CsInR基因影响七星瓢虫滞育时脂肪积累的研究

doi:10.16409/j.cnki.2095-039x.2023.11.019

摘要/Abstract

摘要： 七星瓢虫Coccinella septempunctata作为一种捕食性天敌昆虫，在生物防治中被广泛应用。低温短光照可诱导七星瓢虫进入滞育。滞育态雌虫腹部有大量脂质积累，存活时间显著延长。掌握七星瓢虫滞育调控技术，可大幅度延长产品货架期，促进天敌昆虫产品在生物防治的应用。胰岛素信号在昆虫生理变化过程中具有重要作用，但在七星瓢虫滞育过程的功能尚不清楚。本研究鉴定并分析了七星瓢虫胰岛素受体基因（CsInR）序列及表达特征。RT-qPCR结果表明，与非滞育个体相比，滞育个体CsInR基因表达量在其头部显著下调。CsInR基因在滞育诱导前20 d的表达水平显著低于其他时期。RNA干扰CsInR基因后，七星瓢虫体内脂质含量和脂肪合成通路关键基因（CsFas1，CsFas2和CsFad△11基因）表达量均显著增加。本研究将为进一步探究胰岛素信号调控七星瓢虫滞育时脂肪积累机制奠定基础。

关键词: 七星瓢虫, CsInR, 脂质积累, 滞育

Abstract: The seven-spotted ladybeetle, Coccinella septempunctata L., is a predatory natural enemy widely used as a biological control agent against pests. The diapause of C. septempunctata is induced by low temperature and short light. The diapause females accumulate a large amount of lipid in the abdomen and display a prolonged longevity. The technology of diapause induction is expected to extend the shelf-life of natural enemy insect products and improve the application of the products in biological control programs. Insulin signaling plays an essential role in the physiological changes of insects. Its function on the diapause process of C. septempunctata remains unclear. We identified and analyzed the insulin receptor gene (CsInR) sequence of C. septempunctata. The RT-qPCR results showed that, compared with that in the nondiapause individuals, the CsInR expression was significantly downregulated in the diapause individuals’ heads. The CsInR gene expression of the individuals with diapausing induction for 20 days was significantly lower than that of those individuals induced for diapause for other periods. After RNA interference with the CsInR gene, the lipid content and the expression of essential genes of the lipid synthesis pathway (CsFas1, CsFas2 and CsFad△11 genes) were significantly reduced. This study paves the way for further investigating the mechanism of insulin signaling regulating lipid accumulation during diapause in C. septempunctata..

Key words: Coccinella septempunctata, CsInR, lipid accumulation, diapause

中图分类号:

S476.2

李萍, 陈俊杰, 刘照涵, 郭鹏辉, 刘晓晓, 李玉艳, 张礼生. CsInR基因影响七星瓢虫滞育时脂肪积累的研究[J]. 中国生物防治学报, 2024, 40(4): 834-843.

LI Ping, CHEN Junjie, LIU Zhaohan, GUO Penghui, LIU Xiaoxiao, LI Yuyan, ZHANG Lisheng. The CsInR Gene Regulates Lipid Accumulation in the Diapause Seven-spotted Ladybeetle[J]. Chinese Journal of Biological Control, 2024, 40(4): 834-843.

参考文献

[1] 张礼生, 陈红印, 李保平. 天敌昆虫扩繁与应用[M]. 北京: 中国农业科学技术出版社, 2014.
[2] Hance T, Dixon A F G. Insect predator–prey dynamics: ladybird beetles and biological control[J]. Journal of Insect Conservation, 2006, 10(4): 375-376.
[3] Hodek I, Michaud J P. Why is Coccinella septempunctata so successful? (A point-of-view)[J]. European Journal of Entomology, 2008, 105(1): 1-12.
[4] Honěk A. The distribution of overwintered Coccinella septempunctata L. (Col. Coccinellidae) adults in agricultural crops[J]. Zeitschriftfür Angewandte Entomologie, 1982, 94: 1-5.
[5] 马野萍, 孙洪波, 王瑞霞, 等. 七星瓢虫生物学特性及人工饲养初步研究[J]. 新疆农业科学, 2000(S1): 154.
[6] 任小云. 七星瓢虫滞育的代谢适应及脂代谢的转录组学机理研究[D]. 北京: 中国农业科学院, 2016.
[7] 任小云, 张礼生, 齐晓阳, 等. 滞育七星瓢虫的代谢适应与抗寒性评价[J]. 环境昆虫学报, 2015, 37(6): 1195-1202.
[8] 顾世红, 陈建国. 胰岛素信号转导, 昆虫发育与老化[J]. 昆虫知识, 2009(4): 501-508.
[9] 张龙辉, 王国栋. 无脊椎动物胰岛素样蛋白(Insulin-like/related peptides)研究进展——以昆虫为例[J]. 生物技术通报, 2014(10): 33-42.
[10] Garofalo R S. Genetic analysis of insulin signaling in Drosophila[J]. Trends in Endocrinology & Metabolism, 2002, 13(4): 156-162.
[11] Goberdhan D C, Wilson C. The functions of insulin signaling: size isn’t everything, even in Drosophila[J]. Differentiation, 2003, 71(7): 375-397.
[12] Partridge L, Alic N, Bjedov I, et al. Ageing in Drosophila: the role of the insulin/Igf and TOR signalling network[J]. Experimental Gerontology, 2011, 46(5): 376-381.
[13] Teleman A A. Molecular mechanisms of metabolic regulation by insulin in Drosophila[J]. Biochemical Journal, 2010, 425(1):13-26.
[14] Wu Q, Brown M R. Singnaling and function of insulin-like peptides in insetces[J]. Annual Review of Entomology, 2006, 51: 1-24.
[15] Antonva Y, Arik A J, Moore W, et al. Insulin-like peptides: structure, signaling and function[M]. London: Elsevier Academic Press, 2015, 63-92.
[16] Altintas O, Park S, Lee S J V. The role of insulin/IGF-1 signaling in the longevity of model invertebrates, C. elegans and D. melanogaster[J]. Bmb Reports, 2016, 49(2): 81-92.
[17] Renske E, Amita S. Regulation of insect behavior via the insulin-signaling pathway[J]. Frontiers in Physiology, 2013, 4: 353.
[18] Fruman D A, Meyers R E, Cantley L C. Phophoinositide kinases[J]. Annual Review of Biochemistry, 1998, 67(1): 481-507.
[19] Green D A, Extavour C G. Insulin signaling underlies both plasticity and divergence of a reproductive trait in Drosophila[J]. Proceedings of the Royal Society Biological Sciences, 2014, 281(1779): 20132673.
[20] Nassel D, Liu Y, Luo J. Insulin/IGF singaling and its regulation in Drosophila[J]. General and Comparative Endocrinology, 2015, 221: 266-266.
[21] Fresno Vara J A, Casado E, de Castro J, et al. PI3K/Akt signalling pathway and cancer[J]. Cancer Treatment Reviews, 2004, 30(2): 193-204.
[22] Sheng Z, Xu J, Bai H, et al. Juvenile hormone regulates vitellogenin gene expression through insulin-like peptide signaling pathway in the red flour beetle, Tribolium castaneum[J]. Journal of Biological Chemistry, 2011, 286(49): 41924-41936.
[23] Tatar M, Kopelman A, Epstein D, et al. Mate Choice, sexual conflict, and evolution of senescence[J]. Behavior Genetics, 2001, 292: 107-110.
[24] Meola R W, Petralia R S. Juvenile hormone induction of biting behavior in Culex mosquitoes[J]. Science, 1980, 209: 1548-1550.
[25] Readio J, Chen M H, Meola R. Juvenile hormone biosynthesis in diapausing and nondiapausing Culex pipiens (Diptera:Culicide)[J]. Journal of Medical Entomology, 1999, 36: 355-360.
[26] Belgacem Y H, Martin J R. Disruption of insulin pathways alters trehalose level and abolishes sexual dimorphism in locomotor activity in Drosophila[J]. Neurobiology, 2006, 66: 19-32.
[27] Belgacem Y H, Martin J R. Hmgcr in the corpus allatum controls sexual dimorphism of locomotor activity and body size via the insulin pathway in Drosophila[J]. PLoS ONE, 2007, 2: e187.
[28] Sin C, Denlinger D L. Insulin signaling and FOXO regulate the overwintering diapause of the mosquito Culex pipiens[J]. Proceedings of the National Academy of Science, 2008, 105(18): 6777-6781.
[29] Sin C, Denlinger D L. A shut-down in epression of an insulin-like peptide, ILP-1, halts ovarian maturation during the overwintering diapause of the mosquito Culex pipiens[J]. Insect Molecular Biology, 2009, 18(3): 325-332.
[30] Sin C, Denlinger D L. Transcripition profiling and regulation of fat metabolism genes in diapausing adults of the mosquito Culex pipiens[J]. Physiological Genomics, 2009, 39(3): 202-209.
[31] Sin C, Denlinger D L. Insulin signaling and the egulation of insect dispause[J]. Frontiers in Physiology, 2013, 4: 189.
[32] Broughton S J, Piper M D, Ikeya T, et al. Longer Lifespan,altered metabolism,and stress resistance in Drosophila from ablation of cells maling insulin-like ligands[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(8): 3105-3110.
[33] Ikeya T, Galic M, Belawat P K, et al. Nutrient-dependent expression of insulin-like peptides from neuroendocrine cells in the CNS ontributes to growth regulation in Drosophila[J]. Current Biology, 2002, 12(15): 1293-1300.
[34] Richard D S, Rybczynski R, Wilson T G, et al. Insulin signaling is necessary for vitellogenesis in Drosophila melanogaster independent of the roles of juvenile hormone and ecdysteroids:female sterility of the chicol insulin signaling mutation is autonomous to the ovary[J]. Journal of Insect Physiology, 51(4): 455-464.
[35] 韩艳华. 脂肪酸合酶及酰基-CoAΔ 11去饱和酶在七星瓢虫滞育中的功能研究[D]. 北京: 中国农业科学院, 2018.
[36] 王伟, 张礼生, 陈红印, 等. 北京地区七星瓢虫滞育诱导的温光效应[J]. 中国生物防治学报, 2013, 29(1): 24-30.