Possamai Tyrone, Scota Leonardo, Velasco Riccardo, Migliaro Daniele
CREA-Research Center for Viticulture and Enology, 31015 Conegliano, Italy.
Plants (Basel). 2024 Jul 22;13(14):2001. doi: 10.3390/plants13142001.
Plant breeders utilize marker-assisted selection (MAS) to identify favorable or unfavorable alleles in seedlings early. In this task, they need methods that provide maximum information with minimal input of time and economic resources. Grape breeding aimed at producing cultivars resistant to pathogens employs several resistance loci (, , and ) that are ideal for implementing MAS. In this work, a sustainable MAS protocol was developed based on non-purified DNA (crude), multiplex PCR of SSR markers, and capillary electrophoresis, and its application on grapevine seedlings to follow some main resistance loci was described. The optimized protocol was utilized on 8440 samples and showed high efficiency, reasonable throughput (2-3.2 min sample), easy handling, flexibility, and tolerable costs (reduced by at least 3.5 times compared to a standard protocol). The , , and allelic data analysis did not show limitations to loci combination and pyramiding, but segregation distortions were frequent and displayed both low (undesired) and high rates of inheritance. The protocol and results presented are useful tools for grape breeders and beyond, and they can address sustainable changes in MAS. Several progenies generated have valuable pyramided resistance and will be the subject of new studies and implementation in the breeding program.
植物育种者利用标记辅助选择（MAS）尽早在幼苗中鉴定有利或不利等位基因。在这项工作中，他们需要能以最少的时间和经济资源投入提供最大信息量的方法。旨在培育抗病品种的葡萄育种利用了几个抗病基因座（、和），这些基因座非常适合实施MAS。在本研究中，基于非纯化DNA（粗提物）、SSR标记的多重PCR和毛细管电泳开发了一种可持续的MAS方案，并描述了其在葡萄幼苗上用于追踪一些主要抗病基因座的应用。该优化方案应用于8440个样品，显示出高效、合理的通量（每个样品2 - 3.2分钟）、易于操作、灵活性以及可承受的成本（与标准方案相比至少降低了3.5倍）。对、和等位基因数据分析未显示基因座组合和聚合存在限制，但分离畸变很常见，且呈现出低（不期望的）和高遗传率。所展示的方案和结果是葡萄育种者及其他领域的有用工具，它们可以应对MAS中的可持续变化。所产生的几个后代具有有价值的聚合抗性，将成为育种计划中新研究和实施的主题。
