Rooting and acclimatization to ex vitro conditions of regenerants of frost-resistant members of Rhododendron
Abstract
Rooting and acclimatization are limiting stages in clonal micropropagation, especially of woody plant species. This study aimed to evaluate effect of different treatments with indole-3-butyric acid (IBA) on in vitro and ex vitro rooting and acclimatization of microshoots of six winter-hardy rhododendron genotypes. Two methods of IBA applications: direct cultivation on Anderson’s medium (AM), supplemented with 25.0 μМ IBA and 4-h liquid-pulse treatment with solution of 148.0 μM IBA were tested. After IBA pulse treatment microshoots were transferred either to in vitro on hormone-free AM or ex vitro in a hydroponic system or mixture of peat and sand (1:1). Maximum in vitro rooting after IBA pulse treatment was observed for evergreen cultivars ‘Helsinki University’ (75 %) and ‘Haaga’ (67 %), whereas Far Eastern wild species R. sichotense, R. mucronulatum, and R. schlippenbachii showed 30–40 % rhizogenesis only. Ex vitro rooting in hydroponic system was efficient for evergreen rhododendrons, but R. sichotense microshoots formed roots poorly. The best results for all genotypes studied were obtained under IBA liquid-pulse treatment with ex vitro rooting in the mixture of peat and sand (‘Helsinki University’ – 100 %, R. mucronulatum – 89 %, R. sichotense – 84 %, and R. schlippenbachii – 60 %). Our research revealed genotype differences in rooting percent under all IBA treatment studied. The developed protocol demonstrated some advantages of ex vitro rooting including high quality of root and shoot systems and reduction of acclimatization period. Thus proposed method can be used for breeding and conservation of rhododendron germplasm, as well as for large-scale commercial propagationDownloads
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Almeida R., Goncalves S., Romano A. 2005. In vitro micropropagation of endangered Rhododendron ponticum L. subsp. baeticum (Boissier and Reuter) Handel-Mazzetti. Biodiver. Conserv. 14(5): 1059–1069. DOI:10.1007/s10531-004-8413-3
Anderson W. C. 1984. A revised tissue culture medium for shoot multiplication of rhododendron. J. Amer. Soc. Hort. Sci. 109(3): 43–347.
Benmahioul B., Dorion N., Kaid-Harche M., Daguin F. 2012. Micropropagation and ex vitro rooting of Pistachio (Pistacia vera). Plant Cell Tiss. Organ Cult. 108(2): 353–358. DOI: 10.1007/s11240-011-0040-6
Briggs B. A., McCulloch S. M., Edick L. A. 1988. Micropropagation of azaleas using thidiazuron. Acta Hort. 227: 330–333. DOI: 10.17660/ActaHortic.1988.227.60
Briggs B. A., McCulloch S. M., Caton L. A. 1994. In vitro propagation of Rhododendron. Acta Hort. 364: 21–26. DOI: 10.17660/ActaHortic.1994.364.1
Chamberien D. F., Hyam R., Argent G., Fairweather G., Walter K. S. 1996. The genus Rhododendron, its classification and synonymy. Royal Botanic Garden, Edinburgh, 181 pp.
Chandra S., Bandopadhyay R., Kumar V., Chandra R. 2010. Acclimatization of tissue cultured plantlets: from laboratory to land. Biotechnol. Lett. 32: 1199–1205. DOI: 10.1007/s10529-010-0290-0
Debergh P. C., Maene L. J. 1981. A scheme for commercial propagation of ornamental plants by tissue culture. Sci. Hort. 14(4): 335–345. DOI: 10.1016/0304-4238(81)90047-9
Eeckhaut T., Janssens K., Keyser E., Riek J. 2010. Micropropagation of Rhododendron. In: Protocols for in vitro propagation of ornamental plants: Methods in molecular biology. Eds. Jain S. M., Ochatt S. J. Humana Press, New York, 141–152 pp. DOI: 10.1007/978-1-60327-114-1_14
Erst A. A., Novikova T. I., Karakulov A. V., Zaytseva Y. G. 2012. Adaptation of Rhododendron hybridum regenerants to ex vitro conditions. Belgorod State University Scientific Bulletin: Natural sciences 19(9): 44–49 [In Russian]. (Эрст А. А., Новикова Т. И., Каракулов А. В., Зайцева Ю. Г. Адаптация регенерантов Rhododendron hybridum к условиям ex vitro // Научные ведомости БелГУ: естественные науки, 2012. Т. 19, № 9. С. 44–49).
Filipenia V. L., Garbatsevich V. I., Antipova T. V. 2009. Microclonal propagation of Rhododendron× hybridum hort. Physiology and biochemistry of cultivated plants 41(6): 516–522 [In Russian]. (Филипеня В. Л., Горбацевич В. И., Антипова Т. В. Микроклональное размножение Rhododendron × hybridum hort. // Физиология и биохимия культурных растений, 2009. Т. 41. С. 516–522).
Hazarika B. N. 2006. Morpho-physiological disorders in in vitro culture of plants. Sci. Hort. 108: 105–120. DOI: 10.1016/j.scienta.2006.01.038
Hsia C. N., Korban S. S. 1997. The influence of cytokinins and ionic strength of Anderson’s medium on shoot establishment and proliferation of evergreen azalea. Euphytica 93(1): 11–17. DOI: 10.1023/A:1002945922280
Kharkevich S. S., Katchura N. N. 1981. Redkiye vidy rasteniy sovetskogo Dalnego Vostoka i ikh okhrana [Rare species of plants of the Soviet Far East and their protection]. Nauka Publishers, Moscow, 234 рр. [In Russian]. (Харкевич С. С., Качура Н. Н. Редкие виды растений советского Дальнего Востока и их охрана. М.: Наука, 1981. 234 с.).
Kutas E. N. 2009. Klonalnoye mikrorazmnozheniye rododendronov i ikh prakticheskoye ispolzovaniye [Clonal micropropagation of rhododendrons and their practical use]. Belorusskaya nauka, Minsk, 131 pp. [In Russian]. (Кутас Е. Н. Клональное микроразмножения рододендронов и их практическое использование. Минск: Белорусская наука, 2009. 131 с.).
Leva A. 2012. Innovative protocol for “ex vitro rooting” on olive micropropagation. Cent. Eur. J. Biol. 6(3): 352–358. DOI: 10.2478/s11535-011-0010-3
Nedoluzhko V. A., Koldaeva M. N. 2008. Rhododendron shlippenbachii. In: Krasnaya kniga Primorskogo kraya: rasteniya [The Red Data Book of Primorye Territory: plants]. Apelsin, Vladivostok, 121–122 pp. [In Russian]. (Недолужко В. А., Колдаева М. Н. Рододендрон Шлиппенбаха // Красная книга Приморского края: растения. Владивосток: Апельсин, 2008. C. 121–122).
Nhut D. T., Hahn N. T. M., Tuan P. Q., Nguyet T. M., Tram N. T. H., Chinh N. C., Nguyen N. H., Vinh D. N. 2006. Liquid culture as a positive condition to induce and enhance quality and quantity of somatic embryogenesis of Lilium longiflorum. Sci. Hort. 110(1): 93–97. DOI: 10.1016/j.scienta.2006.05.015
Pavingerova D. 2009. The influence of thidiazuron on shoot regeneration from leaf explants of fifteen cultivars of Rhododendron. Biol. Plant 54:797–799. DOI: 10.1007/s10535-009-0147-3
Pavlova N. S. 2008. Rhododendron shlippenbachii. In: Krasnaya kniga Rossiyskoy Federatsii: rasteniya i griby. [The Red Data Book of the Russian Federation: plants and mushrooms]. Moscow, 204–205 pp. [In Russian]. (Павлова Н. С. Рододендрон Шлиппенбаха // Красная книга Российской Федерации: растения и грибы. М., 2008. C. 204–205).
Petukhova I. P. 2006. Rododendrony na yuge Primorya. Introduktsiya, kultura [Rhododendrons in the south of Primorye]. Vladivostok, 131 pp. [In Russian]. (Петухова И. П. Рододендроны на юге Приморья. Интродукция, культура. Владивосток, 2006. 131 с.).
Phulwaria M., Shekhawat N. S. 2013. An efficient in vitro shoot regeneration from immature inflorescence and ex vitro rooting of Arnebia hispidissima (Lehm). DC. – A red dye (Alkannin) yielding plant. Physiol. Mol. Biol. Plants 19(3): 435–441. DOI: 10.1007/s12298-013-0171-9
Pospisilova J., Ticha I., Kadlecek P., Haisel D., Plzakova S. 1999. Acclimatization of micropropagated plants to ex vitro conditions. Biol. Plant. 42(4): 481–497. DOI: 10.1023/A:1002688208758
Preece J. E., Imel M. R. 1991. Plant regeneration from leaf explants of Rhododendron ‘P. J. M. Hybids’. Sci. Hort. 48: 159–170. DOI: 10.1016/0304-4238(91)90163-S
Van Veen T. 1969. Rhododendron in America. Sweeney Krist Dimm, Portland, 176 pp.
Vasilyeva O. G. 2009. Biologo-morfologicheskiye osnovy klonalnogo mikrorazmnozheniya nekotorykh predstaviteley roda Rhododendron L. [Biology and morphological bases of clonal micropropagation of some members of the genus Rhododendron L.: abstract of the thesis of candidate of biological sciences]. Moscow, 20 pp. [In Russian]. (Васильева О. Г. Биолого-морфологические основы клонального микроразмножения некоторых представителей рода Rhododendron L.: Автореф. дис. ... канд. биол. наук. 2009. М., 20 с.).
Vechernina N. A., Tavartkiladze O. K., Borodulina I. D., Erst A. A. 2008. Adaptatsiya rasteniy-regenerantov s ispolzovaniyem gidroponiki [Adaptation of regenerated plants using hydroponics]. Izvestiya of Altai State University Journal 3: 7–10 [In Russian]. (Вечернина Н. А., Таварткиладзе О. К., Бородулина И. Д., Эрст А. А. Адаптация растений-регенерантов с использованием гидропоники // Известия АлтГУ, 2008. Т. 3. С. 7–10).
Vrishch D. L., Varchenko L. I., Urusov V. M. 2010. Rhododendron L. on Sikhote-Alin: geography, ecology, genesis, economic prospects. The bulletin of KrasGAU 10: 64–71 [In Russian]. (Врищ Д. Л., Варченко Л. И., Урусов В. М. Род рододендрон (Rhododendron L.) на Сихотэ-Алине: география, экология, генезис, хозяйственные перспективы // Вестник КрасГАУ, 2010. Т. 10. С. 64–71).
Zaytseva Y. G., Poluboyarova T. V., Novikova T. I. 2016. Effects of thidiazuron on in vitro morphogenic response of Rhododendron sichotense Pojark. and Rhododendron catawbiense cv. Grandiflorum leaf explants. In Vitro Cell. Dev. Biol.-Plant 52(1): 56–63. DOI: 10.1007/s11627-015-9737-2
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