Convertion and Redevelopment of Molecular Markers of 4 Pleiotropic Disease Resistance Genes in Wheat (Triticum aestivum)
GAO Jie, SONG Guo-Qi, LI Ji-Hu, LI Yu-Lian, ZHANG Shu-Juan, ZHANG Rong-Zhi, GU Tian-Tian, LI Gen-Ying, LI Wei*
1 Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, China; 2 Key Laboratory of Wheat Biology and Genetic Improvement in the North Yellow & Huai River Valley, Ministry of Agriculture/National Engineering Laboratory for Wheat & Maize, Jinan 250100, China
Abstract：It has been more than 20 years since wheat (Triticum aestivum) molecular breeding was introduced to China, only a few commercial cultivars were released. To promote combination of marker assistant selection and traditional breeding, and accelerate molecular breeding development is urgent. There are 4 well characterized pleiotropic disease resistance genes in wheat. It is difficult to select these genes in breeding program through phenotype, marker assistant selection is an option. Markers developed for pleiotropic disease resistance genes have some shortcoming, such as ambiguous genotyping or single marker type, which limit its use. In this study, Focus on the ambiguous genotyping problem of reported kompetitive allele-specific PCR (KASP) markers of pleiotropic disease resistance genes, the basic local alignment search tool (BLAST) was used to align the KASP marker amplification regions and the 'Chinese Spring' reference genome. Except the target regions, several other similar genomic regions were found. Too many similar sequences in the genome may be the reason for genotyping problem of KASP markers. According to the BLAST result, primers were redesigned. The KASP marker genotyping result of improved Lr34K2 and Lr46K3 were better than Lr34_TCCIND for leaf rust resistance gene 34 (Lr34) and Lr46_JF2-2 for Lr46, respectively; KASP markers Sr2K3 for stem rust resistance 2 (Sr2), C6K1C2 and C6K2C1 for Lr34, and Lr46g22K3 for Lr46 were redeveloped, the genotyping results were clear; KASP markers Lr46_JF2-2 for Lr46, TM4 and TM10 for Lr67 were also converted into derived cleaved amplified polymorphic sequences (dCAPS) markers Lr46Rdcaps, TM4dcaps, and TM10dcaps, respectively. dCAPS markers would release the machinery requirement and lower the application threshold. In conclusion, this study enriches marker type and quantity, and provides convenience for marker assistant selection of pleiotropic disease resistance genes. It also has important reference value for KASP marker development and improvement.
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