SSN 2410-7751 (Print)
ISSN 2410-776X (Online)
"Biotechnologia Acta" V. 9, No 4, 2016
https://doi.org/10.15407/biotech9.04.019
Р. 19-27, Bibliography 31, English
Universal Decimal Classification: 581.192: 502.4
N. O. Pushkarova 1, M. S. Kalista 2, M. A. Kharkhota 3, D. B. Rakhmetov 4, M. V. Kuchuk 1
1 Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Kyiv
2 National Museum of Natural History of the National Academy of Sciences of Ukraine, Kyiv
3 Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine, Kyiv
4 Gryshko National Botanical Garden of the National Academy of Sciences of Ukraine, Kyiv
The aim of the study was to establish efficient protocols of seed surface sterilization with further multiplication in vitro for threatened species Crambe koktebelica (Junge) N. Busch and to show the effect of biotechnological approach (in vitro cultivation) of biodiversity conservation on plants biochemical properties. Seed surface sterilization was carried out according to the original method with further microclonal multiplication of aseptic sprouts from lateral buds on the Murashige and Skoog (MS) medium supplemented with different concentrations of growth regulators. Fatty acid content was determined using Gas chromatography-mass spectrophotometry of fatty acid ethers. Antioxidant activity was determined using 2.2-diphenyl-1-picrylhydrazyl radical scavenging method. Total soluble protein content was measured using Bradford method and polyfructan content determination was based upon ketosugars ability to color in the acidic environment with resorcinol. Plants that were grown under in vitro and in vivo conditions and seeds were used in this research. Efficient protocol of surface sterilization that resulted in 45% of aseptic seed material 50% of which has sprouted was elaborated for C. koktebelica as well as fast microclonal multiplication methods that provided with up to 5.25 ± 0.50 new formed plantlets from 1 lateral bud (on the MS medium that contained 1 mg/L of 6-benzylaminopurine). It was also shown that aseptic cultivation benefits to saturated fatty acid accumulation and increases protein content but on the other hand it reduces unsaturated fatty acid amount and polyfructan content as well as antioxidant activity of plant material. Obtained data confirms the prospect of biotechnology approach to biodiversity conservation and suggest the necessity of father in vitro cultivation effect on biochemical composition of plant study.
Key words: Crambe koktebelica, biodiversity conservation, in vitro cultivation.
© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2016
References
1. Prakhova T. Ya. New unconventional oilseed – Abyssinian Crambe. Bulletin of Altai state agricultural university. 2013, 8 (106), 8–10. (In Russian).
2. Gon?alves A. C. Jr., Rubio F., Meneghel A. P., Coelho G. F., Dragunski D. C., Strey L. The use of Crambe abyssinica seeds as adsorbent in the removal of metals from waters. Rev. Bras. Eng. Agr?c. Ambient. 2013, 17 (3), 306–311.
3. Askew M. F. Novel oil, fibre and protein crops in UK – a future perspective. Brighton crop protection conference, weeds Proceedings of an international conference. Brighton, UK, 22?25 November 1993. P. 653–662.
4. Belokurova V. B. Methods of Biotechnology in system of efforts aimed at plant biodiversity preservation. Cytol. Genet. 2010, 44 (3), 174–185. https://doi.org/10.3103/S0095452710030096
5. Al-Qudah T., Shibli R., Alali F. In vitro propagation and secondary metabolites production in wild germander (Teucrium polium L.). In Vitro Cel. Developm. Biol. – Plant. 2011, V. 47, P. 496–505. https://doi.org/10.1007/s11627-011-9352-9
6. Nikolova M., Petrova M., Zayova E. Comparative study of in vitro, ex vitro and in vivo grown plants of Arnica montana – polyphenols and free radical scavenging activity. Acta Bot. Croat. 2013, 71 (1), 13–22. https://doi.org/10.2478/v10184-012-0013-9
7. Xiao-qin S., Hui P., Jian-lin G., Bin P., Ming-ming B., Yue-yu H. Fatty acid analysis of the seed oil in a germplasm collection of 94 species in 58 genera of brassicaceae. Chem. Ind. Forest Prod. 2011, 31 (6), 46–54.
8. Haslam T. M., Kunst L. Extending the story of very-long-chain- fatty acid elongation. Plant Sci. 2013, V. 210, P. 93–107.
9. Taylor D. C., Guo Y., Katavic V., Mietkiewska E., Francis T., Bettger W. New seed oils for improved human and animal health and as industrial feedstocks: Genetic manipulation of the Brassicaceae to produce oils enriched in nervonic acid. Modification of Seed Composition to Promote Health and Nutrition (Krishnan A. B. (Ed.). American Society of Agronomy. 2009, P. 219–233.
10. Medvedkova I., Trudayeva T. Biofuel market: problems and prospects. Mosty. 2013, 6 (3). (In Russian).
11. Los D. A., Mironov K. S., Allakhverdiev S. I. Regulatory role of membrane fluidity in gene expression and physiological functions. Photosynth. Res. 2013, V. 343, P. 489–509. https://doi.org/10.1007/s11120-013-9823-4
12. Kalista M. S., Scherbakova O. F., Popovich A. V. Morphological pecularities of Crambe koktebelica (Junge) N. Busch and Crambe mitridatis Juz. fruits (Brassicaceae). Ukr. Bot. J. 2014, 71 (2), 188–195. (In Ukrainian).
13. Red data book of Ukraine. Plant kingdom, ed. by Didukh Ya. P. Кyiv: Globalconsulting. 2009, 900 p. (In Ukrainian).
14. IUCN Red List of Threatened Species. 2013. Version March 2015, from https://www.iucnredlist.org/details/165277/0.
15. Bilz M., Kell S. P., Maxted N., Landsdown R. V. European Red List of Vascular Plants. Luxemburg: Publications Office of European Union. 2011, 230 p. doi:10.2779/8515.
16. Convention on the Conservation of European Wildlife and Natural Habitats. Appendix I – Strictly Protected Flora Species. Bern, September 19, 1979, from http://rm.coe.int/CoERMPublicCommonSearchServices/DisplayDCTM Con-tent? documentId=0900001680304354.
17. Murashige T., Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 1962, V. 15, P. 473–497.
18. Garces R., Mancha M. One-step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Anal. Biochem. 1993, V. 211, P. 139–143. https://doi.org/10.1006/abio.1993.1244
19. Lyons J. M., Wheaton T. A., Pratt H. K. Relationship between the physical nature of mitochondrial membranes and chilling sensitivity in plant. Plant Physiol. 1964, V. 39, P. 262–268.
20. Jaworski J. G., Stumpf P. K. Fat metabolism in higher plants. Properties of a soluble stearyl-acyl carrier protein desaturase from maturing Carthamus tinctorius. Arch. Biochem. Biophys. 1974, V. 162, P. 158–165. https://doi.org/10.1016/0003-9861(74)90114-3
21. Maznik K. S., Matvieieva N. A. Optimization of fructans extraction from in vitro cultivated chicory „Hairy“ roots. Biotechnol. acta. 2013, 3 (1), 83–88. https://doi.org/10.15407/biotech6.03.083
22. Brand-Williams W., Cuvelier M. E., Berset C. Use of a free radical method to evaluate antioxidant activity. Lebensm. Wiss. Technol. 1995, 28 (1), 25–30. https://doi.org/10.1016/S0023-6438(95)80008-5
23. Bradford M. А rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Аnal. Biochem. 1976, V. 72, P. 248–254.
24. De Winter J. C. F. Using the Student’s t-test with extremely small sample sizes. Pract. Assess. Res. Evaluat. 2013, 18 (10), 1–12.
25. Sakhno L. A. Transgene cruciferous plants as producents of long chain unsaturated fatty acids. Biotekhnologiia. 2010, 3 (2), 9–18. (In Russian).
26. Wang Y. P., Tang J. S., Chu C. Q., Tian J. A preliminary study on the introduction and cultivation of Crambe abyssinica in China, an oil plant for industrial uses. Ind. Crop. Prod. 2000, 12 (1), 47–52.
27. Piovan A., Cassina G., Filippini R. Crambe tataria: actions for ex situ conservation. Biodivers. Conserv. 2011, V. 20, P. 359–371. https://doi.org/10.1007/s10531-010-9949-z
28. Gerhard Knothe. Biodiesel Composition and Its Effect on Fuel Properties. 2012 Collective Biofuels Conference. Temecula, California. Aug. 17–19th, 2012.
29. Campbell B., Han D. Y., Triggs C. M., Fraser A. G., Ferguson L. R. Brassicaceae: nutrient analysis and investigation of tolerability in people with Crohn’s disease in a New Zealand study. Funct. Foods Health Dis. 2012, 2 (2), 460–486.
30. Mirzokhonova G., Aliev K. Synthesis proteins of plants-regenerants of potatoes in process tuberation in vitro. Reports of the Academy of Sciences of the Republic of Tadzhykistan. 2007, 2 (50), 171–176. (In Russian).
31. Kumar V., Kumar Singh S., Bandopadhyay R., Sharma M., Chandra S. In vitro organogenesis secondary metabolite production and heavy metal analysis in Swertia chirayita. Centr. Europ. J. Biol. 2014, V. 9, P. 686–698. https://doi.org/10.2478/s11535-014-0300-7