七星瓢虫雄虫生殖系统精液蛋白基因的表达分析

doi:10.16409/j.cnki.2095-039x.2024.07.008

摘要/Abstract

摘要： 为明确七星瓢虫Coccinella septempunctata雄虫精液蛋白基因在性别间的表达差异并筛选出在雄虫生殖系统特异性表达的基因，从转录组数据库选取10个精液蛋白基因，采用实时荧光定量PCR技术检测其在七星瓢虫不同性别、组织及发育阶段的表达特征。结果显示，在七星瓢虫雄虫10个精液蛋白基因中，有5个精液蛋白基因（PGK、ENO、SDH、PLA2b、SHMT）的mRNA表达水平在雄虫中更高，其中PLA2b具有专一的雄虫生殖系统特异性，在雄虫生殖系统中的表达量是雌虫生殖系统的56.20倍。PGK、ENO、SDH、PLA2b和SHMT在雄虫生殖系统的表达量分别是头部的1.63、5.90、2.63、10.14和1.23倍。PGK、ENO、SDH和PLA2b在成虫期的表达量显著高于低龄幼虫期，其中SDH和PLA2b在成虫期急剧上调表达，分别是卵期表达量的51.01倍和22.22倍，二者具有明显的发育阶段特异性。结果表明，SDH和PLA2b基因可能参与七星瓢虫雄虫生殖作用。

关键词: 七星瓢虫, 精液蛋白基因, 雄虫生殖系统, 表达分析

Abstract: The expression differences of the seminal fluid protein gene of Coccinella septempunctata in different sexes, tissues, and developmental stages and the genes specifically expressed in the male reproductive system were detected using real-time fluorescence quantitative PCR technology from 10 seminal fluid protein genes selected from the transcriptome database. The results showed that among the 10 seminal protein genes in the male ladybug, 5 seminal protein genes (PGK, ENO, SDH, PLA2b, SHMT) had higher mRNA expression levels in males. Among them, PLA2b showed male reproductive system specificity, with expression level 56.20 times higher in the male than in the female. The expression levels of PGK, ENO, SDH, PLA2b and SHMT in the male reproductive system were 1.63, 5.90, 2.63, 10.14 and 1.23 times higher than those in the head, respectively. The expression levels of PGK, ENO, SDH and PLA2b in the adult stage were significantly higher than those in the young larval stage. Among them, SDH and PLA2b were sharply upregulated in the adult stage, with expression levels 51.01 and 22.22 times higher than those in the egg stage, respectively, showing obvious developmental stage specificity. The results indicate that the SDH and PLA2b genes may be involved in the reproductive function of male ladybird beetles.

Key words: Coccinella septempunctata, seminal fluid protein gene, male reproductive tract, gene expression

中图分类号:

S476.2

程英, 周宇航. 七星瓢虫雄虫生殖系统精液蛋白基因的表达分析[J]. 中国生物防治学报, 2025, 41(4): 745-751.

CHENG Ying, ZHOU Yuhang. Expression of Seminal Fluid Protein Genes of the Male Reproductive System in Coccinella septempunctata[J]. Chinese Journal of Biological Control, 2025, 41(4): 745-751.

参考文献

[1] Sarwar M, Saqib S M. Rearing of predatory seven spotted ladybird beetle Coccinella septempunctata L. (Coleoptera: Coccinellidae) on natural and artificial diets under laboratory conditions[J]. Pakistan Journal of Zoology, 2010, 42: 47-51.
[2] Skouras P J, Margaritopoulos J T, Zarpas K D, et al. Development, growth, feeding and reproduction of Ceratomegilla undecimnotata, Hippodamia variegata and Coccinella septempunctata fed on the tobacco aphid, Myzus persicae nicotianae[J]. Phytoparasitica, 2015, 43(2): 159-169.
[3] 周宇航, 程英, 金剑雪, 等. 七星瓢虫规模化生产与释放的应用效果[J]. 西南农业学报, 2017, 30(3): 602-605.
[4] Cartwright B O, Eikenbary R D, Angalet G W, et al. Release and Establishment of Coccinella septempunctata in Oklahoma[J]. Environmental Entomology, 1979, 8(5): 819-823.
[5] 向玉勇, 刘同先, 张世泽. 温湿度、光照周期和寄主植物对小地老虎求偶及交配行为的影响[J]．植物保护学报, 2018, 45(2): 235-242.
[6] 陈洁, 吴春娟, 张青文, 等. 异色瓢虫卵巢发育及卵子发生过程观察[J]. 植物保护学报, 2015, 42(2): 237-243.
[7] 于冰. 褐飞虱精液蛋白鉴定及离子转运肽基因功能分析[D]. 杭州: 浙江大学, 2016.
[8] 高洁, 嵇保中, 刘曙雯, 等. 昆虫雄性附腺功能蛋白研究进展[J]. 生命科学, 2008, 20(4): 618-624.
[9] Neubaum D M, Wolfner M F. Mated Drosophila melanogaster females require a seminal fluid protein, Acp36DE, to store sperm efficiently[J]. Genetics, 1999, 153(2): 845-857.
[10] Lung O, Wolfner M F. Identification and characterization of the major Drosophila melanogaster mating plug protein[J]. Insect Biochemistry and Molecular Biology, 2001, 31(6/7): 543-551.
[11] Koene J M, Sloot W, Montagne-wajer K, et al. Male accessory gland protein reduces egg laying in a simultaneous hermaphrodite[J]. PLoS ONE, 2010, 5(4): e10117.
[12] Avila F W, Sirot L K, Laflamme B A, et al. Insect seminal fluid proteins: identification and function[J]. Annual Review of Entomology, 2011, 56(1): 21-40.
[13] Thailayil J, Magnusson K, Godfray H C J, et al. Spermless males elicit large-scale female responses to mating in the malaria mosquito Anopheles gambiae[J]. Proceedings of the National Academy of Sciences Current issue, 2011, 108(33): 13677-13681.
[14] Baer B, Heazlewood J L, Taylor N L, et al. The seminal fluid proteome of the honey bee Apis mellifera[J]. Proteomics, 2009, 9(8): 2085-2097.
[15] Kamala Jayanthi P D, Sangeetha P, et al. Influence of polyandry on clutch size of the predatory coccinellid, Cryptolaemus montrouzieri (Coleoptera: Coccinellidae)[J]. Florida Entomologist, 2013, 96(3): 1073-1076.
[16] Bayoumy M H, Michaud J. Female fertility in Hippodamia convergens (Coleoptera: Coccinellidae) is maximized by polyandry, but reduced by continued male presence[J]. European Journal of Entomology, 2014, 111(4): 513-520.
[17] Perry J C, Rowe L. Condition-dependent ejaculate size and composition in a ladybird beetle[J]. Proceedings of the Royal Society of London B: Biological Sciences, 2010, 277(1700): 3639-3647.
[18] Xu J, Wang Q. Seminal fluid reduces female longevity and stimulates egg production and sperm trigger oviposition in a moth[J]. Journal of Insect Physiology, 2011, 57(3): 385-390.
[19] Perry J C, Sharpe D M, Rowe L. Condition-dependent female remating resistance generates sexual selection on male size in a ladybird beetle[J]. Animal Behaviour, 2009, 77: 743-748.
[20] Xu J J, Baulding J, Palli S R. Proteomics of Triboliumcas taneum seminal fluid proteins: identification of an angiotensin converting enzyme as a key player in regulation of reproduction[J]. Journal of Proteomics, 2013, 78: 83-93.
[21] Sirot L K, Hardstone M C, Helinski M E H, et al. Towards a semen proteome of the dengue vector mosquito: protein identification and potential functions[J]. PlOS Neglected Tropical Diseases, 2011, 5(3): e989.
[22] Simmons L W, Tan Y F, Millar A H. Sperm and seminal fluid proteomes of the field crickt Teleogryllus oceanicus: identification of novel proteins transferred to females at mating[J]. Insect Molecular Biology, 2013, 22(1): 115-130.
[23] Bonilla M L, Todd C, Erlandson M, et al. Combining RNA-seq and proteomic profiling to identify seminal fluid proteins in the migratory grasshopper Melanoplus sanguinipes (F)[J]. Bmc Genomics, 2015, 16(1): 1096.
[24] 刘梦姚, 王娟, 王曼姿, 等. 七星瓢虫保幼激素环氧水解酶基因克隆及表达分析[J]. 植物保护, 2019, 45(6): 156-162.
[25] Pfaffl M W. A new mathematical model for relative quantification in real-time RT-PCR [J]. Nucleic Acids Research, 2001, 29(9): e45.
[26] Bayram H, Sayadi A, Goenaga J, et al. Novel seminal fluid proteins in the seed beetle Callosobruchus maculatus identified by a proteomic and transcriptomic approach[J]. Insect Molecular Biology, 2017, 26(1): 58-73.
[27] 陈广梅, 高旭渊, 张燕, 等. 广聚萤叶甲雄虫生殖系统特异性精液蛋白基因的筛选及表达分析[J]. 植物保护学报, 2020, 47(6): 1202-1209.
[28] 张帅, 张永军, 苏宏华, 等. 棉铃虫信息素结合蛋白2cDNA的克隆、表达与组织特异性表达[J]. 中国农业科学, 2009, 42(7): 2359-2365.
[29] 张帅, 张永军, 苏宏华, 等. 中红侧沟茧蜂非典型气味受体的克隆及组织特异性表达[J]. 中国农业科学, 2009, 42(5): 1639-1645.
[30] Cheng Y, Zhou Y H, Li C, et al. Cloning and functional analysis of the juvenile hormone receptor gene CsMet in Coccinella septempunctata[J]. Journal of Insect Science, 2024, 24(4): 2; ieae065.
[31] 李鸿波, 刘芳, 符俊洪, 等. 黏虫生殖相关脂蛋白受体基因的克隆与时空表达分析[J]. 植物保护学报, 2022, 49(3): 741-748.
[32] Mccarrey J R, Thomas K. Human testis-specific PGK gene lacks introns and possesses characteristics of a processed gene[J]. Nature, 1987, 326(6112): 501-505.
[33] 姜冬梅, 康波, 马容, 等. 烯醇化酶l的生物学功能[J]. 动物营养学报, 2014, 26(1): 69-73.
[34] 周志雄, 袁江江, 李世香, 等. 柑橘大实蝇滞育蛹海藻糖酶和山梨醇脱氢酶活性的动态变化[J]. 植物保护, 2020, 46(4): 144-148.
[35] 王艇. 山梨醇脱氢酶作用机制及其与滞育的关系[J]. 安徽农学通报, 2012, 18(15): 39-84.
[36] Singh C O, Xin H H, Chen R T, et al. BmPLA2 containing conserved domain WD40 affects the metabolic functions of fat body tissue in silkworm, Bombyx mori[J]. Insect Science, 2016, 23: 28-36.
[37] Ji J Y, Yin Z H, Zhang S S, et al. PLA2 mediates the innate immune response in Asian corn borer, Ostrinia furnacalis[J]. Insect Science, 2022, 29: 245-258.
[38] 余金良, 何俊华. 蜜蜂毒主要过敏原磷脂酶A2研究概述[J]. 昆虫知识, 1999, 36(4): 248-250.