Characteristics of Halloween genes and RNA interference-mediated functional analysis of LmCYP307a2 in Locusta migratoria
Corresponding Author
Xue-Yao Zhang
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
These authors contributed equally to this study.
Correspondence: Xue-Yao Zhang and En-Bo Ma, Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi 030006, China. Tel: +86 351 7018871; email: [email protected], [email protected]
Search for more papers by this authorQi-Hui He
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
These authors contributed equally to this study.
Search for more papers by this authorTing-Ting Zhang
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
These authors contributed equally to this study.
Search for more papers by this authorHai-Hua Wu
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
Search for more papers by this authorJian-Zhen Zhang
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
Search for more papers by this authorEn-Bo Ma
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
Search for more papers by this authorCorresponding Author
Xue-Yao Zhang
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
These authors contributed equally to this study.
Correspondence: Xue-Yao Zhang and En-Bo Ma, Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi 030006, China. Tel: +86 351 7018871; email: [email protected], [email protected]
Search for more papers by this authorQi-Hui He
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
These authors contributed equally to this study.
Search for more papers by this authorTing-Ting Zhang
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
These authors contributed equally to this study.
Search for more papers by this authorHai-Hua Wu
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
Search for more papers by this authorJian-Zhen Zhang
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
Search for more papers by this authorEn-Bo Ma
Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, 030006 China
Search for more papers by this authorAbstract
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.
Supporting Information
| Filename | Description |
|---|---|
| ins12907-sup-0001-FigureS1.pdf18 KB | Fig. S1 Multiple sequence alignment of CYP307A1/CYP307A2 from representative insect. |
| ins12907-sup-0002-FigureS2.pdf17.8 KB | Fig. S2 Multiple sequence alignment of CYP306A1 from representative insect. |
| ins12907-sup-0003-FigureS3.pdf18.9 KB | Fig. S3 Multiple sequence alignment of CYP302A1 from representative insect. |
| ins12907-sup-0004-FigureS4.pdf17.6 KB | Fig. S4 Multiple sequence alignment of CYP315A1 from representative insect. |
| ins12907-sup-0005-FigureS5.pdf17.8 KB | Fig. S5 Multiple sequence alignment of CYP314A1 from representative insect. |
| ins12907-sup-0006-FigureS6.tif131.6 KB | Fig. S6 Tissue-specific expression profiles of Halloween genes from Locusta migratoria. |
| ins12907-sup-0007-FigureS7.tif78.6 KB | Fig. S7 The effect of double-stranded RNA injection on the transcription level of Halloween genes. |
| ins12907-sup-0008-TableS1.docx22 KB | Table S1 Primers used in this study. |
| ins12907-sup-0009-DataS1.docx37.6 KB | Supplementary Data S1 Protein sequence used for phylogenetic analysis in Figure 1. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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