通过弱化黄粉虫幼虫体壁提升摩泽蒲螨繁育效率

doi:10.16409/j.cnki.2095-039x.2024.11.010

摘要/Abstract

摘要： 摩泽蒲螨Pyemotes moseri是一类昆虫体外寄生螨，在蛀干害虫的生物防治方面具有巨大的潜力，虽然可以利用替代寄主黄粉虫Tenebrio molitor来繁育，但黄粉虫体壁过厚影响了繁育效率。本研究利用RNAi技术弱化黄粉虫Tenebrio molitor幼虫体壁的方法，提高了黄粉虫幼虫繁育摩泽蒲螨的效率，取得的主要结果如下：成功构建了黄粉虫酚氧化酶原基因PPO的原核表达载体（TmPPO），诱导表达了dsTmPPO，验证了dsTmPPO能有效降低靶基因的相对表达量，干扰效率达到95.16%。同时能有效降低黄粉虫幼虫体壁的厚度，相比dsEGFP对照组，基因干扰组黄粉虫幼虫头部、胸部、腹部的体壁厚度分别减少了20.9%、23.5%和37.5%。随即饲喂dsTmPPO给黄粉虫幼虫，测定摩泽蒲螨在处理后黄粉虫幼虫上的寄生效率。结果表明，在dsTmPPO处理组幼虫上摩泽蒲螨成熟膨腹体数量为221个，显著多于在dsEGFP对照组黄粉虫上（151个）。综合以上结果得出：饲喂dsTmPPO给黄粉虫幼虫，能够显著弱化幼虫体壁，从而提高替代寄主黄粉虫繁育摩泽蒲螨的效率。

关键词: 摩泽蒲螨, 黄粉虫, RNAi, 体壁弱化, 繁育效率

Abstract: Pyemotes moseri is an ectoparasite mite of insects, showing a great potential in biological control of boring pests. In this study, RNAi technology was applied to weaken larval integument of Tenebrio molitor to improve the efficiency of breeding P. moseri by mealworm. The main results showed that the prokaryotic expression plasmid of PPO gene was successfully constructed to induce the expression of dsTmPPO, which verified that dsTmPPO injection effectively reduced the relative expression level of PPO gene with an interference efficiency of 95.16%, resulting in an effective reduction of integument thickness. The integument thickness of the head, thorax and abdomen of mealworm was reduced by 20.9%, 23.5% and 37.5%, respectively, compared to the dsEGFP control group. The parasitism efficiency of P. moseri measured after feeding the mealworm with dsTmPPO showed that there were 221 physogastric females of P. moseri in the dsTmPPO treatment group, significantly higher than that in the dsEGFP control group (115 physogastric females). In conclusion, feeding dsTmPPO to mealworm can weaken the integument and improve the breeding efficiency of P. moseri effectively.

Key words: Pyemotes moseri, Tenebrio molitor, RNAi, integument softening, breeding efficiency

中图分类号:

S476

史可桂, 苏晓琪, 王敦. 通过弱化黄粉虫幼虫体壁提升摩泽蒲螨繁育效率[J]. 中国生物防治学报, 2025, 41(4): 808-818.

SHI Kegui, SU Xiaoqi, WANG Dun. Improvement of Breeding Efficiency of Pyemotes moseri by Weakening the Integument of Tenebrio molitor[J]. Chinese Journal of Biological Control, 2025, 41(4): 808-818.

参考文献

[1] Luo Y, Huang H, Roques A. Early monitoring of forest wood-boring pests with remote sensing[J]. Annual Review of Entomology, 2023, 68(1): 277-298.
[2] Calderón-Cortés N, Quesada M, Cano-Camacho H, et al. A simple and rapid method for DNA isolation from xylophagous insects[J]. International Journal of Molecular Sciences, 2010, 11(12): 5056-5064.
[3] Tomalski M D, Bruce W A, Travis J, et al. Preliminary characterization of toxins from the straw itch mite, Pyemotes tritici, which induce paralysis in the larvae of a moth[J]. Toxicon, 1988, 26(2): 127-132.
[4] Tomalski M D, Miller L K. Insect paralysis by Baculovirus-mediated expression of a mite neurotoxin gene[J]. Nature, 1991, 352(6330): 82-85.
[5] 张佐双, 熊德平, 程炜. 利用天敌蒲螨控制柏树蛀干害虫双条杉天牛[J]. 中国园林, 2004(2): 80-82.
[6] 刘静, 韩冬银, 张方平, 等. 蒲螨利用研究进展[J]. 热带农业科学, 2008(3): 64-67, 77.
[7] Yu L C, Zhang Z Q, He L M. Two new species of Pyemotes closely related to P. tritici (Acari: Pyemotidae)[J]. Zootaxa, 2019(2723): 1-40.
[8] 胡帅, 周英梅, 苓建强, 等. 中华甲虫蒲螨对寄主的选择性初探[J]. 应用昆虫学报, 2013, 50(2): 460-465.
[9] 张彦龙, 王小艺, 唐艳龙, 等. 一种摩泽蒲螨的人工繁育方法及在防治苹小吉丁虫中的应用. 中国: CN201710636440.X[P]. 2020-06-23.
[10] Zhang D, Chen J, Yao Q, et al. Functional analysis of two chitinase genes during the pupation and eclosion stages of the bett armyworm Spodoptera exigua by RNA interference[J]. Archives of Insect Biochemistry and Physiology, 2012, 79(4/5): 220-234.
[11] Zhang J, Huang W, Yuan C, et al. Prophenoloxidase-mediated ex vivo immunity to delay fungal infection after insect ecdysis[J]. Frontiers in Immunology, 2017, 8: 1445.
[12] Wang Z, Luo J, Feng K, et al. Prophenoloxidase of Odontotermes formosanus (Shiraki) (Blattodea: Termitidae) is a key gene in melanization and has a defensive role during bacterial infection[J]. International Journal of Molecular Sciences, 2022, 24(1): 406.
[13] 王金彦, 张浩, 郭玲, 等. 寄主植物对甜菜夜蛾感染核型多角体病毒后黑化反应关键酶活性的影响[J]. 应用昆虫学报, 2018, 55(5): 882-888.
[14] Cao Z, Cao J, Vlasenko V, et al. Molecular characterization and functional analysis of a beta-1,3-glucan recognition protein from oriental fruit moth Grapholita molesta (Lepidoptera: Tortricidae)[J]. Archives of Insect Biochemistry and Physiology, 2024, 115(1): e22068.
[15] 郭志青, 许曼琳, 张霞, 等. 层生镰刀菌产生的有机挥发物对黄粉虫取食行为的影响[J]. 山东农业科学, 2023, 55(7): 122-127.
[16] 张艺馨, 史俊瑞, 付兴, 等. 白蛾黑棒啮小蜂的生殖力及替代寄主的繁蜂效果[J]. 应用昆虫学报, 2023, 60(3): 872-882.
[17] 胡镇杰, 杨海博, 林晓民, 等. 低温处理替代寄主对管氏肿腿蜂繁殖的影响[J]. 中国生物防治学报, 2017, 33(2): 165-170.
[18] 黄琼, 周祖基, 杨伟, 等. 繁育川硬皮肿腿蜂替代寄主的筛选[J]. 昆虫学报, 2005, 48(3): 375-379.
[19] 熊琳娜, 钱明惠, 范军祥, 等. 广东地区花绒寄甲替代寄主的研究[J]. 环境昆虫学报, 2011, 33(2): 219-224.
[20] Yang S, Ding M, He L, et al. Biodegradation of polypropylene by yellow mealworms (Tenebrio molitor) and superworms (Zophobas atratus) via gut-microbe-dependent depolymerization[J]. Science of the Total Environment, 2021, 756: 144087.
[21] Ganbaatar O, Cao B, Zhang Y, et al. Knockdown of Mythimna separata chitinase genes via bacterial expression and oral delivery of RNAi effectors[J]. BMC Biotechnology, 2017, 17(1): 9.
[22] Schefe J H, Lehmann K E, Buschmann I R, et al. Quantitative Real-time RT-PCR data analysis: Current concepts and the novel “gene expression’s CT difference” formula[J]. Journal of Molecular Medicine, 2006, 84(11): 901-910.
[23] Shah S, Hafeez M, Wu M, et al. Downregulation of chitin synthase A gene by diallyl trisulfide, an active substance from garlic essential oil, inhibits oviposition and alters the morphology of adult Sitotroga Cerealella[J]. Journal of Pest Science, 2020, 93(3): 1097-1106.
[24] 王圣朝, 王敦. 黄粉虫开发利用研究进展[J]. 应用昆虫学报, 2022, 59(4): 703-709.
[25] Zhang X, Fan Z, Zhang R, et al. Bacteria-mediated RNAi for managing fall webworm, Hyphantria cunea: Screening target genes and analyzing lethal effect[J]. Pest Management Science, 2023, 79(4): 1566-1577.
[26] Liu J, Smagghe G, Swevers L. Transcriptional response of BmToll9-1 and RNAi machinery genes to exogenous dsRNA in the midgut of Bombyx mori[J]. Journal of Insect Physiology, 2013, 59(6): 646-654.
[27] 张建珍, 史学凯, 李帅, 等. 昆虫dsRNA吸收机理研究现状与展望[J]. 山西大学学报(自然科学版), 2022, 45(3): 816-825.
[28] 骆旭铭, 陈诗敏, 潘广, 等. 饲喂法RNAi介导的HvCHS1基因沉默抑制茄二十八星瓢虫的存活和发育[J]. 中国生物防治学报, 2022, 38(5): 1083-1093.
[29] Lu A, Zhang Q, Zhang J, et al. Insect prophenoloxidase: the view beyond immunity[J]. Frontiers in Physiology, 2014, 5: 252.
[30] 唐艳龙, 王丽娜, 郑云霄, 等. 利用意大利蜜蜂蛹为替代寄主繁育两种肿腿蜂[J]. 植物保护, 2023, 49(4): 158-162, 184.
[31] 吕朝军, 钟宝珠, 李朝绪, 等. 红脉穗螟寄生蜂寄主选择研究[J]. 生物安全学报, 2022, 31(2): 146-150.
[32] 贺丽敏, 于丽辰, 焦蕊, 等. 人工繁殖中华甲虫蒲螨的替代寄主研究[J]. 中国生物防治学报, 2011, 27(2): 165-170.
[33] 于丽辰, 梁来荣, 敖贤斌, 等. 我国新天敌资源——小蠹蒲螨形态与生物学研究[J]. 蛛形学报, 1997(1): 46-52.