Abstract
Agrobacterium rhizogenes A4M70GUS-mediated transformation of Savoy cabbage (Brassica oleracea L. var. sabauda) and two local lines of cabbage (B. oleracea L. var. capitata) was obtained using hypocotyl and cotyledon explants. The percentage of explants which formed roots was very high in all genotypes: 92.3 % in Savoy Gg-1, 64.4 % in cabbage P22I5, and 87.2 % in P34I5. Spontaneous shoot regeneration of excised root cultures grown on the hormone-free medium occurred in all three genotypes. In cabbage lines P22I5 and P34I5 shoot regeneration was higher (9.3 and 2.6 % respectively) than in Savoy cabbage Gg-1 (1.3 %). Transgenic nature of hairy root-derived plants was evaluated by GUS histological test and PCR analysis. All the tested cabbage shoots were GUS positive whilst in a Savoy cabbage GUS expression was registered only in 55 % of tested clones. PCR analysis demonstrated the presence of the GUS gene in regenerated shoot clones and in T1 progeny.
Similar content being viewed by others
Abbreviations
- BA:
-
6-benzyladenine
- GUS:
-
β-glucuronidase
- IBA:
-
indole-3-butyric acid
- KIN:
-
kinetin (6-phurphuryl aminopurine)
- MS medium:
-
Murashige and Skoog medium
- PCR:
-
polymerase chain reaction
- TDZ:
-
thidiazuron
References
Berthomieu, P., Jouanin, L.: Transformation of rapid cycling cabbage (Brassica oleracea var. capitata) with Agrobacterium rhizogenes.-Plant Cell Rep. 11: 334–338, 1992.
Bijelović, A., Rosić, N., Miljuš-Đukić, J., Ninković, S., Grubušić, D: In vitro regeneration and transformation of Blackstonia perfoliata.-Biol. Plant. 48: 333–338, 2004.
Boulter, M.E., Croy, E., Simpson, P., Shields, R., Croy, R.D.D., Shirsat, A.H.: Transformation of Brasica napus L. (oilseed rape) using Agrobacterium tumefaciens and Agrobacterium rhizogenes-a comparison.-Plant Sci. 70: 91–99, 1990.
Christey, M.C.: Use of Ri-mediated transformation for production of transgenic plants.-In Vitro cell. dev. Biol. Plant 37: 687–700, 2001.
Christey, M.C., Braun, R.H.: Transgenic vegetable and forage Brassica species: rape, kale, turnip and rutabaga (swede).-In: Bajaj, Y.P.S (ed.): Biotechnology in Agriculture and Forestry. Vol. 47. Transgenic Crops II. Pp. 87–101. Springer-Verlag, Berlin-Heidelberg-New York 2001.
Christey, M.C., Braun, R.H., Reader, J.K.: Field performance of transgenic vegetable brassicas (Brassica oleracea and B. rapa) transformed with Agrobacterium rhizogenes.-Sabrao J. Breed. Genet. 31: 93–108, 1999.
Christey, M.C., Sinclair, B.K.: Regeneration of transgenic kale (Brassica oleracea var. acephala), rape (B. napus) and turnip (B. campestris var. rapifera) plants via Agrobacterium rhizogenes mediated transformation.-Plant Sci. 87: 161–167, 1992.
Christey, M.C., Sinclair, B.K., Braun, R.H., Wyke, L.: Regeneration of transgenic vegetable brassicas (Brassica oleracea and B. campestris) via Ri-mediated transformation.-Plant Cell Rep. 16: 587–593, 1997.
David, C., Tempe, J.: Genetic transformation of cauliflower (Brassica oleracea L. var. botrytis) by Agrobacterium rhizogenes.-Plant Cell Rep. 7: 88–91, 1988.
Hamada, M., Hosoki, T., Kusabiraki, Y., Kigo, T.: Hairy root formation and plantlet regeneration from Brussels sprouts (Brassica oleracea var. gemmifera Zenk.) mediated by Agrobacterium rhizogenes.-Plant Tissue Cult. Lett 6: 130–133, 1989.
Henzi, M.X., Christey, M.C., McNeil, D.L.: Factors that influence Agrobacterium rhizogenes-mediated transformation of broccoli (Brasica oleracea L. var. italica).-Plant Cell Rep. 19: 994–999, 2000.
Henzi, M.X., Christey, M.C., McNeil, D.L., Davies, K.M.: Agrobacterium rhizogenes-mediated transformation of broccoli (Brasica oleracea L. var. italica) with an antisense 1-aminocyclopropane-1-carboxylic acid oxidase gene.-Plant Sci. 143: 55–62, 1999.
Hosoki, T., Kanbe, H., Kigo, T.: Transformation of ornamental tobacco and kale mediated by Agrobacterium tumefaciens and A. rhizogenes harbouring a reporter, β-glucuronidase (gus) gene.-J. jap. Soc. hort. Sci. 63: 167–172, 1994.
Hosoki, T. Kigo, T.: Transformation of Brussels sprouts (Brassica oleracea var. gemmifera Zenk.) by Agrobacterium rhizogenes harboring a reporter β-glucuronidase gene.-J. jap. Soc. hort. Sci. 63: 589–592, 1994.
Hosoki, T., Kigo, T., Shiraishi, K.: Transformation and regeneration of broccoli (Brassica oleracea var. italica) mediated by Agrobacterium rhizogenes.-J. jap. Soc. hort. Sci. 60: 71–75, 1991.
Jefferson, R.A., Kavanagh, T.A., Bevan, M.W.: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.-EMBO J. 6: 3901–3907, 1987.
Miljuš-Đukić, J., Vinterhalter, D., Vinterhalter, B., Ćalović, M., Ninković, S.: In vitro propagation and Agrobacterium-mediated transformation of potato cv. Desiree.-Bull. Inst. Bot., Bot. Garden, Univ. Belgrade 29: 115–121, 1996.
Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures.-Physiol. Plant. 15: 473–497, 1962.
Nikolić, R., Mitić, N., Ninković, S., Miljuš-Đukić, J., Nešković, M.: Efficient genetic transformation of Lotus corniculatus L. and growth of transformed plants in field.-Biol. Plant. 47: 137–140, 2003/4.
Petit, A., David, C., Dahl, G.A., Ellis, J.G., Guyon, P., Casse-Delbart, F., Tempe, J.: Further extension of the opine concept: plasmids in Agrobacterium rhizogenes cooperate for opine degradation.-Mol. gen. Genet. 190: 204–214, 1983.
Poulsen, G.B.: Genetic transformation of Brassica.-Plant Breed. 115: 209–225, 1996.
Puddephat, I.J., Riggs, T.J., Fenning, T.M.: Transformation of Brassica oleracea L.: a critical review.-Mol. Breed. 2: 185–210, 1996.
Puddephat, I.J., Robinson, H.T., Fenning, T.M., Barbara, D.J., Morton, A., Pink, D.A.C.: Recovery of phenotypically normal transgenic plants of Brassica oleracea upon Agrobacterium rhizogenes-mediated co-transformation and selection of transformed hairy roots by GUS assay.-Mol. Breed. 7: 229–242, 2001.
Sretenović-Rajičić, T., Ninković, S., Vinterhalter, B., Miljuš-Đukić, J., Vinterhalter, D.: Introduction of resistance to herbicide Basta® in Savoy cabbage.-Biol. Plant. 48: 431–436, 2004.
Tepfer, D., Casse-Delbart, F.: Agrobacterium rhizogenes as a vector for transforming higher plants.-Microbiol. Sci. 4: 24–28, 1987.
Van Larebeke, N., Genetello, C.H., Hernalsteens, J.P., De Picker, A., Zaenen, I., Messens, E., Van Montagu, M., Schell, J.: Transfer of Ti plasmids between Agrobacterium strains by mobilization with the conjugative plasmid RP4.-Mol. gen. Genet. 152: 1119–1124, 1977.
Zhou, X., Cao, G., C., Lin, R., Sun, Y., Li, W.: A rapid and efficient DNA extraction method of genus Fagopyrum for RAPD analysis.-In: Javornik, B., Bohanec, B., Kreft, I. (ed.): Proceedings of Impact of Plant Biotechnology on Agriculture. Pp. 171–175. Biotechnical Faculty, Ljubljana 1994.
Zdravković-Korać, S., Muhovski, Y., Druart, P.H., Ćalić, D., Radojević, Lj.: Agrobacterium rhizogenes-mediated DNA transfer to Aesculus hippocastanum L. and the regeneration of transformed plants.-Plant Cell Rep. 22: 698–704, 2004.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sretenović-Rajičić, T., Ninković, S., Miljuš-Đukić, J. et al. Agrobacterium rhizogenes-mediated transformation of Brassica oleracea var. sabauda and B. oleracea var. capitata . Biol Plant 50, 525–530 (2006). https://doi.org/10.1007/s10535-006-0083-4
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10535-006-0083-4