论文部分内容阅读
【目的】研究刺参养殖环境中蛭弧菌类微生物的多样性及其组成特征。【方法】对刺参养殖用水进行过滤,获得微生物并提取其总DNA,分别采用两对特异性引物Per和Bac扩增蛭弧菌类微生物相应的16S r RNA基因目的片段。通过核糖体DNA扩增片段限制性内切酶分析(ARDRA)方法,使用HaeⅢ和MspⅠ,双酶切各60个目的基因的单克隆,选不同条带克隆进行测序并对测序结果的多重序列分析比对。【结果】确定两对引物分别存在8和9种碱基差异序列,均属于噬菌弧菌属,分属于3个类群中。类群Ⅰ、Ⅱ为优势类群,且均为已知类群,类群Ⅲ为潜在的新类群;Per1(KP214541)和Bac44(KP214551)代表菌株分别为优势种。【结论】刺参养殖环境中蛭弧菌具有较高的多样性,包括已知类型和未知类型的蛭弧菌;可进一步进行蛭弧菌的分离、培养,用于刺参养殖中病害防治研究。
【Objective】 The objective of this study was to investigate the diversity and composition of Bdellovibrio-like microorganisms in the culture environment of sea cucumber. 【Method】 The objective of this study was to filter the microorganisms and extract the total DNA of sea cucumber sea cucumber. Two pairs of specific primers Per and Bac were used to amplify the corresponding 16S rRNA gene fragments of Bdellovibrio. Through the restriction endonuclease analysis (ARDRA) method of ribosomal DNA amplification, using Hae Ⅲ and Msp Ⅰ, double digestion of the 60 gene of each target monoclonal, selected different bands were sequenced and the sequencing results of multiple sequence analysis Comparison. 【Result】 The results showed that there were 8 and 9 different nucleotide sequences in two pairs of primers, belonging to the bacteriophage Vibrio, belonging to three groups. Groups I and II were the dominant groups, and all of them were known groups. Group III was a potential new taxa. Representative strains of Perl (KP214541) and Bac44 (KP214551) were the dominant species. [Conclusion] The Bdellovibrio was highly diverse in the environment of sea cucumber Apostichopus japonicus, including Bdellovibrio of known type and unknown type. The isolation and culture of Bdellovibrio could be further used to prevent and cure the disease in sea cucumber .