Abstract
Internode explants ofin vitro plants ofForsythia x intermedia “Spring Glory” were transformed with thegus andnpt II genes after inoculation with theA. tumefaciens strain EHA 101 harbouring the plasmid pFAJ3000. Shoot organogenesis took place from callused edges of explants. The first transformed buds were detected 4 to 6 weeks after transfer on regeneration medium, containing 25 mg/l kanamycin as selective agent. An average of 1% of explants regenerated transgenic shoots.
β-glucuronidase assays and culture on kanamycin-containing medium provided the first indication of integration and expression of introduced genes in transformants. Southern blot and polymerase chain reaction amplification analyses gave molecular confirmation of genetic transformation. Transgenic plants were acclimatized in the greenhouse. Enzymatic assays on several organs of mature plants still showed β-glucuronidase activity, thus confirming stable integration of T-DNA in the plant genome.
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Abbreviations
- BAP:
-
6-benzyl-aminopurine
- CaMV:
-
Cauliflower Mosaic Virus
- GUS andgus :
-
β-glucuronidase
- IAA:
-
indole-3-acetic acid
- IBA:
-
indole-3-butyric acid
- MS:
-
Murashige and Skoog
- NOS:
-
nopaline synthase
- NPT II andnpt II :
-
neomycin phosphotransferase II
- PCR:
-
polymerase chain reaction
- SDS:
-
sodium dodecyl sulphate
- SSC:
-
sodium chloride-sodium citrate
- X-Gluc:
-
5-bromo-4-cbloro-3-indolyl glucuronide
References
de Bondt A, Eggermont K, Druart P, De Vil M, Goderis I, Vanderleyden J, Broekaert W (1994) Plant Cell Reports 13: 587–593
Dellaporta S, Wood J, Hicks JB (1983) Plant Mol Biol Rep 4: 19–21
Duron M (1977) C R Acad Sci série D 284: 183–185
Duron M (1981) Agronomie l: 865–868
Firoozabady E, Moy Y, Courtney-Gutterson N, Robinson K (1994) Bio/Technology 12: 609–613
Godwin I, Todd G, Ford-Lloyd B, Newbury HJ (1991) Plant Cell Reports 9: 671–675
Guivarc'h A, Caissard J-C, Brown S, Marie D, Dewitte W, Van Onckelen H, Chriqui D (1993) Protoplasma 174: 10–18
Hajela RK, Hajela N, Bolyard MG, Barnes WK Sticklen MB (1993) HortSci 28: 330–332
Hood EE, Helmer GL, Frayley RT, Chilton MD (1986) J Bacteriol 168: 1297–1301
Jefferson RA (1987) Plant Mol Biol Rep 5: 387–405
Morel G, Martin C (1955) C R Acad Agric 41: 472–475
Muraslilge T, Skoog F (1962) Physiol Plant 15: 469–497
Pavingerová D, Bisková R, Kodýtek K (1995) Proc. XVIII Eucarpia Symp, Tel Aviv, p. 23.
Roest S, Bokelman GS (1980) Vakblad voor de Bloemisterij 45: 34–35
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. 2nd edn, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Sampson DR (1971) Can J Genet Cytol 13: 368–371
Schuerman PL, Dandekar A (1993) Sci Hortic 55: 101–124
Vancanneyt G, Schmidt R, O' Connor-Sanchez A, Willmitzer L, RochaSosa M (1990) Mol Gen Genet 220: 245–250
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Rosati, C., Cadic, A., Renou, JP. et al. Regeneration andAgrobacterium-mediated transformation ofForsythia xintermedia “Spring Glory”. Plant Cell Reports 16, 114–117 (1996). https://doi.org/10.1007/BF01275463
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DOI: https://doi.org/10.1007/BF01275463