16SrDNA-RFLP分析繁茂膜海绵可培养放线菌的多样性

16S rDNA-RFLP analysis of actinomycetes associated with marine sponge Hymeniacidon perleve

海绵是迄今为止已知海洋天然产物的最大来源。由于海绵的底栖过滤性摄食和消化选择性等生理特性,其体内蕴藏了丰富的微生物种群。近几年来,发现越来越多的海绵微生物产生很强的生物活性物质,有些并被证明是海绵天然产物的真正生产者。对大连海域繁茂膜海绵中的放线菌进行分离,对于得到的菌株进行16SrDNA扩增和RFLP及测序分析。研究表明海绵中蕴含着丰富的放线菌资源。其中包含链霉菌(Streptomycetes),拟诺卡氏菌(Nocardiopsis),假诺卡氏菌(Pseudonocardia),诺卡氏菌(Nocardia),小单孢菌(Micromonospora),红球菌(Rhodococcus),异壁放线菌(Actinoalloteichus)等属。利用限制性内切酶HhaⅠ对16SrDNA进行的RFLP分析能够对海绵中放线菌的16SrDNA多样性进行有效的分析,结果达到属,部分到种的级别。揭示了繁茂膜海绵中蕴藏着丰富的放线菌资源。

106 culturable symbiotic actinomycetes strains were isolated from intertidal marine sponge Hymeniacidon perleve collected from Chinese Yellow Sea. The 16S rDNA sequences of these strains were amplified and RFLP patterns were analyzed by digestion of 16S rDNA fragments with Hha Ⅰ . 11 different patterns were identified, with confirmation by the RFLP analysis using Vector NTI simulation. Using Hha Ⅰ RFLP patterns, the actinomycetes could be identified at the genus level. Phylogentic trees using 16S rDNA sequences were constructed. Majority of actinomycetes belonged to Streptomycetes （ 78 ）. The other strains belonged to Nocardiopsis （ 18 ）, Pseudonocardia （ 3 ）, Nocardia （ 3 ）, Micromonospora （2）, Rhodococcus （1）, Actinoalloteichus （1）. Strain 177 is the candidate for new species of Actinoalloteichus .