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Conversion of Existing AFLP Markers to SCAR Markers Linked to Globodera rostochiensis and Phytophthora infestans Resistance Could Be Performed Without Using Acrylamide Gel Electrophoresis
Potato Research ( IF 2.3 ) Pub Date : 2021-07-01 , DOI: 10.1007/s11540-021-09499-9
Kengo Ohbayashi

Marker-assisted selection (MAS) is more efficient with single locus marker methods than with the complex multi-locus AFLPTM system. Therefore, a method is described to convert AFLP markers into sequence characterised amplified region (SCAR) markers using arbitrary short primers that were generated based on AFLP markers linked to potato disease resistance genes. In the first step of this method, we synthesised short primers with nucleotides (A, T, G, or C) added to the 3′-ends based on primer sequences of AFLP markers. In the next step, polymerase chain reaction (PCR) analyses were performed with bulked segregant analysis (BSA) by combining the synthesised arbitrary primers with 10-base primers. In the final step, DNA fragments that were associated with disease resistance genes were detected using BSA and were converted into SCAR markers. Using this method, AFLP markers linked respectively to Globodera rostochiensis resistance gene H1 and Phytophthora infestans resistance gene R2 were easily converted to SCAR markers without using AFLP system. The detected SCAR markers were clear and reproducible. Thus, in MAS using SCAR markers, conversion of AFLP markers to SCAR markers using an arbitrary short primer could be an effective breeding tool.



中文翻译:

可以在不使用丙烯酰胺凝胶电泳的情况下将现有的 AFLP 标记转化为与 Globodera rostochiensis 和 Phytophthora infestans 抗性相关的 SCAR 标记

单基因座标记方法的标记辅助选择 (MAS) 比复杂的多基因座 AF​​LP TM更有效系统。因此,描述了一种使用基于与马铃薯抗病基因相关联的 AFLP 标记生成的任意短引物将 AFLP 标记转化为序列特征扩增区域 (SCAR) 标记的方法。在该方法的第一步中,我们根据 AFLP 标记的引物序列合成了在 3' 末端添加核苷酸(A、T、G 或 C)的短引物。在下一步中,通过将合成的任意引物与 10 碱基引物组合,使用批量分离分析 (BSA) 进行聚合酶链反应 (PCR) 分析。在最后一步,使用 BSA 检测与抗病基因相关的 DNA 片段并将其转换为 SCAR 标记。使用这种方法,AFLP 标记分别与Globodera rostochiensis连锁抗性基因H1致病疫霉抗性基因R2 可在不使用 AFLP 系统的情况下轻松转化为 SCAR 标记。检测到的 SCAR 标记清晰且可重复。因此,在使用 SCAR 标记的 MAS 中,使用任意短引物将 AFLP 标记转换为 SCAR 标记可能是一种有效的育种工具。

更新日期:2021-07-01
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