NDLI: Biolistic transformation of grapevine using minimal gene cassette technology

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Biolistic transformation of grapevine using minimal gene cassette technology

Content Provider	Springer Nature Link
Author	Vidal, Jose R. Kikkert, Julie R. Donzelli, Bru D. Wallace, Patricia G. Reisch, Bruce I.
Copyright Year	2006
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 to non-transgenic lines. Moreover, transgenic lines from both MC and CP DNA treatments produced fruit as did the non-transgenic lines in the third year of growth in the greenhouse. Our data confirm the effectiveness of the minimal cassette technology for genetic transformation of grapevine cultivars.
Starting Page	807
Ending Page	814
Page Count	8
File Format	PDF
ISSN	07217714
Journal	Plant Cell Reports
Volume Number	25
Issue Number	8
e-ISSN	1432203X
Language	English
Publisher	Springer-Verlag
Publisher Date	2006-03-10
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Integration events Magainin genes Minimal cassette Transgene-transcription Vitis vinifera Plant Sciences Cell Biology Plant Biochemistry Biotechnology
Content Type	Text
Resource Type	Article
Subject	Plant Science Medicine Agronomy and Crop Science

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