Construction of a 1.2-Mb contig including the citrus tristeza virus resistance gene locus using a bacterial artificial chromosome library of Poncirus trifoliata (L.) Raf.

The citrus tristeza virus resistance gene (Ctv) is a single dominant gene in Poncirus trifoliata, a sexually compatible relative of citrus. To clone this gene, a bacterial artificial chromosome (BAC) library has been constructed from an individual plant that was homozygous for Ctv. This library contains 45 696 clones with an average insert size of 80 kb, corresponding to 9.6 genome equivalents. Screening of the BAC library with five chloroplast DNA probes indicated that 0.58% of the BAC clones contained chloroplast-derived inserts. The chromosome walk across the Ctv locus was initiated using three closely linked genetic markers: C19, AD8, and Z16. The walk has been completed and a contig of ca. 1.2 Mb was constructed. Based on new data, the genetic map in the Ctv region was revised, with Ctv being located between AD8-Z16 and C19 at distances of 1.2 and 0.6 cM, respectively. Utilizing DNA fragments isolated from the contig as RFLP markers, the Ctv locus was further mapped to a region of ca. 300 kb. This contig contains several putative disease-resistance genes similar to the rice Xa21 gene, the tomato Cf-2 gene, and the Arabidopsis thaliana RPS2 gene. This library will therefore allow cloning of Ctv and other putative disease-resistance genes.Key words: Poncirus, citrus tristeza virus, chromosome walk, resistance gene.

A Novel Gene Conferring Citrus Tristeza Virus Resistance in Citrus maxima (Burm.) Merrill

`Chandler' pummelo [Citrus maxima (Burm.) Merrill] was found to be citrus tristeza virus (CTV)–resistant. The inheritance of this resistance in 84 progeny of two crosses derived from `Chandler' pummelo and trifoliate orange [Poncirus trifoliata (L.) Raf.] was controlled by a single dominant gene designated Ctv2. Progeny analysis of four molecular markers closely linked to the Ctv gene, which confers resistance to CTV in trifoliate orange, demonstrated that Ctv2 was an independently assorting gene from Ctv.