Summary
A transient ß-glucuronidase (GUS)-assay was performed to evaluate electroporation parameters and optimize DNA delivery conditions into strawberry protoplasts. Optimal GUS-activity was obtained when protoplasts were subjected to 400 V/cm for 20 ms. GUS-activity could be further increased by the addition of carrier DNA to the electroporation mixture. Callus selected on 10 μg/ml hygromycin produced shoots which exhibited GUS-activity. The transformed nature of the shoots obtained after selection was confirmed by DNA-analysis.
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Abbreviations
- CaMV:
-
cauliflower mosaic virus
- dCTP:
-
deoxycytidine-triphosphate
- EtBr:
-
ethidium bromide
- GUS:
-
ß-glucuronidase
- MES:
-
2(N-morpholino) -ethanesulfonic acid
- X-gluc:
-
5-bromo-4-chloro-3-indolyl glucuronide
References
Bekkaoui F, Pilon M, Laine E, Raju DSS Crosby WL, Dunstan DI (1988) Plant Cell Rep. 7:481–484
Birnboim HC and Doly J (1979) Nucleic Acids Res. 7:1513–1523
Ching-yeh H, Chee PP, Chesney RH, Zhou JH, Miller PD, O'Brien WT (1990) Plant Cell Rep. 9:1–5
Dellaporta SL, Wood J, Hicks JB (1983) Plant Mol. Biol. Rep. 1:19–21
Fromm M, Taylor LP, Walbot V (1985) Proc. Natl. Acad. Sci. 82:5824–5828
Hauptmann RM, Ozias-Akins P, Vasil V, Tabaeizadeh Z, Rogers SG, Horsch RB, Vasil IK, Fraley RT (1987) Plant Cell Rep. 6:265–270
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Fraley RT (1985) Science 227:1229–1231
James DJ, Passey AJ, Barbara DJ (1990) Plant Science 69:79–94
Jefferson RA, Kavanagh TA, Michael WB (1987) EMBO J. 6:3901–3907
Menzel L, Nagy F, Kizz ZsR, Maliga P (1981) Theor. Appl. Genet. 59:191–195
Murashige T, Skoog F (1962) Physiol. Plant. 15:473–497
Negrutiu I, R Shillito, I Potrykus, G Biasini, F Sala (1987) Plant Mol. Biol. 8:363–373
Nehra SN, Chibbar RN, Kartha KK, Datla RSS, Crosby WL, Stushnoff C (1990) Plant Cell Rep. 9:293–298
Nyman M, Wallin A (1988) J. Plant. Physiol. 133:375–377
Paszkowski J, Saul MW, Potrykus I (1989) In Cell Culture and Somatic Cell Genetics of Plants Schell J and Vasil IK (Eds.) Vol. 6:52–68 Academic Press Inc., San Diego
Rhodes CA, Pierce DA, Mettler IJ, Mascarenhas D, Detmer JJ (1988) Science 240:204–207
Séguin A, Lalonde M (1988) Plant Cell Rep. 7:367–370
Shaun LA, Hobbs JA, Jackson A, Baliski DS, DeLong CMO, Mahon JD (1990) Plant Cell Rep. 9:17–20
Shillito RD, Saul MW, Paszkowski J, Muller, Potrykus I (1985) Biotechnology 3:1099–1103
Tautorus TE, Bekkaoui F, Pilon M, Datla RSS, Crosby WL, Fowke LC, Dunstan DI (1989) Theor. Appl. Genet. 78:531–536
Yu-Wen H, Dennis ES (1989) Plant Cell Tissue and Organ culture 18:281–296
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Nyman, M., Wallin, A. Transient gene expression in strawberry (Fragaria x ananassa Duch.) protoplasts and the recovery of transgenic plants. Plant Cell Reports 11, 105–108 (1992). https://doi.org/10.1007/BF00235264
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DOI: https://doi.org/10.1007/BF00235264