Abstract
Nerine (Nerine sarniensis) cv. Salmon Supreme in vitro-grown bulblets, 7–9 mm in diameter, were cut in half longitudinally and used for adventitious bud initiation following dissection of the roots and two-thirds of the upper part of the bulblets. The terminal apex was injured with a hot, sterile microscope dissecting needle. The highest number of buds formed (seven to nine buds per halved bulblet) on a semi-solid Murashige and Skoog (MS) basal salts medium supplemented with 3% sucrose and either 1 μM 6-benzylaminopurine (BA) and 1 μM α-naphthaleneacetic acid (NAA) or 0.5 μM BA and 0.1 μM NAA. Bulblet halves were cultured in the dark for 11–13 weeks with one subculture after 6 weeks. Anatomical studies indicated that the initiation of adventitious buds on the abaxial side of the inner scales of the halved bulblet was adjacent to the basal plate and started from the leaf primordium and a meristematic bulge. Buds developed directly into small bulblets after they were transferred to semi-solid MS basal salts medium supplemented with 6% sucrose, 10 μM indole-3-butyric acid (IBA) and 0.25% activated charcoal. Small bulblets cultured in liquid MS medium supplemented with additional KH2PO4 (170 mg l-1), 6% sucrose and 0.1 μM NAA under a 16/8-h (light/dark) photoperiod for 8 weeks grew into larger bulbs faster than those cultured on semi-solid medium. The bulbs were rooted on a semi-solid medium after 4 weeks and then transferred to the soil. As many as 18 bulblets developed and rooted from one in vitro-grown bulb after 25–27 weeks.
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Abbreviations
- BA :
-
6-Benzylaminopurine
- IBA :
-
Indole-3-butyric acid
- NAA :
-
α-Naphthaleneacetic acid
References
Bryan JE (1989) Bulbs, vol 1. Timber Press, Portland
Chow YN, Selby C, Fraser TW, Harvey BMR (1993) Basal plate tissue in Narcissus bulbs and in shoot clump cultures: its structure and role in organogenic potential of single leaf cultures. Ann Bot 71:437–443
Custers JBM, Bergervoet JHW (1992) Differences between Nerine hybrids in micropropagation potential. Sci Hortic 52:247–256
De Bruyn MH, Ferreira DI, Slabbert MM, Pretorius J (1992) In vitro propagation of Amaryllis belladonna. Plant Cell Tissue Organ Cult 31:179–184
Grootaarts H, Schel JHN, Pierik RLM (1981) The origin of bulblets formed on excised twin scales of Nerine bowdenii W. Watts. Plant Cell Tissue Organ Cult 1:39–46
Hussey G (1975) Totipotency in tissue explants and callus of some members of the Liliaceae, Iridaceae, and Amaryllidaceae. J Exp Bot 26:253–262
Hussey G (1982) In vitro propagation of Narcissus. Ann Bot 49:707–719
Jacobs G, Richard M, Allderman LA, Theron KI (1992) Direct and indirect organogenesis in tissue cultures of Nerine bowdenii W. Watts. Acta Hortic 325:475–479
Langens-Gerrits M, Nashimoto S (1997) Improved protocol for the propagation of Narcissus in vitro. Acta Hortic 430:311–313
Lilien-Kipnis H, Ziv M, Kahany S, Azizbekov N (1992) Proliferation and regeneration of Nerine in liquid culture. Acta Hortic 325:467–473
Macdonald M (1987) Sterilization of Nerines using the twin scaling technique. Comb Proc Int Plant Prop Soc 37:205–210
Mochtak E (1989) Bulblet formation from the explants of Nerine bowdenii W. Watts. Acta Hortic 251:199–204
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–496
Okubo H, Huang CW, Uemoto S (1990) Role of outer scale in twin-scale propagation of Hippeastrum hybridum and comparison of bulblet formation from single- and twin-scales. Acta Hortic 266:59–66
Pierik RLM, Ippel BJ (1977) Plantlet formation from excised bulb scale segments of Nerine. Acta Hortic 78:197–202
Pierik RLM, Steegmans HHM (1986) Adventitious plantlet regeneration from floral stem explants of Nerine bowdenii W. Watts. Neth J Agric Sci 34:217–223
Pierik RLM, Blokker JS, Dekker MWC, de Does H, Kuip AC, van der Made TA, Menten YMJ, de Vetten NCMH (1990) Micropropagation of Hippeastrum hybrids. In: de Jong J (ed) Integration of in vitro techniques in ornamental plant breeding. Eucarpia, Section Ornamentals, pp 21–27
Rees AR (1985) Nerine. In: Halevy AH (ed) Handbook of flowering, vol 1. CRC Press, Boca Raton, pp 297–299
Sass JE (1958) Botanical microtechnique. Iowa State College Press, Ames
Slabbert MM, Niederwieser JG (1999) In vitro production of Lachenalia. Plant Cell Rep 18:620–624
Slabbert MM, De Bruyn MH, Ferreira DI, Pretorius J (1993) Regeneration of bulblets from twin scales of Crinum macowanii in vitro. Plant Cell Tissue Organ Cult 33:133–141
Van Brenk G, Benschop M (1993) Nerine. In: De Hertogh A, Le Nard M (eds) The physiology of flower bulbs. Elsevier, Amsterdam, pp 559–588
Vishnevetsky J, Lilien-Kipnis H, Azizbekova N, Ziv M (1997) Bulb and inflorescence development in Nerine sarniensis. Isr J Plant Sci 45:13–18
Vishnevetsky J, Zamski E, Ziv M (2000) Carbohydrate metabolism in Nerine sarniensis bulbs developing in liquid culture. Physiol Plant 108:361–369
Ziv M (1997) The contribution of biotechnology to breeding, propagation and disease resistance in geophytes. Acta Hortic 430:247–258
Ziv M, Lilien-Kipnis H (2000) Bud regeneration from inflorescence explants for rapid propagation of geophytes in vitro. Plant Cell Rep 19:845–850
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Communicated by K.K. Kamo
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Vishnevetsky, J., Zamski, E. & Ziv, M. Enhanced bud and bulblet regeneration from bulbs of Nerine sarniensis cultured in vitro. Plant Cell Rep 21, 645–650 (2003). https://doi.org/10.1007/s00299-003-0580-2
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DOI: https://doi.org/10.1007/s00299-003-0580-2