硇洲岛牡蛎相关细菌抗菌筛选及系统发育分析

Screening & Phylogenetic Analysis of Oyster Related Bacterial Antibiosis in Naozhou Island

以8个敏感菌株作为指示菌,采用管碟法对分离自湛江硇洲岛(20°52'N^20°56'N,110°33'E^110°38'E)潮汐带香港巨牡蛎(Crassostrea hongkongensis)中的72株细菌的发酵液进行抗菌筛选,并对阳性菌株进行基因组DNA提取、16S rRNA基因PCR扩增和序列测定,继而进行系统发育分析。抗菌实验结果表明,受试菌株中有23株菌的发酵产物具有抗菌活性(阳性率31.9%),其中有5个菌株(JSM 111024、JSM 111027、JSM111029、JSM 111076、JSM 111083)具有较强的抗菌活性。基于16S rRNA基因序列的系统发育分析表明,这23株菌具有较高的类群多样性和物种多样性,属于3个系统发育群/门(Alpha-Proteobacteria、Gamma-Proteobacteria、Bacteroidetes)中的8个科(Aeromonadaceae、Flavobacteriaceae、Halomonadaceae、Idiomarinaceae、Phyllobacteriaceae、Pseudoalteromonadaceae、Shewanellaceae、Xanthomonadaceae)的8个属(Idiomarina、Halomonas、Myroides、Nitratireductor、Oceanimonas、Pseudoalteromonas、Shewanella、Wohlfahrtiimonas),可分为11个物种。优势类群为Gamma-Proteobacteria亚门(14株),其中优势属为Oceanimonas属(6株);第二大类群为Bacteroidetes门(7株),都属于Flavobacteriaceae科的Myroides属。具有较强抗菌活性的5个菌株中,有4个菌株(JSM 111024、JSM 111027、JSM 111029、JSM 111083)属于Alpha-Proteobacteria亚门Phyllobacteriaceae科Oceanimonas属,而菌株JSM 111076属于Gamma-Proteobacteria亚门Aeromonadaceae科Nitratireductor属。

The antibiosis screening of the fermentation broths of 72 marine bacteria,isolated from Hongkong giant oyster（Crassostrea hongkongensis） collected the tidal flat in Naozhou Island of Zhanjiang city（20°52＇ N ~ 20°56＇ N,110°33＇E ~ 110°38＇E） Guangdong Province,was carried out using cylinder-plate method with 8 sensitive strains as indicators（3 Gram-positive and 3 Gram-negative bacteria,and 2 fungi）,and extract DNA from positive strains＇ genome,carried out 16 S rRNA genes PCR amplification and sequencing,and then systematic phylogenetic analysis.The results of antibiotic experiments showed that among them fermentation broth of 23 tested strains possessed antibiotic activity（positive rate at 31.9%）,of which 5 strains（JSM111024,JSM111027,JSM111029,JSM111076,JSM111083） exhibited strong antibiotic activity.Based on 16 S rRNA sequence and phylogenetic analysis suggested the 23 strains possessed group diversity and species diversity,they belonged to 8 genera（Idiomarina,Halomonas,Wohlfahrtiimonas,Pseudoalteromonas,Shewanella,Oceanimonas,Nitratireductor,Myroides） of 8 families（Aeromonadaceae,Flavobacteriaceae,Halomonadaceae,Idiomarinaceae,Phyllobacteriaceae,Pseudoalteromonadaceae,Shewanellaceae,Xanthomonadaceae） in 3 phylogenetic groups / phyla（Alpha-Proteobacteria,Gamma-Proteobacteria,Bacteroidetes）,and divided into 11 species.And the dominant groups were Gamma-Proteobacteria subphylum（14strains）,among them the dominant genus was Oceanimonas（6 strains）,and the second large groups was the phylum of Bacteroidetes（7 strains）,all belonged to the genus of Myroides of the family of Flacobacteriaceae.There were four strains（JSM 111024,JSM 111027,JSM 111029,JSM 111083） among 5 strains possessing strong antibiotic activity,belonged to the genus Oceanimonas of the family of Phyllobacteriaceae,subphylum of Alpha-Proteobacteria,and strain JSM 111076 belonged to the genus Nitratireductor of the family of Aeromonadaceae,subphylum of Gamma-Proteobacteria.