Biologia plantarum 55:414-422, 2011 | DOI: 10.1007/s10535-011-0105-8
Variability in shoot cultures regenerated from hairy roots of Gentiana punctata
- 1 Plant Physiology Department, Institute for Biological Research "S. Stanković", Belgrade University, Belgrade, Serbia
- 2 Institute for Medicinal Plants Research "Josif Pančić", Belgrade, Serbia
- 3 Institute of Molecular Genetics and Genetic Engineering, Belgrade, Serbia
Differences among three clones of Gentiana punctata L. hairy root shoot regenerants were investigated in relation to their growth patterns, production of secondary metabolites and 2D protein profiles. Prominent differences in growth parameters were stable thus qualifying regenerant clones as true somaclones. Marked differences in protein spots were registered among the regenerant clones but not in comparison with the non-transformed control. Southern blot hybridization of regenerants showed the absence of rolA, B and C genes, initially present in the main hairy root lines. Orf13 and rolD were present and orf8 was missing in all three regenerant clones whereas orf3 was missing only in clone 2. Although lacking the three major rol genes, plants of regenerant clones retained characteristics of the hairy root phenotype.
Keywords: Agrobacterium rhizogenes; 2D-electrophoresis; rol genes; secondary metabolites; somaclonal variation
Received: March 18, 2010; Accepted: June 9, 2010; Published: September 1, 2011Show citation
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References
- Batra, J., Dutta, A., Singh, D., Kumar, S., Sen, J.: Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri T-DNA gene integration. - Plant Cell Rep. 23: 148-154, 2004.
Go to original source...
- Bauer, N., Kiseljak, D., Jelaska, S.: The effect of yeast extract and methyl jasmonate on rosmarinic acid accumulation in Colleus blumei hairy roots. - Biol. Plant. 53: 650-656, 2009.
Go to original source...
- Birot, A.-M., Bouchez, D., Casse-Delbart, F., Durand-Tardif, M., Jouanin, L., Pautot, V., Robaglia, C., Tepfer, D., Tepfer, M., Tourneur, J., Vilaine, F.: Studies and uses of the rRi plasmids of Agrobacterium rhizogenes. - Plant Physiol. Biochem. 25: 323-335, 1987.
- Chaudhury, K.N., Ghosh, B., Tepfer, D., Jha, S.: Spontaneous plant regeneration in transformed roots and calli from Tylophora indica: changes in morphological phenotype and tylophorine accumulation associated with transformation by Agrobacterium rhizogenes. - Plant Cell Rep. 25: 1059-1066, 2006.
Go to original source...
- Christey, M.C.: Transgenic crop plants using Agrobacterium rhizogenes mediated transformation. - In: Doran, P.M. (ed.): Hairy Roots: Culture and Applications. Pp. 99-111. Harwood Academic Publishers, Amsterdam 1997.
- Christey, M.C.: Use of Ri mediated transformation for production of transgenic plants. - In Vitro cell. dev. Biol. Plant. 36: 687-700, 2001.
Go to original source...
- 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.
- David, C., Tempe, J.: Genetic transformation of cauliflower (Brassica oleracea L. var. botrytis) by Agrobacterium rhizogenes. - Plant Cell Rep. 7: 88-91, 1988.
Go to original source...
- Dmitrović, S., Mitić, N., Zdravković-Korać, S., Vinterhalter, B., Ninković, S., Ćulafić Lj.: Hairy roots formation in recalcitrant-to-transform plant Chenopodium rubrum. - Biol. Plant. 54: 566-570, 2010.
Go to original source...
- Filipecki, M., Malepszy, S.: Unintended consequences of plant transformation: a molecular insight. - J. appl. Genet. 47: 277-286, 2006.
Go to original source...
- Gangopadhyay, M., Sircar, D., Mitra, A., Bhattacharya, S.: Hairy root culture of Plumbago indica as a potential source for plumbagin. - Biol. Plant. 52: 533-537, 2008.
Go to original source...
- Gheysen, G., Dhaese, P., Van Montagu, M., Zambrzsky, P.: Integration of Agrobacterium tumefaciens transfer DNA (T-DNA) involves rearangements of target plant DNA sequences. - Proc. nat. Acad. Sci. USA 84: 6169-6173, 1987.
Go to original source...
- Guivarc'h, A., Boccara, M., Prouteau, M., Chriqui, D.: Instability of phenotype and gene expression in long term culture of carrot hairy root clones. - Plant Cell Rep. 19: 43-50, 1999.
Go to original source...
- Hosokawa, K., Matsuki, R., Oikawa, Y., Yamamura, S.: Genetic transformation using wild-type Agrobacterium rhizogenes. - Plant Cell Tissue Organ Cult. 51: 137-140, 1997.
Go to original source...
- Jouanin, L.: Restriction map of an agropine-type Ri plasmid and its homologies with Ti plasmid. - Plasmid 12: 92-102, 1984.
Go to original source...
- Jouanin, L., Bouchez, D., Drong, R.F., Tepfer, D., Slightom, J.L.: Analysis of TR-DNA / plant junctions in the genome of Concolvulus arvensis clone transformed with Agrobacterium rhizogenes strain A4. - Plant mol. Biol. 12: 75-85, 1989.
Go to original source...
- Lemke, K., Schmülling, T.: Gain of function assays identify non-rol genes from Agrobacterium rhizogenes TL-DNA that alter plant morphogenesis or hormone sensitivity. - Plant J. 15: 423-433, 1998.
Go to original source...
- Levesque, H., Delepelaire, P., Rouzé, P., Slightom, J., Tepfer, D.: Common evolution origin of the central portions of the Ri TL-DNA of Agrobacterium rhizogenes and Ti T-DNAs Agrobacterium tumefaciens. - Plant mol. Biol. 11: 731-744, 1988.
Go to original source...
- Lloyd, G.B., McCown, B.H.: Commercially feasible micropropagation of mountain laurel - Kalmia latifolia by use of shoot-tip culture. - Proc. int. Plant Propag. Soc. 30: 421-427, 1980.
- Mauro, M.L., Trovato, M., De Paolis, A., Gallelli, A., Costantino, P., Altamura, M.M.: The plant oncogene rolD stimulates flowering in transgenic tobacco plants. - Dev. Biol. 180: 693-700, 1996.
Go to original source...
- Menković, N., Šavikin-Fodulović, K., Vinterhalter, B., Vinterhalter, D., Grubišić, D.: Secoiridoid content of naturally grown and in vitro cultured Gentiana punctata. - Pharm. Pharmacol. Lett. 8: 110-111, 1998.
- Menković, N., Šavikin-Fodulović, K., Vinterhalter, B., Vinterhalter, D., Janković, T., Krstić, D.: Secoiridoid content in hairy roots of Gentiana punctata. - Pharm. Pharmacol. Lett. 10: 73-75, 2000.
- Mishiba, K.-I., Nishihara, M., Abe, Y., Nakatsuka. T., Kawamura, H., Kodama, K., Takesawa, T., Abe, J., Yamamura, S.: Production of dwarf potted gentian using wild-type Agrobacterium rhizogenes. - Plant Biotechnol. 23: 33-38, 2006.
Go to original source...
- Mishiba, K.-I., Nishihara, M., Nakatsuka, T., Abe, Y., Hirano, H., Yokoi, T., Kikuchi, A., Yamamura, S.: Consistent transcriptional silencing of 35S' driven transgenes in gentian. - Plant J. 44: 541-556, 2005.
Go to original source...
- Momčilović, I., Grubišić, D., Kojić, M., Nešković, M.: Agrobacterium rhizogenes-mediated transformation and plant regeneration of four Gentiana species. - Plant Cell Tissue Organ Cult. 50: 1-6, 1997.
Go to original source...
- Momčilović, I., Grubišić, D., Nešković, M.: Transgenic Gentiana species (Gentian). - In: Bajaj, Y.P.S. (ed.): Biotechnology in Agriculture and Forestry. Vol. 48. Transgenic crops III. Pp. 123-138. Springer-Verlag, Berlin - Heidelberg - New York 2001.
- Murray, M.G., Thompson, W.F.: Rapid isolation of high molecular weight plant DNA. - Nucl. Acids Res. 8: 4321-4325, 1980.
Go to original source...
- Nacry, P., Camilleri, C., Courtial, B., Caboche, M., Bouchey, D.: Major chromosomal rearangements induced by T-DNA transformation in Arabidopsis. - Genetics 149: 641-650, 1998.
- Nillson, O., Olsson, O.: Getting to the root: The role of the Agrobacterium rhizogenes rol genes in the formation of hairy roots. - Physiol. Plant. 100: 463-473, 1997.
Go to original source...
- 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.
- Piispanen, R., Aronen, T., Chen, X., Saranpää, P., Häaggman, H.: Silver birch (Betula pendula) plants with aux and rol genes show consistent changes in morphology, xylem structure and chemistry. - Tree Physiol. 23: 721-733, 2003.
Go to original source...
- Rosić, N., Momčilović, I., Kovačević, N., Grubišić D.: Genetic transformation of Rhamnus fallax and hairy roots as a source of anthraquinones. - Biol. Plant. 50: 514-518, 2006.
- Sevón, N., Dräger, B., Hiltunen, R., Oksman-Caldentey, K.M.: Characterization of transgenic plants derived from hairy roots of Hyosciamus muticus. - Plant Cell Rep. 16: 605-611, 1997.
Go to original source...
- Sevón, N., Oksman-Caldentey, K.M.: Agrobacterium rhizogenes- mediated transformation: root cultures as a source of alkaloids. - Planta med. 68: 859-868, 2002.
Go to original source...
- Slightom, J.L., Durand-Tardif, M., Jouanin, L., Tepfer, D.: Nucleotide sequence analysis of TL-DNA of Agrobacterium rhizogenes agropine type plasmid. - J. biol. Chem. 261: 108-121, 1986.
- Spano, L., Mariotti, D., Cardarelli, M., Branca, C., Constantino, P.: Morphologenesis and auxin sensitivity of transgenic tobacco with different complements of Ri T-DNA. - Plant Physiol. 87: 479-483, 1988.
Go to original source...
- Spena, A., Schmulling, T., Koncz, C., Schell, J.S.: Independent and synergistic activity of rolA, B and C loci in stimulating abnormal growth in plants. - EMBO J. 6: 3891-3899, 1987.
Go to original source...
- Sretenović-Rajičić, T., Ninković, S., Miljuš-Đukić, J., Vinterhalter, B., Vinterhalter, D.: Agrobacterium rhizogenes-mediated transformation of Brassica oleracea var. sabauda and B. oleracea var. capitata. - Biol. Plant. 50: 525-530, 2006.
Go to original source...
- Subotić, A., Budimir, S., Grubišić, D., Momčilović, I.: Direct regeneration of shoots from hairy root cultures of Centaurium erythraea inoculated with Agrobacterium rhizogenes. - Biol. Plant. 47: 617-619, 2003.
- Suginuma, C., Akihama, T.: Transformation of Gentian with Agrobacterium rhizogenes. - Acta Hort. 392: 153-160, 1995.
Go to original source...
- Tax, F.E., Vernon, D.M.: T-DNA-associated duplication/ translocation in Arabidopsis. Implications for mutant analysis and functional genomics. - Plant Physiol. 126: 1527-1538, 2001.
Go to original source...
- Teixeira da Silva, J.A.: Chrysanthemum: advances in tissue culture, cryopreservation, postharvest technology, genetics and transgenic biotechnology. - Biotechnol. Adv. 21:715-766, 2003.
Go to original source...
- Tepfer, D.: Genetic transformation using Agrobacterium rhizogenes. - Physiol. Plant. 79: 140-146, 1990.
Go to original source...
- Tepfer, M., Casse-Delbart, F.: Agrobacterium rhizogenes as a vector for transforming higher plants. -Microbiol. Sci. 4: 24-28, 1987.
- Tiwari, R.K., Trivedi, M., Guang, Z.-C., Guo, G.-Q., Yang, G.-C.: Genetic transformation of Gentiana macrophylla with Agrobacterium rhizogenes: growth and production of secoiridoid glucoside gentiopicroside in transformed hairy root cultures. - Plant Cell Rep. 26: 199-210 2007.
Go to original source...
- Tiwari, R.K., Trivedi, M., Guang, Z.-C., Guo, G.-Q., Zheng, G-C.: Agrobacterium rhizogenes mediated transformation of Scutellaria baicalensis and production of flavonoids in hairy roots. - Biol. Plant. 52: 26-35, 2008.
Go to original source...
- Veena, V., Taylor, G.C.: Agrobacterium rhizogenes: recent developments and promissing applications. - In Vitro cell. Dev. Biol. Plant 43: 383-403, 2007.
Go to original source...
- Vinterhalter, B., Momčilović, I., Vinterhalter, D.: High biomass producing root cultures of Gentiana punctata L. transformed with Agrobacterium tumefaciens C58C1 (pArA4b). - Arch. Biol. Sci. Belgrade 52: 85-90, 2000.
- Vinterhalter, B., Ninković, S., Cingel, A., Vinterhalter D.: Shoot and root culture of Hypericum perforatum L. transformed with Agrobacterium rhizogenes A4M70GUS. - Biol. Plant. 50: 767-770, 2006.
Go to original source...
- Vinterhalter, B., Orbović, V., Vinterhalter, D.: Transgenic root cultures of Gentiana punctata L. - Acta Soc. Bot. Polon. 68: 275-280, 1999.
- Vinterhalter, B., Savić, J., Platiša, J., Raspor, M., Ninković, S., Mitić, N., Vinterhalter, D.: Nickel tolerance and hyperaccumulation in shoot cultures regenerated from hairy root cultures of Alyssum murale Waldst et Kit. - Plant Cell Tissue Organ Cult. 94: 299-303, 2008.
Go to original source...
- Vinterhalter, B., Vinterhalter, D.: In vitro propagation of spotted gentian Gentiana punctata L. - Arch. Biol. Sci. Belgrade 50: 177-182, 1998.
- White, F.F., Taylor, B.H., Huffman, A., Gordon, M.P., Nester, E.W.: Molecular and genetic analysis of the transferred DNA regions of the root inducing plasmid of Agrobacterium rhizogenes. - J. Bacteriol. 164: 33-44, 1985.
- Zdravković-Korać, S., Muhovski, Y., Druart, P., Ćalić, D., Radojević, L.: Agrobacterium rhizogenes-mediated DNA transfer to Aesculus hippocastamum L. and the regeneration of transformed plants. - Plant Cell Rep. 22: 698-704, 2004.
Go to original source...