葱地种蝇幼虫伴生细菌多样性及两种培养基分离效果比较

Diversity and Isolation of Symbiotic Bacteria on Delia antiqua Larvae

【目的】探索蒜蛆幼虫伴生细菌的多样性,并比较两种常用培养基对其伴生细菌的分离效果,为开展蒜蛆伴生细菌研究提供技术指导。【方法】采用高通量扩增子测序方法检测蒜蛆幼虫体表和肠道的细菌多样性;使用LB和TSB从蒜蛆幼虫的体表和肠道分离细菌,并比较两种培养基分离结果、测序结果和两种培养基分离效果的差异。【结果】高通量测序从蒜蛆幼虫体表和肠道共检测到4个门、66个科、115个属的细菌,其中绝大部分不能够鉴定到种。使用LB培养基从蒜蛆幼虫体表和肠道共分离得到463株细菌,鉴定后共得到29种细菌,分别属于11个科18个属。使用TSB培养基从蒜蛆幼虫体表和肠道分离得到391株细菌,经测序后共鉴定到44种细菌,分别属于19个科28个属。总体来说,TSB分离到的细菌种类更多,部分细菌可同时用LB与TSB分离到;此外,与高通量测序结果相比,两种培养基均只能分离到很小一部分种类的细菌。【结论】在蒜蛆伴生细菌的分离试验中,最好同时使用两种培养基对细菌进行分离,以获得更多种类的细菌。

【Objective】Diversity of the symbiotic bacteria on Delia antiqua larvae was studied,and effectiveness of two commonly used culture media in isolating them compared.【Methods】Diversity of the bacterial population on the surface and in the guts of D.antiqua larvae were determined using the high-throughput amplicon sequencing technique.The effectiveness of the LB and TSB media to culture and isolate the bacteria were compared based on the sequencing results.【Results】The inhabitation bacteria on the larvae were from 115 genera,66 families,and 4 phyla according to the sequencing.Most of them could not be annotated to species level.On LB medium,463 strains were isolated that belonged to 29 species,18 genera,and 11 families,while on TSB,391 strains from 44 species,28 genera,and 19 families.TSB isolated more species than LB,and some of the species could be isolated by either method.However,the high-throughput amplicon sequencing revealed that these culture methods could provide only a minute portion of the bacteria population that coinhabited on the host.【Conclusion】It was necessary to use both TSB and LB media in order to maximize the isolation of all symbiotic bacteria on D.antiqua larvae.