AFLP标记在蔷薇遗传检测中的应用

采用单酶切和双酶切AFLP技术分别对5种不同蔷薇品种基因组DNA的遗传多态性进行了检测,比较了两种方法的多态性比率.单酶切采用10条人工设计的与接头序列相识别的AFLP选择性引物,用Pst Ⅰ酶切,共获得108个AFLP标记,片段大小在100 2 000 bp,得到33个多态位点,多态性位点百分率为30.56%;双酶切采用25对双酶切AFLP引物,在50-600bp扩增出清晰条带658条,得到182个多态位点,占总扩增带的27.6%.对多态性片段进行统计,计算出不同品种间的相似系数和遗传距离.结果表明:硕苞蔷薇同其它4种蔷薇的遗传距离偏远,且其它4种蔷薇的遗传距离均较近,这与蔷薇品种来源和培育史相符.该研究表明两种AFLP标记均可用于蔷薇品种基因组DNA的遗传多态性的检测,准确评价尚待和其它品种对比研究后确定.

Abstract：

Single and double restriction digestion methods were used to detect the genetic polymorphism of the genomic DNA of five rose varieties, and the results of the two methods were compared. In single digestion, 10 human-designed AFLP selective primers different from the adaptor sequence were cut by Pst Ⅰ enzyme and 108 AFLP markers were obtained with a fragment size of 100-2 000 bp. Meanwhile, 33 polymorphic loci were gained and the proportion of polymorphic loci was 30.56%. In double digestion, 25 pairs of double enzyme primers were adopted, 658 clear bands were amplified in 50-600 bp and 182 polymorphic loci were gained, which were 27.6% of the total amplified bands. Based on the statistical analysis of polymorphic bands, the similarity coefficient and genetic distance among the different varieties were estimated.The results indicated that macartney rose (Rosa bracteaea) was genetically distant from the other four rose varieties, which were genetically close among themselves, a fact which is in agreement with the origins of the varieties and their cultivation history. Thus, the research results showed that the two types of AFLP markers can be used in detecting the genetic polymorphism of the genomic DNA of rose varieties, but more accurate evaluation will depend on comparison with more varieties.

Application of AFLP in the Genetic Detection of Rose