贵州黔南喀斯特洞穴放线菌抗菌活性筛选及差异研究
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摘要
目的:探索贵州黔南荔波喀斯特地区洞穴放线菌资源,筛选具有抗菌活性的放线菌菌株。方法:采集贵州荔波茂兰喀斯特森林无名洞穴内土壤样品,采用纯培养的方法分离菌株,并扩增其16S rRNA基因、筛选其中放线菌;使用3种培养基发酵,管碟法对放线菌进行抗菌活性检测。结果:共分离出35株放线菌,分属于3个目、5科、9个属、22个种,红球菌属为优势菌属(45.7%);42.8%的菌株对至少一种测试菌具有抗菌活性;使用AM3及ISP5培养基获得具有抗性发酵液的比例均占53.3%,RA培养基只有33.3%的发酵液具有抗性。结论:贵州黔南喀斯特无名洞穴内存在丰富的具备潜在抗菌活性的放线菌资源,不同的发酵培养条件影响放线菌抗菌活性。
Objective: To explore local actinobacteria resources, and screen actinobacteria strains with antibacterial activity in Southern Guizhou China. Methods: Soil samples from unknown caves in Maolan forest were collected and isolated by pure culture. The 16S rRNA gene was amplified and the actinomycetes were screened. Five kinds of culture media were used for fermentation, and the antimicrobial activity of three clinical pathogens was detected by cylinder platr method. Results: A total of 35 actinomycetes from Karst caves were isolated, belonging to 3 orders, 5 families, 9 genera and 22 species. The antimicrobial activity of these actinomycetes were tested aganist pathogenic bacteria. The result suggested that 42.8% of these strains showed antimicrobial activity against at least one tested bacterium. The media used for the growth played a crucial role in the antimicrobial activity. The rate of the antimicrobial activity was highest in AM3 and ISP5(55.3%) followed by R2A(33.3%). Conclusion: There are abundant actinobacteria with antimicrobial activity in Karst caves. Fermentation media have influence on the antimicrobial activity.
Objective: To explore local actinobacteria resources, and screen actinobacteria strains with antibacterial activity in Southern Guizhou China. Methods: Soil samples from unknown caves in Maolan forest were collected and isolated by pure culture. The 16S rRNA gene was amplified and the actinomycetes were screened. Five kinds of culture media were used for fermentation, and the antimicrobial activity of three clinical pathogens was detected by cylinder platr method. Results: A total of 35 actinomycetes from Karst caves were isolated, belonging to 3 orders, 5 families, 9 genera and 22 species. The antimicrobial activity of these actinomycetes were tested aganist pathogenic bacteria. The result suggested that 42.8% of these strains showed antimicrobial activity against at least one tested bacterium. The media used for the growth played a crucial role in the antimicrobial activity. The rate of the antimicrobial activity was highest in AM3 and ISP5(55.3%) followed by R2A(33.3%). Conclusion: There are abundant actinobacteria with antimicrobial activity in Karst caves. Fermentation media have influence on the antimicrobial activity.
引文
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[22]唐彤彤,孙星,董元华,等.对番茄枯萎病具防效的海洋源赤杆菌YH-07的发酵条件探究[J].中国生物防治学报,2018,34(5):108-116.
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[24]张莉,倪孟祥.海洋放线菌AM8发酵条件的优化及抑菌活性物质的初步研究[J].化学与生物工程,2017(6):759-762.
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[2] LUO W,WANG S,LIU X.Research progresses and prospect of chimney effect about carbon cycle in the karst cave system[J].Advances in Earth Science,2014,29(12):1333-1340.
[3] 金方,刘涛华,牟丽丽,等.贵州地区土壤中需氧放线菌的分离鉴定[J].贵州农业科学,2015(3):114-117.
[4] 刘倩瑛.喀斯特地区微生物生态系统及其与气候因子之间的关系——以湖北清江和尚洞为例[D].武汉:中国地质大学,2010.
[5] 左俊辉,罗群,郭喜梅.中国地貌三大阶梯对喀斯特发育的若干影响——以中国西南岩溶区为例[J].科技信息,2012(18):210-211.
[6] LANE,D J.16S/23S rRNA sequencing [M].NewYork:John Wileyand Sons Inc,1991,115-175.
[7] MENG Y C,LIU H C,YANG L L,et al.Microbacterium sorbitolivorans sp.nov.a novel member of Microbacteriaceae isolated from fermentingbed in pigpen[J].Int J Syst Evol Microbiol,2016,66(12):5556.
[8] THOMPSON J D,GIBSON T J,PLEWNIAK F,et al.The CLUSTAL_X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Res,1997,25:4876-4882.
[9] YOON S H,HA S M,KWON S,et al.Introducing EzBio-Cloud:a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies[J].Int J Syst Evol Microbiol,2017,67:1613-1617.
[10]李青,刘华梅,陈振民,等.管碟法测定苏云金杆菌上清液中增效物质含量的研究[J].微生物学通报,2002,29(3):73-74.
[11]戴素娟,李静,刘少伟,等.河南神仙洞放线菌资源勘探及抗菌活性筛选[J].中国抗生素杂志,2015,40(9):1112-1115.
[12]程娟.云南喀斯特洞穴可培养放线菌多样性及其酶活筛选研究[D].昆明:云南大学,2014.
[13]祖健,刘少伟,庹利,等.湖北利川喀斯特洞穴放线菌多样性及抗菌活性研究[J].中国抗生素杂志,2016,41(3):378-381.
[14]KANG Y Q,MING H,GONOI T,et al.Gordonia iterans sp.nov.isolated from a patient with pneumonia[J].International Journal of Systematic & Evolutionary Microbiology,2014,64(10):3520-3525.
[15]王梅竹,王颜颜,曹煜,等.贵州地区149株临床白念珠菌对抗真菌药物的敏感性[J].贵阳医学院学报,2014,39(5):617-620.
[16]XIAO M,FAN X,CHEN C A,et al.Antifungal susceptibilities of Candida glabrata species complex,Candida krusei,Candida parapsilosis species complex and Candida tropicalis causing invasive candidiasis in China:3 year national surveillance[J].Journal of Antimicrobial Chemotherapy,2015,70(3):802-810.
[17]王梅竹,曹煜,赵亮,等.贵州地区临床酵母菌种类组成和分布[J].山东大学学(医学版),2014,52(8):94-97.
[18]刘波.临床患者深部真菌感染菌群分布情况及耐药性分析[J].中国当代医药,2017,3:135-137.
[19]AFSARIAN S M,BADALI H,SHOKOHI T,et al.Molecular diversity of Candida albicans isolated from immunocompromised patients,based on MLST method[J].Iran J Public Health,2015,44(9):1262-1269.
[20]GIBBERD R.Globocan 1:cancer incidence and mortality worldwide[J].Statistics in Medicine,2015,19(19):2714-2715.
[21]詹萍,李福连.产抗菌活性红树林放线菌G15发酵条件的优化[J].中国酿造,2012,31(7):77-80.
[22]唐彤彤,孙星,董元华,等.对番茄枯萎病具防效的海洋源赤杆菌YH-07的发酵条件探究[J].中国生物防治学报,2018,34(5):108-116.
[23]倪孟祥,毕启瑞,郭润妮,等.南极放线菌N-1产抑茵成分的响应面优化[J].中国海洋药物,2013,32(3):11-20.
[24]张莉,倪孟祥.海洋放线菌AM8发酵条件的优化及抑菌活性物质的初步研究[J].化学与生物工程,2017(6):759-762.
[25]姚蓉.海洋来源放线菌Kocuria sp.的次级代谢产物研究[D].杭州:浙江工业大学,2017.
[26]SA-UTH C,RATTANASENA P,CHANDRAPATYA A,et al.Modification of medium composition for enhancing the production of antifungal activity from,xenorhabdus stockiae,pb09 by using response surface methodology[J].International Journal of Microbiology,2018,2018:1-10.
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