ISHS Acta Horticulturae 603: VIII International Conference on Grape Genetics and Breeding
EVALUATION OF DIFFERENT GENE CONSTRUCTS FOR PRODUCTION OF RESISTANT GRAPEVINES AGAINST GRAPEVINE FANLEAF AND ARABIS MOSAIC VIRUSES |
| Authors: | R. Jardak-Jamoussi, B. Bouamama, T. Wetzel, A. Mliki, G.M. Reustle, A. Ghorbel |
| Keywords: | Vitis, nepovirus, virus resistance, mouvement protein, inverted repeat, somatic embryogenesis, Tunisian grapevines. |
| Abstract:
Grapevine fanleaf virus (GFLV) and arabis mosaic virus (ArMV) are two nepoviruses causing Fanleaf degeneration, the major disease of grapevines. With the aim to obtain a protection against this viruses, we developed four constructs, containing non-translatable sequences of the movement protein (MP) and the coat protein (CP) of GFLV from a Tunisian isolate and of the movement protein of ArMV from a German isolate. This constructs including sense, antisense and inverted repeat sequences have been first introduced into the herbaceous host Nicotiana bethamiana by Agrobacterium tumefaciens. Transformed tissues were selected by continued proliferation on Kanamycin or Phosphinotricin containing media. Integration of transgenes in tobacco vitroplants was verified by PCR. Homozygous transgenic tobacco lines were obtained by selection of the T2 seeds in corresponding selection medium. Challenge inoculation of T2 progeny tobacco lines with GFLV and ArMV yielded plants showing immunity, delayed infection, recovery and susceptibility. Molecular analysis are in process to investigate possible gene silencing events in the resistant plants. Anther derived embryogenic calli from Tunisian autochthonous varieties were co-cultured with LBA 4404 harboring the construct carrying a GFLV movement protein sequence in inverted repeat orientation. Putative transformed plantlets were regenerated under Kanamycin selection. | |
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