Agrobacterium-mediated transformation of five wild Solanum species using in vitro microtubers |
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Authors: | A Kumar M Miller P Whitty J Lyon P Davie |
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Institution: | (1) Scottish Crop Research Institute, DD2 5DA Invergowrie, Dundee, Scotland, UK;(2) Potato Marketing Board, Broad Field House, 4 Between Town Road, OX4 3NA Cowley, Oxford, UK |
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Abstract: | Summary A genetic transformation method, using in vitro microtubers and Agrobacterium-mediated transformation has been developed for five wild Solanum species: S. verrucosum, S. hjertingii, S. papita, S. stoloniferum, S. demissum, which range in ploidy from diploid to hexaploid. A disarmed A. tumefaciens strain, C58 harbouring the co-integrate vector pGV3580::pKU2 with the genes of neomycin phosphotransferase (NPTII) and hygromycin phosphotransferase (HPTII) as selectable markers, was used. Selection of putative transformants was based on their ability to grow and produce roots on a medium containing 150 mg/l kanamycin. The transgenic nature of the putative transformants was confirmed by Polymerase Chain Reaction analysis and by NPTII dotblot assay to show the expression of the NPTII gene. Additionally, the transmission of transgenes, NPTII and HPTII in selfed-sexual progeny of some transgenic plants was also determined.Abbreviations MS
Murashige and Skoog
- NPTII
neomycin phosphotransferase
- HPTII
hygromycin phosphotransferase
- PCR
polymerase chain reaction
- GA3
gibberellic acid
- CCC
chlorocholine chloride
- BAP
benzylaminopurine
- NAA
naphthalene acetic acid
- ZR
zeatin riboside
- IAA
indole acetic acid
- LB
Luria Broth |
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