Fan L, Zhang M-Y, Liu Q-Z, Li L-T, Song Y, Wang L-F, Zhang S-L, Wu J
Plant Mol Biol Report. 2013;31(6):1271-1282. doi: 10.1007/s11105-013-0586-z.
A set of 120 simple sequence repeats (SSRs) was developed from the newly assembled pear sequence and evaluated for polymorphisms in seven genotypes of pear from different genetic backgrounds. Of these, 67 (55.8 %) primer pairs produced polymorphic amplifications. Together, the 67 SSRs detected 277 alleles with an average of 4.13 per locus. Sequencing of the amplification products from randomly picked loci NAUPy31a and NAUpy53a verified the presence of the SSR loci. When the 67 primer pairs were tested on 96 individual members of eight species in the Rosaceae family, 61.2 % (41/67) of the tested SSRs successfully amplified a PCR product in at least one of the Rosaceae genera. The transferability from pear to different species varied from 58.2 % (apple) to 11.9 % (cherry). The ratio of transferability also reflected the closer relationships within Maloideae over Prunoideae. Two pear SSR markers, NAUpy43c and NAUpy55k, could distinguish the 20 different apple genotypes thoroughly, and UPGMA cluster analysis grouped them into three groups at the similarity level of 0.56. The high level of polymorphism and good transferability of pear SSRs to Rosaceae species indicate their promise for application to future molecular screening, map construction, and comparative genomic studies among pears and other Rosaceae species.
从新组装的梨序列中开发了一组120个简单序列重复(SSR),并在7种不同遗传背景的梨基因型中评估了其多态性。其中,67对(55.8%)引物对产生了多态性扩增。这67个SSR共检测到277个等位基因,每个位点平均有4.13个。对随机挑选的位点NAUPy31a和NAUpy53a的扩增产物进行测序,验证了SSR位点的存在。当在蔷薇科8个物种的96个个体成员上测试这67对引物对时,61.2%(41/67)的测试SSR在至少一个蔷薇科属中成功扩增出PCR产物。从梨到不同物种的可转移性从58.2%(苹果)到11.9%(樱桃)不等。可转移性的比例也反映了苹果亚科内比李亚科内的亲缘关系更近。两个梨SSR标记NAUpy43c和NAUpy55k能够完全区分20种不同的苹果基因型,并且UPGMA聚类分析在相似性水平为0.56时将它们分为三组。梨SSR的高度多态性和对蔷薇科物种的良好可转移性表明它们有望应用于未来的分子筛选、图谱构建以及梨和其他蔷薇科物种之间的比较基因组研究。