Cliviaminiata is an ornamental plant with pharmaceutical potential, but its cultivation is costly. The plant’s slow growth has prompted the search for alternative cultivation methods aimed at producing phenolic compounds, such as the in vitro culture. This study investigated the effect of sucrose concentration and fruit maturity on the in vitro germination (IG) of C.miniata seeds and subsequent callus induction (CI). Sucrose was added to the MS medium: 0 - 40 g？L？1. During IG, the concentration of sucrose affected the speed of germination, and at 0 and 10 g？L？1, seeds from ripe fruit had a higher germination percentage (63% and 67%). Mature seeds showed significantly higher percentages than immature ones. When inducing callus, a high concentration of growth regulators (GR) resulted in significant mortality of the explants. Combinations such as 2.4-D mg？L？1 + 0.52 mg？L？1 PIC + 2 mg？L？1 BAP and 4 mg？L？1 BAP + 4 mg？L？1 ANA showed high survival rates and CI. The results demonstrate that sucrose concentrations > 5.1 g？L？1 reduce germination in mature seeds, likely due to osmotic stress, while lower doses (0 - 10 g？L？1) optimize germination (up to 67%), indicating limited reliance on exogenous carbon sources. The cytokinin BAP proved to be essential for callogenesis, regardless of the auxin used.
References
[1]
Koopowitz, H. (2023) Clivia. 2nd Edition, Timber Press.
[2]
Musara, C., Aladejana, E.B. and Aladejana, A.E. (2021) Cliviaminiata (Lindl.) Bosse, (Amaryllidaceae): Botany, Medicinal Uses, Phytochemistry and Pharmacological Properties. Journal of Applied Pharmaceutical Science, 11, 12-18.
[3]
Zhou, L.G. and Wu, J.Y. (2006) Development and Application of Medicinal Plant Tissue Cultures for Production of Drugs and Herbal Medicinals in China. Natural Product Reports, 23, 789-810. https://doi.org/10.1039/b610767b
[4]
Zaker, A., Sykora, C., Gössnitzer, F., Abrishamchi, P., Asili, J., Mousavi, S.H., et al. (2015) Effects of Some Elicitors on Tanshinone Production in Adventitious Root Cultures of Perovskia Abrotanoides Karel. Industrial Crops and Products, 67, 97-102. https://doi.org/10.1016/j.indcrop.2015.01.015
[5]
Cui, X., Murthy, H.N., Jin, Y., Yim, Y., Kim, J. and Paek, K. (2011) Production of Adventitious Root Biomass and Secondary Metabolites of Hypericum perforatum L. in a Balloon Type Airlift Reactor. Bioresource Technology, 102, 10072-10079. https://doi.org/10.1016/j.biortech.2011.08.044
[6]
Yildirim, A.B. and Turker, A.U. (2014) Effects of Regeneration Enhancers on Micropropagation of Fragaria vesca L. and Phenolic Content Comparison of Field-Grown and in Vitro-Grown Plant Materials by Liquid Chromatography-Electrospray Tandem Mass Spectrometry (LC-ESI-MS/MS). Scientia Horticulturae, 169, 169-178. https://doi.org/10.1016/j.scienta.2014.01.038
[7]
Attaran Dowom, S., Abrishamchi, P., Radjabian, T. and Salami, S.A. (2017) Enhanced Phenolic Acids Production in Regenerated Shoot Cultures of Salvia virgata Jacq. after Elicitation with Ag+ Ions, Methyl jasmonate and Yeast Extract. Industrial Crops and Products, 103, 81-88. https://doi.org/10.1016/j.indcrop.2017.03.043
[8]
Anand, S. (2010) Various Approaches for Secondary Metabolite Production through Plant Tissue Culture. Pharmacia, 1, 1-7.
[9]
Matkowski, A. (2008) Plant in Vitro Culture for the Production of Antioxidants—A Review. BiotechnologyAdvances, 26, 548-560. https://doi.org/10.1016/j.biotechadv.2008.07.001
[10]
Sagharyan, M., Ganjeali, A., Cheniany, M. and Mousavi Kouhi, S.M. (2020) Optimization of Callus Induction with Enhancing Production of Phenolic Compounds Production and Antioxidants Activity in Callus Cultures of Nepetabinaloudensis Jamzad (Lamiaceae). IranianJournalofBiotechnology, 18, e2621.
[11]
Efferth, T. (2019) Biotechnology Applications of Plant Callus Cultures. Engineering, 5, 50-59. https://doi.org/10.1016/j.eng.2018.11.006
[12]
George, E.F., Hall, M.A. and Klerk, G.D. (2008) Plant Tissue Culture Procedure—Background. In: Plant Propagation by Tissue Culture, Springer, 1-28. https://doi.org/10.1007/978-1-4020-5005-3_1
[13]
Deng, L., Zhang, Y., Wang, M.M. and Zhao, S.W. (2012) Study on Non-Asymbiotic Germination of Cypripediummacranthos Seed. Seed, 31, 31-39.
[14]
Huh, Y.S., Lee, J.K., Nam, S.Y., Hong, E.Y., Paek, K.Y. and Son, S.W. (2016) Effects of Altering Medium Strength and Sucrose Concentration on in Vitro Germination and Seedling Growth of Cypripedium macranthossw. Journal of Plant Biotechnology, 43, 132-137. https://doi.org/10.5010/jpb.2016.43.1.132
[15]
Murthy, H.N., Lee, E. and Paek, K. (2014) Production of Secondary Metabolites from Cell and Organ Cultures: Strategies and Approaches for Biomass Improvement and Metabolite Accumulation. Plant Cell, Tissue and Organ Culture, 118, 1-16. https://doi.org/10.1007/s11240-014-0467-7
[16]
Xiao, Y., Niu, G. and Kozai, T. (2010) Development and Application of Photoautotrophic Micropropagation Plant System. Plant Cell, Tissue and Organ Culture, 105, 149-158. https://doi.org/10.1007/s11240-010-9863-9
[17]
Gibson, S.I. (2000) Plant Sugar-Response Pathways. Part of a Complex Regulatory Web. Plant Physiology, 124, 1532-1539. https://doi.org/10.1104/pp.124.4.1532
[18]
Modarres, M., Esmaeilzadeh Bahabadi, S. and Taghavizadeh Yazdi, M.E. (2018) Enhanced Production of Phenolic Acids in Cell Suspension Culture of Salvia Leriifolia Benth. Using Growth Regulators and Sucrose. Cytotechnology, 70, 741-750. https://doi.org/10.1007/s10616-017-0178-0
[19]
Fortini, E.A., Batista, D.S., Mamedes-Rodrigues, T.C., Felipe, S.H.S., Correia, L.N.F., Chagas, K., et al. (2021) Gas Exchange Rates and Sucrose Concentrations Affect Plant Growth and Production of Flavonoids in Vernonia condensata Grown in Vitro. Plant Cell, Tissue and Organ Culture, 144, 593-605. https://doi.org/10.1007/s11240-020-01981-5
[20]
Zha, X., Luo, J., Jiang, S. and Wang, J. (2007) Enhancement of Polysaccharide Production in Suspension Cultures of Protocorm-Like Bodies from Dendrobium huoshanense by Optimization of Medium Compositions and Feeding of Sucrose. Process Biochemistry, 42, 344-351. https://doi.org/10.1016/j.procbio.2006.09.008
[21]
Badr, A., Angers, P. and Desjardins, Y. (2015) Comprehensive Analysis of in Vitro to Ex Vitro Transition of Tissue Cultured Potato Plantlets Grown with or without Sucrose Using Metabolic Profiling Technique. Plant Cell, Tissue and Organ Culture, 122, 491-508. https://doi.org/10.1007/s11240-015-0786-3
[22]
Zahara, M., Datta, A. and Boonkorkaew, P. (2016) Effects of Sucrose, Carrot Juice and Culture Media on Growth and Net CO2 Exchange Rate in Phalaenopsis Hybrid ‘Pink’. Scientia Horticulture, 205, 17-24. https://doi.org/10.1016/j.scienta.2016.04.011
[23]
Hilae, A. and Te-Chato, S. (2005) Effects of Carbon Sources and Strength of MS Medium on Germination of Somatic Embryos of oil Palm (Elaeisquineensis Jacq.). SongklanakarinJournalofScienceandTechnology, 27, 629-635.
[24]
Zeng, S., Zhang, Y., Teixeira da Silva, J.A., Wu, K., Zhang, J. and Duan, J. (2013) Seed Biology and in Vitro Seed Germination of Cypripedium. CriticalReviewsinBiotechnology, 34, 358-371. https://doi.org/10.3109/07388551.2013.841117
[25]
Hernández, M.I.P., Lobo, M.A., Medina, C.I.C., Cartagena, J.R.V. and Delgado, O.A.P. (2009) Comportamiento de la germinación y categorización de la latencia en semillas de mortiño (Vacciniummeridionale Swartz). AgronomíaColombiana, 27, 15-23.
[26]
Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum, 15, 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
[27]
Udomdee, W., Wen, P., Lee, C., Chin, S. and Chen, F. (2013) Effect of Sucrose Concentration and Seed Maturity on in Vitro Germination of Dendrobium Nobile Hybrids. Plant Growth Regulation, 72, 249-255. https://doi.org/10.1007/s10725-013-9856-x
[28]
Rasmussen, H.N. (1995) Terrestrial Orchids. Cambridge University Press. https://doi.org/10.1017/cbo9780511525452
[29]
Gould, A.R., Everett, N.P., Wang, T.L. and Street, H.E. (1981) Studies on the Control of the Cell Cycle in Cultured Plant Cells. Protoplasma, 106, 1-13. https://doi.org/10.1007/bf02115957
[30]
Wu, C., Dewir, Y.H., Hahn, E. and Paek, K. (2006) Optimization of Culturing Conditions for the Production of Biomass and Phenolics from Adventitious Roots of Echinacea angustifolia. Journal of Plant Biology, 49, 193-199. https://doi.org/10.1007/bf03030532
[31]
Deb, C.R. and Pongener, A. (2011) Asymbiotic Seed Germination and in Vitro Seedling Development of Cymbidium aloifolium (L.) Sw.: A Multipurpose Orchid. Journal of Plant Biochemistry and Biotechnology, 20, 90-95. https://doi.org/10.1007/s13562-010-0031-4
[32]
Shim S.W., Hahn E.J., Paek K.Y. (2003). In vitro and ex vitro growth of grapevine rootstock 5BB as influenced by number of air exchanges and the presence or absence of sucrose in culture media. PlantCell, TissueandOrganCulture, 75: 57–62.
[33]
Shohael, A.M., Chakrabarty, D., Ali, M.B., Yu, K.W., et al. (2006) Enhancement of Eleutherosides Production in Embryogenic Cultures of Eleutherococcussessiliflorus in Response to Sucrose-Induced Osmotic Stress. ProcessBiochemistry, 41, 512-518.
[34]
Johnson, T.R., Kane, M.E. and Pérez, H.E. (2010) Examining the Interaction of Light, Nutrients and Carbohydrates on Seed Germination and Early Seedling Development of Bletia Purpurea (Orchidaceae). Plant Growth Regulation, 63, 89-99. https://doi.org/10.1007/s10725-010-9516-3
[35]
Cintra de Souza, G., Laene Dias, L., Alves D’Abadia, A.C., Zildo da Silva, G., Gomes Machado, C., Fialho do Reis, E., et al. (2021) Sucrose Concentration Alters the Physiological Potential of in Vitro Germinated Gabiroba Seeds. Plant Cell Culture & Micropropagation, 17, 1-8. https://doi.org/10.46526/pccm.2021.v17.167
[36]
Lee, Y., Lee, N., Yeung, E.C. and Chung, M. (2005) Embryo Development of Cypripedium formosanum in Relation to Seed Germination in Vitro. Journal of the American Society for Horticultural Science, 130, 747-753. https://doi.org/10.21273/jashs.130.5.747
[37]
Hartmann, H.T., Kester, D.E., Davies, Jr. and Geneve, R. L. (2002) Plant Propagation: Principles and Practices. 7th Edition, Prentice Hall.
[38]
Nambara, E., Okamoto, M., Tatematsu, K., Yano, R., Seo, M. and Kamiya, Y. (2010) Abscisic Acid and the Control of Seed Dormancy and Germination. Seed Science Research, 20, 55-67. https://doi.org/10.1017/s0960258510000012
[39]
Priscila, O.S., Leonardo, M.R., Maria, O.V.M., Paulo, S.R.N.L., Teddy, M.F. and Queila, S.G. (2013) Fruit Maturation and in Vitro Germination of Macaw Palm Embryos. African Journal of Biotechnology, 12, 446-452. https://doi.org/10.5897/ajb12.2529
[40]
Barbedo, C.J. (2018) A New Approach Towards the So-Called Recalcitrant Seeds. Journal of Seed Science, 40, 221-236. https://doi.org/10.1590/2317-1545v40n3207201
[41]
Pereira, J.E.S., Maciel, T.M.S., Costa, F.H.D.S. and Pereira, M.A.A. (2006) Germinação in Vitro de embriões zigóticos de murmuru (Astrocaryumulei). Ciência e Agrotecnologia, 30, 251-256. https://doi.org/10.1590/s1413-70542006000200009
[42]
Neves, S., Ribeiro, L.M., Silva, P.O. and Andrade, I.G. (2010) Germinação in Vitro de embriões de coquinho-azedo [Butiacapitata (Mart.) Becc. (Arecaceae)] obtidos de frutos com diferentes graus de maturação. JournalofNeotropicalBiology, 7, 47-54.
[43]
Pan, Z., Zhu, S., Guan, R. and Deng, X. (2010) Identification of 2,4-D-Responsive Proteins in Embryogenic Callus of Valencia Sweet Orange (Citrus Sinensis Osbeck) Following Osmotic Stress. Plant Cell, Tissue and Organ Culture, 103, 145-153. https://doi.org/10.1007/s11240-010-9762-0
[44]
Long, Y., Yang, Y., Pan, G. and Shen, Y. (2022) New Insights into Tissue Culture Plant-Regeneration Mechanisms. Frontiers in Plant Science, 13, Article 926752. https://doi.org/10.3389/fpls.2022.926752
[45]
Gallo-Meagher, M. and Gerrn, J. (2002) Somatic Embryogenesis and Plant Regeneration from Immature Embryos of Saw Palmetto, an Important Landscape and Medical Plant. PlantCell, TissueandOrganCulture, 68, 253-256.
[46]
Steinmacher, D.A., Clement, C.R. and Guerra, M.P. (2007) Somatic Embryogenesis from Immature Peach Palm Inflorescence Explants: Towards Development of an Efficient Protocol. Plant Cell, Tissue and Organ Culture, 89, 15-22. https://doi.org/10.1007/s11240-007-9207-6
[47]
Kodaz, S., Haliloğlu, K., Pour, A.H. and Aydin, M. (2021) Culture Media and Growth Regulators Influence Callus Induction and Plant Regeneration of Mature Embryos of Orchardgrass (Dactylis glomerata L.). Turkish Journal of Agriculture and Forestry, 45, 454-464. https://doi.org/10.3906/tar-1910-61
[48]
Gaj, M.D. (2004) Factors Influencing Somatic Embryogenesis Induction and Plant Regeneration with Particular Reference to Arabidopsis thaliana (L.) Heynh. Plant Growth Regulation, 43, 27-47. https://doi.org/10.1023/b:grow.0000038275.29262.fb
[49]
Siatka, T. (2019) Effects of Growth Regulators on Production of Anthocyanins in Callus Cultures of Angelica archangelica. Natural Product Communications, 14, 1-4. https://doi.org/10.1177/1934578x19857344
