Veilleux R E, Shen L Y, Paz M M
Department of Horticulture, Virginia Polytechnic Institute and State University, Blacksburg 24061-0327, USA.
Genome. 1995 Dec;38(6):1153-62. doi: 10.1139/g95-153.
Randomly amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) analyses were used to characterize the genetic composition of anther-derived plants of a diploid potato clone, CP2 (Solanum chacoense 80-1 x S. phureja 1-3). The ploidy of anther-derived plants was first determined by flow cytometry. A total of 44 decamer primers was screened for RAPD polymorphism. The loci that segregated were selected and scored. The monoploids had less than half as many loci carrying RAPD markers compared with the anther donor. Among 14 anther-derived diploids, 5 were identified as homozygous by marker frequency similar to monoploids and 9 as heterozygous. Five of seven SSRs obtained from published potato sequences were polymorphic in CP2. CP2 was found to be heterozygous with two alleles at four SSR loci (TC/TA, AAG, AGA, CTT) and three alleles at a ACTC locus. Primer pairs flanking each of the five polymorphic SSRs revealed that monoploids had only the allele contributed by S. chacoense 80-1. Homozygous diploids had only one band per SSR locus, whereas heterozygous diploids displayed more than one allele for at least one SSR locus. Results of the SSR analysis supported the findings based on RAPD markers; the same five diploid clones were characterized as homozygous by both SSR and RAPD markers.
采用随机扩增多态性DNA(RAPD)和简单序列重复(SSR)分析方法,对二倍体马铃薯克隆CP2(Solanum chacoense 80 - 1×S. phureja 1 - 3)花药培养植株的遗传组成进行了表征。首先通过流式细胞术确定花药培养植株的倍性。共筛选了44个十聚体引物用于RAPD多态性分析。对分离的位点进行选择和评分。与花药供体相比,单倍体携带RAPD标记的位点数量不到其一半。在14个花药培养的二倍体中,5个通过与单倍体相似的标记频率被鉴定为纯合子,9个为杂合子。从已发表的马铃薯序列中获得的7个SSR中有5个在CP2中具有多态性。发现CP2在4个SSR位点(TC/TA、AAG、AGA、CTT)为杂合子,有两个等位基因,在一个ACTC位点有3个等位基因。侧翼位于5个多态性SSR每个位点的引物对显示,单倍体仅具有由S. chacoense 80 - 1贡献的等位基因。纯合二倍体在每个SSR位点只有一条带,而异合二倍体在至少一个SSR位点显示多个等位基因。SSR分析结果支持基于RAPD标记的发现;相同的5个二倍体克隆通过SSR和RAPD标记均被表征为纯合子。