石刁柏雄性连锁的AFLP及SCAR标记的建立

目的:对石刁柏雌雄株基因组差异进行分析,筛选雄性或雌性连锁的分子标记.方法:利用限制性片段长度多态性技术,设计了多个引物组合,分别对石刁柏雌雄株基因组进行扩增.结果:在使用的72个引物组合中,引物组合E-AAG+M-CAT从雄性基因组中扩增出了一个雄性连锁的标记(MLDA555),该序列长度为555bp,AT含量为59%.Blast检索未发现相似序列.根据该片段序列设计的引物将该标记转化为雄性连锁的大小为523bp的稳定的SCAR标记,经过不同基因型雄性个体的验证证明该标记广泛存在于不同基因型石刁柏雄性个体中.结论:通过AFLP扩增筛选得到了石刁柏雄性连锁的AFLP和SCAR标记,为石刁柏性别决定机制的理解及石刁柏的分子标记辅助育种提供理论资料和技术支持.

Construction of male-specific AFLP and SCAR Markers in Asparagus of ficinalis L

Objective:To analyze genome difference between Asparagus male and female plants for screening male-or female-specific molecular markers.Method:Using amplified fragment length polymorphism(AFLP)technique,genomes of asparagus male and female plants were amplified respectively,by designing multiple primer combinations.Result:A total of 72selective primer pairs were screened,and only one primer pair(E-AAG/M-CAT)amplified a 555bp band(MLDA555)in male plants.The marker was AT rich in sequence(59%)and there was no sequence showed significant similarity to this marker by Blast test.This male-specific AFLP marker MLDA555 was converted into sequence-characterized amplified region(SCAR)marker using primers designed according its sequence.As expected,the SCAR primers were able to amplify a single DNA band of 523bp and the SCAR marker was efficient in the identification of male individuals from different asparagus populations.Conclusion:Male-specific AFLP and SCAR markers were identified from A.officinalis by AFLP technique,which could provide theoretical information and technical support for the understanding of sex determination mechanism and molecular marker assistant breeding of A.officinalis.