MUTAGENESIS IN MICROSPORE CULTURE OF BRASSICA NAPUS
Main Article Content
Authors
K.Zh. Zhambakin
Institute of Plant Biology and Biotechnology, 45 Timiryazev str., Almaty, 050040, Kazakhstan
A.K. Zatybekov
Institute of Plant Biology and Biotechnology, 45 Timiryazev str., Almaty, 050040, Kazakhstan
D.V. Volkov
Institute of Plant Biology and Biotechnology, 45 Timiryazev str., Almaty, 050040, Kazakhstan
M.Kh. Shamekova
Institute of Plant Biology and Biotechnology, 45 Timiryazev str., Almaty, 050040, Kazakhstan
Abstract
The influence of chemical mutagens on a culture of isolated microspores of oilseed rape, Brassica napus, was studied. Microspores and the embrioids derived from them were subjected to mutagenesis. It was observed that the mutagens sodium azide (NaN3) and ethylmethanesulfonate significantly influenced the induction of embryogenesis within an hour of application to isolated microspores. When compared to the control group, the lines with a higher seed weight and a weight of 1000 seeds could be isolated from the plant material. The best results were observed by the lines that were obtained with 25mM NaN3 after a 4-hour exposure. There is a significant increase of oleic acid levels due to the decrease in concentration of linoleic acid in the first mutant generation. A significant drop in oleic acid content is seen in all cultured lines in the second generation M2, with oleic acid levels being higher in the first generation M1. At the same time, linoleic acid levels are increased in those lines. Dwarf and ripening forms was observed during the process of growth and development of plants in the field conditions. Most of the obtained mutant strains were sterile. We also observed an average decrease in the unsaturated fatty acid content in the mutants of the second generation. Mutant double haploids with signs of high productivity and high quality of seed oil were obtained in this study.
Keywords
Brassica napus, mutagenesis, haploids, microspores, embryos
Article Details
References
Downey R.K., Rakow G.F.W. Rapeseed and Mustard. In: Principles of cultivar development. Crop Species. Macmillan Publishing Company, 1987, vol. 2, pp. 437-486.
Jambhulkar S.J. Mutagenesis: Generation and Evaluation of Induced Mutations, in M. Delseny J.-C. Kader (Editors-in-Chief). Advances in Botanical Research Incorporating Rapeseed Breeding. Academic Press is an imprint of Elsevier, 2007, vol. 45, pp. 417-434. doi: 10.1016/S0065-2296(07)45014-5.
Improvement of new and traditional industrial crops by induced mutations and related biotechnology. IAEA, Vienna, 2003, pp. 1-164.
Siyuan Tan, Richard R. Evans, Mark L. Dahmer, Bijay K. Singh, Dale L. Shaner. Imidazolinone - tolerant crops: history, current status and future. Pest Manag. Sci., 2005, vol. 61, pp. 246-257. doi: 10.1002/ps.993.
Emrani S.N., Arzani A., Saeidi G. Seed viability, germination and seedling growth ofcanola (Brassica napus L.) as influenced by chemical mutagens. African Journal of Biotechnology, 2011, vol. 10(59), pp. 12602-12613.7
Maluszynski M., Szarejko Y., Sigurbjurnsson B. Haploidy and mutation techniques. In: Jain S.M., Sopory S.K. Veilleux R.E. (eds.). In vitro haploid production in higher plants. Kluwer, Dordrecht, 1996, vol. 1, pp. 67-93. doi: 10.1007/978-94-017-1860-8_5.
Szarejko I., Forster B.P. Doubled haploidy and induced mutation. Euphytica, 2007, vol. 158, pp. 359-370. doi: 10.1007/s10681-006-9241-1.
Scott L. McClinchey, Laima S. Kott. Production of mutants with high cold tolerance in spring canola (Brassica napus). Euphytica, 2008, vol. 162, pp. 51-67. doi: 10.1007/s10681-007-9554-8.
Sakhno L.O. Variability in the Fatty Acid Composition of Rapeseed Oil: Classical Breeding and Biotechnology. Cytology and Genetics., 2010, vol. 44, no. 6, pp. 389-397. doi: 10.3103/s0095452710060101.
Alison M.R. Ferrie, Christian Mollers. Haploids and doubled haploids in Brassica spp. for genetic and genomic research. Plant Cell Tissue Organ Culture, 2011, vol. 104, pp. 375-387. doi: 10.1007/s11240-010-9831-4.
Choung P.V., Beversdorf W.B. High frequency embryogenesis through isolated microspore culture in Brassica napus L. and B. carinata. Braun. Plant Sci., 1985, vol. 39, pp. 219-226. doi: 10.1016/0168-9452(85)90178-5.
Lichter R. From microspores to rape plants: A tentativeway to low glucosinolate strains in world crops. In: Sorensen H. (ed). Production, utilization, description, Nijhoff, Junk, Dordrecht, 1985, vol. 11, pp. 268-277.
Swanson E.B., Coumans M.P., Wu S.C., Barsby T.L., Beversdorf W.D. Efficient isolation of microspores and the production of microspore-derived embryos in Brassica napus L. Plant Cell Rep., 1987, vol. 6, pp. 94-97.
Swanson E.B., Coumans M.P., Brown G.L., Patel J.D., Beversdorf W.D. The characterization of herbicide tolerant plants in Brassica napus L. after in vitro selection of microspores and protoplasts. Plant Cell Rep., 1988, vol. 7, pp. 83-87. doi: 10.1007/bf00270110. 8
Zhambakin K.Zh., Shamekova M.H., Volkov D.V., Zatybekov A.K., Daurov D.L., Zhorabekova A.K., Halikov A.R. Getting doubling haploids of rape. KazNU Bulletin. Biology series., 2012, vol. 3(55), pp. 47-57.
Ferrie A.M.R., Taylor D.C., Mackenzie S.L., Rakow G., Raneyand J.P., Keller W.A.. Microspore mutagenesis of Brassica species for fatty acid modifications: a preliminary evaluation. Plant Breeding, 2008, vol. 127, pp. 501-506. doi: 10.1111/j.1439-0523.2008.01502.x.
Foster B.P. Technical Research on Microspore Mutation with EMS in Brassica napus. URL
He Jiangming, Wang Jingqiao, Chen Wei, Li Genze, Cun Shouxian. Effect of EMS treatment on microspore embryogenesis in vitroin Brassica napus. Southwest China Journal of Agricultural Sciences, 2004, vol. 17(6), pp. 690-693.
Wan G.L. In Vitro Mutagenesis for Selection of Novel Germplasm in Brassica Napus. Crop Sciense, 2008, vol. 194, pp. 169-247.
Report on the research project №0109RK00320 02.02.02.r3 "Development of biotechnology produce genetically modified canola with increased resistance to salinity," made in the framework of STP 0.0489. Almaty, 2009.
Eric B. Swanson. Microspore Culture in Brassica. Plant Cell and Tissue Culture, 1990, vol. 6, pp. 159-169. doi: 10.1385/0-89603-161-6:159.
Gamborg O.L., Miller R.A. & Ojima K. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res., 1968, vol. 50. 151-158.
Murashige T., Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physol Plantarum, 1962, vol. 15, pp. 473-497.
GOST 51483-99. Vegetable oils and animal fats. Determination by gas chromatography mass of methyl esters of individual fatty acids to their sum, the State Standard of the Russian Federation. Moscow, 1999, pp. 151-159.