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从32对叶绿体DNA通用引物中筛选出8对引物用于评价辽宁省梨属种质的遗传多样性,其中4对引物所扩增的叶绿体DNA片段存在突变位点。合并后的叶绿体基因片段(3 688 bp)含有单一突变位点9个、简约信息性位点3个以及插入/缺失(INDEL)2个。在37个样品中,trnL-trnF-487区域、trnL-trnF-413区域、rbcL区域和trnS-psbC区域的单倍型分别为2个、4个、2个和3个,合并之后的叶绿体基因片段的单倍型为4个。非编码区trnL-trnF-413多态性最高,具有最高的单倍型数、最高的单倍型(基因)多样性、最高的核苷酸多样性、最高的单倍型多样性方差和最高的单倍型多样性标准差。辽宁省37份梨种质的单倍型(基因)多样性(Hd)、核苷酸多样性(Pi)分别为0.577、0.00055。Tajima’s D检验中除了rbcL区域的D值在P<0.05水平上显著外,其余区域的D值未达显著水平,表明自然选择对rbcL区域的突变位点产生了作用。秋子梨和白梨种质共有单倍型HAP_1、HAP_2和HAP_3,显示出一定的亲缘关系;西洋梨则单独共享HAP_5。运用中介邻接网络(Median-Joining network,MJ)算法绘制的中介网络图显示HAP_1与HAP_2、HAP_3的亲缘关系较远,而HAP_5则与上述3个单倍型的亲缘关系较远。
Eight pairs of primers were selected from 32 pairs of chloroplast DNA universal primers to evaluate the genetic diversity of Liaoning Province pear genus. Among them, four pairs of primers amplified the chloroplast DNA fragment with mutation sites. The combined chloroplast gene fragment (3 688 bp) contained 9 single mutation sites, 3 parsimony informative sites and 2 INDELs. In 37 samples, the haplotypes of trnL-trnF-487, trnL-trnF-413, rbcL and trnS-psbC were 2, 4, 2 and 3 respectively. The combined chloroplast genes Fragment haplotype is 4. The trnL-trnF-413 in the noncoding region had the highest polymorphism with the highest haplotype number, the highest haplotype (genetic) diversity, the highest nucleotide diversity, and the highest variance in haplotype diversity Haplotype diversity standard deviation. The haplogroup (gene) diversity (Hd) and nucleotide diversity (Pi) of 37 pear cultivars in Liaoning Province were 0.577 and 0.0005. In Tajima’s D test, except for the value of r in rbcL region being significant at the level of P <0.05, the D value of other regions did not reach significant level, indicating that natural selection had a role in the mutation site of rbcL region. There were some haplogroups HAP_1, HAP_2 and HAP_3 in the seeds of autumn pear and pear cultivars, which showed that HAP_5 shared with pear. The intermediary network map drawn by the Median-Joining network (MJ) algorithm shows that HAP_1 is more distantly related to HAP_2 and HAP_3, while HAP_5 is more distantly related to the above three haplotypes.