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Characteristics of halloween genes and RNA interference-mediated functional analysis of LmCYP307a2 in Locusta migratoria

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Insect Science 2022年第1期29卷 51-64页

作者： Xue-Yao Zhang Qi-Hui He Ting-Ting Zhang Hai-Hua Wu Jian-Zhen Zhang En-Bo Ma Research Institute of Applied Biology Shanxi UniversityTaiyuanShanxi030006China

halloween genes are involved in the biosynthesis of the molting hormone, which plays a key role in insect ecdysis, development, metamorphosis, and reproduction. Our previous work identified five halloween genes from Locusta migratoria, but their functions are currently unknown. In this study, the sequences of these five halloween genes were analyzed and characterized. LmCYP307a2, LmCYP306a1, LmCYP302a1, and LmCYP315a1 were primarily expressed in the prothoracic glands, while LmCYP314a1 was universally expressed in peripheral tissues, especially in the ovaries and Malpighian tubules. All five halloween genes were mainly expressed from the 5th to the 7th d in 5th-instar nymphs. RNA interference (RNAi) silencing of LmCYP307a2 resulted in severe molting delays and molting failure, which could be rescued by supplementary 20-hydroxyecdysone. A hematoxylin and eosin staining analysis suggested that the RNAi of LmCYP307a2 inhibited the ecdysis process by inhibiting the apolysis and degradation of the old cuticle, and by promoting the synthesis of a new cuticle. Quantitative reverse transcription polymerase chain reaction results showed that the expressions of LmE74, LmCht5, and LmCht10 were dramatically down-regulated, while that of LmChsI was substantially up-regulated, after knockdown of LmCYP307a2. The results suggest that LmCYP307a2 is related to the molt process via regulation of chitin synthesis and degradation.

关键词： halloween genes Locusta migratoria RNAi silencing Spookier

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Ecdysteroids： the overlooked sex steroids of insects？ Males： the black box

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Insect Science 2006年第5期13卷 325-338页

作者： ARNOLD DE LOOF Laboratory for Developmental Physiology Genomics and Proteomics Zoological Institute K.U. Leuven Leuven Belgium

The paradigm, still around in textbooks, that ＇in insects sex is strictly genetic, thus that they do not have sex hormones＇, is mainly based on a wrong interpretation of the ＇gynandromorph argument＇. It is no longer tenable. Given the fact that vertebrates and invertebrates probably had a common, sexually reproducing ancestor, there is no reason to assume that only vertebrates need sex hormones. The major function of sex hormones is to inform the somatoplasm about developmental changes that take place in the gonads. In contrast to juvenile hormone and neuropeptides, ecdysteroids meet all criteria to act as sex hormones, which was probably their ancient role. Their much better documented role in moulting and metamorphosis was a secondary acquisition that enabled arthropods to cope with growth problems, imposed by a rigid cuticle. Female insects use 20-hydroxyecdysone （20E）, secreted by the follicle cells of the ovary, in a similar way as females of egg-laying vertebrates use estrogens. For a variety of reasons, the possibility that ecdysteroids, in particular ecdysone （E）, might also act as sex hormones in male insects, thus as the counterpart of testosterone of vertebrates, has been very much overlooked. Thanks to the recent discovery of the molecular basis of the haploid-diploid system of sex determination in the honeybee, the characterization of halloween genes, proteomics, RNAi and so on, it now becomes possible to verify whether in insects, as with vertebrates, males are the endocrinologically default gender form.

关键词： ecdysone gynandromorph halloween genes 20-hydroxylase juvenilehormone sex determination

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