摘要
【目的】分离鉴定对亮斑扁角水虻Hermetia illucens产卵具有诱导作用卵携带细菌,探索提高亮斑扁角水虻人工繁育过程中卵收集效率途径。【方法】使用脑心浸出液肉汤(BHI)培养基对亮斑扁角水虻卵携带细菌分离纯培养;以灭菌BHI液体培养基为对照,使用筛选到的单菌株摇瓶发酵液对亮斑扁角水虻成虫进行产卵诱导试验。【结果】筛选纯培养到15株细菌,编号分别为EEAM-1,EEAM-2,EEAM-3,EEAM-5,EEAM-6,EEAM-7,EEAM-8,EEAM-9,EEAM-10A,EEAM-10B,EEAM-11,EEAM-12,EAM-13,EEAM-14和EEAM-15。经发酵液产卵诱导试验,发现有5株能显著提高亮斑扁角水虻成虫产卵量的菌株,分别为EEAM-3,EEAM-6,EEAM-7,EEAM-10A和EEAM-10B。经菌落培养特征、菌体形态比较、16S rDNA序列测定和发育树比对分析鉴定这5株细菌分别为:特基拉芽孢杆菌Bacillus tequilensis、粘质沙雷氏菌Serratia marcescens、粪肠球菌Enterococcus faecalis、短杆菌属Brevibacterium sp.和甲基营养芽孢杆菌Bacillus methylotrophicus。其中,EEAM-7粪肠球菌E.faecalis发酵液诱导的亮斑扁角水虻成虫产卵量最高,是对照组的7.400倍;其次为EEMA-10B甲基营养芽孢杆菌B.methylotrophicus,诱导的产卵量是对照组的6.531倍;再次是EEMA-6粘质沙雷氏菌S.marcescens、EEAM-10A短杆菌Brevibacterium sp.和EEMA-3特基拉芽孢杆菌B.tequilensis,诱导的产卵量分别是是对照组的5.100,4.187和2.636倍。【结论】亮斑扁角水虻卵携带细菌能显著影响繁殖期成虫产卵量,为建立稳定水虻人工养殖体系提供理论依据。
[ Aim] To screen and identify bacterial strains isolated from Hermetia illucens eggs inducing the oviposition of Hermetia illucens adults and to discover the way of improving the egg collection efficiency in the process of artificial breeding of this insect. [ Methods ] Separation and pure culture of egg-associated bacteria were fulfilled by using brain heart infusion broth （BHI） medium, and the screened solo bacteria strain was fermented in shaking flask to verify the effects of these fermentation broths on adult oviposition behavior of H. illucens with the sterilized BHI medium as the negative control. [Results] Fifteen strains were obtained and numbered as EEAM-1, EEAM-2, EEAM-3, EEAM-5, EEAM-6, EEAM-7, EEAM-8, EEAM-9, EEAM-10A, EEAM-10B, EEAM-11, EEAM-12, EEAM-13,EEAM-14 and EEAM-15, respectively. It was found that five strains including EEAM-3, EEAM-6, EEAM-7, EEAM-10A and EEAM-10B significantly improved the fecundity of H. illucens adults, and they were identified as Bacillus tequilensis, Serratia marcescens, Enterococcus faecalis, Brevibacterium sp. and Bacillus methylotrophicus, respectively, through the morphology comparison, characteristics of colonies, 16S rDNA sequence analyses and phylogenetic tree construction. Especially, the fermentation broth of Enterococcus faecalis （ EEAM-7 ） had the highest inducing effect on adult oviposition, 7. 400-fold as high as that of the control group. The oviposition amounts in the treatment groups of the fermentation broths of B. methylotrophicus （ EEMA-10B）, S. marcescens （ EEMA-6）, Brevibacterium sp. （ EEAM- 10A） and B. tequilensis （EEMA-3） were 6. 531-, 5. 100-, 4. 187- and 2. 636-fold as high as that of the control group, respectively. [ Conclusion] The results indicate that the egg-associated functional bacteria of H. iUucens affect the fecundity of H. illuceus adults. This study will provide a theoretical basis for establishing the stable feeding system of H. illucens.
出处
《昆虫学报》
CAS
CSCD
北大核心
2017年第2期163-172,共10页
Acta Entomologica Sinica
基金
国家自然科学基金项目(31400433)
河南省重点科技攻关项目(152102210060)
郑州市农委科技创新项目
