阿合买提, 郭文超. 马铃薯甲虫LdATPaseE调控幼虫蜕皮的分子机制[J]. 新疆农业科学, 2021, 58(6): 1124-1137.

LIAO Lanlan, WANG Jun, FU Kaiyun, DING Xinhua, HE Jiang, Tursun Ahmat, GUO Wenchao, . LdATPaseE in the Regulation of Larval Molting Mechanism in Leptinotarsa decemlineata[J]. Xinjiang Agricultural Sciences, 2021, 58(6): 1124-1137.

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马铃薯甲虫LdATPaseE调控幼虫蜕皮的分子机制

LdATPaseE in the Regulation of Larval Molting Mechanism in Leptinotarsa decemlineata

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