In vitro direct organogenesis of the medicinal single-mountain local prioritized vulnerable Greek endemic Achillea occulta under different medium variants
Keywords:auxins, cytokinins, ex situ conservation, gelling agents, Greek yarrow, micropropagation, sterilization methods
Achillea species are of high medicinal value based on their use in traditional medicine and their content in secondary metabolites with potential application in dermatology and cosmetic industry. Achillea occulta is a single-mountain local prioritized vulnerable Greek endemic encountering propagation problems through seeds and root division. In vitro propagation can be used for the rapid and mass production of clonal plants complying with phytochemical and sanitary quality criteria. In this study, several culture medium variants were tested including the effect of different indole-3-butyric acid (IBA) concentrations with 6-benzyladenine (BA), different cytokinin types, agar concentrations, and zeatin (ZT) sterilization methods on in vitro shoot proliferation, as well as the effect of different auxin types and concentrations on in vitro rooting and ex vitro survival rate of shoot-tip explants. The results showed BA was the most preferable cytokinin type as evidenced by all proliferation attributes and autoclaved ZT provided longer shoots. Therefore, the two more promising and cost-effective treatments for proliferation were 2.2 μM BA + 0.25 μM IBA or 4.4 μM BA + 0.5 μM IBA after 21 days of culture on modified Murashige-Skoog (MS) medium (x2 Fe) enriched with 20 g L-1 sucrose and 6-7 g L-1 Agar (90-93.8% shoot formation, 2.7-2.9 shoots 0.4-0.6 cm long, 2.5-2.6 proliferation rates). Explants rooted better after 21 days of culture on plain MS medium with 2 μM IBA or 1 μM α-naphthalene acetic acid (NAA) (13.3-20% rooting, 1-1.5 roots 1.8-2 cm long). NAA gave more roots, IBA longer roots, while both auxins exhibited highest rooting (13.33-20%) and ex vitro survival rates (50-66.7%).
Abbasi F, Khadivi A, Taghizadeh M, Valizadehkaji B (2019). Micropropagation of Prunus scoparia, a suitable rootstock for almond under drought conditions. International Journal of Fruit Science 19:221-230. https://doi.org/10.1080/15538362.2018.1539695
Aghaye RNM, Yadollahi A (2012). Micropropagation of GF677 rootstock. Journal of Agricultural Science 4:131-138. http://dx.doi.org/10.5539/jas.v4n5p131
Ali A, Ahmad T, Abbasi NA, Hafiz IA (2009). Effect of different media and growth regulators on in vitro shoot proliferation of olive cultivar ‘Moraiolo’. Pakistan Journal of Botany 41:783-795.
Alvarenga ICA, Silva ST, Bertolucci SKV, Pinto JEBP, Pacheco FV (2015) Application of thidiazuron (TDZ) for in vitro multiplication of yarrow (Achillea millefolium L.) and profile of volatile compounds. Australian Journal of Crop Science 9:948-953.
Baker CM, Wetzstein HY (2004). Influence of auxin type and concentration on peanut somatic embryogenesis. Plant Cell, Tissue and Organ Culture 36:361-368. https://doi.org/10.1007/BF00046094
Biswas A, Bari MA, Roy M, Bhadra SK (2011). In vitro propagation of Stemona tuberosa Lour. ‐ A rare medicinal plant through high frequency shoot multiplication using nodal explants. Plant Tissue Culture & Biotechnology 21:151‐159. https://doi.org/10.3329/ptcb.v21i2.10238
Buah JN, Danso E, Taah KJ, Abole EA, Bediako EA, Asiedu J, Baidoo R (2010). The effects of different concentrations cytokinins on the in vitro multiplication of plantain (Musa sp). Biotechnology 9:343-347. https://doi.org/10.3923/biotech.2010.343.347
Cellarova E, Grelakova K, Repcak M, Honcariv R (1982). Morphogenesis in callus tissue cultures of some Matricaria and Achillea species. Biologia Plantarum 24:430-433. https://bp.ueb.cas.cz/pdfs/bpl/1982/06/08.pdf
Chaturvedi HC, Jain M, Kidwai NR (2007). Cloning of medicinal plants through tissue culture - A review. Indian Journal of Experimental Biology 45:937-948.
Constantinidis T, Kalpoutzakis E (2009). Achillea occulta Constantin. & Kalpoutz. Vulnerable (VU). In: Phitos D, Constantinidis T, Kamari G (Eds). The red data book of rare and threatened plants of Greece. Vol 1 (A–D). Hellenic Botanical Society, Patras pp 40-42.
Dimopoulos P, Raus T, Bergmeier E, Constantinidis Th, Iatrou G, Kokkini S, Strid A, Tzanoudakis D (2016). Vascular plants of Greece ‒ An annotated checklist. Supplement. Willdenowia 46:301-347. https://doi.org/10.3372/wi.46.46303
Dobranszki J, Silva JAT (2010). Micropropagation of apple - A review. Biotechnology Advances 28:462-488. https://doi.org/10.1016/j.biotechadv.2010.02.008
Elmongy MS, Cao Y, Zhou H, Xia Y (2018). Root development enhanced by using indole-3-butyric acid and naphthalene acetic acid and associated biochemical changes of in vitro azalea microshoots. Journal of Plant Growth Regulation 37:813-825. https://doi.org/10.1007/s00344-017-9776-5
Evenor D, Reuveni M (2004). Micropropagation of Achillea filipendulina cv. ‘Parker’. Plant Cell, Tissue and Organ Culture 79:91-93. https://doi.org/10.1023/B:TICU.0000049441.48404.70
Figueiredo ACS, Pais MSS (1991). Achillea millefolium (yarrow) cell suspension cultures: Establishment and growth conditions. Biotechnology Letters 13:63-68. https://doi.org/10.1007/BF01033519
Ghasempour H, Dabiri S, Cheraghi M (2012). Composition of the volatile oil, tissue culture and micro-regeneration optimization of wild yarrow (Achillea biebersteinii). Iranian Journal of Plant Physiology 2:539-546.
Grigoriadou K, Krigas N, Maloupa E (2011). GIS-facilitated in vitro propagation and ex situ conservation of Achillea occulta. Plant Cell, Tissue and Organ Culture 107:531-540. https://doi.org/10.1007/s11240-011-0004-x
Grigoriadou K, Vasilakakis M, Eleftheriou EP (2002). In vitro propagation of the Greek olive cultivar ‘Chondrolia Chalkidikis’. Plant Cell, Tissue and Organ Culture 71:47-54. https://doi.org/10.1023/A:1016578614454
Hart DH, Keightley A, Sappington D, Nguyen PTM, Chritton C, Seckinger GR, Torres KC (2016). Stability of adenine-based cytokinins in aqueous solution. In Vitro Cellular and Developmental Biology - Plant 52:1-9. https://doi.org/10.1007/s11627-015-9734-5
Higuchi M, Pischke MS, Mahonen AP (2004). In planta functions of the Arabidopsis cytokinin receptor family. Scientia Horticulturae 45:366-388. https://doi.org/10.1073/pnas.0402887101
Holobiuc I, Catană R, Cogălniceanu G, Cristea V (2018). Biotechnological approach for ex-situ conservation of the vulnerable species Moehringia jankae. Romanian Biotechnological Letters 23:954-963.
Iliev I, Gajdošová A, Liviaková G, Jain SM (2010). Plant micropropagation. In: Davey MR, Anthony P (Eds). Plant cell culture: Essential methods. Wiley-Blackwell, West Sussex, UK pp 1-24.
IUCN (2001). IUCN red list categories and criteria. Version 3.1. IUCN Species Survival Commission. Gland and Cambridge. Retrieved 2023 March 16 from http://www.iucnredlist.org/documents/redlist_cats_crit_en.pdf
Kadota M, Niimi Y (2003). Effects of cytokinin types and their concentrations on shoot proliferation and hyperhydricity on in vitro pear cultivar shoots. Plant Cell, Tissue and Organ Culture 72:261-265. https://doi.org/10.1023/A:1022378511659
Kahrizi D, Kakaei M (2010). Effect of 6-benzylaminopurine, 2,4-dichlorophenoxyacetic acid and indole-3-butyric acid on micropropagation stages of Achillea biebersteinii. Asian Journal of Chemistry 22:2383-2386.
Kim KH, Park HK, Park MS, Yea UD (2001). Effects of auxin and cytokinin on organogenesis of soybean Glycine max L. Journal of Plant Biotechnology 3:95-100.
Liu J, Moore S, Chen C, Lindsey K (2017). Crosstalk complexities between auxin, cytokinin, and ethylene in Arabidopsis root development: from experiments to systems modeling, and back again. Molecular Plant 10:1480-1496. https://doi.org/10.1016/j.molp.2017.11.002
Manole-Păunescu A (2014). Biotechnology for endangered plant conservation. In: Ahuja MR, Ramawat KG (Eds). Biotechnology and biodiversity. Springer, Switzerland pp 181-202. https://doi.org/10.1007/978-3-319-09381-9_10
Mansseri-Lamrioui A, Louerguioui A, Bonaly J, Yakoub-Bougdal S, Allili N, Gana-Kebbouche S (2011). Proliferation and rooting of wild cherry: The influence of cytokinin and auxin types and their concentration. African Journal of Biotechnology 10:8613-8624. https://doi.org/10.5897/AJB11.450
Marcu D, Halmágyi A, Besenyei E, Clapa D, Fira A, Cristea V (2014). In vitro conservation of Achillea pyrenaica Sibth. ex Godr., a Pyrenean endemic species. Muzeul Olteniei Craiova Oltenia Studii şi comunicări Ştiinţele Naturii 30:55-60. http://biozoojournals.ro/oscsn/cont/30_2/08_Marcu_pp55-60.pdf
Miller CO (1974). Ribosyl-trans-zeatin, a major cytokinin produced by crown gall tumor tissue. Proceedings of the National Academy of Sciences of the United States of America 71:334-338. https://doi.org/10.1073/pnas.71.2.334
Miller CO, Skoog F, Von Saltza MHV, Strong FM (1955). Kinetin, a cell division factor from deoxyribonucleic acid. Journal of the American Chemical Society 77:1392. https://doi.org/10.1021/ja01610a105
Mohammadhosseini M, Sarker SD, Akbarzadeh A (2017). Chemical composition of the essential oils and extracts of Achillea species and their biological activities: A review. Journal of Ethnopharmacology 199:257-315. https://doi.org/10.1016/j.jep.2017.02.010
Monteiro ACBA, Higashi EN, Goncalves AN, Rodriguez PM (2000). A novel approach for the definition of the inorganic medium components for micropropagation of yellow passion fruit (Passiflora edulis Sims. f. flavicarpa Deg.). In Vitro Cellular and Developmental Biology - Plant 36:527-531. https://doi.org/10.1007/s11627-000-0094-3
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Panaia M, Bunn E, McComb J (2011). Primary and repetitive secondary somatic embryogenesis of Lepidosperma drummondii (Cyperaceae) and Bloskion tetraphyllum (Restionaceae) for land restoration and horticulture. In Vitro Cellular and Developmental Biology - Plant 47:379-386. https://doi.org/10.1007/s11627-010-9335-2
Pasternak T, Miskolczi P, Ayaydin F, Mészáros T, Dudits D, Feher A (2000). Exogenous auxin and cytokinin dependent activation of CDKs and cell division in leaf protoplast-derived cells of alfalfa. Plant Growth Regulation 32:129-141. https://doi.org/10.1023/A:1010793226030
Păunescu A (2009). Biotechnology for endangered plant conservation: A critical overview. Romanian Biotechnological Letters 14:4095-4103. https://doi.org/10.1007/978-3-319-09381-9_10
Phillips GC, Garda M (2019). Plant tissue culture media and practices: An overview. In Vitro Cellular and Developmental Biology - Plant 55:242-257. https://doi.org/10.1007/s11627-019-09983-5
Radmann EB, Gallo CM, Ritterbusch CW, Bianchi VJ, Fernando JA, Peters JA (2014). In vitro rooting and acclimatization of ‘Mr. S. 2/5’ plum rootstock. Plant Cell & Micropropagation 10:21-31.
Rogova M, Ganeva T, Stefanova M, Koleva D, Toteva VK (2015). Morphoanatomical study of in vitro propagated and ex vitro adapted Achillea thracica Velen. plants. Bulgarian Journal of Agricultural Science 21:947-950.
Rugini E, Gutierrez-Pesce P, Sampinato PL (1999). New perspective for biotechnologies in olive breeding: Morphogenesis, in vitro selection and gene transformation. Acta Horticulturae 474:107-110. https://doi.org/10.17660/ActaHortic.1999.474.18
Shatnawi MA (2013). Multiplication and cryopreservation of yarrow (Achillea millefolium L., Asteraceae). Journal of Agriculture, Science and Technology 15:163-173.
Smith MAL, Spomer LA (1995). Vessels, gels, liquid media, and support systems. In: Aitken-Christie J, Kozai T, Smith ML (Eds). Automation and environmental control in plant tissue culture. Springer, Dordrecht pp 371-404. https://doi.org/10.1007/978-94-015-8461-6_16
Srivastava A, Joshi AG (2009). In vitro behavior of nodal explants of Portulaca grandiflora under the influence of cytokinins. Acta Universitatis Latviensis Biology 753:43-48.
Stojanovic G, Radulovic N, Hashimoto T, Palic R (2005). In vitro antimicrobial activity of extracts of four Achillea species: The composition of Achillea clavennae L. (Asteraceae) extract. Journal of Ethnopharmacology 101:185-190. https://doi.org/10.1016/j.jep.2005.04.026
Strzępek-Gomółka M, Gaweł-Bęben K, Kukula-Koch W (2021). Achillea species as sources of active phytochemicals for dermatological and cosmetic applications. Hindawi Oxidative Medicine and Cellular Longevity 2021:6643827. https://doi.org/10.1155/2021/6643827
Torres KC (1989). Tissue culture techniques for horticultural crops. Springer, New York. https://doi.org/10.1007/978-1-4615-9756-8
Turker AU, Yucesan B, Gurel E (2009). In vitro regeneration of Achillea millefolium L. from shoot-tips and root segments of seedlings. Journal of Plant Biochemistry and Biotechnology 18:65-69. https://doi.org/10.1007/BF03263297
Wawrosch C, Kopp B, Kubelka W (1994). In vitro propagation of Achillea asplenifolia Vent. through multiple shoot regeneration. Plant Cell Reports 14:161-164. https://doi.org/10.1007/BF00233782
Wolella EK (2017). Surface sterilization and in vitro propagation of Prunus domestica L. cv. Stanley using axillary buds as explants. Journal of Biotech Research 8:18-26.
Yadav MK, Gaur AK, Garg GK (2003). Development of suitable protocol to overcome hyperhydricity in carnation during micropropagation. Plant Cell, Tissue and Organ Culture 72:153-156. https://doi.org/10.1023/A:1022236920605
Yamamoto M, Yamamoto KT (1998). Differential effects of 1-napthaleneacetic acid, indole-3-acetic acid and 2,4-dichlorophenoxyacetic acid on the gravitropic response of roots in an auxin-resistant mutant of Arabidopsis aux1. Plant Cell Physiology 39:660-664. https://doi.org/10.1093/oxfordjournals.pcp.a029419
Younes LS, Shibli RA, Al-Qudah TS (2015). In vitro propagation and acclimatization of Achillea fragrantissima Frossk Sch. Bip. Jordan Journal of Agricultural Sciences 11:339-351.
Zebarjadi AR, Najafi SH, Ghasempour HR, Motamedi J (2011). Establishment of a practical tissue culture for producing hairy roots of Valeriana officinalis L. via Agrobacterium rhizogenes. Journal of Medicinal Plants Research 5:4984-4992.
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