利用16s rDNA方法检测刺参消化道细菌种类

Study on the bacterial diversity in the digestive tract of Apostichopus japonicus by 16s rDNA method

以2007年7月采自大连瓦房店养殖厂的刺参为实验材料,通过对刺参肠道中的细菌16s rDNA V3区基因进行扩增、克隆测序及序列同源性分析,对其种类进行初步研究。本研究测序获得11条16s rDNA V3区基因序列,并进行Blast同源比对,确定序列同源性,分析出11种细菌,三种梭菌属(Clostridium)细菌,两种为假单胞菌属细菌(Pseudomonas),一种产丙酸菌属细菌(Propionigenium),一种为金黄色葡萄球菌(Staphylococcus aureus),有六种细菌分别与4种不可培养的未命名细菌同源,并根据测序结果建立系统发生树。本研究提供了一种检测海洋微生物的方法,为刺参病害诊断提供科学依据,为海洋微生物资源的开发和新种的发现提供了资料。

In this work,by amplifying and analyzing the 16s rDNA genes based on the obtained eleven sequences,the bacteria were identified in the digestive tract of Apostichopus japonicus. And eleven bacteria were obtained, among which three samples belong to Clostridium bacteria, two are Pseudomonas,one is Propionigenium, and the rest one is Staphylococcus aureus. Six and 4 different kinds of uncultured bacteria have homologous. Based on these results ,a phylogenetie tree was constructed. The results provide helpful information for the Apostichopus japonicus disease diagnosis ,and even for the exploitation of maline microbe resource.