Gazendam Inge, Mojapelo Pinkie, Bairu Michael W
Agricultural Research Council-Vegetable, Industrial and Medicinal Plants (ARC-VIMP), Roodeplaat, Pretoria 0001, South Africa.
Faculty of Natural & Agricultural Sciences, School of Agricultural Sciences, Food Security and Safety Niche Area, North-West University, Private Bag X2046, Mmabatho 2735, South Africa.
Plants (Basel). 2022 Jun 10;11(12):1546. doi: 10.3390/plants11121546.
DNA fingerprinting is a molecular technique applied to identify genetic differences between plant cultivars or lines and is used for genetic purity testing. The suitability of single nucleotide polymorphism (SNP) panels for the fingerprinting of tetraploid potato were investigated as a new high throughput, objective, and cost-effective method instead of simple sequence repeats (SSRs) and polyacrylamide gel electrophoresis (PAGE). One-hundred and ninety (190) potato cultivars, including various cultivars currently important in South Africa, were genotyped at 500 SNP positions utilising SeqSNP by LGC Biosearch Technologies. An optimal panel of 25 SNP markers was identified that could discriminate between South African potato cultivars on genetic allele dosage. The genotypes of these SNPs were validated on selected potato genotypes using KASP (Kompetitive Allele Specific PCR) SNP assays. A database of SNP genotype profiles was compiled for all the entries of the germplasm database. The panel of 21 successful SNPs accurately identified the unique potato cultivars in the database. The KASP SNP assays of the successful SNP panel are therefore available for potato DNA fingerprinting as new germplasm, or purity test requests are submitted to ARC-VIMP. This panel provides an objective method for assigning putative cultivar identity to unknown samples submitted for fingerprinting.
DNA指纹图谱是一种用于识别植物品种或品系间遗传差异的分子技术,用于遗传纯度检测。研究了单核苷酸多态性(SNP)面板用于四倍体马铃薯指纹图谱分析的适用性,作为一种取代简单序列重复(SSR)和聚丙烯酰胺凝胶电泳(PAGE)的新的高通量、客观且经济高效的方法。利用LGC生物搜索技术公司的SeqSNP对190个马铃薯品种进行了基因分型,其中包括南非目前重要的各种品种,在500个SNP位点上进行了检测。确定了一个由25个SNP标记组成的最佳面板,该面板能够根据遗传等位基因剂量区分南非马铃薯品种。使用竞争性等位基因特异性PCR(KASP)SNP检测方法在选定的马铃薯基因型上验证了这些SNP的基因型。为种质数据库的所有条目编制了SNP基因型图谱数据库。由21个成功的SNP组成的面板准确地识别了数据库中独特的马铃薯品种。因此,当有新的种质或纯度检测请求提交给ARC-VIMP时,成功的SNP面板的KASP SNP检测方法可用于马铃薯DNA指纹图谱分析。该面板为将提交进行指纹图谱分析的未知样品的假定品种身份进行鉴定提供了一种客观方法。