安佳婷,徐海轩,杨浩成,闫春财,孙泽阳,2023,红裸须摇蚊幼虫肠道可培养细菌多样性及耐受重金属功能初步研究[J].环境昆虫学报,(4):942-951
红裸须摇蚊幼虫肠道可培养细菌多样性及耐受重金属功能初步研究
Preliminary analysis of culturable bacteria in the guts of chironomid larvae(Propsilocerus akamusi Tokunaga) and their potential roles in tolerating heavy metals
  
DOI:
中文关键词:  红裸须摇蚊  肠道细菌  重金属  分离培养  16S rRNA
英文关键词:Propsilocerus akamusi  gut bacteria  heavy metal  isolation  16S rRNA
基金项目:天津市教委科研计划项目(自然科学)(2021KJ183)
作者单位
安佳婷,徐海轩,杨浩成,闫春财,孙泽阳 天津师范大学生命科学学院天津市动物多样性保护与利用重点实验室天津 300387 
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中文摘要:
      红裸须摇蚊Propsilocerus akamusi为华北地区常见耐污种,广泛分布于各种水体中。本研究采用传统培养法对4龄红裸须摇蚊幼虫肠道内细菌种类进行分离,利用16S rRNA基因序列分析比对,结合菌落形态、系统进化分析等方法进一步鉴定菌株种类,同时探究可培养菌株对重金属(铜、镍)的耐受情况。结果表明,红裸须摇蚊幼虫在重金属铜暴露下的24 h-LC50、48 h-LC50、72 h-LC50、96 h-LC50分别为6304.86、403.59、29.33以及14.43 mmol/L,在重金属镍暴露下的24 h-LC50、48 h-LC50、72 h-LC50、96 h-LC50分别为339.52、182.18、102.59以及60.94 mmol/L。利用LB、NA、BHI、MC 4种培养基,从幼虫肠道分离纯化到16种不同遗传型细菌,分属2门 9科16属,其中优势菌门为变形菌门Proteobacteria。另外,丛毛单胞菌科Comamonadaceae丛毛单胞菌属Comamonas丛毛单胞菌Comamonas sp.对重金属铜和镍有良好耐受性。通过研究红裸须摇蚊幼虫肠道可培养细菌群落组成以及重金属胁迫下摇蚊幼虫肠道细菌群落生长情况,可为水生昆虫肠道菌群资源库的开发提供了一定的思路,也可为筛选出具备重金属代谢功能的细菌资源奠定基础。
英文摘要:
      Propsilocerus akamusi is a common species in North China and characterized by their tolerant ability to deal with environmental stressors as well as ubiquitous distribution in various water environments. In this study, traditional culture method was employed to isolate the bacterial species in chironomid larval guts. 16S rRNA gene sequence analysis together with colony morphology and phylogenetic determination were adopted to further identify the bacterial species. Meanwhile, the tolerance of culturable strains to heavy metals(e.g. copper and nickel)was investigated. Our results showed that the concentrations of 24 h-LC50、 48 h-LC50、 72 h-LC50 and 96 h-LC50 to copper were 6304.86、403.59、29.33 and 14.43 mmol/L, respectively whereas those for nickle were 339.52、182.18、102.59 and 60.94 mmol/L, respectively. 16 different genotypes of bacteria were successfully isolated and purified from the intestinal tracts of fourth instar larvae, and belonged to 16 genera, 9 families and 2 phyla, among which Proteobacteria was proved to be the predominant phyla. Comamonas sp., belonging to Comamonadaceae, had relatively strong tolerance to both copper and nickel. Exploring the culturable microbes and potentially heavy metal-resistant strains in chironomid guts would provide clues for the development of aquatic insect microbial resources and lay a preliminary foundation for discovering certain strains endowed with bioremediation capability of heavy metals.
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