In vitro reproduction, ex situ growth and development of a rare species Lilium pensylvanicum Ker.-Gawl. (Liliaceae)

The possibility of applying the methods and approaches of biotechnology of higher plants for the conservation and reproduction of Lilium pensylvanicum is shown. The use of seeds as primary explants allowed introducing the species into an in vitro culture without causing damage to the donor mother plants. Subculture of bulbs aseptic plants on the Hamburg medium with half the composition of salts micro and macro elements enriched in kinetin (5 mg/l) and indolylacetic acid (1 mg/l) caused the formation of three bulbs per explant. Subsequent passages on four media options allowed to isolate Murashige-Skoog medium supplemented with 6-benzylaminopurine (0.4 mg/l), which stimulated the best development of bulbs and shoot growth. Rooting was performed on hormone-free media. For the first time, long-term observations of the acclimatization of ex situ regenerated plants were carried out. Five-year field observations in the conditions of the collection nursery of the Yakutsk Botanical Garden for the survival, growth and development of regenerated plants (n = 30) showed that 77% of individuals successfully acclimatized. Mass flowering and fruiting were noted in the fourth year. Ontogenetic features of the development of young plants were revealed, which were expressed by the morphological polyvariance of the flowers and the high variability of the stem height. The developed protocol for the clonal micropropagation of Lilium pensylvanicum the results of long-term observations of the growth of regenerated plants allow us to recommend this method for propagating endangered cenopopulations.

1. Stanilova M.I., Ilcheva V.P., Zagorska N.A. Morphogenetic potential and in vitro micropropagation of endangered plant species Leucojum aestivum L. and Lilium rhodopaeum Delip. // Plant Cell Rep. 1994. Vol. 13. N 8. Р. 451–453.

2. LingFei X., FengWang M., Dong L. Plant regeneration from in vitro cultured leaves of Lanzhou lily (Lilium davidii var. unicolor) // Sci. Hortic-Amsterdam. 2009. Vol. 119. N 4. P. 458–461.

3. Yarmolinsky L., Zaccai M., Ben-Shabat Sh., Mills D., Huleihel M. Antiviral activity of ethanol extracts of Ficus binjamina and Lilium candidum in vitro // New Biotechnol. 2009. Vol. 26. N 6. P. 307–313.

4. Chang Ch., Chen Ch-T., Tsai Y-Ch., Chang W-Ch. A tissue culture protocol for propagation of a rare plant, Lilium speciosum Thunb. var. gloriosoides Baker // Bot. Bull. Acad. Sinica. 2000. Vol. 41. N 2. P. 139–142.

5. Joshi S.K., Dhar U. In vitro propagation from axenic explants of Lilium oxypetalum (D. Don) Baker, an endemic bulbous plant of high altitude Himalaya // Acta Physiol. Plant. 2009. Vol. 31. P. 833–838.

6. Sahoo M.R., Devi M.P., Dasgupta M., Prakash N., Ngachan Sh.V. An efficient protocol for in vitro regeneration and conservation of Shirui lily (Lilium mackliniae Sealy): a lab-to-land approach to save the rare endangered Asiatic lily species // In Vitro Cell. Dev.-Pl. 2018. Vol. 54. N 6. P. 701–710.

7. Wu Y., Ma Y-D., Li Y., Zhang L., Xia Y-P. Plantlet regeneration from primary callus cultures of Lilium brownii F. E. Br. ex Miellez var. giganteum G.Y. Li & Z.H. Chen, a rare bulbous germplasm // In Vitro Cell. Dev.-Pl. 2019. Vol. 55. N 1. P. 44–59.

8. Bakhshaie M., Khosravi S., Azadi P., Bagheri H., Tuyl J.M. Biotechnological advances in Lilium // Plant Cell Rep. 2016. Vol. 35. N 9. P. 1799–1826.

9. Ault J.R., Siqueira S.S. Morphogenetic response of Lilium michiganense to four auxin-type plant growth regulators in vitro // Hortscience. 2008. Vol. 43. N 6. P. 1922–1924.

10. Dhyani A., Sharma G., Nautiyal B.P., Nautiyal M.Ch. Propagation and conservation of Lilium polyphyllum D. Don ex Royle // J. Appl. Res. Med. Aromat. Plants. 2014. Vol. 1. N 4. P. 144–147.

11. Красная книга Республики Саха (Якутия). Т. 1: Редкие и находящиеся под угрозой исчезновения виды растений и грибов / Отв. ред. Н.С. Данилова. М.: Изд-во Реарт, 2017. 412 с.

12. Gambourg O.L., Elevegh D. Culture methods and detection of glucanases in suspension cultures of weat and parleys // Can. J. Biochem. 1968. Vol. 46. P. 417–421.

13. Murashige T., Skoog F. A revised medium for rapid growth and bioassay with tobacco tissue cultures // Physiol. Plantarum. 1962. Vol. 15. N 3. P. 473–476.

14. Бейдеман И.Н. Методика изучения фенологии растений и растительных сообществ. Новосибирск: Изд-во Наука, 1974. 139 с.

15. Пугачева Г.М., Соколова М.А. Клональное микроразмножение лилий // Вестн. МичГАУ. 2010. № 1. С. 35–37.

16. Степанова А.Ю., Ильина В.С., Староверов В.В., Терешонок Д.В. Микроклональное размножение лилии азиатской // Плодоводство и ягодоводство России. 2011. Т. 26. С. 237–243.

17. Ikeda N., Niimi Y., Han D. Production of seedlings from ovules excised at the zygote stage in Lilium spp. // Plant Cell Tiss. Organ Cult. 2003. Vol. 73. N 2. P. 159–166.

18. Takayama S., Misawa M. A scheme for mass propagation of Lilium in vitro // Sci. Hortic-Amsterdam. 1983. Vol. 18. N 4. P. 353–362.

19. Osmanova G.O. Biomorhology of Actaea erythrocarpa Fisch. individuals and ecological–demographic characterization of its cenopopulations // Biology Bull. 2016. Vol. 43. N 5. P. 468–473.