洋桔梗品种遗传多样性的AFLP分析  

AFLP Analysis of Genetic Diversity among Cultivars in Eustoma grandiflorum (Raf.) Shinn.

作　　者：王瑞鑫[1] 韦银凤[1] 李琳[1] 付晓佳 李爽[1] 陈崇顺[1] 

机构地区：[1]南京师范大学生命科学学院/江苏省生物多样性与生物技术重点实验室,南京210023

出　　处：《西北植物学报》2014年第7期1325-1330,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家基础科学人才培养基金(J1103507)

摘　　要：该研究以洋桔梗（Eustoma grandi florum）2个品种‘玛丽艾基粉色’和‘圣剑白底紫边’为试材,提取叶片DNA,经过EcoRI /Mse Ⅰ双酶切、连接、预扩增、选择性扩增,建立了洋桔梗的AFLP最佳反应体系;并以64个常用引物组合进行扩增,得到154个多态性条带,从中筛选出扩增条带较多且多态性较好的4个引物组合（E-ACA/M-CTC,E-ACC/M-CAC,E-AGC/M-CTT,E-ACT/M-CAG）,其多态位点百分率均值为24.36％.利用上述4个引物组合,以最佳反应体系为基础,构建了7个常见洋桔梗品种的AFLP指纹图谱,统计7个品种各4个引物组合在1 000～300 bp区间7个区段的扩增条带,并将各个品种的AFLP指纹图谱转换成各品种4组7位数构成的28位特异数字指纹,极大地方便了种质比较及鉴定;7个品种间的遗传相似系数介于0.683 5～0.860 8之间,平均值为0.774 6.研究结果为进一步进行洋桔梗的种质研究及利用奠定了基础.Taking Eustoma grandiflorum cultivars ‘Double Mariachi Pink’ and ‘Excalibur Blue Picotee’ as experimental material,we extracted genomic DNA from the leaves.The optimized AFLP reaction system was established,after EcoR Ⅰ/Mse Ⅰ double enzyme digestion,ligation,pre amplification and selective am plification.154 polymorphic bands were acquired with 64 primer combinations commonly used,from which 4 selective amplification primer combinations （E-ACA/M-CTC,E-ACC/M-CAC,E-AGC/M-CTT,E-ACT/ M-CAG） with more polymorphic amplified bands were selected.The average percentage of polymorphic bands was 24.36 ％.Using the 4 selective amplification primer combinations,through the optimized reaction system,the AFLP fingerprints of 7 common E.grandiflorum cultivars were constructed.Subsequently,the AFLP fingerprints were transformed into the specific digital fingerprints of 28 digits,based on the counting of the amplified bands in the 7 sections of 1 000～300 bp interval of the 4 primer combinations,greatly fa cilitating the comparison and identification of the germplasm.The genetic similarity coefficient between the cultivars ranged 0.683 5～0.860 8,and the average value was 0.774 6.The results of that study have laid an important foundation for the further research on E.grandiflorum.

关 键 词：洋桔梗 AFLP 最佳反应体系 引物筛选 指纹图谱 特异数字指纹 遗传相似系数 

分 类 号：Q789[生物学—分子生物学]