Osipova E S, Koveza O V, Troitskiĭ A V, Dolgikh Iu I, Shamina Z B, Gostimskiĭ S A
Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, 127276 Russia.
Genetika. 2003 Dec;39(12):1664-72.
RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter Simple Sequence Repeats) markers were used to analyse the genetic divergence between the regenerated plants derived from callus cultures and the original maize line A188. Analysis of polymorphism by using 38 RAPD- and 10 ISSR-oligonucleotide primers showed that the differences between eight examined somaclones and the original line ranged from 6.5 to 23%. As confirmed using new primers, the regenerants derived from callus cultures grouped into two clusters according to their origin. The regenerants isolated from calluses grown for eight months differed from one another and the original line to a larger extent than the regenerants obtained from two-month callus cultures. In some somaclones, molecular marking of the regenerants revealed specific RAPD and ISSR fragments that were absent in other somaclones or the original maize line. On the basis of six specific fragments (five RAPD and one ISSR), SCAR (Sequence Characterized Amplified Region) markers were developed. Specific polymorphism revealed with random primers was completely confirmed using five SCAR markers. Polymorphism of one SCAR marker differed from that revealed with random primers. Five SCAR fragments were inherited as simple dominant traits. One SCAR fragment displayed codominant inheritance.
利用随机扩增多态性DNA（RAPD）和简单序列重复区间（ISSR）标记分析了愈伤组织培养再生植株与原始玉米自交系A188之间的遗传差异。使用38个RAPD寡核苷酸引物和10个ISSR寡核苷酸引物进行多态性分析，结果表明，8个检测的体细胞无性系与原始自交系之间的差异在6.5%至23%之间。使用新引物证实，愈伤组织培养获得的再生植株根据其起源分为两个聚类。从生长8个月的愈伤组织中分离出的再生植株彼此之间以及与原始自交系的差异程度大于从2个月愈伤组织培养物中获得的再生植株。在一些体细胞无性系中，再生植株的分子标记显示出其他体细胞无性系或原始玉米自交系中不存在的特定RAPD和ISSR片段。基于6个特定片段（5个RAPD和1个ISSR）开发了序列特异性扩增区域（SCAR）标记。使用5个SCAR标记完全证实了随机引物揭示的特异性多态性。一个SCAR标记的多态性与随机引物揭示的不同。5个SCAR片段表现为简单显性遗传。一个SCAR片段表现为共显性遗传。
