Tissue culture of Pyrethrum (Tanacetum cinerariifolium) and associated microbial contamination
DOI:
https://doi.org/10.14720/abs.53.1.15369Keywords:
tissue culture, pyrethrum, Tanacetum cinerariifolium, callus culture, microbial contamination, PCRAbstract
Microbial contamination was observed on several subcultures of Pyrethrum (Tanacetum cinerariifolium) (Trevir.) Schultz-bip. callus lines. The presence of microorganisms- sms was detected by isolation of contaminants in pure culture from 7 out of 34 callus lines and direct ampliication of eubacterial 16S rDNA in the pyrethrum callus and plants and isolated bacteria. Altogether 16 contaminants were further analyzed, observing their morphology on several media and restriction of ampliied 16S rDNA. Analysis revealed presence and persistence of morphologically and genetically diverse bacteria in pyrethrum tissue culture. Due to cross-reactivity of 16S rDNA primers with DNA of plant origin, no conclusions could be drawn on the origin of contaminants.
References
Altschul, S.F., Gish, W., Miller W, Myers, E.W., Lipman, D.J., 1990. Basic local alignment search tool. Journal of Molecular Biology, 215, 403–410.
Altschul, S.F., Madden, T.L., Schaer, A.A., Zhang, J., Zhang, Z., Miller, W., Lipman, D.J., 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research, 25, 3389–3402.
Ambrožič-Dolinšek, J., Kovač, M., Žel, J., Camloh, M., 2007. Pyrethrum (Tanacetum cinerariifo- lium) from Northern Adriatic as a potential source of natural insecticide. Annales Serie Historia Naturalis, 17(1), 39–46.
Ambrožič-Dolinšek, J., Camloh, M., Žel, J., Kovač, M., Ravnikar, M., Carraro, L., Petrovič, N., 2008. Phytoplasma infection may affect morphology, regeneration and pyrethrin content in pyrethrum shoot culture. Scientia Horticulturae, 116(2), 213–218.
Bergant, M., Ambrožič-Dolinšek, J., Demšar, T., Dreo, T., Ravnikar, M., Žel, J., Camloh, M., 2005. Effect of antibiotic on contaminated callus cultures of pyrethrum. Phyton (Horn, Austria), 45(1), 197–206.
Bianciotto, V., Bandi, C. Minerdi, D., Sironi, M., Tichy, H.V., Bonfante, P., 1996. An obligately mycorrhizal fungus itself harbors obligately intracellular bacteria. Applied and Environmental Microbiology, 62(8), 3005–3010.
Cankar, K., Kraigher, H., Ravnikar, M., Rupnik, M., 2005. Bacterial endophytes from seeds of Norway spruce (Picea abies L. Karst). FEMS Microbiology Letters, 244, 341–345.
Dent, K.C., Stephen, J.R., Finch-Savage, W.E., 2004. Molecular proiling of microbial communities associated with seeds of Beta vulgaris subsp. Vulgaris (sugar beet) Journal of Microbiological Methods, 56, 17–26.
Herman, E.B., 2004: Microbial »Contaminants« in Plant Tissue Cultures: Solutions and Opportunities 1996–2003. In: Herman EB (ed) Recent Advances in Plant Tissue Culture Volume 8. Agricell Report, New York.
Herman, E.B., 2007: Microbial Contaminants in Plant Tissue Cultures: 2003–2007. In: Herman, E.B. (ed) Recent Advances in Plant Tissue Culture Volume 11. Agritech Agricell Report, New York.
Hitmi, A., Coudret, A., Barthomeuf, C., 2000. The Production of pyrethrins by plant cell and tissue cultures of Chrysanthemum cinerariaefolium and Tagetes species. Critical Reviews on Bioche- mistry and Molecular Biology, 35, 317–337.
Gspan, M., Vrtačnik, M., Ambrožič-Dolinšek, J., Kovač, M., Camloh, M., Žel, J., 2004. Tissue cul- ture of pyrethrum (Tanacetum cinerariifolium) (Trevir.) Schultz Bip.). Acta Biologica Slovenica, 47(2), 45–56.
Jovetić, S., de Gooijer, C.D., 1995. The production of pyrethrins by in vitro systems. Critical Review in Biotechnology, 15(2), 125–138.
Keskitalo, M.K., Pohto, A., Savela, M.L., Valkonen, J.P.T., Simon, J., Pehu, E., 1998b. Alteration in growth of tissue-cultured tansy (Tanacetum vulgare L.) treated with antibiotics. Annals of Applied Biology, 133, 281–294.
Leifert, C., Cassells, A.C., 2001. Microbial hazards in plant tissue and cell cultures. In vitro Cellular and Developmental Biology – Plant, 37(2), 133–138.
Misaghi, I.J., Donndelinger, C.R., 1990. Endophytic bacteria in symptom-free cotton plants. Phyto- pathology, 80, 808–811.
Roberts, R.J., Vincze, T., Posfai, J., Macelis, D., 2007. REBASE – enzymes and genes for DNA restriction and modiication. Nucleic Acids Research, 35, 269–270.
Rosenblueth, M., Martinez-Romero, E., 2006. Bacterial endophytes and their interactions with hosts. Molecular Plant Microbe Interaction, 19(8), 827–837.
Sessitsch, A., Reiter, B., Pfeifer, U., Wilhelm, E., 2002. Cultivation-independent population analysis of bacterial endophytes in three potato varieties based on eubacterial and Actinomycetes- speciic PCR of 16S RNA genes. FEMS Microbiology Ecology , 39(1), 23–32.
Thomas, P., Kumari, S., Swarna, G.K., Praskash, D.P., Dinesh, M.R., 2007. Ubiquitous presence of fastidious endophytic bacteria in ield shoots and index-negative apparently clean shoot-tip cultures of papaya. Plant Cell Reports 26, 1491–1499.
Webster, K.D., Barker, H., 1994: Molecular biological techniques. Dundee, Scottish Crop Research Institute, pp 23–26.
Wozniak, G., 1997: Molecular methods for typing of Clostridium dificile strains. Graduation thesis, University of Ljubljana, Biotechnical faculty, Ljubljana, pp 12.
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.