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PCR-based DNA markers linked to a gall midge resistance gene, Gm4t, has potential for marker-aided selection in rice


Tag. Theoretical and Applied Genetics. Theoretische und Angewandte Genetik 92(6): 660-665
PCR-based DNA markers linked to a gall midge resistance gene, Gm4t, has potential for marker-aided selection in rice
Rice DNAs from a gall midge resistant variety, 'Abhaya', a susceptible variety, 'Tulsi' and their F3 progeny were screened using 500 random primers in conjunction with bulked-segregant analysis in a polymerase chain reaction (PCR) with a view to detecting random amplified polymorphic DNAs (RAPDs) linked to the gene, Gm4t, which confers resistance to gall midge, a dipteran insect pest of rice. A total of 454 primers were able to produce a distinct amplification pattern, and 3695 bands/loci were amplified between the phenotypically different parents. Of these, 304 bands were polymorphic between the parents, with 19 being phenotype-specific. One of these primers, E20, amplified 2 bands, E20(570) and E20(583), which are tightly linked to resistance and susceptibility, respectively. These specific bands were cloned and sequenced, and a 94% sequence homology was found between the two fragments. Two specific 20-mer oligonucleotides were synthesized, based on the sequence information of E20(583), for use in PCR amplification directly from genomic DNAs. These PCR primers were able to amplify phenotype-specific bands, a 583-bp fragment in susceptible F3 lines and a 570-bp fragment in resistant F3 lines that had been derived from a cross between the parents, indicating their potential and utility for marker-aided selection of the Gm4t gene in rice. Its use would facilitate the early and efficient selection of resistant genes in plant breeding programmes and even in those areas where the insect is not known to occur. These phenotype-specific bands are single-copy sequences and are being mapped to ascertain their chromosomal location in rice.

Accession: 002914452

PMID: 24166388

DOI: 10.1007/BF00226086

Download PDF Full Text: PCR-based DNA markers linked to a gall midge resistance gene, Gm4t, has potential for marker-aided selection in rice



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