ITS1 rDNA序列用于瘤胃原虫研究可行性的探讨

文章主要探讨瘤胃原虫rDNA序列的ITS1区段用于其群体研究的可行性和有效性。试验1由3只瘘管山羊提供瘤胃液、以不同结构碳水化合物底物(可溶性淀粉/滤纸纤维:A,100:0、B,70:30、C,50:50、D,30:70、E,0:100)进行体外培养;试验2以4只瘘管山羊为试验动物,以A(鱼粉)、B(羽毛粉)、C(玉米蛋白粉)和D(豆粕)等蛋白质饲料配制的4种等氮日粮进行4×4拉丁方试验,采用ITS1rDNA序列的遗传指纹检测及细胞计数技术研究原虫群体组成的变化。遗传指纹检测结果表明,在试验1不同碳水化合物结构底物培养条件下或在试验2不同蛋白日粮条件下,原虫群体组成都发生了变化。细胞计数的研究结果与之一致。依据本研究结果可见,ITS1rDNA序列是原虫研究的良好素材。

The feasibility of ITS1 rDNA sequence on the research of ruminal protozoa

The objective of this paper was to investigate the feasibility and validity of ITS1 rDNA for the research of protozoal community in the rumen. The experiments included 2 trails, in vitro and in vivo trails. In vitro culture was carried out using 3 goats fisted with cannulas, and the treatment were designed by varying the rations of starch to filter paper ratio here: A (100: 0), B (70: 30), C (50: 50), D (30: 70), and E (0: 100) respectively. 4 goats were assigned in a 4×4 Latin square design in the in vivo experiment, and isonitrogen formula diets were divided into 4 groups according to their nitrogen source, which was, respectively, A (Fish meal), B (Feather meal), C (Corn gluten meal), and D (Soybean meal). The genetics analysis of ITS1 rDNA and cell -counting technique were introduced in current article to determine protozoal community.The results showed that: protozoa populations were manipulated by substrates, either the in vitro or the in vivo trail experiment, through the analysis of genetic fingerprint of ITS1 rDNA. And the same results were also obtained by cell -counting technique, which agreed with the genetic analysis of ITS1 rDNA.It was therefore concluded that, ITS1 rDNA has a potential for the research on characterizing rumen protozoa.