FACTORS INFLUENCING SUCCESSFUL AGROBACTERIUMMEDIATED GENETIC TRANSFORMATION OF MUSKMELON | ||||
Fayoum Journal of Agricultural Research and Development | ||||
Article 10, Volume 20, Issue 2, July 2006, Page 111-120 PDF (639.06 K) | ||||
Document Type: Research articles. | ||||
DOI: 10.21608/fjard.2006.197590 | ||||
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Authors | ||||
Eissa Ahmed Eissa1; Velich I.2 | ||||
1Department of Genetics, Faculty of Agriculture, Fayoum University, Egypt | ||||
2Department of Genetics and Plant Breeding, Faculty of Horticultural Sciences, Szent István University, Budapest, Hungary | ||||
Abstract | ||||
The development of a robust Agrobacterium-mediated transformation protocol for muskmelon (Cucumis melo L.) requires the identification and optimization of the factors affecting DNA delivery and plant regeneration. We have used pieces of mature cotyledon from cultivar Hales Best Jumbo and the Agrobacterium tumefaciens strain LBA4404, to investigate and optimize regeneration and transformation. Agrobacterium strain harboring the binary vectors which contains phosphino-thricin acetyl transferase gene (bar) as selectable marker for herbicide resistance (glufosinat ammonium) and β-glucuronidase gene (uidA) as reporter. Factors that produced differences in DNA delivery and regeneration included bacterial concentration, length of exposure, the time allowed for co-cultivation and antibiotic concentration (claforan, cefotaximum). These factors combined as a whole led to successful transformation. Bacterial concentration, time of exposure of the explants to the bacteria, co-cultivation period and antibiotic concentration were found to affect regeneration and transformation efficiency. Analysis of these parameters allowed the development of an optimized protocol for Agrobacterium-mediated transformation of Cucumis melo cv. Hales Best Jumbo cotyledon explants. We fully describe a protocol that allowed efficient DNA delivery and gave rise stable transgenic muskmelon plants. Selectable marker gene expression and reporter gene assay demonstrated that transgenes were integrated into the muskmelon genome. Genetic transformation of calli and regenerated plantlets was confirmed by histochemical β- glucuronidase assays. | ||||
Keywords | ||||
Transformation; muskmelon; Agrobacterium-mediated; factors; herbicide; β-glucuronidase | ||||
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