caa a22 4500 467901066 CHVBK 20180406152818.0 cr unu---uuuuu 170328e20060801xx s 000 0 eng 10.1007/s00299-006-0139-0 doi (NATIONALLICENCE)springer-10.1007/s00299-006-0139-0 Improved methods in Agrobacterium -mediated transformation of almond using positive (mannose/ pmi ) or negative (kanamycin resistance) selection-based protocols [Elektronische Daten] [Sunita Ramesh, Brent Kaiser, Tricia Franks, Graham Collins, Margaret Sedgley] A protocol for Agrobacterium-mediated transformation with either kanamycin or mannose selection was developed for leaf explants of the cultivar Prunus dulcis cv. Ne Plus Ultra. Regenerating shoots were selected on medium containing 15μM kanamycin (negative selection), while in the positive selection strategy, shoots were selected on 2.5g/l mannose supplemented with 15g/l sucrose. Transformation efficiencies based on PCR analysis of individual putative transformed shoots from independent lines relative to the initial numbers of leaf explants tested were 5.6% for kanamycin/nptII and 6.8% for mannose/pmi selection, respectively. Southern blot analysis on six randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene in each, and five randomly chosen lines identified to contain the pmi transgene by PCR showed positive hybridisation to a pmi DNA probe. The positive (mannose/pmi) and the negative (kanamycin) selection protocols used in this study have greatly improved transformation efficiency in almond, which were confirmed with PCR and Southern blot. This study also demonstrates that in almond the mannose/pmi selection protocol is appropriate and can result in higher transformation efficiencies over that of kanamycin/nptII selection protocols. Springer-Verlag, 2006 Almond nationallicence Mannose nationallicence pmi nationallicence Prunus nationallicence Transformation nationallicence Ramesh Sunita School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut Kaiser Brent School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut Franks Tricia School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut Collins Graham School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut Sedgley Margaret Faculty of Sciences, University of New England, NSW 2351, Armidale, Australia aut Plant Cell Reports Springer-Verlag 25/8(2006-08-01), 821-828 0721-7714 25:8<821 2006 25 299 https://doi.org/10.1007/s00299-006-0139-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-006-0139-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ramesh Sunita School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Kaiser Brent School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Franks Tricia School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Collins Graham School of Agriculture and Wine, Waite Campus, University of Adelaide, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Sedgley Margaret Faculty of Sciences, University of New England, NSW 2351, Armidale, Australia aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/8(2006-08-01), 821-828 0721-7714 25:8<821 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer