Utilization of bacterial artificial chromosomes (BACs) subclones in saturating Pi-2(T) locus on the rice chromosome
1997
Baraoidan, M.R. | Hei Lung | Sharma, R.C. | Guo-liang Wang | Nelson, R.J. (International Rice Research Inst., P.O. Box 933, 1099 Manila (Philippines). Entomology and Plant Pathology Div.)
Understanding the allelic diversity for molecular markers in the vicinity of blast resistance genes can be helpful in diagnosing blast resistance genes present in unknown germplasm. To increase the number of molecular markers linked to blast resistance genes, a BAC clone (28-A6), carrying the Pi-2(+)-linked RFLP marker, RG64, was subcloned in pBLUESCRIPT. A total of 129 sub-clones carrying fragments ranging in size from 0.75-2.8 kb were obtained. Twenty four of the 82 clones analyzed appeared to carry single-copy sequences. These single-copy probes were used for marker assisted genetic analysis in rice at the 28-A6 locus together with RG64. Using RG64/Dra 1,2 alleles were detected for the 13 entries. Using RG64/6 enzymes, three composite haplotypes were detected. The donor of the RG64 locus could be restricted to six of the parents. Using four 28-A6 subclones and only one enzyme (Dra 1), seven haplotypes were detected among 13 entries. The origin of the 28-A6 locus were traced in IR41431, a rainfed elite line, and its parents. The donor of the 28-A6 subclones could be restricted to three of the parents. Results demonstrate the feasibility of using BAC subcloning as a strategy to enrich the target loci with polymorphic markers
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