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本研究利用RAPD和ISSR分子标记对常山胡柚的优良基因型进行鉴定,并探讨常山胡柚的起源。从100个RAPD引物中筛选出12个多态性引物用于正式扩增,共得到117条DNA带,其中多态性DNA带64条,占扩增片段的54．7%；从105个ISSR引物中筛选出11个多态性引物用于正式扩增,共得到94条DNA带,其中多态性DNA带58条,占扩增片段的61．7%。RAPD和ISSR分析揭示了常山胡柚及其近缘种的一些特异性条带。ISSR共产生了15条特异条带,RAPD共产生12特异性条带。实验数据用AMOVA软件计算遗传距离,用NTSYS-pc软件构建UPGMA聚类树状图。结果显示,所有的基因型及不同种之间均能够彼此区分,分析得到的指纹图谱对常山胡柚种和基因型的鉴定具有潜在的应用价值,可用于优良基因型的鉴定。聚类分析结果显示常山胡柚和甜柚聚为一枝,确定了甜柚是杂交亲本之一,但是常山胡柚和柚的遗传距离较远,说明常山胡柚可能是甜橙、柚和柑桔属其他种的多重自然杂交的结果。
In this study, RAPD and ISSR markers were used to identify the excellent genotypes of Changshan Huyou, and the origin of Changshan Huyou was explored. Twelve polymorphic primers were screened from 100 RAPD primers for formal amplification. A total of 117 DNA bands were obtained, of which 64 polymorphic DNA bands accounted for 54.7% of the amplified fragments. From 105 ISSR Eleven polymorphic primers were screened out for amplification. A total of 94 DNA bands were obtained, of which 58 were polymorphic DNA bands, accounting for 61.7% of the amplified fragments. RAPD and ISSR analysis revealed some specific bands of Changshan Huyou and its related species. ISSR produced a total of 15 specific bands, RAPD co-produce 12 specific bands. Experimental data The genetic distance was calculated using AMOVA software and the UPGMA cluster dendrogram was constructed using NTSYS-pc software. The results showed that all the genotypes and the different species could be distinguished from each other. The fingerprints obtained from the analysis showed potential application value for the identification of Changshan Huyou species and genotype, and could be used for the identification of excellent genotypes. The results of cluster analysis showed that Changshan grapefruit and sweet grapefruit clustered into one branch, and identified sweet grapefruit as one of the hybrid parents. However, the genetic distance between Changshan grapefruit and grapefruit was far, indicating that Changshan grapefruit may be sweet orange, grapefruit and citrus Is the result of multiple natural crosses of other species.