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Fiziol. rast. genet. 2017, vol. 49, no. 6, 495-505, doi: https://doi.org/10.15407/frg2017.06.495


Likhanov A.F., Overchenko O.V., Kostenko S.M., Subin O.V.

  • Institute for evolutionary ecology, national academy of sciences of Ukraine 37 Acad. Lebedeva St., Kyiv, 03143, Ukraine
  • National University of Life and Environmental Sciences of Ukraine 15 Heroiv Oborony St., Kyiv, 03041, Ukraine

This paper present researches of callusogenesis specificity and cell differentiation of resistant (RP) and non-resistant (NRP) forms to horse-chestnut leaf miner (HCLM) of Aesculus hippocastanum according peculiarities of plant tissue primary and secondary metabolism. The most active callusogenesis was observed on DKW medium, with the addition of 0.5 mg/L kinetin and 3.0 mg/L 2,4-D. Structurally, three areas in calluses of RP and NRP forms could be recognized. The superficial part (area III) of callus in RP form was formed by cells filled with condensed tannins, with thick cell walls. A layer of parenchyma with thin cell walls underlined the outer layer of callus (area II). Numerous tracheate elements, capable of accelerated transportation of nutrients into tissues supporting cellular nutrition and differentiation, were formed among them. The internal area (area I) consisted of parenchymal cells. Many of them had in protoplasts the amorphous structures with polysaccharide and tannin complexes. The lignification of the parenchyma cell walls in callus tissues occurred under increasing of anionic peroxidases activity. This rate was five or more times higher than for the NRP form. Concerning to the RP form of Aesculus hippocastanum the viscosity of leaf cell juice may be the key factor limiting HCLM larval development. The amount of phenols in the leaves is not related with it. The kinematic viscosity of the RP form leaves (1,889 mm2/s) was 1.53 times higher than that of NRP form (1.214 mm2/s). In contrast, the content of phenolic substances was twice higher in the NRP form. The confirmed metabolic specificity of RP form can be explained by the relatively richer quantitative and qualitative composition of free amino acids in its tissues, compared to the NRP form. In general, the metabolism specificity of RP form callus tissues is a convenient model for studying the mechanisms of resistance against pathogens and pests of common horse chestnut.

Keywords: Aesculus hippocastanum L., common horse chestnut, callus, cells, regeneration, morphogenesis

Fiziol. rast. genet.
2017, vol. 49, no. 6, 495-505

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1. Andreeva, V.A. (1988). Peroxidase Enzyme: Participation in the plant defense mechanism. Moscow: Nauka [in Russian].

2. Gorshkova, T.A. (2007). Plant cell wall as a dynamic system. Moscow: Nauka [in Russian].

3. Zerova, M., Nikitenko, G & Narolskiy, N. (2007). Chestnut Mining Moth in Ukraine. Kiev [in Russian].

4. Lihanov, A.F., Pentelyuk, O.S., Grigoryuk, I.P. & Kostenko, S.M. (2016). Spatial specificity of accumulation of phenols in leaves of plants of bitter-chestnut common (Aesculus hippocastanum L.). Bioresursy i pryrodokorystuvannya, 8, No. 3-4, pp. 5-13 [in Ukrainian].

5. Sibgatullina, G.V., Haertdinova, L. R. & Gumerova, E. A. (2011). Methods for determining the redox status of cultured plant cells. Kazan: Kazan (Volga Region) Federal University [in Russian].

6. Ermakov, A.Y. (Ed.) (1972). Methods of biochemical research of plants. Leningrad: Kolos [in Russian].

7. Furst, G.G. (1979). Methods of anatomic-histochemical studies of plant tissues. Moscow: Nauka [in Russian].

8. Checheneva, T.M., Shavanova, K.E. & Mashkovska, S.P. (2010). Introduction to in vitro culture of different species of bitter chestnut (genus Aesculus L.). Fiziologiya i biokhimiya kult. rastenij, 42(2), pp. 132-136 [in Ukrainian].

9. Bueno, M.A., Gomez, A. & Manzanera, J.A. (2000). Somatic and gamatic embryogenesis in Quercus suber L. Somatic Embryogenesis in Woody Plants, 6, pp. 479-508. https://doi.org/10.1007/978-94-017-3030-3_16

10. D'Costa L.E. (2014). Resistance and susceptibility to the invasive leaf miner Cameraria ohridella within the genus Aesculus. (Extended abstract of Doctor Thesis). Royal Holloway, University of London.

11. Gastaldo, P., Carli, S. & Profumo, P.(1994). Somatic embryogenesis from stem Aesculus hippocastanum. Plant Cell Tiss. Organ Cult., 39, pp. 97-99. https://doi.org/10.1007/BF00037597

12. Jorgensen, J. (1989). Somatic embryogenesis in Aesculus hippocastanum L. by culture of filament callus. Journal of Plant Physiology, 135, pp. 240-241. https://doi.org/10.1016/S0176-1617(89)80185-3

13. Kiss, J., Heszky, L.E., Kiss, E. & Gyulai, G.(1992). High efficiency adventive embryogenesis on somatic embryos of anther, filament and immature proembryo origin in horse-chestnut (Aesculus hippocastanum L.) tissue culture. Plant Cell Tissue and Organic Culture, 30, pp. 59-64. https://doi.org/10.1007/BF00040001

14. Oszmianski, J., Kalisz, S. & Aneta, W. (2014). The content of phenolic compounds in leaf tissues of white (Aesculus hippocastanum L.) and red horse chestnut (Aesculus carnea H.) colonized by the horse chestnut leaf miner (Cameraria ohridella Deschka & Dimic). Molecules,19, pp. 625-636. https://doi.org/10.3390/molecules190914625

15. Profumo, P., Dameri, R.M. & Modenesi, O.P. (1980). Aescin content in calluses from explants of Aesculus hippocastanum cotyledons grown in vitro. Giorn. Bot. Ital.,114, pp. 25-28. https://doi.org/10.1080/11263508009426430

16. Profurno, P., Caviglia, A.M., Gastaldo, P. & Damer, R.M. (1991). Aescin content in embryogenic callus and in embryoids from leaf explants of Aesculus hippocastanum. Planta Med., 57, pp. 50-52. https://doi.org/10.1055/s-2006-960016

17. Radojevic, L. (1988). Plant regeneration of Aesculus hippocastanum L. (horse chestnut) through somatic embryogenesis. Journal of Plant Physiology, 132, pp. 322-326. https://doi.org/10.1016/S0176-1617(88)80114-7

18. Sarwar, M. & Skirvin, R.M. (1997). Effect of thidiazuron and 6-benzylaminopurine on adventitious shoot regeneration from leaves of three strains of 'McIntosh' apple (Malus x domestica Borkh.) in vitro. Sci. Horticul., 68, pp. 95-100. https://doi.org/10.1016/S0304-4238(96)00971-5

19. Saito, A.(1980). In vitro differentiation of embryoid from somatic callus tissues in Aesculus. Jap. For. Soc., 62(8), pp. 308-310.

20. Sediva, A., Vlasinova, H. & Mertelik, J. (2013). Shoot regeneration from various explants of horse chestnut (Aesculus hippocastanum L.). Sci. Horticul., 161, 223-227. https://doi.org/10.1016/j.scienta.2013.06.030

21. Welander, M. (1988).Plant regeneration from leaf and stem segments of shoots raised in vitro from mature apple trees. Journal of Plant Physiology,132, pp.738-744. https://doi.org/10.1016/S0176-1617(88)80238-4