Delimitation of the PSH1(t) gene for rice purple leaf sheath to a 23.5 kb DNA fragment.

Leaf sheath color plays an important role as a marker for rice genetic improvement. A recombinant inbred line (RIL) population consisting of 220 individuals was developed from a cross between an Oryza sativa subsp. indica variety, IRBB60, and an Oryza sativa subsp. japonica variety, 9407. Within the RIL population, a line, RI51, was found to have purple leaf sheath (PSH). To map the gene governing PSH, RI51 was crossed with 9407 green leaf sheath (GSH) to develop an F2 segregating population. The distribution of F2 plants with PSH and GSH fitted a segregation ratio of 3:1, indicating that the PSH was controlled by a major dominant gene. The gene locus for PSH, tentatively designated as PSH1(t), was identified by surveying two bulks made of the respective 40 individuals with PSH and GSH with SSR markers covering the entire rice genome. The survey indicated that the PSH1(t) region was located on chromosome 1. Further confirmation was made using a large random sample of 360 individuals from the same F2 population and the PSH1(t) locus was then mapped on chromosome 1 between SSR markers RM3475 and RM7202 with genetic distances of 2.0 and 1.1 cM, respectively. For fine mapping of PSH1(t), a large F(2:3) segregating population with 3300 individuals from the seven heterozygous F2 plants in the RM3475-RM7202 region was constructed. Analysis of recombinants in the PSH1(t) region anchored the gene locus to an interval of 23.5 kb flanked by the left marker L03 and the right marker L05. Sequence analysis of this fragment predicted six open reading frames encoding a putative trans-sialidase, a putative Plastidic ATP/ADP-transporter, and four unknown proteins. The detailed genetic and physical maps of the PSH1(t) locus will be very useful in molecular cloning of the PSH1(t) gene.