龙须菜RSAP分析及其SCAR标记的转化  

RSAP Analysis on Gracilariopsis lemaneiformis and Conversion of SCAR Marker

作　　者：王津果[1] 隋正红[1] 周伟[1] 马金华[1] 张淑[1] 常连鹏[1] 

机构地区：[1]中国海洋大学海洋生物遗传学与育种教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2014年第4期47-53,共7页Periodical of Ocean University of China

基　　金："十二五"农村领域国家科技计划项目(2012AA100811-3);农业部公益性项目(200903030)资助

摘　　要：初步探讨限制性位点扩增多态性（Restriction site amplified polymorphism，RSAP）分子标记技术在龙须菜遗传多样性检测和种质鉴定上的应用。利用所优化的适合于龙须菜RSAP分析的PCR反应体系，对青岛湛山湾野生群体和栽培品系981、07-2进行遗传多样性分析和比较。试验采用15对引物组合在3个群体14个龙须菜个体中共扩增出669个位点，其中多态位点数为146个，多态性比例22％，平均每对引物产生10条多态性条带，片段大小在100～1000bp之间。湛山湾野生群体的Na（1．0075）、Ne（1．0071）、H（O．0036）、I（0．0051）与栽培品系981的Na（1．003O）、NP（1．0021）、H（0．0012）、I（0．0018），以及栽培品系07-2的Na（1．009O）、Ne（1．0063）、H（0．0037）、I（0．0054）相比较可知3个群体的遗传多样性是有差异的。种群间的遗传多样性分析表明，在所有检测的样品中，遗传多样性多来自不同群体间的多样性。群体遗传结构分析表明，2个栽培品系981、07—2间遗传距离不大，但栽培品系与湛山湾野生群体间的遗传距离相对较大，而UPGMA聚类分析也明显的将湛山湾野生群体与栽培品系981、07—2区分开来，表明野生群体与栽培品系间已产生一定的遗传隔离。通过分析湛山湾野生群体及栽培品系981、07—2的RSAP指纹图谱，从中筛选栽培品系981的特异条带，并将其转化为稳定性好、特异性高的序列特征化扩增区（Sequence characterized amplified regions，SCAR）分子标记。Restriction Site Amplified Polymorphism （RSAP） technique was preliminary applied on the de- tection of genetic diversity and germplasm identification of Gracilariopsis lemaneiformis. Three popula- tions, wild from Zhanshan Bay, Qingdao and two cultivar （981 and 07-2 cultivar） were analyzed and com- pared using suitable RSAP PCR amplification system. 15 primer pairs produced a total of 669 reproducible bands within 14 individual used, 146 of which （22%） were polymorphic. Each primer pairs generated 10 polymorphic bands, with fragment size ranged from 100 to 1 000 bp. The Na, Ne, H and I was 1. 007 5, 1. 007 1, 0. 003 6 and 0. 005 1 for wild population of Zhanshan Bay and 1. 003 0, 1. 002 1, 0. 001 2 and 0. 001 8 {or cultivar 981, respectively. The above genetic index was 1. 009 0, 1. 006 3, 0.003 7 and 0.005 4 for cuhivar 07-2, respectively Genetic diversity between different populations was revealed by the compari- son of genetic index. The cultivar 981 showed lower genetic variation than both the cultivar 07-2 and wild population of Zhanshan Bay. The analysis of genetic diversity between populations showed that genetic di- versity in all samples was from the diversity between the different populations largely. In addition, popu- lation genetic structure revealed small genetic distance and high genetic similarity between the three groups of G. lemaneiformis. However, the genetic distance between the cultivated population and the wild popu- lation was higher than that between 981 and 07-2 cuhivar. The UPGMA analysis revealed two main clus- ters, which obviously separated the cultivated population from the wild population. After analyzing the RSAP fingerprinting among three populations, the specific bands of cultivar 981 were determined and con- verted into sequence characterized amplified regions （SCAR） molecular marker with high stability and spe- cificity.

关 键 词：龙须菜 RSAP 遗传多样性 栽培品系981 SCAR 

分 类 号：Q785[生物学—分子生物学]