【摘要】 应用PCR/DGGE技术对山羊采食前后瘤胃原虫的多样性随时间的变化进行了研究。采食前和采食后1、2、4和8h分别采集瘤胃内容物样品,对瘤胃原虫特异性的18S rRNA基因经PCR扩增、DGGE电泳分析。结果显示,采食前后DGGE图谱由10条左右清晰可辨的谱带组成,图谱上的泳带反映了瘤胃内的优势原虫,泳带数量和位置的复杂性说明了瘤胃原虫的多样性。并对其中两个优势条带切胶测序,基因序列分析分属于毛口目和内毛目。瘤胃原虫区系相似性分析,采食前为90.5%～94.1%;采食后相似性发生改变,且存在个体差异。多样性指数由采食前的0.82逐渐上升到采食后8h的0.90(P>0.05)。DGGE指纹技术有效地反映了不同样品中原虫优势种的组成,并可通过此技术跟踪研究瘤胃原虫区系在不同条件下的动态变化。更多还原

【Abstract】 The goats rumen protozoal communities were investigated,and the change of which were monitored by using PCR/DGGE technique based on 18S rRNA gene.The protozoal diversity and similarity were determined on the base of the position and number of bands in DGGE gel.The results showed that over ten bright bands were showed and differences were observed for four goats immediately prior to feeding and after feeding at 1,2,4 and 8 hour.Different profiles in gel were observed,the amount and position of bands represented respectively the rumen predominant protozoal species and community variation of different samples.The similarity indices of the protozoa were between 90.5%~94.1%,while variation of the similarity were found after feeding.These indicate that the components and the changes of the rumen predominant protozoa can be reflected effectively by using PCR/DGGE technique,tracing and researching the variation of the protozoal community at different situation.更多还原