Yunnan Provincial Key Laboratory of Animal and Feed Science, Yunnan Agricultural University, Kunming, 650201, China.
Mol Biol Rep. 2011 Jan;38(1):577-85. doi: 10.1007/s11033-010-0143-x. Epub 2010 Apr 1.
Library of ruminal protozoal 18S rRNA of Yunnan Yellow Cattle has been constructed in the present study. Phylogenic analysis of sequences was meanwhile employed to reveal the diversity of protozoa in the rumen of Yunnan Yellow Cattle. One Yellow Cattle was fed malt meal (YCRPB) and the other was fed wheat straw (YCRPS). A protozoa-specific primer (P-SSU-342f) and a eukarya-specific primer (Medlin B) were used to amplify a 1,360-bp fragment of DNA encoding protozoal small subunit (SSU) ribosomal RNA from rumen fluid. The results showed as follows: A total of 121 clones were obtained and fell into four genera identified as Entodinium (66.9%), Dasytricha (5.8%), Isotricha (9.1%), and Diplodinium (18.2%). Within the genus Entidinium, 48 of the YCRPB sequences and 33 of the YCRPS sequences clustered with the Entodinium caudatum. 7 of the YCRPB sequences were phylogenetically placed within the genus Dasytricha. 11 of the YCRPB sequences were related with high confidence to Isotricha intestinalis. 22 of the YCRPS sequences were phylogenetically placed within the genus Diplodinium. The predominant protozoal genus identified in the rumen fluid belonged to the Entodinium group, and the divergences between two cattle may due to diet and individual differences.
本研究构建了云南黄牛瘤胃原生动物 18S rRNA 文库。同时，对序列进行系统发育分析，揭示了云南黄牛瘤胃原生动物的多样性。一头黄牛饲喂麦芽（YCRPB），另一头饲喂麦秸（YCRPS）。使用原生动物特异性引物（P-SSU-342f）和真核生物特异性引物（Medlin B）从瘤胃液中扩增编码原生动物小亚基（SSU）核糖体 RNA 的 1360bp DNA 片段。结果如下：共获得 121 个克隆，分为 4 个属，分别为 Entodinium（66.9%）、Dasytricha（5.8%）、Isotricha（9.1%）和 Diplodinium（18.2%）。在 Entodinium 属中，YCRPB 序列的 48 个和 YCRPS 序列的 33 个与 Entodinium caudatum 聚类。YCRPB 序列中有 7 个系统发育上位于 Dasytricha 属内。YCRPB 序列中有 11 个与 Isotricha intestinalis 高度相关。YCRPS 序列中有 22 个系统发育上位于 Diplodinium 属内。瘤胃液中主要的原生动物属属于 Entodinium 组，两种牛之间的差异可能与饮食和个体差异有关。
