PCR-DGGE技术分析断奶仔兔肠道微生物菌群结构及多样性

利用PCR-DGGE技术对断奶后10、20、30 d仔兔的十二指肠、空肠、回肠、盲肠和结肠16S rDNA V3区基因进行图谱分析,通过对清晰的特异性和共性条带切胶回收DNA和克隆测序,对不同日龄和不同肠段内容物细菌群落的结构和多样性进行比较,并鉴定其群落成员,分析断奶仔兔不同日龄和肠段部位对微生物群落结构与多样性的影响.肠道菌群DGGE图谱显示断奶仔兔肠道部位和不同日龄对细菌种群结构和多样性影响很大,随着日龄增加,十二指肠和盲肠段微生物种类趋于复杂,空肠和结肠段经历由简单到复杂再到简单的过程,而回肠段肠道微生物则出现由复杂到简单再到复杂的过程.各肠段的优势细菌有厚壁菌门、拟杆菌、梭状杆菌、博德氏菌属、毛螺旋菌科,以及各种不可培养的细菌等.

Using PCR-DGGE technical analysis weaned rabbit structure and di versity of intestinal microflora

In this paper,mapping analysis of the genes that are exsisted in the 16S rDNA V3 districts of duodena,jejuna,ilea,caeca and cola of the rabbits which have been weaned 10,20 and 30 days respectively,is carried out by using PCR-DGGE thechnology.Through the clear specificity and generality of bands cutting rubber recycling of DNA and cloning sequencing,the comparisons of the structure and the diversity between bacterial communities,which are in different parts of the intestines and belong to the rabbits of differdent days have been done.Also,their community members have been ascertained.More importantly,it analyzed the influence of the days that the rabbits had been weaned and the different parts of the intestines on the structure and diversity of the micropopulation.On the basis of the DGGE map of the bacterial community in the intestines,it could be seen obviously that different parts of the intestines,as well as the days that the rabbits had been weaned,had great effects on the structure and diversity of the bacterial community.Specifically,there were varying degrees of dynamic changes: with the increase of the days,the types of the bacterial communities in the duodena and caeca were more varied;those in the jejuna and cola had experienced a transversion of "scarce to abundant to scarce",while those in the ilea was on the contrary.From the DGGE map,we could conclude that the dominant bacteria in different parts of the intestines were Firmicutes,Bacteroidetes,Clostridia,Azohydromonas sp,Lachnospiraceae and many uncultured bacteria and so on.In conclution,the research results indicated: the days that the rabbits had been weaned and the different parts of their intestines did have effects on the structure and diversity of the bacterial community.