In vitro rhizogenesis of sugar beet microclones

  • V. V. Polishchuk Uman National University of Horticulture
  • L. M. Karpuk Bila Tserkva National Agrarian University
  • V. P. Mykolaiko Pavlo Tychyna Uman State Pedagogical University
  • A. A. Polishchuk Uman National University of Horticulture
  • I. I. Mykolaiko Pavlo Tychyna Uman State Pedagogical University
Keywords: auxins, cytokinins, ex vitro adaptation, micropropagation

Abstract

The features of rhizogenesis of male-sterile simple hybrids, maternal components of sugar beet heterozosis hybrid and O-types of Darinka variety grown from seeds were investigated. The paper presents the results of studying the main basic nutrient media (B5–A1, B5–A2, B5–A3, B5–A4, B5–A5 and B5–A6) for rhizogenesis, in which the content of macro- and microelements has been reduced by three times, the content of sucrose is reduced by almost 6 times, the amount of agar is reduced by almost five times, while the vitamin РР content is increased by 1.6 times, and the BAP is completely removed. The largest number of rooted microclones was obtained with NAAs by the high indexes of average number of roots and medium indexes of average roots length. To the second, by the number of rooted microclones is assigned, media of 2,4–D, 2,4–DB containing, potassium salt of NAA and IBA were classified, while IAA was characterized by a lower efficiency. The possibilities of regulation of growth and development processes of sugar beet in vitro explants in favor of undifferentiated growth at the stage of proliferation or organogenesis (hemo- and rhizogenesis) have been determined. To improve the quality of rooting of sugar beet plants-regenerants, we have developed the method for dimming agarized nutrient medium with methylene blue at a 0.05% concentration, which made it possible to reduce the inhibitory effect of light on the development of lateral roots. The average index of micro-roots rooting for MS-forms was 81.5 %. The technology of adaptation of micro-roots rooting was developed, in which the plant material at the initial stage of growth had slight morphological changes in the puffer apparatus and stem, but by the end of the vegetation, the plants acquired the appearance characteristic of the donor plants of explants. The clone’s resiliency is reached about 100% in the soil. According to the variability of morphological characteristics in reproduced in vitro sugar beet plants of the MS-forms of the Darynka hybrid, all the plants grown from the regenerants were more, than seed progeny from seed sowing of the same component obtained as a result of the attachment of sterility to the corresponding O-type. It has been found that in vitro cultivation has not weakened, and in some cases even increased the mitotic index of meristem cells of young roots of sugar beet. Most genotypes have reduced the number of pathological mitoses, apparently due to the activation of reparative systems, but did not extend the duration of individual phases of mitosis; without increasing the number of anaphase with bridges and fragments, but changed the number of anaphases with other chromosomal abnormalities. In all variants of the experiment, where activation of growth processes has been observed, the lowest variability of the signs is observed, that is, under the influence of stimulants, which are part of nutrient media, obviously there is unification of cell populations. 

References

Beyaz, R., Alizadeh, B., Gürel, S., Özcan, S. F., & Yildiz, M. (2013). Sugar beet (Beta vulgaris L.) growth at different ploidy levels. Caryologia, 66(1), 90–95.

Dhooghe, E., Van Laere, K., Eeckhaut, T., Leus, L., & Van Huylenbroeck, J. (2011). Mitotic chromosome doubling of plant tissues in vitro. Plant Cell, Tissue and Organ Culture, 104(3), 359–373.

Gürel, E., & Wren, M. J. (1995). In vitro development from leaf explants of sugar beet (Beta vulgaris L). Rhizogenesis and the effect of sequential exposure to auxin and cytokinin. Annals of Botany, 75(1), 31–38.

Gurel, E., Gurel, S., & Lemaux, P. G. (2008). Biotechnology applications for sugar beet. Critical Reviews in Plant Sciences, 27(2), 108–140.

Joersbo, M. (2007). Sugar beet. In: Pua, E. C., & Davey, M. R. (Eds.). Biotechnology in Agriculture and Forestry, Transgenic Crops, Springer-Verlag, Berlin, 59, 355–378.

Kagami, H., Taguchi, K., Arakawa, T., Kuroda, Y., Tamagake, H., & Kubo, T. (2016). Efficient callus formation and plant regeneration are heritable characters in sugar beet (Beta vulgaris L.). Hereditas, 153(1), 12.

Klyachenko, O. L., Lichanov, A. F., & Krylovskaya, S. A. (2012). Cell regulation of morphogenic processes in the callus tissues of sugar beet in vitro conditions. Bulletin of Agrarian Science, 12, 48–50 (in Ukrainian).

Kolomiets, Y. V. (2008). Clonal microprosthetics of sugar beet in vitro: possibilities for reproduction and preservation of biological diversity. Scientific Support of NAU, 1(9), 1–14.

Kosenko, I. S., Opalko, A. I., & Nebykov, M. V. (2008). Regeneration of plants in the process of micropropagation. Autochthonous and Alien Plants, 3–4, 57–67 (in Ukrainian).

Krylovskaya, S. A. (2012). Features of in vitro sugar beet morphogenesis (Beta vulgaris L.). Collection of scientific works of Institute of Bioenergetic Cultures and Sugar Beets, 14, 461–465 (in Ukrainian).

Mezei, S., Kovacev, L., & Nagl, N. (2006). Sugar beet micropropagation. Biotechnology and Biotechnological Equipment, 20, 9–14.

Miguel, C., & Marum, L. (2011). An epigenetic view of plant cells cultured in vitro: Somaclonal variation and beyond. Journal of Experimental Botany, 62(11), 3713–3725.

Mishutkina, Y. V., & Gaponenko, A. K. (2006). Study of the influence of nutrient medium composition, explant type and genotype on the frequency of regeneration of sugar beet plants (Beta vulgaris L.) in vitro. Genetics, 42(2), 210–218 (in Ukrainian).

Mishutkina, Y. V., & Gaponenko, A. K. (2006). Sugar beet (Beta vulgaris L.) morphogenesis in vitro: Effects of phytohormone type and concentration in the culture medium, type of explants, and plant genotype on shoot regeneration frequency. Russian Journal of Genetics, 42(2), 150–157.

Nagl, N., Maksimovic, I., Curcic, Z., Putnik-Delic, M., & Kovacev, L. (2010). Effect of induced water deficit on sugar beet micropropagation. In: Proceedings of 72nd IIRB Congress. (22th–24th Jun, 2010, Copenhagen, Denmark). International Institute for Beet Research (IIRB), Bruxelles, Belgium. pp. 179–185.

Neelwarne, B. (2013). Cell and tissue culture studies in Beta vulgaris L. In: Neelwarne, B. (Ed.). Red beet biotechnology. Springer Science + Business Media, New York, pp. 175–198.

Pasqual, M., Pio, L. A. S., Oliveira, A. C. L., & Soares, J. D. R. (2012). Flow cytometry applied in tissue culture. Leva, A., & Rinaldi, L. M. R. (Eds.): Recent advances in plant in vitro culture (Ch. 6), In Tech Prepress, Novi Sad, pp. 109–122.

Penna, S., Vitthal, S. B., & Yadav, P. V. (2012). In vitro mutagenesis and selection in plant tissue cultures and their prospects for crop improvement. Bioremediation, Biodiversity and Bioavailability: Global Science Books, 6(1), 6–14.

Pillen, K., Steinrücken, G.,Wricke, G., Herrmann, R. G., & Jung, C. (1992). A linkage map of sugar beet (Beta vulgaris L.). Theoretical and Applied Genetics, 84(1), 129–135.

Polischuk, V. V., Polischuk, O. V., & Karpuk, L. M. (2012). Selection of nutrient medium for sugar beet cms component introduction and proliferation. Autochthonous and Introduced Plants of Ukraine, 8, 123–128 (in Ukrainian).

Polishchuk, V. (2012). Seedlings sterilisation of parental forms of sugar beets hybrids for input in vitro. Agrobiology, 9, 45–48 (in Ukrainian).

Putnik-Delic, M., Maksimovic, I., Venezia, A., & Nagl, N. (2013). Free proline accumulation in young sugar beet plants and in tissue culture explants under water deficiency as tools for assessment of drought tolerance. Romanian Agricultural Research, 30, 141–148.

Ritchie, G. A, Short, K. C., & Davey, M. R. (1989). In vitro shoot regeneration from callus, leaf axils and petioles of sugar beet (Beta vulgaris L.). Journal of Experimental Botany, 40, 277–283.

Rizkalla, A. A., Badr-Elden, A. M., Ottai, M. E. S., Nasr, M. I., & Esmail, M. N. (2012). Development of artificial seed technology and preservation in sugar beet. Sugar Tech, 14(3), 312–320.

Ryabovol, L. O. Ryabovol, Y. S., & Lyubchenko, A. I. (2012). Nutrient medium for rooting homozygous material of sugar beet. Collection of scientific works of Uman National University of Horticulture, 81(1), 196–199.

Sullivan, C. F., Finch, I., Dix, P. J., & Burke, J. I. (1993). Studies of in vitro propagation systems for sugar beet. Irish Journal of Agricultural and Food Research, 32, 27–35.

Taski-Ajdukovic, K., Nagl, N., Kovacev, L., Curcic, Z., & Danojevic, D. (2012). Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit. Electronic Journal of Biotechnology, 15(6), 10.

Tomaszewska-Sowa, M. (2012). Effect of growth regulators and other components of culture medium on morphogenesis of sugar beet (Beta vulgaris L.) in unfertilised ovule in vitro cultures. Acta Agrobotanica, 65(4), 91–100.

Trush, A. O., Perfenyuk, S. G., & Tatarchuk, V. M. (2015). The efficiency of different methods of cytological control of the ployidity of multi-growth tetraploid plasters of sugar beet in the selection of triploid hybrids on the basis of the CMF. Collection of Scientific Works of Uman National University of Horticulture, 87(1), 176–182 (in Ukrainian).

Us-Camas, R., Rivera-Solís, G., Duarte-Aké, F., & De-la-Pena, C. (2014). In vitro culture: An epigenetic challenge for plants. Plant Cell Tissue and Organ Culture, 118(2), 187–201.

Wright, T. R., & Penner, D. (1998). Cell selection and inheritance of imidazolinone resistance in sugarbeet (Beta vulgaris). Theoretical and Applied Genetics, 96(5), 612–620.

Yildiz, M., Alizadeh, B., & Beyaz, R. (2013). In vitro explant growth and shoot regeneration from petioles of sugar beet (Beta vulgaris L.) lines at different ploidy levels. Journal of Sugar Beet Research, 50, 22–36.

Zhang, C. L., Chen, D. F., Elliott, M. C., & Slater, A. (2001). Thidiazuron-induced organogenesis and somatic embryogenesis in sugar beet (Beta vulgaris L). In Vitro Cellular and Developmental Biology – Plant, 37(2), 305–310.

Published
2017-10-29
How to Cite
Polishchuk, V. V., Karpuk, L. M., Mykolaiko, V. P., Polishchuk, A. A., & Mykolaiko, I. I. (2017). In vitro rhizogenesis of sugar beet microclones. Regulatory Mechanisms in Biosystems, 8(4), 616–622. https://doi.org/10.15421/021794