桃花粉可育基因连锁的RAPD 标记与SCAR标记的转化

 桃品种以重阳红(花粉不育) ×大久保(花粉可育) 的52株F₁代群体为试材, 应用RAPD技术, 结合集群分组分析法(Bulked Segregate Analysis, BSA) 构建花粉可育/不育基因池, 利用180个随机引物, 筛选在花粉可育基因池中稳定扩增的一个RAPD标记OPW03-900。经过重复性验证和群体单株验证, 该标记仅在花粉可育个体(重组型除去) 中出现, 与桃花粉可育/不育位点的连锁距离为5.80cM。将该特异性片段回收、克隆、测序, 并设计一对特异SCAR引物, 再对F1代个体进行PCR扩增, 发现该特异带的位点及重组型个体的数目与RAPD扩增结果一致, 片段大小为906 bp, 命名为SCW03-906,表明RAPD标记已成功转化为与桃花粉可育基因连锁的SCAR标记。该序列已被GenBank收录, 登录号为DQ659676。

Identification of RAPD Marker Linked to Pollen Fertility Gene and Conversion to SCAR Marker in Peach

In this study, a RAPD marker (OPW03-900) linked to pollen fertility gene was screenedfrom 180 RAPD arbitrary p rimers in 52 F₁ individuals from the cross‘Chongyanghong’ בOkuba’, pollenfertility/ sterility poolwas constructed respectively by using RAPD technique and BSA method. After repeatingtests and checking up individuals of population, thismarker could only be amp lified in the pollen fertility individuals ( excep t recombinants) , the linkage distance was 5.80 cM with the pollen fertility/ sterility locus. Thisband was collected and sequenced to synthesize a pair of SCAR p rimerwhich was used to amp lify the individuals of the F1 p rogeny, the locus of the special band and the number of recombinantswere the same as the resultof RAPD amp lification, the size of thismarkerwas 906 bp, which was designated as SCW03-906, indicatingthe RAPD marker has been successfully converted into a SCAR markerwhich linked to pollen fertility gene inpeach. This sequence was adop ted by GenBank, the logging number wasDQ659676.