Agrobacterium-mediated transformation and shoot regeneration in elite breeding lines of western shipper cantaloupe and honeydew melons (Cucumis melo L.) Academic Article uri icon

abstract

  • A reliable Agrobacterium-mediated transformation and shoot regeneration protocol was developed for breeding lines of commercially important western-shipper cantaloupe and honeydew melons, 'F39' and '150', respectively. Different media were tested to select a shoot regeneration system for each of three elite breeding lines 'F39', '141' and 'TMS'. Murashige & Skoog (MS) basal medium supplemented with 1 mg l-1 benzyladenine (BA), 0.26 mg l-1 abscisic acid (ABA) and 0.8 mg l-1 indole-3-acetic acid (IAA) was used for shoot regeneration from cotyledonary explants in 'F39' and '150'. Kanamycin sensitivity as well as Timentin and Clavamox were evaluated using wild-type 'F39' and '150' cotyledons. Kanamycin concentrations of 200 and 150 mg l-1 were chosen as the threshold levels for 'F39' and '150', respectively. No significant differences were found between Timentin and Clavamox in 'F39'; however, Clavamox reduced the incidence of vitrification and increased the frequency of shoot elongation in '150'. A. tumefaciens strain EHA105, harboring pCNL56 carrying neomycin phosphotransferase II (nptII) and gusA reporter genes, was selected to establish a transformation protocol for 'F39' and '150'. Putative transformants were evaluated using -glucuronidase (GUS) histochemical assay, polymerase chain reaction (PCR) and Southern blot analyses. Based on these parameters, the transformation efficiency for cantaloupe 'F39' was 0.3% and that for honeydew '150' was 0.5%. 2011 Springer Science+Business Media B.V.

published proceedings

  • Plant Cell, Tissue and Organ Culture (PCTOC)

author list (cited authors)

  • Ren, Y., Bang, H., Curtis, I. S., Gould, J., Patil, B. S., & Crosby, K. M.

citation count

  • 15

complete list of authors

  • Ren, Yan||Bang, Haejeen||Curtis, Ian S||Gould, Jean||Patil, Bhimanagouda S||Crosby, Kevin M

publication date

  • January 2012