不同发育阶段马铃薯块茎蛾可培养内生细菌多样性及对消化作用的影响

Diversity of culturable endophytic bacteria at different developmental stages of Phthorimaea operculella( Zeller) and its effects on digestion

  • 摘要: 【目的】 明确马铃薯块茎蛾Phthorimaea operculella(Zeller)可培养内生细菌随宿主生长发育的变化动态及对取食消化功能的影响,为昆虫肠道微生物功能研究和微生物资源的开发利用打下理论基础。【方法】 以不同发育阶段的马铃薯块茎蛾为研究对象,通过传统分离培养技术对不同发育阶段的马铃薯块茎蛾体内(卵、1龄和2龄幼虫)或肠道(3龄和4龄幼虫、雌雄蛹、雌雄成虫)进行可培养内生细菌分离培养,通过16S rDNA序列分析对分离菌株进行种属鉴定;通过重饲喂优势细菌和常驻细菌,采用重量法测定马铃薯块茎蛾营养指标,分析优势细菌和常驻细菌对马铃薯块茎蛾幼虫取食及消化的影响。【结果】 马铃薯块茎蛾不同发育阶段的可培养内生细菌组成及多样性不同,其中从卵中分离到的可培养内生细菌种类数量最少,幼虫可培养内生细菌种类最丰富,且随着龄期的增加其可培养内生细菌数量逐渐增多,化蛹后开始下降。马铃薯块茎蛾可培养内生细菌共有3门13科17属30种,其中卵可培养内生细菌有2门3科3属3种,1~4龄幼虫、蛹和成虫可培养内生细菌分别有2门2科2属3种、2门4科5属6种、3门9科10属11种、3门10科10属11种、3门5科5属7种、3门8科8属10种。不同发育阶段的马铃薯块茎蛾可培养内生细菌优势种不同,其中1龄幼虫可培养细菌优势种为路氏肠杆菌(Enterobacter ludwigii),而卵、2~4龄幼虫、雌雄蛹和雌雄成虫可培养内生细菌优势种均为蒙氏肠球菌(Enterococcus mundtii)。蒙氏肠球菌在不同发育阶段马铃薯块茎蛾中均稳定存在,深红沙雷氏菌(Serratia rubidaea)在3龄和4龄幼虫、雌雄蛹和雌雄成虫中存在。蒙氏肠球菌、路氏肠杆菌和深红沙雷氏菌能促进马铃薯块茎蛾消化。【结论】 马铃薯块茎蛾可培养内生细菌种类丰富,不同发育阶段马铃薯块茎蛾可培养内生细菌组成结构不同,幼虫期可培养内生细菌种类明显高于卵和蛹,但优势种无明显差异。内生细菌优势种能通过促进幼虫的消化而促进宿主的生长发育。

     

    Abstract: 【Objective】 To clarify the dynamic changes of culturable endophytic bacteria of Phthorimaea operculella(Zeller) with the growth and development of the host and their effects on feeding and digestive function, which could provide theoretical basis for exploring the interaction between symbiotic bacteria and hosts. 【Method】 Taking Phthorimaea operculella(Zeller) at different developmental stages as the research object, the culturable endophytic bacteria were isolated and cultured in the body(eggs, 1st and 2nd instar larvae) or intestine(3rd and 4th instar larvae, male and female pupae, male and female adults) of Phthorimaea operculella(Zeller) at different developmental stages by traditional isolation and culture technology. The isolated strains were identified by 16S rDNA sequence analysis, through refeeding dominant bacteria and resident bacteria, the nutritive indexes of Phthorimaea operculella(Zeller) were detected by gravimetric method, and effects of dominat bacteria and resident bacteria on the feeding and digestion of Phthorimaea operculella(Zeller) larvae were analyzed.【Result】The composition and diversity of culturable endophytic bacteria at different developmental stages of Phthorimaea operculella(Zeller) were different. The number of culturable endophytic bacteria isolated from eggs was the least, and the number of culturable endophytic bacteria isolated from larvae was the most abundant. With the increase of age, the number of culturable endophytic bacteria gradually increased, and began to decline after pupation. There were 3 phyla, 13 families, 17 genera and 30 species of culturable endophytic bacteria in Phthorimaea operculella(Zeller). Among them, there were 2 phyla, 3 families, 3 genera and 3 species of culturable endophytic bacteria in eggs. There were 2 phyla, 2 families, 2 genera and 3 species; 2 phyla, 4 families, 5 genera and 6 species; 3 phyla, 9 families, 10 genera and 11 species; 3 phyla, 10 families, 10 genera and 11 species; 3 phyla, 5 families, 5 genera and 7 species; 3 phyla, 8 families, 8 genera and 10 species of culturable endophytic bacteria in 1st-4th instar larvae, pupae and adults respectively. The dominant species of endophytic bacteria in Phthorimaea operculella(Zeller) at different developmental stages were different. The dominant species of culturable bacteria of the 1st instar larvae was Enterobacter ludwigii, while the dominant species of culturable bacteria of eggs, 2nd-4th instar larvae, pupae, and adults were Enterococcus mundtii. Enterococcus mundtii was stable at all developmental stages of the Phthorimaea operculella(Zeller), and Serratia rubidaea was found in the 3rd and 4th instar larvae, pupae and adults. Enterococcus mundtii, Enterobacter ludwigii and Serratia rubidaea can promote the digestion of Phthorimaea operculella(Zeller).【Conclusion】 The species of culturable endophytic bacteria in Phthorimaea operculella(Zeller) are abundant. The composition and structure of culturable endophytic bacteria in Phthorimaea operculella(Zeller) at different developmental stages are different. The species of culturable endophytic bacteria in larvae are greatly higher than those in eggs and pupae, but there is no obvious difference in dominant species. The dominant species of endophytic bacteria can promote the growth and development of the host by promoting the digestion of larvae.

     

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