Biolistic transformation of Carrizo citrange (Citrus sinensis Osb. × Poncirus trifoliata L. Raf.)

Publication Overview
TitleBiolistic transformation of Carrizo citrange (Citrus sinensis Osb. × Poncirus trifoliata L. Raf.)
AuthorsWu H, Acanda Y, Jia H, Wang N, Zale J
TypeJournal Article
Journal NamePlant cell reports
Year2016
CitationWu H, Acanda Y, Jia H, Wang N, Zale J. Biolistic transformation of Carrizo citrange (Citrus sinensis Osb. × Poncirus trifoliata L. Raf.). Plant cell reports. 2016 Jun 8.

Abstract

KEY MESSAGE
The development of transgenic citrus plants by the biolistic method. A protocol for the biolistic transformation of epicotyl explants and transgenic shoot regeneration of immature citrange rootstock, cv. Carrizo (Citrus sinensis Osb. × Poncirus trifoliata L. Raf.) and plant regeneration is described. Immature epicotyl explants were bombarded with a vector containing the nptII selectable marker and the gfp reporter. The number of independent, stably transformed tissues/total number of explants, recorded by monitoring GFP fluorescence 4 weeks after bombardment was substantial at 18.4 %, and some fluorescing tissues regenerated into shoots. Fluorescing GFP, putative transgenic shoots were micro-grafted onto immature Carrizo rootstocks in vitro, confirmed by PCR amplification of nptII and gfp coding regions, followed by secondary grafting onto older rootstocks grown in soil. Southern blot analysis indicated that all the fluorescing shoots were transgenic. Multiple and single copies of nptII integrations were confirmed in five regenerated transgenic lines. There is potential to develop a higher throughput biolistics transformation system by optimizing the tissue culture medium to improve shoot regeneration and narrowing the window for plant sampling. This system will be appropriate for transformation with minimal cassettes.

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Bf0026Bf0026genetic_marker
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Additional details for this publication include:
Property NameValue
Publication ModelPrint-Electronic
ISSN1432-203X
eISSN1432-203X
Publication Date2016 Jun 8
Journal AbbreviationPlant Cell Rep.
LanguageEnglish
Language AbbrENG
Publication TypeJournal Article
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PMID: PubMedPMID:27277128