中国农业科学 ›› 2009, Vol. 42 ›› Issue (6): 2126-2132 .doi: 10.3864/j.issn.0578-1752.2009.06.032

• 畜牧·资源昆虫 • 上一篇    下一篇

蒙古绵羊瘤胃固、液相附着微生物的RT-PCR检测

杨金丽,侯先志,高爱武   

  1. (内蒙古农业大学动物科学与医学学院)
  • 收稿日期:2008-09-23 修回日期:2008-12-08 出版日期:2009-06-10 发布日期:2009-06-10
  • 通讯作者: 侯先志

Quantity of Microorganisms Associated with Solid and Liquid Phases of Rumen Contents Using RT- PCR

YANG Jin-li, HOU Xian-zhi, GAO Ai-wu   

  1. (内蒙古农业大学动物科学与医学学院)
  • Received:2008-09-23 Revised:2008-12-08 Online:2009-06-10 Published:2009-06-10
  • Contact: HOU Xian-zhi

摘要:

【目的】应用Real-time PCR(RT-PCR)对蒙古绵羊瘤胃内容物固、液相附着微生物的生态分布进行定量研究。【方法】采集3只蒙古绵羊瘤胃内容物,固液分离后对各相中所附着的微生物进行定量检测,包括总细菌、总厌氧真菌和3种主要纤维降解菌-白色瘤胃球菌Ruminococcus albus、黄色瘤胃球菌Ruminococcus flavefaciens、产琥珀酸拟杆菌Fibrobacter succinogenes。【结果】固相中总细菌、总厌氧真菌和3种纤维降解菌的16S或18S rRNA 基因考贝数显著高于液相,分别是液相附着微生物的7.22倍(P<0.05)、56.85倍(P<0.01)、2.69倍(P<0.05)、4.19倍(P<0.01)和7.59倍(P<0.01)。瘤胃固相中F.succinogenes 菌的丰度为0.55%,高于R.flavefaciens 菌(0.53%)和R.albus菌(0.09%)。瘤胃液相中R.flavefaciens菌的丰度最高,为0.91%,高于F.succinogenes菌(0.52%)和R.albus 菌(0.24%)。【结论】RT-PCR方法能够准确对瘤胃固、液相附着微生物进行定量分析。

关键词: Real-time PCR, 蒙古绵羊, 瘤胃, 微生物, 生态分布

Abstract:

【Objective】 Using Real-time PCR (RT-PCR)assays to quantify the microbes associated with particular phases of rumen contents. 【Method】 Whole rumen contents were sampled from three Mongolian sheep and partitioned into solid- and liquid-associated microorganisms. 【Result】 On the basis of 16S or 18S rRNA gene copy number analysis, the number of general bacteria, general anaerobic fungi and three representative ruminal cellulolytic bacteria (Ruminococcus albus, Ruminococcus flavefaciens, Fibrobacter succinogenes) associated with rumen solid content were 7.22-fold (P<0.05), 56.85-fold (P<0.01), 2.69-fold (P<0.05), 4.19-fold (P<0.01) and 7.59-fold (P<0.01) as those associated with rumen liquid content. The abundance of F. succinogenes in the solid phase was 0.55%, which was higher than that of R. flavefaciens (0.53%) and R. albus (0.09%). However in the liquid phase R. flavefaciens accounted for 0.91% of the bacterial rRNA gene copies, which was more abundant than F. succinogenes (0.52%) and R. albus (0.24%). 【Conclusion】 RT-PCR method could accurately detect the distribution and abundance of microorganisms associated with particular phase of rumen content.

Key words: Real-time PCR, Mongolia sheep, rumen, microorganisms, distribution