GENETIC AND PHYSICAL LOCALIZATION OF THE ROOT-KNOT NEMATODE RESISTANCE LOCUS MI IN TOMATO

As part of a map-based cloning strategy designed to isolate the root-k not nematode resistance gene Mi, tomato F2 populations were analyzed i n order to identify recombination points close to this economically im portant gene. A total of 21089 F2 progeny plants were screened using m orphological markers. An additional 1887 F2 were screened using PCR-ba sed flanking markers. Fine-structure mapping of recombinants with newl y developed AFLP markers, and RFLP markers derived from physically map ped cosmid subclones, localized Mi to a genomic region of about 550 kb . The low frequency of recombinants indicated that recombination was g enerally suppressed in these crosses and that crossovers were restrict ed to particular regions. To circumvent this problem, a population of Lycopersicon peruvianum, the species from which Mi was originally intr ogressed, that was segregating for resistance was developed. Screening of this population with PCR, RFLP and AFLP markers identified several plants with crossovers near Mi. Recombination frequency was approxima tely eight-fold higher in the Mi region of the L. peruvianum cross. Ho wever, even within the wild species cross, recombination sites were no t uniformly distributed in the region. By combining data from the L. e sculentum and L. peruvianum recombinant analyses, it was possible to l ocalize Mi to a region of the genome spanning less than 65 kb.