萝卜品种指纹图谱SRAP与AFLP分析(英文)

Cultivar Fingerprinting in Radish(Raphanus sativus) with SRAP and AFLP Marker

应用SRAP与AFLP两种分子标记技术进行了萝卜品种鉴定分析。对萝卜基因组DNA的SRAP-PCR反应体系中引物、Mg2+、dNTPs浓度进行优化,确定最优体系为引物0.3μmol.L-1,dNTP 0.2 mmo.lL-1,Mg2+3.0 mmo.lL-1。对SRAP-PCR中的退火温度(50℃)设置了12个梯度处理,以em2-me2为引物时带型无明显差异。7个供试萝卜材料的SRAP和AFLP指纹图谱分析表明,供试材料均可被SRAP和14个AFLP引物准确鉴定,每对引物组合都产生独特的指纹图谱。11个SRAP引物组合共产生155条带,多态性条带84条。聚类分析与相对遗传距离(GD)表明,供试材料聚为4类,CB-03-2与SHCB-02-1亲缘关系最近(GD=0.054 9);齐虹大连和Heiseng的亲缘关系最远(GD=0.203 4)。基于16个AFLP标记引物组合分析结果表明,供试材料聚为3类,CB-03-2和SHCB-02-1的亲缘关系最近。SRAP与AFLP综合分析结果表明,供试材料可聚为3类,其中CB-03-2与SHCB-02-1亲缘关系最近(GD=0.047 6)。

Two molecular marker systems, SRAP and AFLP, were employed to fingerprint the radish cuhivars. The SRAP-PCR system was optimized with the concentration of primer at 0. 3 μmol · L^-1, dNTPs at 0.2 mmol · L^-1 and Mg^2＋ at 3.0 mmol · L^-1. Similar patterns were obtained from 12 gradient annealing temperature for 50℃ with primer em2-me2. The fingerprinting and genetic diversity of seven radish cuhivars was analyzed using the optimized SRAP-PCR and AFLP marker system. All cultivars can be distinguished clearly with SRAP primer combinations and 14 AFLP primer combinations. Total of 155 bands were observed with 11 SRAP primer combinations, of which 84 were polymorphic bands. Cluster analysis and relatively genetic distance indicated that all cuhivars were clustered into four groups. There was the closest relationship between ‘CB-03-2＇ and ‘SHCB-02-1 ＇ with the genetic distance （GD） of 0. 054 9 and a distant relationship between ‘ Qihongdalian＇ and ‘Heiseng＇. The cluster analysis of AFLP data showed that all cuhivars clustered into three groups with the closest relationship between ‘ CB-03-2＇ and ‘ SHCB-02-1 ＇ The cluster analysis of combination of SRAP and AFLP marker data indicated that ‘ CB-03-2＇ and ‘SHCB-02-1 ＇ had the closest relationship （ GD = 0. 047 6） and all cultivars clustered into three clusters.