Actinidia, a genus of kiwifruit, has 54 species, with the commercially dominant species being one with two subspecies: Actinidia chinensis var. chinensis and A. chinensis var. deliciosa. The global kiwifruit industry has used a single variety for decades, but in the last 20 years, new varieties have been introduced, with breeders and propagation rights concerned about protecting them from fraudulent use. The genome of A. chinensis var. chinensis ‘Red5’ was searched for perfect microsatellites with repeat motifs of no less than two to ten bases. Out of 216,456 possible perfect microsatellite loci, 82 were chosen to be spread across all 29 chromosomes. Twenty microsatellite loci with repeat motifs over two have been developed for the varietal characterization of kiwifruit. The markers consist of repeating motifs of at least three bases that suffer less from problems in interpreting electrophoretic profiles due to stuttering. A single amplification protocol valid for all loci was developed. The markers were tested on a sample of 100 genotypes that included diploid, tetraploid, hexaploid, and octoploid individuals. The selected markers were able to clearly discriminate all genotypes except for two clonal mutations. This indicates that the microsatellite loci are highly effective in distinguishing between different kiwifruit varieties, regardless of their ploidy level. The ability to accurately characterize kiwifruit genotypes using these markers can greatly benefit breeding programs and conservation efforts for this important fruit crop.

中文翻译：

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改进猕猴桃分子表征的标记物


猕猴桃属 Actinidia 有 54 种，其中商业优势种是 1 个亚种：Actinidia chinensis var. chinensis var. chinensis 和 A. chinensis var. deliciosa。几十年来，全球猕猴桃行业一直使用单一品种，但在过去 20 年中，新品种被引入，育种者和繁殖权担心保护它们免受欺诈使用。在 A. chinensis var. chinensis 'Red5' 的基因组中寻找具有不少于 2 至 10 个碱基重复基序的完美微卫星。在 216,456 个可能的完美微卫星基因座中，选择了 82 个分布在所有 29 条染色体上。已经开发了 20 个具有 2 个重复基序的微卫星基因座，用于猕猴桃的品种表征。标记物由至少三个碱基的重复基序组成，这些基序由于口吃而在解释电泳曲线时受到的问题较少。开发了一种对所有基因座有效的单一扩增方案。这些标记物在 100 种基因型的样本上进行了测试，其中包括二倍体、四倍体、六倍体和八倍体个体。选定的标记能够清楚地区分除两个克隆突变之外的所有基因型。这表明微卫星基因座在区分不同的猕猴桃品种方面非常有效，无论它们的倍性水平如何。使用这些标记准确表征猕猴桃基因型的能力可以极大地有利于这种重要水果作物的育种计划和保护工作。