Biolistic transformation of grapevine using minimal gene cassette technology

Plant Cell Reports
Jose R VidalBruce I Reisch

Abstract

The use of minimal gene cassettes (MCs), which are linear DNA fragments (promoter+open reading frame+terminator) lacking the vector backbone sequence, was compared to the traditional use of whole circular plasmids (CPs) for transformation of grapevine. Embryogenic cell suspensions of 'Chardonnay' (Vitis vinifera L.) were transformed via particle co-bombardment using two nonlinked genes in either MCs or CPs. One construct contained the npt-II selectable marker and the second construct contained the MSI99 antimicrobial peptide gene. A total of five lines each from MC and CP treatments that showed positive signals by PCR for both the npt-II and MSI99 genes were selected. Southern blot analyses revealed up to five integration events in the DNA treatments. Transcription levels determined by semi-quantitative RT-PCR varied among transgenic lines. No significant differences were found in transgene transcription between lines from MC and CP transformation. The correlation between npt-II and MSI99 transcription levels was positive (P<0.05), however, no correlation between the transcription level and the number of integration events was observed. Transgenic lines presented a similar phenotype in leaf morphology and plant vigor compared t...Continue Reading

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Citations

Mar 3, 2011·Genetics and Molecular Research : GMR·G R ViannaE L Rech
Dec 12, 2012·Journal of Biotechnology·Laura SanjurjoFrancisco de la Torre
Nov 29, 2014·Plant Biotechnology Journal·Noémie S JellyPascale Maillot
Sep 22, 2007·Journal of Genetics and Genomics = Yi Chuan Xue Bao·Yan ZhaoDanian Huang
Jun 19, 2012·Physics of Life Reviews·Ana Leonor RiveraAchim M Loske
May 26, 2017·PloS One·Ikuko NakajimaMasaki Endo
Oct 1, 2008·Phytochemistry Reviews : Proceedings of the Phytochemical Society of Europe·Georg SchmidtFatima Ferreira
Mar 12, 2008·Journal of Experimental Botany·Denise V Duclos, Thomas Björkman
May 5, 2021·Plant Biotechnology Journal·Gaetano GiudiceWalter Chitarra

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