Articles
APPLICATION OF RANDOM AMPLIFIED POLYMORPHIC DNA MARKERS FOR DETECTION OF RESISTANT CULTIVARS OF MELON (CUCUMIS MELO) AGAINST CUCURBITACEAE VIRUSES
Article number
588_52
Pages
321 – 329
Language
English
Abstract
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, Yamatouri, 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 Yamatouri 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.
Since no cultivars showed resistance to CGMMV, 4 cultivars, Shinjong (Inner Mongolian cultivar), Mawatauri, Yamatouri, 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 Yamatouri 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.
Publication
Authors
B.S. Daryono, K.T. Natsuaki
Keywords
RAPDs, resistant cultivars, Cucumis melo L., CMV, CGMMV.
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