桃花粉可育基因连锁的RAPD标记与SCAR标记的转化

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

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906bp,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。

In this study, a RAPD marker （OPW03-900） linked to pollen fertility gene was screened from 180 RAPD arbitrary primers in 52 F1 individuals from the cross ＇ Chongyanghong＇ × ＇ Okuba＇ , pollen fertility/sterility pool was constructed respectively by using RAPD technique and BSA method. After repeating tests and checking up individuals of population, this marker could only be amplified in the pollen fertility individuals （ except recombinants）, the linkage distance was 5.80 cM with the pollen fertility/sterility locus. This band was collected and sequenced to synthesize a pair of SCAR primer which was used to amplify the individuals of the F1 progeny, the locus of the special band and the number of recombinants were the same as the result of RAPD amplification, the size of this marker was 906 bp, which was designated as SCW03-906, indicating the RAPD marker has been successfully converted into a SCAR marker which linked to pollen fertility gene in peach. This sequence was adopted by GenBank, the logging number was DQ659676.