Application of Molecular Biology for Identification of Virus Resistance Gene in Melon
https://doi.org/10.22146/ijbiotech.15265 Budi Setiadi Daryono(1*)
(1) Laboratory of Genetics and Breeding, Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia
(*) Corresponding Author
Abstract
Source of resistance to an Indonesia isolate of Cucumber mosaic virus (CMV-B2) in melon cultivar
Yamatouri has been reported. Moreover, Creb-2, a locus that confers resistance to CMV-B2 in Yamatouri has
been determined as a single dominant gene. To elucidate the resistance mechanism conferred by Creb-2 in
more detail, it is necessary to clone the Creb-2 gene and determine its molecular structure. One approach is
by amplification and cloning of melon resistance gene analogs (MRGAs) based on degenerated PCR primers
designed from conserved amino acids in the NBS-LRR motifs (P-loop, Kinase-2, and the GLPL) and Toll/
Interleukin-1 receptor-like region (TIR). This study was aimed to identify and characterize the resistance gene
analogs from Cucumis melo L. cv. Yamatouri by employing polymerase chain reactions (PCR) as a molecular
biology tools with degenerate primers based on conserved motifs of cloned R genes. The application of molecular
biology such as DNA isolation, degenerate primers and PCR condition, cloning, sequencing, linkage analysis
and mapping of resistance gene analogs to Creb-2 gene in melon will be widely discussed in this paper
Yamatouri has been reported. Moreover, Creb-2, a locus that confers resistance to CMV-B2 in Yamatouri has
been determined as a single dominant gene. To elucidate the resistance mechanism conferred by Creb-2 in
more detail, it is necessary to clone the Creb-2 gene and determine its molecular structure. One approach is
by amplification and cloning of melon resistance gene analogs (MRGAs) based on degenerated PCR primers
designed from conserved amino acids in the NBS-LRR motifs (P-loop, Kinase-2, and the GLPL) and Toll/
Interleukin-1 receptor-like region (TIR). This study was aimed to identify and characterize the resistance gene
analogs from Cucumis melo L. cv. Yamatouri by employing polymerase chain reactions (PCR) as a molecular
biology tools with degenerate primers based on conserved motifs of cloned R genes. The application of molecular
biology such as DNA isolation, degenerate primers and PCR condition, cloning, sequencing, linkage analysis
and mapping of resistance gene analogs to Creb-2 gene in melon will be widely discussed in this paper
Keywords
resistance gene analogs; Cucumber mosaic virus; Cucumis melo L.
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PDF DOI: https://doi.org/10.22146/ijbiotech.15265
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