摘要
"新吉富"(NEW GIFT)尼罗罗非鱼是以1994年引进的"吉富"(GIFT)品系尼罗罗非鱼为基础群体,经过14年9代系统选育后获得的新品种,该品种已被国家审定为良种,已在全国推广,迫切需要有效的分子遗传标记来鉴别该新品种并实施科学管理。该研究对良种群体开展了RAPD分析,并进一步将特异性的RAPD标记转化成了稳定的SCAR标记。通过对"新吉富"群体的RAPD分析,找到了两个品种特异性的RAPD条带(S304624bp和S36568bp)。对品种特异性的RAPD条带进行了胶回收、克隆和序列测定。根据测序结果设计了两对位点特异性引物来扩增这两个品种特异性条带(简称SCAR标记Ⅰ和Ⅱ),检测这两个SCAR标记在"新吉富"、"吉富"以及国内外7个养殖品系中的出现频率。SCAR标记Ⅰ(553bp)在"新吉富"群体中的出现频率为85.7%,而在其基础群体("吉富")中的出现频率仅为16.7%;SCAR标记Ⅱ(558bp)在"新吉富"群体中的出现频率达到91.4%,但在其它7个养殖品系中的出现频率仅在0%～70%之间。为验证这两个标记的可靠性,检测了这两个标记在一个埃及罗非鱼野生群体中的出现频率,发现SCAR标记Ⅰ和SCAR标记Ⅱ的出现频率分别为10%和70%,远低于在"新吉富"群体中的出现频率。这两个SCAR标记在"新吉富"良种群体中的高出现频率,预示它们可能与生长性能相关QTL位点间存在连锁关系,显示SCAR标记技术对选育世代间的纵向追溯、以及选育良种群体与其它养殖品系间的横向鉴别有较好应用前景。
The NEW GIFT Nile tilapia （Oreochromis niloticus niloticus L.） is a nationally certificated new strain selected over 14 years and 9 generations from the base strain of GIFT Nile tilapia, introduced in 1994. This new variety has been extended in most of areas of China. The management of genetically improved strains, including the genetic markers for identification is needed urgently. RAPD analysis was conducted and their conversion to SCAR markers was developed. From NEW GIFT Nile tilapia, two strain-specific RAPD bands, S304624 bp and S36568 bp were identified. The strain-specific RAPD bands were gel-purified, cloned, and sequenced. Locus-specific primers were then designed to amplify the strain-specific bands. PCR amplification was conducted to test the variations in allele frequencies of two converted SCAR markers among the NEW GIFT Nile tilapia and its base strains, as well as 7 additional farmed strains worldwide. The frequency of SCAR marker Ⅰ （553 bp） was 85.7% in NEW GIFT Nile tilapia, but 16.7% in the base strain. The frequency of SCAR marker Ⅱ （558 bp） was 91.4% in NEW GIFT Nile tilapia, but 0% 70% in the 7 other strains. In order to confirm the utility of these two markers, an examination was conducted for a wild population from Egypt, resulted the frequency of SCAR Ⅰ and Ⅱ was 10% and 70%, respectively, much lower than that of New GIFT strain. The increase in allele frequency of these two SCAR markers suggests that these markers might be genetically linked to the quantitative trait loci （QTL） underlining the performance traits by long term selection, and indicate the bright potential of SCAR marker technology for tracking generations during selection progress and for distinguishing among genetically improved strain and other strains.
基金
National Tilapia Industry Technical System(nycytx-48-3)
National Tilapia Seed Program(nyhyzx07-044-01)
