深海沉积物中产淀粉酶细菌的rep-PCR基因指纹分析

The rep-PCR Fingerprints Analysis of Amylase-producing Bacteria Isolated from Deep-sea Sediments

利用淀粉酶筛选培养基平板从27个深海沉积物样品中筛选得到30个具有胞外淀粉酶活性的菌株.其中革兰氏阴性细菌有22株,阳性细菌有8株.对这些细菌的基因组DNA进行ERICPCR和BOXPCR扩增,指纹图谱显示大部分菌株均存在数目不等的各自独特的带型,各特异性扩增的主带型能重复稳定出现.比较两种引物的指纹图谱,显示ERICPCR比BOXPCR具有较丰富的图谱多态性.对产生的指纹图谱进行聚类学分析,30株细菌可分为10组,其中5株细菌分别独立成组,最多的第Ⅱ组包含有8株细菌,而在同一聚类组内的细菌可能是处于进化上亲缘关系较近的位置.研究显示,30株菌具有丰富的种属多样性,揭示了深海微生物资源的丰富和潜力.ERICPCR和BOXPCR技术可用于对深海微生物群落组成和多样性的研究.

Thirty strains with the activity of extracellular amylase were screened from 27 deep-sea sediment samples using agar plating.Twenty two of these strains are Gram negative,and the remains are Gram positive.These bacteria were genotyped using ERIC-PCR and BOX-PCR techniques.The fingerprinting data showed that these strains had different banding patterns with various specificities,and the main banding pattern of each strain has a steady reproducibility.The fingerprints in ERIC-PCR using multiple primers showed more polymorphism than those of BOX-PCR using a single primer.Based on the fingerprints,a dendrogram was plotted.These 30 strains were classified into 10 groups.Of which,5 strains each formed a unique group respectively,while others clustered together in various groups.The largest groupⅡ included 8 strains.The strains in the same group might have close evolutionary relationship.Our studies demonstrated that these 30 strains had great diversity at genus and species level,indicating that there is rich and potential resource of microorganism in the deep sea bed.ERIC-PCR and BOX-PCR can be used for the research of population and diversity of deep-sea bed microorganism community.