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Agrobacterium -mediated transformation of Solanum phureja

Agrobacterium -mediated transformation of Solanum phureja A population of transgenic plants was produced by the transformation of internodal explants of Solanum phureja, DB337/37 (the cultivar Mayan Gold) using an Agrobacterium tumefaciens LBA4404-based vector containing a phytoene synthase gene ( crtB ). The regeneration strategy utilised a two-step protocol, with a 12-day callus induction stage mediated by 1.07 μM α-napthaleneacetic acid (NAA), 7.10 μM zeatin riboside and 0.06 μM gibberellic acid (GA3), followed by a prolonged (up to 90 day) shoot induction stage on medium containing 0.11 μM NAA, 7.10 μM zeatin riboside and 0.06 μM GA3 supplemented with kanamycin at 50 mg l −1 as the selection agent. Southern analysis of the transgenic population revealed that the transgene copy number varied between one and five in the lines tested. Northern blot analysis showed significant expression of the introduced crtB gene in some lines during tuber development. Cytological analysis of the material showed a high incidence of chromosome doubling in the transgenic population with over 80% of all lines tested having doubled their chromosome complement during the transformation process. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Cell Reports Springer Journals

Agrobacterium -mediated transformation of Solanum phureja

Plant Cell Reports , Volume 24 (1) – Apr 1, 2005

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References (20)

Publisher
Springer Journals
Copyright
Copyright © 2005 by Springer-Verlag
Subject
LifeSciences
ISSN
0721-7714
eISSN
1432-203X
DOI
10.1007/s00299-004-0902-z
pmid
15666166
Publisher site
See Article on Publisher Site

Abstract

A population of transgenic plants was produced by the transformation of internodal explants of Solanum phureja, DB337/37 (the cultivar Mayan Gold) using an Agrobacterium tumefaciens LBA4404-based vector containing a phytoene synthase gene ( crtB ). The regeneration strategy utilised a two-step protocol, with a 12-day callus induction stage mediated by 1.07 μM α-napthaleneacetic acid (NAA), 7.10 μM zeatin riboside and 0.06 μM gibberellic acid (GA3), followed by a prolonged (up to 90 day) shoot induction stage on medium containing 0.11 μM NAA, 7.10 μM zeatin riboside and 0.06 μM GA3 supplemented with kanamycin at 50 mg l −1 as the selection agent. Southern analysis of the transgenic population revealed that the transgene copy number varied between one and five in the lines tested. Northern blot analysis showed significant expression of the introduced crtB gene in some lines during tuber development. Cytological analysis of the material showed a high incidence of chromosome doubling in the transgenic population with over 80% of all lines tested having doubled their chromosome complement during the transformation process.

Journal

Plant Cell ReportsSpringer Journals

Published: Apr 1, 2005

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