新疆高盐极端环境可培养放线菌的的种群及其生物活性初步研究

Preliminary studies on population and bioactivity of educable actinomycetes from hyperhaline extreme environment in xinjiang

目的积累微生物资源,研究从土壤中选择性分离放线菌的种群及其次生代谢产物的生物活性评价。方法采用两种分离方法,4种高盐培养基对来自新疆高盐环境-阿勒泰福海县盐场和罗布泊地区等采集的土壤样品中的放线菌进行了选择性分离纯化;以3种致病真菌灰葡萄孢菌(Botrytis cinerea)、番茄早疫病菌(Alternaria solani)、稻瘟病菌(Magnaporth grisea)和4种致病细菌大肠埃希氏(Escherichia coli)、金黄色葡萄球菌(Staphy lococcus aureus)、枯草芽孢杆菌(Bacillus subtilis),草分枝杆菌(Mycobacterium phlei)为指示菌,对其中分离的100株放线菌发酵液及其乙酸乙酯提取物进行抗菌活性筛选。同时利用一种秀丽隐杆线虫(Caenorhabditis elegans)进行抗线虫活性实验,并采用16SrDNA基因序列对部分具有抗活性的放线菌以及具有特殊的菌落形态的放线菌进行系统发育分析。结果通过选择性分离方法共分离到123株放线菌,其中47株放线菌来自阿勒泰福海县盐场土壤样品,67株来自罗布泊地区土壤样品。选择100株具有代表性的菌株进行次生代谢产物的生物活性筛选及抗线虫活性追踪,其结果表现为43株菌株在其中一个或两个活性筛选模型中都显示为阳性,总阳性率为43%;其对具有特殊的菌落形态特征及其次生代谢产物具有活性的61株放线菌进行16SrRNA分析,结果表明,覆盖7属;32株属于拟诺卡氏菌属(Nocardiopsis),17株属于链霉菌属(Streptomyces),7株属于普氏菌属(Prauserella),2株属于嗜盐糖多孢菌属(Saccharopolyspora Halophilic),1株属于假诺卡氏(Pseudonocardia),1株属于放线菌属(Actinomyces)和1株属于高温单孢菌属(Thermomonospora);有1株为潜在新种。结论新疆高盐环境-阿勒泰福海县盐场和罗布泊地区中存在较为丰富的放线菌资源并能产生多种活性次级代谢产物,值得进一步的研究。

Objective To study the population and bioactivity of actinomycetes from soil in hyperhaline region of Xingjang ,China and accumulate microbial resources .Methods Using two isolation methods ,four different kinds of cultural media ,123 isolates were purified from twenty e soil samples in salt field of Fuhai county of Altay and Lopnor area .100 among 123 strains were cultured and both of their cultural broths and its organic extracts were assayed by antibacterial andnematicide model .Based on the morphological characteristics and biological activity ,61 strains were selected to carry out the 16S rRNA sequences test .Results 100 isolates were purified ,43strains exhibi-ted positive Results in one or two screening model ,the total positive rate was 43% .the 16S rRNA sequences test showed that the 61 strains belongs to 7 different genera ,including Streptomyces ,Nocardiopsis ,Pseudonocardia ,Prauserella ,Saccharopolyspora Halophilic , Actinomyces and Thermomonospora .The sequence similarity of strain L-068 may be a potential new species .Conclusion The studies dem-onstrated there are untapped actinomycetal resources e in hyperhaline region of Xingjang ,China and its bioactive secondary metabolites de-served further exploration for microbial drugs .