Modification of nutriculture medium for inducing callusogenesis in vitro of Camelina sativa
DOI:
https://doi.org/10.31548/dopovidi2018.01.011Abstract
Camelina sativa is a promising agricultural crop of multi-use. Introduction of new high-quality varieties is important to increase the volume of crop production. The use of biotechnological methods makes it possible to reduce significantly the duration of selection process and increase the efficiency of creating new plant forms with desirable economic and valuable features. One of the first stages of in vitro research is to obtain callus biomass in sufficient quantities with high morphogenic parameters as the main source material for the creation of new genotypes. A number of factors influence the process of callusogenesis.
The purpose of our research is to determine the optimal composition of nutriculture medium and its modifications to induce callusogenesis in vitro of camelina sativa.
The work was carried out in the biotechnological laboratory of Uman National University of Horticulture. Segments of camelina sativa seedlings of Stepovyi 1 variety were used as explants. Murashige and Skoog, Schenck and Hildebrant and Gamborg nutriculture media were modified with growth regulators of auxin (2.4-dichlorophenoxyacetic acid) and cytokinin (6-benzylaminopurine) at concentrations of 0.1; 0.5; 1.0; 1.5 mg/l.
The effectiveness of each variant of the nutriculture medium was evaluated by the activity of proliferation and morphogenic characteristics of callus tissues.
There is no initiation of callus tissues on the non hormonal media. The most intense callusogenesis of camelina sativa was carried out for introduction of 0.1 mg/l 2.4-D and 1.0 mg/l 6-BAP to the nutritional substrate. In Murashige and Skoog nutriculture medium, the proportion of explants with dedifferentiation and proliferation of callus tissues was 76.6 %. In Schenck and Hildebrant and Gamborg nutriculture media, the indicator was 27–45 % lower.
Calluses were average filled with water, white or light green, having loose or semi-solid consistency with a large number of morphogenically active cells.
The increase in the concentration of 2.4-dichlorophenoxyacetic acid caused the inhibition of explant tissues of camelina sativa, significantly reduced the intensity of callusogenesis and morphogenetic characteristics of calluses.
On the basis of the dispersion analysis, it is found that 2.4-D concentration (65.8 %) and the ratio of growth regulators in the nutriculture substrate (17.9 %) had the greatest influence on the induction of callusogenesis. The share of influence of the nutriculture medium was only 5.6 %.
Consequently, during the research, the nutriculture medium was modified to obtain the callus tissue of camelina sativa with high morphogenic characteristics.
Key words: camelina sativa, plant growth regulators, callusogenesis, in vitro
References
Кomarova, I. B., Rozhkovan, V. V. (2003). Ryzhij — al'ternatyvna olijna kul'tura ta perspektyvy joho vykorystannia [Camelina is an alternative oil-bearing culture and prospects of her development]. Proposycia. 1, 3–12.
Moskva, I. S. (2016). Stan ta perspektyvy vyroschuvannia ryzhiiu iaroho na pivdni Stepu Ukrainy [The state and prospects of growing сamelina in the south of the Ukrainian steppe]. Bulletin of the agrarian science of the black sea region. 1, 99–109.
Liakh, V. O., Komarova, I. B. (2010). Vmist ta zhyrnokyslotnyj sklad olii ryzhiiu iaroho [The content and fatty acid composition of camelina oil]. Bulletin of the Institute of grain farming. 38, 137–142.
Kalens'ka, S. M., Yunyk, A. V. (2011). Rol' olijnykh kul'tur u vyrishenni enerhetychnoi bezpeky Ukrainy [Oilseeds role in addressing the energy security of Ukraine]. Collection of scientific works of the Institute of bioenergetic plants and sugar beet. 2, 90–96.
Babikova, A. V., Gorpenchenko, T. Ju., Zhuravlev, Ju. N. (2007). Rastenie kak ob"ekt biotekhnologii [Рlants аs object biotechnology]. Komarovskie readings. LV, 184–211.
Kunakh, V. A. (2005) Biotekhnolohiia likars'kykh roslyn. Henetychni ta fizioloho-biokhimichni osnovy [Biotechnology of medicinal plants. Genetic and physiological and biochemical basis]. Logos, 730.
Checheneva, T. N. (2006). Izmenchivost' zlakov v kul'ture in vitro i v processe regeneracii rastenij [Variability of cereals in culture in vitro and in the process of plant regeneration]. Physiology and biochemistry of cultivated plants. 38, 2, 163–175.
Konstantinov, Yu. M., Rivkin, M. I. (1991).Vozmozhnyj svobodnoradikal'nyj mekhanizm vozniknoveniya somaklonal'noj izmenchivosti rastenij [Possible free-radical mechanism of occurrence of somaclonal plant variability]. Collected papers of the All-Union Symposium «Molecular mechanisms of genetic processes». 166–185.
Ashapkin, V. V., Kutuev, L. I., Vanyushin, B. F. (2016). Ehpigeneticheskaya izmenchivost' u rastenij: nasleduemost', adaptivnost', ehvolyucionnoe znacheniee [Epigenetic variability in plants: heritability, adaptability, evolutionary significanc]. Physiology of plants. 63, 2, 191–204. https://doi.org/10.1134/S1021443716020059
Larkin, P. G. (1981). Somaklonal variation — source of variability from cell cultures for improvement. Theoretical and Applied Genetics. 4., 197–214. https://doi.org/10.1007/BF02342540
Ozigit, I. I., Gozukirmizi, N., Ozigit, I. I., Semiz, B. D. (2006). Indukciya kallusa i regeneraciya rastenij iz zrelyh zarodyshej podsolnechnika [Induction of callus and plant regeneration from mature sunflower embryos]. Physiology of plants. 53, 4. 621–624.
https://doi.org/10.1134/S1021443706040194
Teplickaya, L. M., Yurkova, I. N., Sidyakin, A. I., Zhupanov, I. V. (2011). Poluchenie kallusnoj kul'turi donnika lekarstvennogo (Melilotus officinalis L.) i ee citomorfologicheskie osobennosti [Obtaining of callus culture of the drug (Melilotus officinalis L.) and its cytomorphological features]. Uchenye zapiski Tavricheskogo National University im. V.I. Vernadskogo. 24 (63), 2. 284–290.
Konstantinov, A. V. (2013). Vliyanie sostava gormonov v srede prekul'tivirovaniya na regeneraciyu v kallusnyh kul'turah berezy povisloj (Betula pendula Roth.) [Influence of the composition of hormones in the precultivation medium on regeneration in callus cultures of birch (Betula pendula Roth.)]. Modern Phytomorphology. 4, 241–244.
Sel'dimirova, O. A., Kruglova, N. N. (2015). Balans ehndogennyh i ehkzogennyh gormonov i puti morfogeneza v androklinnyh kallusah pshenicy in vitro [Balance of endogenous and exogenous hormones and pathways of morphogenesis in androclinic callus of wheat in vitro]. Izvestiya Ufimskogo NC RAN. 1. 33–39.
Kolomiiets', Yu. V. (2008). Klonal'ne mikrorozmnozhennia tsukrovykh buriakiv in vitro: mozhlyvosti dlia rozmnozhennia i zberezhennia biolohichnoho riznomanittia [Clonal micropropagation of sugar beets in vitro: opportunities for the reproduction and conservation of biological diversity]. Scientific reports of NAU. 1 (9). http://www.nbuv.gov.ua/e-Journals/nd/2008-1/08kjvobv.pdf
Downloads
Published
Issue
Section
License
Relationship between right holders and users shall be governed by the terms of the license Creative Commons Attribution – non-commercial – Distribution On Same Conditions 4.0 international (CC BY-NC-SA 4.0):https://creativecommons.org/licenses/by-nc-sa/4.0/deed.uk
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
