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Application of random amplified polymorphic DNA markers for detection of resistant cultivars of melon (Cucumis melo) against Cucurbitaceae viruses
- Daryono, B.S., Natsuaki, K.T.
- Acta horticulturae 2002 no.588 pp. 321-329
- Cucumber green mottle mosaic virus, Cucumber mosaic virus, Cucumis melo, DNA, cultivars, enzyme-linked immunosorbent assay, genetic markers, loci, marker-assisted selection, operon, parents, random amplified polymorphic DNA technique, reciprocal crosses, viruses, China, Japan, Turkmenistan
- Twenty melon cultivars collected mostly in Asia were tested their resistance against an Indonesian isolate of cucumber mosaic virus (CMV-B2) and a Kagoshima-Japan isolate of cucumber green mottle mosaic virus (CGMMV-K) by manual inoculation and scored as resistant or susceptible using DAS ELISA analysis. Since no cultivars showed resistance to CGMMV, 4 cultivars, 'Shinjong' (Inner Mongolian cultivar), 'Mawatauri', 'Yamatour'i, ‘Miyamauri' (three cultivars from Japan) were shown to be CMV resistant. Forty-six RAPD markers were evaluated to detect DNA bands specific to resistant cultivars. Two specific bands (550 bp and 1050 bp, respectively) by OPE-E14 primer (Operon Technologies Inc.) and APB-05 primer (Amersham Pharmacia Biotech Inc.) were obtained in the 4 resistant cultivars. Among them, bulked segregant analysis of 'Yamatouri' (a resistant cultivar) and its family were selected and constructed by crossing between 'Yamatour' and 'Vakharman' (a susceptible cultivar from Turkmenistan) to identify RAPD markers linked to CMV-B2 resistance. Specific DNA bands which linked to CMV-B2 resistance were obtained in P1 'Yamatouri', F1, some F2 and BCs. On the other hand, the result of reciprocal crossing showed that all of the tested F1 plants had one specific DNA band derived from both parents, respectively. Thus, we concluded that the resistance to CMV-B2 in 'Yamatouri' family plants depends on the existence of the DNA locus that segregated in a single dominant manner. The results also showed that RAPD markers could be used as a tool for marker-assisted selection (MAS) linked to CMV resistance in 'Yamatouri' family.