江苏盐城沿海滩涂湿地放线菌多样性及功能基因的筛选

Diversity and biosynthetic gene of culturable Actinobacteria isolated from beach in Yancheng of Jiangsu province

目的研究江苏盐城沿海滩涂湿地中可培养放线菌的多样性,并比较不同分离培养基对该批样品放线菌的分离效果以及进行NRPS、PKS I、PKS Ⅱ功能基因筛选。方法使用6种分离培养基,对采集的4份土壤样品中的放线菌进行选择性分离。通过16S rRNA 基因序列分析所分离得到的放线菌多样性。运用PCR技术检测菌株合成相关基因的情况。结果菌株去重复后,分离得到放线菌共134株,主要分布于放线菌域的4个目7个科12个属中,除优势菌链霉菌属( Streptomyces )外,还分离到微杆菌属( Microbacterium )、节杆菌属( Arthrobacter )、 Paenarthrobacter 等稀有放线菌属;菌株BY5-10与最近有效发表菌株 Agromyces binzhouensis Oact353^T (GenBank 登录号:KC493987)的16S rRNA基因序列相似率为 98.35%,可能为放线菌科壤霉菌属的潜在新种;菌株EY6-7与最近有效发表菌株 Mycetocola manganoxydans MB1-14^T (GenBank 登录号:GU217690)的16S rRNA基因序列相似率为99.14%,可能为诺卡氏菌科 Mycetocola 的潜在新种;53株菌存在至少1种所探测的生物合成基因簇,阳性率为87.3%。结论江苏盐城沿海滩涂湿地中有着多样性丰富的放线菌资源,具有从中发现放线菌新物种及新抗生素的潜力。

Objective The diversity of cultured Actinobacteria isolated from beach in Yancheng of Jiangsu province and their potential application were investigated by this study. Methods Actinobacteria were isolated and purified from a total of 4 beach samples by traditional plate dilution method using 6 isolation media.To determine the diversity of cultivable Actinobacteria in beach 16S rRNA gene sequences of strains were obtained by PCR amplification and direct sequencing,then compared with EzBioCloud database and the phylogenetic tree was established by the neighbor-joining method.Screening of PKSI,PKSⅡ and NRPS genes was carried out by PCR and agarose electrophoresis. Results A total of 134 strains were Actinobacteria and affiliated to 12 genera of 7 families,including genera Streptomyces , Microbacterium , Arthrobacter , Paenarthrobacter , Agromyces , Curtobacterium , Micromonospora , Dietzia , Pseudarthrobacter , Rhodococcus , Isoptericola , Mycetocola ,in which Streptomyces was the dominant genus. Conclusion The study demonstrates that the abundance of culturable actinobacteria isolated from beach in Yancheng of Jiangsu province could provide further exploration and research for new Actinobacteria and new antibiotics.