芽孢杆菌的引入对褐飞虱微生物群和生长发育的影响  

Effects of introduction of Bacillus spp.on the microbiota and growth and development of the brown planthopper,Nilaparvata lugens(Hemiptera:Delphacidae)

作　　者：王渭霞[1] 朱廷恒[2] 赖凤香[1] 魏琪 万品俊 何佳春[1] 傅强[1] WANG Wei-Xia;ZHU Ting-Heng;LAI Feng-Xiang;WEI Qi;WAN Pin-Jun;HE Jia-Chun;FU Qiang(State Key Laboratory of Rice Biology and Breeding,China National Rice Research Institute,Hangzhou 311401,China;College of Biotechnology and Bioengineering,Zhejiang University of Technology,Hangzhou 310014,China)

机构地区：[1]中国水稻研究所,水稻生物育种全国重点实验室,杭州311401 [2]浙江工业大学生物工程学院,杭州310014

出　　处：《昆虫学报》2023年第10期1289-1301,共13页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(32272538);水稻生物学国家重点实验室开放课题(20210302);稻飞虱灾变机制与可持续防控技术研究(2021YFD1401100);青海省重点研发与转化计划(2022-SF-147)。

摘　　要：【目的】明确褐飞虱Nilaparvata lugens体内可培养共生细菌的种类,探索其中芽孢杆菌类共生菌对褐飞虱微生物群和生长发育的影响。【方法】通过离体培养的方法,从褐飞虱两种不同致害性种群(TN1敏感种群和IR56高致害种群)中分离可培养共生细菌。通过16S rDNA测序对获得的可培养共生菌进行鉴定,在此基础上利用原位杂交研究了共生细菌在褐飞虱体内的分布。利用人工饲料添加抗生素和回补共生细菌的方式,研究了减菌和回补芽孢杆菌的处理对褐飞虱生长发育以及共生菌丰度的影响。比较了通过饲喂和显微注射对芽孢杆菌引入的影响,考察了芽孢杆菌的定殖与褐飞虱TN1种群致害性之间的相关性。【结果】利用离体培养法从褐飞虱中获得了15株不同的共生细菌,其中包括2株源于IR56高致害种群的芽孢杆菌类共生细菌BPH-S36和BPH-S33。原位杂交结果表明共生细菌在褐飞虱成虫唾液腺、肠道、脂肪体和雌虫内生殖器中均有分布,而在雄虫内生殖器中鲜有分布。体内共生细菌对褐飞虱的生长发育至关重要,共生细菌的减少导致褐飞虱存活率在第3和6天时显著下降,而回补共生芽孢杆菌BPH-S36或BPH-S33可使其存活率在第6天时显著上升。褐飞虱通过取食添加共生芽孢杆菌的人工饲料可使芽孢杆菌的相对丰度增加23.2~24.5倍,而通过若虫显微注射并不能够提高芽孢杆菌的相对丰度。随着共生芽孢杆菌的增加,类酵母共生菌(yeast-like symbiont,YLS)的相对丰度也显著增加5~12倍。芽孢杆菌BPH-S36的引入使褐飞虱敏感种群TN1在抗性水稻IR56上的存活率由52.1%±1.5%显著提高至64.2%±3.0%。【结论】共生细菌在褐飞虱唾液腺、肠道和卵巢中广泛分布。抗生素处理能够有效降低其体内共生细菌的丰度,从而导致存活率下降。人工饲喂法可以使外源共生细菌在褐飞虱体内成功定殖。源于致害性强�【Aim】To identify the species of culturable symbiotic bacteria in the brown planthopper,Nilaparvata lugens,explore the effects of the symbiotic Bacillus spp.on the microbiota and growth and development of N.lugens.【Methods】The culturable symbiotic bacteria were isolated from two different virulent populations of N.lugens(the susceptible population TN1 and the virulent population IR56)by in vitro culture.The obtained cuttural symbiotic bacteria were identified by 16S rDNA sequencing technology.On this basis,the distribution of symbiotic bacteria in N.lugens was studied by in situ hybridization.The effects of reducing symbionts and supplementation of Bacillus spp.on the growth and development of N.lugens,as well as the abundance of symbionts,were studied using artificial diet added with antibiotics or symbiotic Bacillus spp.The effects of introducing symbiotic Bacillus spp.through feeding and microinjection methods were compared,and the correlation between Bacillus spp.colonization and the virulence of TN1 population was investigated.【Results】In total 15 strains of different symbiotic bacteria were obtained from N.lugens by in vitro culture,including 2 strains of the symbiotic Bacillus spp.(BPH-S36 and BPH-S33)from the highly virulent population IR56.The result of in situ hybridization showed that symbiotic bacteria were distributed in the salivary glands,gut,fat body and female internal genitalia of N.lugens,but rarely in the male internal genitalia.Symbionts are crucial for the growth and development of N.lugens.The reduction of symbiotic bacteria led to a significant decrease in the survival rate of N.lugens on the 3rd and 6th day,while supplementing the symbiotic Bacillus BPH-S33 or BPH-S36 resulted in a significant increase in the survival rate of N.lugens on the 6th day.The relative abundance of Bacillus spp.in N.lugens increased by 23.2-24.5-fold by feeding on the artificial diet supplemented with the symbiotic Bacillus spp.,but not increased through microinjection into nymphs.With the increase of

关 键 词：褐飞虱 共生细菌 类酵母共生菌 致害性 芽孢杆菌 丰度 

分 类 号：Q965.8[生物学—昆虫学]