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解析克氏同源盒基因1在棉铃虫变态发育及雌性生殖中的作用

Dissecting the role of Krüppel homolog 1 in the metamorphosis and female reproduction of the cotton bollworm, Helicoverpa armigera.

作者信息

Zhang W-N, Ma L, Liu C, Chen L, Xiao H-J, Liang G-M

机构信息

Institute of Entomology, Jiangxi Agricultural University, Nanchang, China.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.

出版信息

Insect Mol Biol. 2018 Aug;27(4):492-504. doi: 10.1111/imb.12389. Epub 2018 May 2.

Abstract

In insects, metamorphosis and reproduction are controlled by juvenile hormone (JH) and 20-hydroxyecdysone (20E). Krüppel homolog 1 (Kr-h1), a transcription factor, is regarded as a JH-early inducible gene responsible for the repression of metamorphosis. However, the role of Kr-h1 in reproduction of holometabolic insects is relatively less understood. In this study, we studied the role of Kr-h1 in larvae-pupae transition and female reproduction in the major agricultural pest Helicoverpa armigera. Two HaKr-h1 isoforms (HaKr-h1α and HaKr-h1ß) were identified, with HaKr-h1α predominant in the cotton bollworm. In larvae, HaKr-h1 was predominately expressed in the epidermis and markedly up-regulated during the moult stage, whereas in adults HaKr-h1 was mainly expressed in females and the highest transcription was detected in the ovaries. Considering the function of hormones in larval metamorphosis, we examined the modulation of gene expression in response to hormones, which showed that HaKr-h1 was significantly induced by both JH analogue (JHA) and 20E. Knockdown of HaKr-h1 in fifth-instar larvae resulted in precocious metamorphosis from larvae to pupae. Moreover, a fluorescence immunoassay coupled with heterologous expression revealed that HaKr-h1 was localized in the nucleus of oocyte membrane. In female adults, depletion of HaKr-h1 severely repressed the transcription of vitellogenin, disrupted oocyte maturation and reduced the number of eggs laid, suggesting that HaKr-h1 is required for vitellogenesis and egg production in H. armigera. The present study provides insight into the roles of HaKr-h1 in JH-mediated reproduction and highlights HaKr-h1 as a target for suppression of lepidopteran pests.

摘要

在昆虫中,变态和繁殖受保幼激素(JH)和20-羟基蜕皮酮(20E)的控制。转录因子Krüppel同源物1(Kr-h1)被认为是一种JH早期诱导基因,负责抑制变态。然而,Kr-h1在全变态昆虫繁殖中的作用相对了解较少。在本研究中,我们研究了Kr-h1在主要农业害虫棉铃虫幼虫-蛹转变和雌性繁殖中的作用。鉴定出两种HaKr-h1异构体(HaKr-h1α和HaKr-h1β),其中HaKr-h1α在棉铃虫中占主导地位。在幼虫中,HaKr-h1主要在表皮中表达,在蜕皮阶段显著上调,而在成虫中,HaKr-h1主要在雌性中表达,在卵巢中检测到最高转录水平。考虑到激素在幼虫变态中的作用,我们检测了基因表达对激素的响应调节,结果表明HaKr-h1受到JH类似物(JHA)和20E的显著诱导。敲低五龄幼虫中的HaKr-h1导致幼虫提前变态为蛹。此外,荧光免疫测定结合异源表达表明,HaKr-h1定位于卵母细胞膜的细胞核中。在雌性成虫中,HaKr-h1的缺失严重抑制了卵黄原蛋白的转录,破坏了卵母细胞成熟并减少了产卵数量,这表明HaKr-h1是棉铃虫卵黄发生和产卵所必需的。本研究深入了解了HaKr-h1在JH介导的繁殖中的作用,并突出了HaKr-h1作为鳞翅目害虫防治靶点的地位。

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