In vitro direct organogenesis of the Cretan dittany (Origanum dictamnus L.), an important threatened Greek endemic species

Authors

  • Virginia SARROPOULOU Hellenic Agricultural Organization (HAO)-DEMETER, Institute of Plant Breeding and Genetic Resources, Laboratory of Protection and Evaluation of Native and Floriculture Species, Balkan Botanic Garden of Kroussia, P.C. 570 01 Thermi, Thessaloniki, P.O. Box 60458 (GR)
  • Eleni MALOUPA Hellenic Agricultural Organization (HAO)-DEMETER, Institute of Plant Breeding and Genetic Resources, Laboratory of Protection and Evaluation of Native and Floriculture Species, Balkan Botanic Garden of Kroussia, P.C. 570 01 Thermi, Thessaloniki, P.O. Box 60458 (GR)
  • Katerina GRIGORIADOU Hellenic Agricultural Organization (HAO)-DEMETER, Institute of Plant Breeding and Genetic Resources, Laboratory of Protection and Evaluation of Native and Floriculture Species, Balkan Botanic Garden of Kroussia, P.C. 570 01 Thermi, Thessaloniki, P.O. Box 60458 (GR)

DOI:

https://doi.org/10.15835/nbha50212715

Keywords:

basal tissue culture media, conservation priority species, ex situ conservation, medicinal-aromatic plants, micropropagation, plant growth regulators, seed germination

Abstract

Dittany of Crete (Origanum dictamnus L.) is a threatened medicinal-aromatic plant of Lamiaceae family which is a local endemic to the Island of Crete, Greece. Its high culinary use and increasing demand in the pharma, perfumery, cosmetic and food industry along with its overexploitation from its natural habitat has threatened this species and has necessitated its large-scale production for industrial exploitations using advanced technologies. Micropropagation is considered a good tool for ex situ conservation of endangered species with reduced populations in the wild, low germination rates and low seed production. In this study, moderate germination percentages (40-41.38%) were exhibited for seeds after 40 days of culture at 21-23 oC in MS medium regardless of photoperiod regime (16h light/ 8h dark, 24h darkness), without significance difference. In the proliferation and rooting stage, three basal culture media (MS, WPM, Gamborg B5) were tested in combination with two concentrations of 6-benzyladenine (BA) (1.1, 2.2 μM) and indole-3-butyric acid (IBA) (0.125, 0.25 μM), all supplemented with 0.3 μM gibberellic acid (GA3), 20 g L-1 sucrose and 6 g L-1 Plant Agar (pH: 5.8). The results showed that the MS medium + 2.2 μM BA + 0.25 μM IBA was the most effective treatment for micropropagation of shoot nodal explants in a single stage within a 30-day culture period, exhibiting 85% shoot formation, 1.8 new shoots/ explant 2.8 cm long with a 3.2 proliferation rate, 100% rooting, 16.5 roots/ rooted explant 2.1 cm long. Rooted plants obtained in vitro from MS medium enriched with 2.2 μM BA + 0.25 μM IBA gave 100% ex vitro survival rate on a peat: perlite (1:1 v/v) substrate mixture after 2 months in the greenhouse mist. In this study, an efficient in vitro propagation system of O. dictamnus is described for the first time through optimization of direct organogenesis stages (seed germination, proliferation, rooting, ex vitro acclimatization), as a means to facilitate domestication procedure, ex-situ conservation and future sustainable exploitation strategies, thus promoting wider usability of this local endemic with significant commercial potential.

Metrics

Metrics Loading ...

References

Abdallah SAS, Yakoup MYA, Abdalla MYH (2017). Micropropagation of oregano (Origanum syriacum L.) through tissue culture technique. Journal of Plant Production 8(5):635-639. https://doi.org/10.21608/JPP.2017.40497 DOI: https://doi.org/10.21608/jpp.2017.40497

Ahmed NR (2018). Seed germination for Origanum syriacum in laboratory. Journal of Science, Special Issue for the 2nd Annual Conference on Theories and Applications of Basic and Biosciences, 1st September 2018, pp 770-776.

Arafeh RM, Mahmoud MS, Shibli RA (2003). In vitro seed propagation of wild Syrian marjoram (Origanum syriacum L.) Advances in Horticultural Science 17(4):241-244. https://www.jstor.org/stable/42883371

Atar H, Çölgeçen H (2019). Regeneration in Origanum onites L. by plant tissue culture. Karaelmas Fen ve Mühendislik Dergisi 9(2):177-180. https://doi.org/10.7212/zkufbd.v9i2.1237 DOI: https://doi.org/10.7212/zkufbd.v9i2.1237

Bilz M, Kell SP, Maxted N, Lansdown RV (2011). European Red List of Vascular Plants. Publications Office of the European Union, Luxembourg.

Buah JN, Danso E, Taah KJ, Abole EA, Bediako EA, Asiedu J, Baidoo R (2010). The effects of different concentration cytokinins on the in vitro multiplication of plantain (Musa sp.). Biotechnology 9(3):343-347. https://doi.org/10.3923/biotech.2010.343.347 DOI: https://doi.org/10.3923/biotech.2010.343.347

Chatzopoulou A, Karioti A, Gousiadou C, Vivancos VL, Kyriazopoulos P, Golegou S, Skaltsa H (2010). Depsides and other polar constituents from Origanum dictamnus L. and their in vitro antimicrobial activity in clinical strains. Journal of Agricultural and Food Chemistry 58(10):6064-6068. https://doi.org/10.1021/jf904596m DOI: https://doi.org/10.1021/jf904596m

Chinou I (2013). Assessment report on Origanum dictamnus L., herba. European Medicines Agency 2013, EMA/HMPC/200431/2012, Committee on Herbal Medicinal Products (HMPC).

Chishti S, Kaloo ZA, Sultan P (2013). Medicinal importance of genus Origanum: A review. Journal of Pharmacognosy and Phytotherapy 5(10):170-177. https://doi.org/10.5897/JPP2013.0285

El-Beyrouthy M, Elian G, Abou-Jaoudeh C, Chalak L (2015). In vitro propagation of Origanum syriacum and Origanum ehrenbergii. Acta Horticulturae 1083(1083):169-172. https://doi.org/10.17660/ActaHortic.2015.1083.19 DOI: https://doi.org/10.17660/ActaHortic.2015.1083.19

EMA/HMPC (2013). Final assessment report on Origanum dictamnus L., herba (200431/2012). European Medicines Agency/ Committeeon Herbal Medicinal Products, London. Retrieved 2013 July 9 from http://www.ema.europa.eu

Farashah HD, Afshari RT, Sharifzadeh F, Chavoshinasab S (2011). Germination improvement and α-amylase and β-1,3-glucanase activity in dormant and non-dormant seeds of Oregano (Origanum vulgare). Australian Journal of Crop Science 5(4):421-427.

Figuérédo G, Cabassu P, Chalchat J, Pasquier B (2005). Studies of Mediterranean oregano populations -V. Chemical composition of essential oils of oregano: O. syriacum L. var. bevanii (Holmes) Ietswaart, O. syriacum L. var. sinaicum (Boiss) Ietswaart, and O. syriacum L. var. syriacum from Lebanon and Israel. Flavour and Fragrance Journal 20(2):164-168. https://doi.org/10.1002/ffj.1408 DOI: https://doi.org/10.1002/ffj.1408

Fournaraki C (2010). Conservation of threatened plants of Crete – Seed ecology, operation and management of a gene bank. PhD Thesis, National and Kapodistrian University of Athens, Faculty of Biology, Department of Botany, Athens, Greece.

Gamborg OL, Miller RA, Ojima K (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research 50:151-158. https://doi.org/10.1016/0014-4827(68)90403-5 DOI: https://doi.org/10.1016/0014-4827(68)90403-5

Goleniowski ME, Flamarique C, Bima P (2003). Micropropagation of oregano (Origanum vulgare x aplii) from meristem tips. In Vitro Cellular and Developmental Biology - Plant 39:125-128. https://doi.org/10.1079/IVP2002361 DOI: https://doi.org/10.1079/IVP2002361

Hanlidou E, Karouspu R, Kleftoyanni V, Kokkini S (2004). The herbal market of Thessaloniki (N. Greece) and its relation to the ethnobotanical tradition. Journal of Ethnopharmacology 91(2-3):281-299. https://doi.org/10.1016/j.jep.2004.01.007 DOI: https://doi.org/10.1016/j.jep.2004.01.007

IUCN (2001). IUCN Red List Categories and Criteria: Version 3.1. IUCN Species Survival Commission, IUCN, Gland, Switzerland and Cambridge, UK.

Korkor AM, Mohamed SA, Abd El-kafie OM, Gohar AA (2017). Adaptation of the in vitro culture of Origanum majorana L. for production of phenolic acids. IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) 12(2):30-38. https://doi.org/10.9790/3008-1202013038 DOI: https://doi.org/10.9790/3008-1202013038

Kougioumoutzis K, Kokkoris IP, Panitsa M, Strid A, Dimopoulos P (2021). Extinction risk assessment of the Greek endemic flora. Biology 10(3):195. https://doi.org/10.3390/biology10030195 DOI: https://doi.org/10.3390/biology10030195

Krigas N, Lazari D, Maloupa E, Stikoudi M (2015). Introducing Dittany of Crete (Origanum dictamnus L.) to gastronomy: A new culinary concept for a traditionally used medicinal plant. International Journal of Gastronomy and Food Science 2:112-118. https://doi.org/10.1016/j.ijgfs.2015.02.001 DOI: https://doi.org/10.1016/j.ijgfs.2015.02.001

Kumar M, Bhardwaj D (2020). The underexploited biotechnology of overexploited Origanum species: Status, knowledge gaps, prospects and potential. Plant Science Today 7(4):512-522. https://doi.org/10.14719/pst.2020.7.4.816 DOI: https://doi.org/10.14719/pst.2020.7.4.816

Kumari N, Saradhi PP (1992). Regeneration of plants from callus cultures of Origanum vulgare L. Plant Cell Reports 11(9):476-479. https://doi.org/10.1007/BF00232694 DOI: https://doi.org/10.1007/BF00232694

Leelavathi D, Kuppan N (2013). Callus induction and regeneration of multiple shoots from in vitro apical bud explant of Origanum vulgare, an important medicinal plant. International Journal of Research in Pharmacy and Chemistry 3(4):898-903.

Liolios CC, Graikou K, Skaltsa E, Chinou I (2010). Dittany of Crete: a botanical and ethnopharmacological review. Journal of Ethnopharmacology 131(2): 229-241. https://doi.org/10.1016/j.jep.2010.06.005 DOI: https://doi.org/10.1016/j.jep.2010.06.005

Lloyd G, McCown B (1980). Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. International Plant Propagation Society Proceedings 30:421-427.

Minas GJ (2001). Certain dittany apical meristem micropropagation in vitro. Agricultural Research Institute Ministry of Agriculture, Natural Resources and the Environment, Nicosia, Cyprus. Miscellaneous-Reports 80, pp 7.

Montezuma-De-Carvalho J, Paiva J, Pimenta M, Celestina M (1984). Effect of cold storage on seed viability of aromatic plants from the Portuguese flora. In: Proceedings of Eucarpia International Symposium on Conservation of Genetic Resources of Aromatic and Medicinal Plants. Oeiras, Portugal pp 111-116.

Murashige T, Skoog F (1962). A revised method for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:472-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x DOI: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Nitsch JP, Nitsch C (1969). Haploid plants from pollen grains. Science 163:85-87. https://doi.org/10.1126/science.163.3862.85 DOI: https://doi.org/10.1126/science.163.3862.85

Olivas NA, Bejarano CV, Soto GA, Ortega MZ, Salas FS, Chávez ES, Hernández Ochoa L (2020). Bioactive compounds and antioxidant activity of essential oils of Origanum dictamnus from Mexico. AIMS Agriculture and Food 5(3):387-394. https://doi.org/10.3934/agrfood.2020.3.387 DOI: https://doi.org/10.3934/agrfood.2020.3.387

Oluk EA, Çakır A (2009). Micropropagation of Origanum sipyleum L., an endemic medicinal herb of Turkey. African Journal of Biotechnology 8(21):5769-5772. https://doi.org/10.5897/AJB09.1216 DOI: https://doi.org/10.5897/AJB09.1216

Özkum D (2007). In vitro shoot regeneration of oregano (Origanum minutiflorum O. Schwarz & Davis). Hacettepe Journal of Biology and Chemistry 35(2):97-100.

Phillips GC, Collins GB (1979). In-vitro tissue culture of selected legumes and plant regeneration from callus of Red clover. Crop Science 19:59-64. https://doi.org/10.2135/cropsci1979.0011183X001900010014x DOI: https://doi.org/10.2135/cropsci1979.0011183X001900010014x

Sajina A, Geetha SP, Minoo D, Rema J, Nirmalbabu K, Sadanandan AK, Ravindran PN (1997). Micropropagation of some important herbal species. In: Proceedings of Biotechnology of Spices, Medicinal and Aromatic Plants. Indian Society for Spices, Calicut, India pp 79-86.

Sevindik B, Izgu T, Simsek O, Tutunku M, Curuk P, Yilmaz O, … Mendi YY (2017). In vitro culture of Turkish Origanum sipyleum L. American Journal of Plant Biology 2(5-1):32-36. https://doi.org/10.11648/j.ajpb.s.2017020501.16

Socorro O, Tarrega I, Rivas F (1998). Essential oils from wild and micropropagated plants of Origanum bastetanum. Phytochemistry 48(8):1347-1349. DOI: https://doi.org/10.1016/S0031-9422(97)00926-6

Takano T, Oki K, Kawabata M (1990). Germination characteristics of herb seeds in Labiatae. Scientific Reports of the Faculty of Agriculture, Meijo University, Japan 26:17-24.

Tejvathi DH, Padma AV (2012). In vitro multiplication of Majorana hortensis Moench - An aromatic medicinal herb. Indian Journal of Plant Sciences 1(1):48-56.

Thanos CA, Doussi MA (1995). Ecophysiology of seed germination in endemic Labiates of Crete. Israel Journal of Plant Sciences 43:227-237. https://doi.org/10.1080/07929978.1995.10676607 DOI: https://doi.org/10.1080/07929978.1995.10676607

Thanos CA, Kadis CC, Skarou F (1995). Ecophysiology of germination in the aromatic plants thyme, savory and oregano (Labiatae). Seed Science Research 5(3):161-170. https://doi.org/10.1017/S0960258500002786 DOI: https://doi.org/10.1017/S0960258500002786

Theophrastus (1926). Enquiry into Plants. Vol. II. (A.F. Hort, translator). Harvard University Press and William Heinemann Ltd., Cambridge, Mass., London.

Yildirim MU (2013). Micropropagation of Origanum acutidens (HAND.-MAZZ.) IETSWAART using stem node explants. The Scientific World Journal 2013:276464. http://dx.doi.org/10.1155/2013/276464 DOI: https://doi.org/10.1155/2013/276464

Zayova EG, Geneva MP, Miladinova-Georgieva KD, Hristozkova MG, Stancheva IV (2019). Impact of plant growth regulators on Greek oregano micropropagation and antioxidant activity. Biosciences, Biotechnology Research Asia 16(2):297-305. https://doi.org/10.13005/bbra/2746 DOI: https://doi.org/10.13005/bbra/2746

Published

2022-06-30

How to Cite

SARROPOULOU, V., MALOUPA, E., & GRIGORIADOU, K. (2022). In vitro direct organogenesis of the Cretan dittany (Origanum dictamnus L.), an important threatened Greek endemic species. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 50(2), 12715. https://doi.org/10.15835/nbha50212715

Issue

Section

Research Articles
CITATION
DOI: 10.15835/nbha50212715