黑腹果蝇和斑翅果蝇肠道可培养的细菌多样性分析
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摘要
为了了解斑翅果蝇Drosophila suzukii和黑腹果蝇D. melanogaster可培养的肠道细菌多样性。本研究将2种果蝇卵中及幼虫、蛹和成虫肠道中的细菌分别分离纯化,并利用16srDNA基因分析的方法鉴定。结果显示,在2种果蝇的不同发育阶段中,卵中的细菌数量最少,而幼虫肠道中最多,且斑翅果蝇中细菌的数量显著多于黑腹果蝇。2种果蝇共分离出31种细菌,以变形菌门细菌为主。斑翅果蝇和黑腹果蝇中细菌的种类分别为20和21种,其中相同种类的细菌为13种。斑翅果蝇肠道内比较稳定的细菌种类为弗氏柠檬酸杆菌Citrobacter freundii、产酸克雷伯菌Klebsiella oxytoca、金黄杆菌属Chryseobacterium sp.、形赖氨酸芽孢杆菌Lysinibacillus fusiformis,黑腹果蝇肠道内比较稳定的细菌种类为醋酸菌Acetobacter thailandicus、类芽孢杆菌Paenibacillus taichungensis、雷氏普罗威登斯Providencia rettgeri、摩氏摩根菌Morganella morganii。因此,不同发育阶段的斑翅果蝇和黑腹果蝇肠道中细菌的数量和种类均有差别。
To understand the intestinal bacterial diversity in Drosophila suzukii and D. melanogaster, the bacteria in eggs, the intestine of larvae, pupae and adults of two Drosophila species were separated and purified, and identified with 16 S rDNA genetic analysis. The results showed that, the number of bacteria was the lowest in eggs and the highest in larval intestine at different developmental stages of the two Drosophila species. And the number of bacteria in D. suzukii was more than that in D. melanogaster significantly. 31 species of bacteria were isolated from the two Drosophila species, the bacteria of Proteobacteria was the dominant population. The species of bacteria were 20 and 21 in D. suzukii and D. melanogaster, respectively.Therein, there were 13 species in both the two Drosophila species. The stable bacteria in intestine of D. suzukii were Citrobacter freundii, Klebsiella oxytoca, Chryseobacterium sp. and Lysinibacillus fusiformis. While,Acetobacter thailandicus, Paenibacillus taichungensis, Providencia rettgeri and Morganella morganii were stable bacteria in intestine of D. melanogaster. It is concluded that, the number and species of bacteria in D.suzukii and D. melanogaster are different at different development stages.
To understand the intestinal bacterial diversity in Drosophila suzukii and D. melanogaster, the bacteria in eggs, the intestine of larvae, pupae and adults of two Drosophila species were separated and purified, and identified with 16 S rDNA genetic analysis. The results showed that, the number of bacteria was the lowest in eggs and the highest in larval intestine at different developmental stages of the two Drosophila species. And the number of bacteria in D. suzukii was more than that in D. melanogaster significantly. 31 species of bacteria were isolated from the two Drosophila species, the bacteria of Proteobacteria was the dominant population. The species of bacteria were 20 and 21 in D. suzukii and D. melanogaster, respectively.Therein, there were 13 species in both the two Drosophila species. The stable bacteria in intestine of D. suzukii were Citrobacter freundii, Klebsiella oxytoca, Chryseobacterium sp. and Lysinibacillus fusiformis. While,Acetobacter thailandicus, Paenibacillus taichungensis, Providencia rettgeri and Morganella morganii were stable bacteria in intestine of D. melanogaster. It is concluded that, the number and species of bacteria in D.suzukii and D. melanogaster are different at different development stages.
引文
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[3]Bolda M P,Goodhue R E,Zalom F G.Spotted wing Drosophila:potential economic impact of a newly established pest[J].Journal of Agricultural and Resource Economics,2010,13(3):5-8.
[4]林清彩,王圣印,周成,等.铃木氏果蝇研究进展[J].江西农业学报,2013,25(6):45-47.
[5]Mitsui H,Takahashi H K,Kimura M T.Spatial distributions and clutch sizes of Drosophila species ovipositing on cherry fruits of different stages[J].Population Ecology,2006,48:233-237.
[6]David J R,Vanherrewege J.Adaptation to alcoholic fermentation in Drosophila species:Relationship between alcohol tolerance and larval habitat[J].Comparative Biochemistry Physiology,1983,74:283-288.
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[8]Ryu J H,Kim S H,Lee H Y.Innate immune homeostasis by the homeobox gene caudal and commensal-gut mutualism in Drosophila[J].Science,2008,319:777-782.
[9]Shi S C,Kim S H,You H.Drosophila microbiome modulates host developmental and metabolic homeostasis via insulin signaling[J].Science,2011,334:670-674.
[10]Fischer C,Trautman E P,Crawford J M.Metabolite exchange between microbiome members produces compounds that influence Drosophila behavior[J].elife,2017,136(1):1-11.
[11]Poisot T,Bever J D,Nemri A.A conceptual framework for the evolution of ecological specialisation[J].Ecolgy Letters,2011,14:841-851.
[12]Becher P G,Flick G,Rozpedowska E.Yeast,not fruit volatiles mediate Drosophila melanogaster attraction,oviposition and development[J].Functional Ecology,2012,26:822-828.
[13]Atallah J,Teixeira,L,Salazar R.The making of a pest:the evolution of a fruit-penetrating ovipositor in Drosophila suzukii and related species[J].Proceedings of the Royal Society of London Series B.Biological Science,2014,281:2013-2840.
[14]Begon M.Yeasts and Drosophila.The Genetics and Biology of Drosophila[M].Academic Press,U.K.,1983:345-384.
[15]Douglas A E.The microbial dimension in insect nutritional ecology[J].Functional Ecology,2009,23:38-47.
[16]高欢欢,翟一凡,王海艳,等.韭菜迟眼蕈蚊体内及韭菜根际微生物的多样性分析[J].应用昆虫学报,2015,52(4):1006-1013.
[17]刘莉,王中康,俞和伟,等.贡嘎蝠蛾幼虫肠道细菌多样性分析[J].微生物学报,2008,48(5):616-622.
[18]Xiang H,Wei G F,Jia S,et al.Microbial community in the larval midgut of laboratory and fieldpopulations of cotton bollworm(Helicoverpaarmigera)[J].Canadian Journal of Microbiology,2006,52(11):1085-1092.
[19]相辉,李木旺,赵勇,等.家蚕幼虫中肠细菌群落多样性的PCR-DGGE和16SrDNA文库序列分析[J].昆虫学报,2007,50(3):222-233.
[20]刘玉升,李明立.东亚飞蝗肠道细菌的研究[J].中国微生态杂志,2007,19(1):34-39.
[21]Bourtzis K,Miller T A.Insect symbiosis,vol.3.Contemporary topics in entomology[M].Boca Raton,CRC Press,2009.