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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
agricultural and biological sciences
Development and evaluation of a GFLV inverted repeat construct for genetic transformation of grapevine
Plant Cell, Tissue and Organ Culture, Volume 97, No. 2, Year 2009
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Description
Grapevine Fanleaf virus (GFLV) is one of the most damaging and widespread nepovirus in grapevine, causing the fanleaf disease. Here, we report the development of inverted repeat (IR) constructs, consisting of GFLV-derived sequences, for genetic transformation of grapevine to induce GFLV-specific silencing. The IR construct was designed with fragments of a conserved region of the GFLV movement protein (MPc) gene from a Tunisian isolate (GFLV tun) and with cassettes containing the neomycin phosphotransferase or bar gene as selectable marker. For proof of concept, the IR construct was transferred into Nicotiana benthamiana by Agrobacterium mediated transformation. Challenge inoculation of T1 transgenic plant lines with the relevant virus resulted in plants showing resistance, recovery, retarded infection and susceptibility. Northern blot analysis, using a virus-derived sequence (MPc) as probe, could detect transgene specific small interfering RNA in resistant transgenic N. benthamiana plants after GFLV inoculation. Embryogenic cells of grapevine (Vitis vinifera cv Arich dressé) were transformed with the IR construct in combination with the bar gene using A. tumefaciens strain LBA4404. After molecular characterization of regenerated putative transgenic grapevine plants, Northern blot analysis detected different levels of IR transcripts in independent transgenic grapevine lines indicating a post-transcriptional gene silencing. © 2009 Springer Science+Business Media B.V.
Authors & Co-Authors
Jardak, Rahma
Tunisia, Hamam Lif
Centre de Biotechnologie de Borj Cédria
Winterhagen, Patrick
Germany, Neustadt/weinstrasse
Rlp Agroscience Gmbh
Bouamama, Badra
Tunisia, Hamam Lif
Centre de Biotechnologie de Borj Cédria
Dubois, Cornelia
Germany, Neustadt/weinstrasse
Rlp Agroscience Gmbh
Mliki, A.
Tunisia, Hamam Lif
Centre de Biotechnologie de Borj Cédria
Wetzel, Thierry
Germany, Neustadt/weinstrasse
Rlp Agroscience Gmbh
Ghorbel, Abdelwahed
Tunisia, Hamam Lif
Centre de Biotechnologie de Borj Cédria
Reustle, Goetz M.
Germany, Neustadt/weinstrasse
Rlp Agroscience Gmbh
Statistics
Citations: 30
Authors: 8
Affiliations: 2
Identifiers
Doi:
10.1007/s11240-009-9514-1
ISSN:
01676857
Research Areas
Genetics And Genomics