Year-end Sale % OFF 
Abstract
The goal of this research was to develop an efficient transformation system for 'Marshall McIntosh' apple. To determine the optimum combination of agar and Gelrite gelling agents in the media to maximize regeneration and minimize hyperhydicity (vitrification), the following combinations of agar (A)+Gelrite (G) in g l-1 were tested: 7.0 A+0 G; 5.2 A+0.6 G; 3.5 A+2.5 G; 1.7 A+1.8 G; and 0 A+2.5 G. Both 5.2 A+0.6 G and 3.5 A+1.2 G provided greatest regeneration of healthy non-hyperhydric shoots. To determine the optimal concentration of aminoglycoside for the selection and regeneration of transgenic 'Marshall McIntosh' on agar-Gelrite-based media, kanamycin was tested at 0, 10, 25, 50, 75 and 100 mg l-1, and paromomycin was tested at 0, 50, 100, 150, 200 and 250 mg l-1. Kanamycin was more effective than paromomycin in the initial selection of transgenics. For selection of transformants of 'Marshall McIntosh', the use of kanamycin at 25 mg l-1on 5.2 A+0.6 G solidified medium is suggested. By optimizing the medium and selection conditions, a protocol was developed that resulted in four transgenic lines as confirmed by a GUS assay, NPT II ELISA, PCR, and Southern analysis. In repeated experiments with this protocol, transformation efficiencies of 3.1 and 2.6% were obtained.
Published Version
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
