Prospects for the conservation of Fraxinus sogdiana through micropropagation and slow growth storage approaches
Article Sidebar
Abstract Views: 130
File Views: 72
Main Article Content
Abstract
Abstract. Rakhimzhanova A, Kali B, Zhumabay N, Tussipkan D, Manabayeva S. 2026. Prospects for the conservation of Fraxinus sogdiana through micropropagation and slow growth storage approaches. Biodiversitas 27 (4): d270421. https://doi.org/10.13057/biodiv/d270421. Fraxinus sogdiana is a rare species listed in the Red Book of Kazakhstan, highlighting the urgent need for effective conservation strategies. This study aims to develop an effective micropropagation protocol, slow growth storage approaches and analyze phylogenetic relationships by comparing F. sogdiana from Kazakhstan with species from the NCBI database. The results of this study demonstrate that the DKW medium outperformed the MS medium in promoting shoot development. A DKW-based MP-V medium supplemented with 1.0 mg/L BAP, 0.1 mg/L NAA, and 0.5 mg/L gibberellic acid was found to be optimal for shoot proliferation. This resulted in maximum shoot lengths of 11.54 cm and an average of seven shoots per explant. High concentrations of BAP alone inhibited shoot formation. In slow-growth storage experiments, the addition of 0.5 mg/L CCC provided the best balance between shoot survival and growth retardation. In contrast, mannitol and ABA suppressed shoot development and decreased the survival rate. Based on their ITS patterns, five main groups were identified according to the sections of genus Fraxinus, including Fraxinus, Sciadanthus, Melioides, Pauciflorae, and Ornus. Notably, seven sequences from the species Fraxinus angustifolia, Fraxinus obliqua, Fraxinus sogdiana, and Fraxinus turkestanika, previously considered synonyms, were grouped together with strong bootstrap support. The results of the matK and rbcL gene patterns did not correspond to clearly defined taxonomic groups according to sections. The main finding of the matK gene region analysis was that the populations from Kazakhstan and China were closely related, as indicated by a high bootstrap value. These findings provide an effective protocol for preserving the genetic resources of F. sogdiana, as well as a phylogenetic framework to support future biotechnological and molecular studies.
Article Details
Issue
Section
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
References
Aldibekova A, Kurmanbayeva M, Aksoy A, Permitina V, Dimeyeva L, Zverev N. 2023. Anatomical structure and phytochemical composition of a rare species Fraxinus sogdiana Bunge (Oleaceae) growing in different soils in Kazakhstan. Diversity 15 (6): 769. https://doi.org/10.3390/d15060769.
Benelli C, Tarraf W, Izgu T, De Carlo A. 2022. In vitro conservation through slow growth storage technique of fruit species: An overview of the last 10 years. Plants 11 (23): 3188. https://doi.org/10.3390/plants11233188.
Bussmann RW, Paniagua-Zambrana NY, Kikvidze Z, Müller L, Mehdiyeva NP, Mursal N, Batsatsashvili K, Khutsishvili M, Maisaia I, Sikharulidze S, Tchelidze D, Alizade VM, Salimov R, Fayvush G, Aleksanyan A. 2025. Fraxinus angustifolia Vahl, Fraxinus excelsior L., Fraxinus ornus L., Fraxinus oxycarpa M. Bieb. ex Willd. Oleaceae. In: Bussmann RW (eds). Ethnobotany of the Caucasus. Ethnobotany of Mountain Regions. Springer, Cham. https://doi.org/10.1007/978-3-319-50009-6_320-1.
Capuana M. 2012. In vitro propagation of ash (Fraxinus excelsior L.) by somatic embryogenesis. In: Lambardi M, Ozudogru E, Jain S (eds). Protocols for Micropropagation of Selected Economically-Important Horticultural Plants. Methods in Molecular Biology, vol 994. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-074-8_16.
CBOL Plant Working Group, Hollingsworth PM, Forrest LL et al. 2009. A DNA barcode for land plants. Proc Natl Acad Sci USA 106 (31): 12794-12797. https://doi.org/10.1073/pnas.0905845106.
Cremon T, Dresch DM, Scalon SPQ, Masetto TE. 2018. Drying and reduction in sensitivity to desiccation of seeds of Alibertia edulis: The influence of fruit ripening stage. An Acad Bras Ciênc 90 (2): 1481-1491. https://doi.org/10.1590/0001-3765201820170664.
Dancheva D, Iliev N, Iliev I. 2013. In vitro propagation of Fraxinus excelsior L. Oltenia J Stud Nat Sci 29 (1): 78-84.
Doyle JJ, Doyle JL. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19: 11-15.
Driver JA, Kuniyuki AH. 1984. In vitro propagation of Paradox walnut rootstock. HortScience 19 (4): 507-509. https://doi.org/10.21273/hortsci.19.4.507.
El-Bahr MK, Abd El-Hamid A, Matter MA, Shaltout A, Bekheet SA, El-Ashry AA. 2016. In vitro conservation of embryogenic cultures of date palm using osmotic mediated growth agents. J Genet Eng Biotechnol 14 (2): 363-370. https://doi.org/10.1016/j.jgeb.2016.08.004.
El-Dawayati MM, Baker E-I, Gomaa AH, E.Zayed Z. 2013. In vitro conservation of date palm shoot-tip explants under minimal growth conditions. Egypt J Agric Res 91 (3): 1043-1062. https://doi.org/10.21608/ejar.2013.167078.
Fenning T, O’Donnell M, Preedy K, Bézanger A, Kenyon D, Lopez G. 2022. The rooting ability of in vitro shoot cultures established from a UK collection of the common ash (Fraxinus excelsior L.) and their ex vitro survival. Ann For Sci 79: 30. https://doi.org/10.1186/s13595-022-01146-8.
Gabr AMM, Sayed SS. 2010. Slow growth conservation and molecular characterization of Deutzia scabra Thunb. Afr J Plant Sci 4 (10): 409-416. https://doi.org/10.5897/ajps.9000223.
Garcia RO, Pacheco G, Vianna MG, Mansur E. 2011. In vitro conservation of Passiflora suberosa L.: Slow growth and cryopreservation. Cryo Lett 32 (5): 377-388.
Govaerts RHA. 1996. World Checklist of Seed Plants 2. MIM, Deurne.
Hammatt N. 1994. Shoot initiation in the leaflet axils of compound leaves from micropropagated shoots of juvenile and mature common ash (Fraxinus excelsior L.). J Exp Bot 45 (6): 871-875. https://doi.org/10.1093/jxb/45.6.871.
Kali B, Bekkuzhina S, Tussipkan D, Manabayeva S. 2024. A first approach for the in vitro cultivation, storage, and DNA barcoding of the endangered endemic species Euonymus koopmannii. Plants 13 (16): 2174. https://doi.org/10.3390/plants13162174.
Khan AM, Bhadauria S. 2019. Molecular characterization of keratin degrading fungi isolated from semi-arid soil by PCR using ITS4 and ITS5 primers. J King Saud Univ-Sci 31 (4): 1418-1423. https://doi.org/10.1016/j.jksus.2018.04.014.
Kress WJ, Erickson DL. 2008. DNA barcodes: Genes, genomics, and bioinformatics. Proc Natl Acad Sci USA 105 (8): 2761-2762. https://doi.org/10.1073/pnas.0800476105.
Kuzmina ML, Johnson KL, Barron HR, Hebert PD. 2012. Identification of the vascular plants of Churchill, Manitoba, using a DNA barcode library. BMC Ecol 12: 25. https://doi.org/10.1186/1472-6785-12-25.
Lebedev V, Schestibratov K. 2013. Effect of natural and synthetic growth stimulators on in vitro rooting and acclimatization of common ash (Fraxinus excelsior L.) microplants. Nat Sci 5 (10): 1095-1101. https://doi.org/10.4236/ns.2013.510134.
Lee JH, Pijut PM. 2017. Adventitious shoot regeneration from in vitro leaf explants of Fraxinus nigra. Plant Cell Tiss Organ Cult 130: 335-343. https://doi.org/10.1007/s11240-017-1228-1.
Librado P, Rozas J. 2009. DnaSP v5: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25 (11): 1451-1452. https://doi.org/10.1093/bioinformatics/btp187.
Lu G, Moriyama EN. 2004. Vector NTI, a balanced all-in-one sequence analysis suite. Brief Bioinform 5 (4): 378-388. https://doi.org/10.1093/bib/5.4.378.
Masi M, Di Lecce R, Calice U, Linaldeddu BT, Maddau L, Superchi S, Evidente A. 2022. Diplofuranoxin, a disubstituted dihydrofuranone, was produced together with sphaeropsidin A and epi-sphaeropsidone by Diplodia subglobosa, an emerging ash (Fraxinus excelsior L.) pathogen in Europe. Phytochemistry 202: 113302. https://doi.org/10.1016/j.phytochem.2022.113302.
Meger J, Kozioł C, Pałucka M, Burczyk J, Chybicki IJ. 2024. Genetic resources of common ash (Fraxinus excelsior L.) in Poland. BMC Plant Biol 24: 186. https://doi.org/10.1186/s12870-024-04886-z.
Mitras D, Kitin P, Iliev I, Dancheva D, Scaltsoyiannes A, Tsaktsira M, Nellas C, Rohr R. 2009. In vitro propagation of Fraxinus excelsior L. by epicotyls. J Biol Res-Thessaloniki 11: 37-48.
Mitrofanova OV, Lesnikova-Sedoshenko NP, Ivanova NN, Smykova NV, Mitrofanova IV. 2020. In vitro propagation and preservation of promising chrysanthemum cultivars and hybrid forms. Acta Hortic 1285: 139-145. https://doi.org/10.17660/actahortic.2020.1285.22.
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15 (3): 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x.
Nurtaza A, Dyussembekova D, Islamova S, Samatova I, Zhanybekova Z, Umirzakova A, Magzumova G, Muranets A, Kakimzhanova A. 2024. In vitro conservation and genetic diversity analysis of rare species Ribes janczewskii. Sci Rep 14: 31117. https://doi.org/10.1038/s41598-024-82320-y.
Pan XJ, Zhang WE, Li X. 2014. In vitro conservation of native Chinese wild grape (Vitis heyneana Roem. & Schult.) by slow growth culture. Vitis 53 (4): 207-214. https://doi.org/10.5073/vitis.2014.53.207-214.
Pérez-Parrón M, González-Benito M, Pérez C. 1995. Micropropagation of Fraxinus angustifolia from mature and juvenile plant material. Plant Cell Tiss Organ Cult 37: 297-302. https://doi.org/10.1007/bf00042343.
Preece JE, Bates SA, Van Sambeek J. 1995. Germination of cut seeds and seedling growth of ash (Fraxinus spp.) in vitro. Can J For Res 25 (8): 1368-1374. https://doi.org/10.1139/x95-149.
Ramazanova MB, Tussipkan D, Manabayeva SA. 2025. DNA barcoding and phylogenetic analysis of Brassicaceae species. Ġylym ža̋ne bìlìm 3 (2): 214-222. https://doi.org/10.52578/2305-9397-2025-2-3-214-222.
Richins M, Montes C, Merkle SC. 2024. Conservation of green and white ash germplasm using the cryopreservation of embryogenic cultures. Plants 13 (3): 352. https://doi.org/10.3390/plants13030352.
Sadyrova G, Taskuzhina A, Yanin K, Kerimbek N, Nurmakhanova A, Shaganbek K, Bekenova N, Orazbekova K, Gritsenko D. 2025. Ecological, anatomical, and genomic insights into the rare tree species Fraxinus sogdiana, Celtis caucasica, and Betula jarmolenkoana from the Northern Tien Shan. Forests 16 (8): 1340. https://doi.org/10.3390/f16081340.
Salmurzauly R, Myrzagaliyeva A, Irsaliyev S, Samarkhanov T, Orazov A, Nurtazin S, Altynbek M, Sagynysh B. 2024. Fraxinus sogdiana Bunge forests in Charyn Canyon, Kazakhstan. Caspian J Environ Sci 22 (5): 1117-1131. https://doi.org/10.22124/cjes.2024.8290.
Sutula M, Kakanay A, Tussipkan D, Dzhumanov S, Manabayeva S. 2024. Phylogenetic analysis of rare and endangered Tulipa species (Liliaceae) of Kazakhstan based on universal barcoding markers. Biology 13 (6): 365. https://doi.org/10.3390/biology13060365.
Tonon G, Capuana M, Di Marco A. 2001. Plant regeneration of Fraxinus angustifolia by in vitro shoot organogenesis. Sci Hortic 87 (4): 291-301. https://doi.org/10.1016/S0304-4238(00)00178-3.
Trejgell A, Kamińska M, Tretyn A. 2015. In vitro slow growth storage of Senecio macrophyllus shoots. Acta Physiol Plant 37: 234. https://doi.org/10.1007/s11738-015-1983-8.
Turdiyev TT, Kovalchuk IY, Kabylbekova BZ, Chukanova NI, Frolov SN. 2020. In vitro germplasm cold storage of fruit and berry plants of Kazakhstan. EurAsian J Biosci 14 (1): 1213-1219.
Wallander E. 2008. Systematics of Fraxinus (Oleaceae) and evolution of dioecy. Plant Syst Evol 273: 25-49. https://doi.org/10.1007/s00606-008-0005-3.
Wong TKF, Ly-Trong N, Ren H, Baños H, Roger AJ, Susko E, Bielow C, De Maio N, Goldman N, Hahn MW, Huttley G, Lanfear R, Minh BQ. 2025. IQ-TREE 3: Phylogenomic inference software using complex evolutionary models. EcoEvoRxiv 2025: 1-11. https://doi.org/10.32942/x2p62n.
Zhang CY, Wang FY, Yan HF, Hao G, Hu CM, Ge XJ. 2012. Testing DNA barcoding in closely related groups of Lysimachia L. (Myrsinaceae). Mol Ecol Resour 12 (1): 98-108. https://doi.org/10.1111/j.1755-0998.2011.03076.x.