Institute of Field and Vegetable Crops, Maksima Gorkog 30, 21000 Novi Sad, Serbia.
Department of Plant Genetics, Institute of Biological Sciences, University of Rostock, Albert-Einstein-Str. 3, D-18059 Rostock, Germany.
Genes (Basel). 2022 Mar 5;13(3):465. doi: 10.3390/genes13030465.
Single nucleotide polymorphisms (SNPs) were significantly associated with fertility restoration of cytoplasmic male sterility (CMS) PET1 by the restorer gene . For these SNPs, four Kompetitive allele-specific PCR (KASP) markers were successfully designed. The KASP markers cover the fertility restorer locus , spanning about 3 Mb, and clearly differentiate restorer and maintainer lines. For genetic purity testing in sunflower hybrid production, the efficiency for detecting contaminations in samples was simulated using mixtures of hypocotyls or leaves. Contaminations of restorer lines with 1%, 3%, 5%, 10%, and 50% of maintainer lines were screened with all four KASP markers. Contaminations of 10% could be clearly detected in pools of 100 plants. Contaminations below this level require detection on a single plant level. For single plant detections, ethyl methanesulfonate-treated sunflower F1 hybrids, which had been phenotypically evaluated for male sterility (potential mutation in the gene) were screened. Nine identified either partially male-sterile or male-sterile plants were analyzed with all four KASP markers and only one proved to be a hybrid with a mutation, seven were male-sterile contaminants in the F1 seeds used (1.6%) and one a recombinant plant. The four KASP markers should be valuable tools for marker-assisted selection (MAS) in sunflower breeding regarding the restorer locus .
单核苷酸多态性 (SNPs) 与细胞质雄性不育 (CMS) PET1 的恢复基因显著相关。针对这些 SNPs，成功设计了四个 Kompetitive allele-specific PCR (KASP) 标记。这些 KASP 标记涵盖了育性恢复基因座，跨度约 3 Mb，可清楚地区分恢复系和保持系。为了在向日葵杂交生产中进行遗传纯度检测，使用下胚轴或叶片混合物模拟了检测样品污染的效率。用所有四个 KASP 标记筛选了恢复系中含有 1%、3%、5%、10%和 50%保持系的污染情况。可以在 100 株植物的混合物中清楚地检测到 10%的污染。低于此水平的污染需要在单株水平上进行检测。对于单株检测，筛选了经乙基甲磺酸处理的向日葵 F1 杂种，这些杂种已经进行了雄性不育表型评估（基因中的潜在突变）。用所有四个 KASP 标记分析了 9 株部分雄性不育或雄性不育的植株，其中只有一株被证明是带有突变的杂种，7 株是用于 F1 种子的雄性不育污染物（1.6%），一株是重组植株。这四个 KASP 标记应该是向日葵育种中关于恢复基因座的标记辅助选择 (MAS) 的有价值的工具。
