甘蓝型油菜EST分子标记技术体系的建立  

Establishment of EST molecular marker system in Brassica napus L.

作　　者：杨畅[1] 郑普英[1] 张学昆[1] 邹崇顺[1] 程勇[1] 李桂英[1] 

机构地区：[1]中国农业科学院油料作物研究所

出　　处：《中国油料作物学报》2008年第1期17-24,共8页Chinese Journal of Oil Crop Sciences

基　　金：国家863计划(2006AA10Z1C2);国家科技支撑计划(2006BAD07A04-1)

摘　　要：根据从NCBI上随机选取的67条油菜和拟南芥等植物的表达序列标签(Expressed sequence tag,EST)设计EST标记,以14个不同地理来源或亲缘关系的甘蓝型油菜作为验证对象,系统研究油菜EST分子标记技术体系及其在分析油菜遗传多样性和构建指纹图谱中的可行性。在67对EST引物中,有63对扩增出DNA片段,其中48对具有多态性,占总引物的71.6%。用聚丙烯酰胺凝胶电泳共检测出392条带,其中293条带具有多态性,多态率达74.7%,平均每对引物获得8.2条带,多态性带6.1条。聚类分析表明,这些EST标记可较准确地反映14个材料间的不同亲缘关系或地理关系,并且其中2对多态性较好的EST引物(ESTP36和ESTP37)可用于构建14个甘蓝型油菜的指纹图谱。研究结果表明,EST作为一种新型的分子标记,具有信息量大、多态性和重复性好、设计简便等优点,可以应用于油菜遗传多样性鉴定和构建指纹图谱。67 EST-markers were randomly selected from expressed sequence tags originated from Brassica napus L. and Arabidopsis thaliana in NCBI website. With 67 EST-markers, 14 rapeseed cultivars （B. napus L.） with different phylogenesis and biogeography were used to analyze genetic diversity. The DNA bands could be amplified using 63 EST-markers. Polymorphic DNA bands could be amplified using 48 EST-markers, the polymorphic rate was 71.6% and 392 bands were detected by polyacrylamide gel electrophoresis （PAGE）. Among the 392 bands, 293 bands were polymorphic and the polymorphic rate was 74.7%, the average EST-marker produced 8.2 bands and 6.1 polymorphic bands. The dendrogram analysis by EST-markers accurately revealed the relationship of different phylogenesis and biogeography of these 14 rapeseed cultivars. Fingerprinting of 14 varieties was constructed with two polymorphic EST-markers （ESTP36 and ESTP37）. The results indicated that EST molecular marker can provide great amount of information, and it also has advantage of high polymorphism, highly repetitive and easy application, it can be used to identify the genetic diversity and to construct a fingerprint map.

关 键 词：甘蓝型油菜 EST分子标记 遗传多样性 指纹图谱 

分 类 号：S565.403[农业科学—作物学]