Abstract
Transgenic plants of hyacinth (Hyacinthus orientalis L.) cvs. Edisson and Chine Pink have been obtained by Agrobacterium-mediated transformation. Leaf explants of the both hyacinth cultivars regenerated shoots on MS medium containing 2.2 μM BAP and 0.3 μM NAA at a frequency of 95%. A. tumefaciens strain CBE21 carrying binary vector pBIThau35 was used for transformation. Plasmid pBIThau35 has been produced by cloning preprothaumatin II cDNA into pBI121 instead of uidA gene. Inoculated leaf explants formed calli and shoots at high frequency on selective medium with 100 mg l−1 kanamycin. Four hyacinth transgenic lines of cv. Chine Pink and one line of cv. Edisson have been selected on medium containing 200 mg l−1 kanamycin. The insertion of thaumatin II gene into hyacinth genome has been confirmed by PCR-analysis. All transgenic plants expressed substantial amounts of thaumatin II (between 0.06 and 0.28% of the total soluble protein). Hyacinth transgenic lines were assayed for resistance to the pathogenic fungi Fusarium culmorum and Botrytis cinerea. There were no significant differences between nontransformed control and transgenic leaves of both cultivars. At the same time the bulbs of the transgenic line Н7401 cv. Chine Pink showed the higher level of resistance to B. cinerea, the bulbs of the transgenic line Н7404 were more resistant to F. culmorum. In both cases the signs of the fungal disease were developed more slowly. The resistance of the bulbs cv. Edisson line to these fungi was not changed. All transgenic hyacinth plant were successfully transferred to soil for further evaluation.
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Abbreviations
- BAP:
-
6-benzylaminopurine
- CaMV:
-
Cauliflower mosaic virus
- IBA:
-
3-indolebutyric acid
- NAA:
-
Naphthaleneacetic acid
- uidA :
-
β-glucuronidase gene
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Acknowledgements
We are grateful to Dr. A.M. Ledeboer for kindly provided pUR528:thaumatin II and Dr. V.S. Golubeva (Central botanical garden NАS of Belarus) for fungal cultures and help with fungal infection assays.
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Popowich, E.A., Firsov, A.P., Mitiouchkina, T.Y. et al. Agrobacterium-mediated transformation of Hyacinthus orientalis with thaumatin II gene to control fungal diseases. Plant Cell Tiss Organ Cult 90, 237–244 (2007). https://doi.org/10.1007/s11240-007-9254-z
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DOI: https://doi.org/10.1007/s11240-007-9254-z