In Vitro Production of Bulblet in Galanthus transcaucasicus Fomin, an Endangered Medicinal Plant

Document Type : Research Paper

Authors

1 Department of Horticultural Science, Maragheh Branch, Islamic Azad University, Maragheh, Iran

2 Department of Plant Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran

3 Department of Biology, Ahar Branch, Islamic Azad University, Ahar, Iran

Abstract

To optimize conditions for micropropagating Galanthus transcaucasicus Fomin, the effect of explant type, different concentrations of sucrose, type and different concentrations of auxin were examined on bulblet production. For the induction of bulblets, three types of bulb scales were usesd as explants. A two-step sterilization procedure was applied. Assessments were made after incubation for 20 weeks at 23±1ºC with 16h photoperiods. Highest number of bulblets were proliferated culturing bulb scales on  modified MS medium supplemented with 2.0 mg.l-1 BA and 2.0 mg.l-1 IBA. About  27% of bulblets were bigger than 5 mm in diameter, suitable for acclimatization. With the increase of sucrose concentration, number of bulblets decreased, but their size increased. This protocol provided a basis for future study on large-scale multiplication system for commercial nurseries of Galanthus transcaucasicus.
 
 

Keywords


Article Title [فارسی]

تولید درون شیشه‌ای پیازچه‌های گیاه در حال انقراض Galanthus transcaucasicus Fomin

Abstract [فارسی]

جنس گالانتوس متعلق به تیره آماریلیداسه بوده که پراکنش آن در اروپا، آسیای صغیر و منطقه قفقاز است. آلکالوئیدهای خاصی در گیاهان این تیره وجود دارد که به دلیل داشتن خواص دارویی توجه پژوهشگران را به خود جلب نموده است. در پژوهش حاضر اثر غلظت‌های مختلف ساکاروز و چند نوع اکسین بر روی تولید پیازچه درون شیشه‌ای مورد آزمایش قرار گرفت. پیازهای گالانتوس با روش دو مرحله‌ای ضدعفونی شدند. پیازها بعد از تیمار با اتانول 70 درصد به مدت یک دقیقه، به مدت 20 دقیقه در هیپوکلریت سدیم 5 درصد قرار گرفته و سپس پیازهای نصف شده به مدت 10 دقیقه در هیپوکلریت سدیم 5/2 درصد قرار داده شدند. بر اساس نتایج حاصل، بیشترین پیازچه در محیط کشت MS تغییر یافته حاوی 2 میلی‌گرم در لیتر بنزیل آمینوپورین (BAP) و 2 میلی‌گرم در لیتر ایندول بوتیریک اسید (IBA) به دست آمد که در این حالت 27 درصد پیازچه ­ها بزرگتر از 5 میلی‌متر بوده و مناسب سازش به شرایط برون‌شیشه‌ای بودند. همچنین با افزایش غلظت ساکاروز، تعداد پیازچه‌ها کمتر شد ولی به اندازه آن­ها افزوده شد. این تحقیق دستورالعملی را جهت تولید گونه خزری جنس گالانتوس در خزانه در مقیاس بزرگ فراهم می‌نماید.
 

Keywords [فارسی]

  • ریزازدیادی
  • کشت بافت
  • گل برفی
Ault JR,1995. In vitro propagation of Eucomis autumnalis, E. comosa andE. zambesiaca by twin-scaling. HortScience 30: 1441–1442.
Berkov S, Cuadrado M, Osorio E, Viladomat F, Codina C and Bastida J, 2009a. Three new alkaloids from Galanthus nivalis and Galanthus elwesii. Planta Medica75: 1351-1355.
Berkov S, Pavlov A, Georgiev V, Bastida J, Burrus M, Ilieva M and Codina C, 2009b. Leucojum aestivum in vitro cultures: variation in the alkaloid patterns. Natural Product Communications 4: 359-364.
Chow YN, Selby C, Fraser TW and Harvey BMR, 1992. Stimulation by sucrose of Narcissus bulbil formation in vitro. Journal of Horticultural Sciences 62: 289–293.
Davis AP, 1999. The genus Galanthus. In: Mathew B (Ed.) A Botanical Magazine Monograph. Timber Press Inc., Oregon 17: 55– 140.
Davis AP and Ozhatay N, 2001. Galanthus trojanus: a new species of Galanthus (Amaryllidaceae) from northwestern Turkey. Botanical Journal of the Linnean Society137: 409–412.
Fennell CW and Van-Staden J, 2004. Biotechnology of southern African bulbs. South African Journal of Botany 70: 37–46.
Fennell CW, Crouch NR  and Van-Staden J, 2001. Micropropagation of the River Lily, Crinum variable (Amaryllidaceae). South African Journal of Botany 67: 74–77.
Girmen M, 1986. Untersuchungen zur in vitro kultur von geopheten. Dissertation der Univ. Hannover, 43: 382-388.
Jalili A and  Jamzad Z, 1999. Red data book of Iran. Research Institute of Forests & Ranglands Publication, 215p In Persian).
Kim EK, Hahn EJ, Murthy HN and Paek KY, 2003. High frequency of shoot multiplication and bulblet formation of garlic in liquid cultures. Plant Cell, Tissue and Organ Culture 73: 231– 236.
Larsen MM, Adsersen AA, Davis AP,  Lledo MD, Jager AK and Ronsted N, 2010. Using a phylogenetic approach to selection of target plants in drug discovery of acetylcholinesterase inhibiting alkaloids in Amaryllidaceae tribe Galantheae. Biochemical Systematics and Ecology 38: 1026–1034.
Meerow AW, Francisco-Ortega J, Kuhn DN and Schnell RA, 2006. Phylogenetic relationships and biogeography within the Eurasian clade of Amaryllidaceae based on plastid ndhF and nrDNA ITS sequences: lineage sorting in a reticulate area. Systematics Botany 31: 42–60.
Mirici S, Parmaksiz I, Ozcan S, Sacak C, Uranbey S, Sarinhan EO, Gumuscu A, Gurbuz B and Arsalan N, 2005. Efficient in vitro bulblet regeneration from immature embryos of endangered Sternbergia fischeriana. Plant Cell, Tissue and Organ Culture 80: 239-246.
Murashige T and Skoog FA, 1962. A revised medium for rapid growth and bioassay with Tobacco tissue cultures. Plant Physiology 15: 473-479.
Ozel CA, Khawar KM and Unal F, 2015. Factors affecting efficient in vitro micropropagation of Muscari muscarimi Medicus using twin bulb scale. Saudi Journal of Biological Sciences 22 (2): 132-138.
Paek KY and Murthy HN, 2002. High frequency of bulblet regeneration from bulb scale sections of Fritillaria thunbergii. Plant Cell, Tissue and Organ Culture 68: 247–252.
Pavlov A and Bley T, 2006. Betalains biosynthesis by Beta vulgaris L. hairy root culture in a temporary immersion cultivation system. Process Biochemistry 41: 848-852.
Rice LJ, Finnie JF and Van-Staden J, 2011. In vitro bulblet production of Brunsvigia undulate from twin-scales. South African Journal of Botany 77: 305-312.
Ronsted N, Zubov D, Bruun-Lund and Davis AP, 2013. Snowdrops falling slowly into place: an improved phylogeny for Galanthus (Amaryllidaceae). Molecular Phylogenetics and Evolution 69: 205-217.
Santos J, Santos I and Salema R, 1998. In vitro production of bulbs of Narcissus bulbocodium flowering in the first season of growth. Scientia Hoticulturea 76: 205-217.
Staikidou I, Watson S, Harvey BMR and Selby C, 2005. Narcissus bulblet formation in vitro: effects of carbohydrate type and osmolarity of the culture medium. Plant Cell, Tissue and Organ Culture 80: 313–320.
Tipirdamaz R, 2003. Rooting and acclimatization of in vitro micropropagated snowdrop (Galanthus ikariae Baker.) bulblets. Ziraat Fakultesi Dergisi 16: 121-126.
Ulrich MR, Davies FT, Koh YC, Duray SA and Egilla JN, 1999. Micropropagation of Crinum ‘Ellen Bosanquet’ by triscales. Scientia Horticulturea 82: 95–102.
Uranbey S, 2011. In vitro bulblet regeneration from immature embryos of endangered and endemic Muscari azureum. Archive of Biological Science 63 (1): 209-215.
Wawrosch C, Malia PR and Kopp B, 2001. Clonal propagation of Lilium nepalense D. Don, a threatened medicinal plant of Nepal. Plant Cell Reports 20: 285–288.
Yeoman MM, 1986. Plant cell culture technology. Botanical Monographs, vol. 23. Blackwell Scientific Publications, Oxford. 253pp.
Zayed R, El-Shamy H and Berkov S, 2011. In vitro micropropagation and alkaloids of Hippeastrum vittatum. In Vitro Cellular & Developmental Biology - Plant 47: 695-701.
Zubov DA and Davis AP, 2012. Galanthus panjutinii sp. Nov: a new name for an invalidly published species of Galanthus (Amaryllidaceae) from the northern Colchis area of Western Transcaucasia. Phytotaxa 50: 55–63.