Genomic and cDNA fragments with homology to known disease resistance genes (RGH fragments) were cloned from Cucumis melo using degenerate-primer PCR. Fifteen homologues of the NBS-LRR gene family have been isolated. The NBS-LRR homologues show high divergence and, based on the partial NBS-fragment sequences, appear to include members of the two major subfamilies that have been described in dicot plants, one that possesses a TIR-protein element and one that lacks such a domain. Genomic organization of these sequences was explored by DNA gel-blot analysis, and conservation among other Cucurbitaceae was assessed. Two mapping populations that segregate for several disease and pest resistance loci were used to map the RGH probes onto the melon genetic map. Several NBS-LRR related sequences mapped to the vicinity of genetic loci that control resistance to papaya ringspot virus, Fusarium oxysporum race 1, F. oxysporum race 2 and to the insect pest Aphis gossypii. The utility of such markers for breeding resistant melon cultivars and for cloning the respective R-genes is discussed.

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A new linkage map of Cucumis melo, derived from the F2 progeny of a cross between PI 414723 and C. melo 'TopMark' is presented. The map spans a total of 1421 cM and includes 179 points consisting of random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), inter-simple sequence repeats (ISSRs), simple sequence repeats (SSRs), and restriction fragment length polymorphism (RFLP) markers. The map also includes an aphid resistance trait (Vat) and the sex type gene, andromonoecious (a), the two of which are important in resistance breeding and the control of hybrid seed production, as well as a seed-color gene, Wt-2. Most RFLPs represent sequence-characterized cDNA probes from C. melo and Cucumis sativus. These include resistance gene homologues and genes involved in various aspects of plant development and metabolism. A sub-set of our SSR and RFLP markers were also mapped, as part of this study, on additional mapping populations that were published for this species. This provides important reference points ("anchors"), enabling us to identify several linkage groups with respect to other melon maps.Key words: Cucumis melo, melon, genetic map, molecular markers, resistance gene homologues.

Members of the MADS-box gene family control reproductive development in higher plants. In cucumber, floral development exhibits several interesting features related to a genetically determined sex-expression mechanism, that affects the differentiation of male and female flowers. In this study, three cDNA homologues of the homeotic gene AGAMOUS have been cloned from early-stage floral buds of Cucumis sativus and fully sequenced. Their expression was studied by Northern analysis using two contrasting sex genotypes, an androecious line and a gynoecious one. The three genes are expressed at low levels at earlier bud stages, the levels rising as the bud matures. Two of the clones, CAG1 and CAG3, are expressed in the third and fourth whorl of mature flowers, while CAG2 is restricted to the carpel; none is expressed in leaves. The transcript levels do not appear to be modulated by gibberellin or ethephon, two treatments that alter sex expression in cucumber. While MADS-box genes probably play an essential role in cucumber floral development, as they do in other plants, our findings may imply that the pathway leading to reproductive organ arrest in cucumber unisexual buds acts independently of MADS-box gene expression.

Resistance to cotton-melon aphid (Aphis gossypii Glover) segregated as a single dominant gene in a melon (Cucumis melo L.) mapping population derived from the cross `Top Mark' × PI 414723. Sixty-four F2-derived F3 families were used to map the aphid resistance locus, Vat, with respect to randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers. RFLP markers NBS-2 and AC-39 flanked Vat at distances of 3.1 cM and 6.4 cM, respectively. NBS-2 is homologous to the nucleotide binding site-leucine-rich repeat (NBS-LRR) superfamily of plant resistance genes. Another homolog of this superfamily, NBS-5, was positioned ≈16.8 cM from Vat, raising the possibility that Vat resides in a cluster of NBS-LRR paralogs. RFLP marker AC-8, which has similarity to plant lipoxygenases, was positioned at ≈5.5 cM from Vat. Monogenic resistance to A. gossypii has been identified in two sources of melon germplasm, Indian accession PI 371795 (progenitor of PI 414723) and Korean accession PI 161375. To test for an allelic relation between the genes controlling aphid resistance in these two distinct germplasm sources, melon plants of a backcross population from a cross between two resistant lines having Indian- or Korean-derived resistance were infested with aphids. At least 90 out of 92 segregating progeny were aphid resistant, suggesting that the same resistance gene, Vat, is present in both sources of melon germplasm.