Multiplex SSR-PCR approaches for semi-automated genotyping and characterization of loci linked to blast disease resistance genes in rice

作者：Ashkani Sadegh^*; Rafii Mohd Yusop; Shabanimofrad Mahmoodreza; Foroughi Majid; Azizia Parisa; Akhtar Mohd Sayeed; Sahebi Mahbod; Harun Abd Rahim; Nasehi Abbas

来源：Comptes Rendus Biologies, 2015, 338(11): 709-722.

DOI：10.1016/j.crvi.2015.07.007

• Rice (Oryza sativa L.)
• Rice blast (Magnaporthe oryzae)
• Multiplex PCR
• Fluorescent-labelled
• SSR genotyping
• Capillary electrophoresis
• Linkage map construction
• Segregation distortion analysis

In the present study, 63 polymorphic microsatellite markers related to rice blast resistance genes were fluorescently labelled at the 5'-end with either 6-FAM or HEX using the G5 dye set and incorporated into a multiplex SSR-PCR for the detection of fragments using an automated system. For rice F-3 families obtained from crosses between Pongsu Seribu 2 (Malaysian blast resistant cultivar) and Mahsuri (a susceptible rice cultivar), the genotypes for 13 designated multiplex SSR panels were determined. The genotyping assays were performed using a capillary-based ABIPRISM 3100 genetic analyser. The sizes of the SSRs alleles observed in the range from 79 to 324 bp. The observed marker segregation data were analysed using the Chi(2) test. A genetic linkage map covering ten chromosomes and comprising 63 polymorphic SSR markers was constructed, and the distorted loci were localised to linkage groups. The results indicated that distorted loci are presented on eight chromosomes.

• 出版日期2015-11