Enhanced bud and bulblet regeneration from bulbs of Nerine sarniensis cultured in vitro

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 mM 6-benzylaminopurine (BA) and 1 mM anaphthaleneacetic acid (NAA) or 0.5 mM BA and 0.1 mM 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 mM indole-3-butyric acid (IBA) and 0.25% activated charcoal. Small bulblets cultured in liquid MS medium supplemented with additional KH 2 PO 4 (170 mg l -1 ), 6% sucrose and 0.1 mM 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.

Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis inLeucojum aestivumL. plants cultured in RITA®bioreactor

“…However, the results of the research conducted by Ivanov et al (2012) and Georgiev et al (2014) on shoot cultures of L. aestivum, as well as Pancratium maritimum, with the use of standard amount of sucrose in the medium, showed that the optimal conditions for growth in the RITA Ò bioreactor (immersion frequency and stand-by periods) provided both the optimal biomass accumulation and the best sugar utilization from the medium. Vishnevetsky, Zamski & Ziv (2003) reported that in the case of Nerine sarniensis bulbs developing in a liquid culture, sucrose in the medium was hydrolyzed to glucose and fructose and total soluble sugars after 8 weeks decreased from 60 g/L to approximately 39 g/L. On the other hand, according to Ziv (2005), sucrose is reduced or rapidly removed from the medium in both agar-gelled and liquid cultures.…”

Biochar successfully replaces activated charcoal for in vitro culture of two white poplar clones reducing ethylene concentration

“…AC, produced from wood, wood waste, paper-mill waste liquors and peat (El-Hendawy et al 2001) is often used in plant tissue culture: it is well known that it may improve cell growth and development (Pan and Van Staden 1998) and better growth responses of plant tissues are generally associated with the addition of AC to both liquid and semi-solid media (Anagnostakis 1974;Fridborg and Eriksson 1975;Horner et al 1977;Weatherhead et al 1978;Peck and Cumming 1986;Bon et al 1988;Zaghmout and Torello 1988). AC has been used to promote somatic embryogenesis (Buccheim et al 1989;Andrade and Merkle 2005;Zouine et al 2005), androgenesis (Johansson 1986;Nomizu et al 2004;Laura et al 2006), rooting (Sul and Korban 2004;Loc et al 2005), shoot and root elongation (Mohamed-Yasseen 2001;Sul and Korban 2005), bulb formation (Peck and Cumming 1986;Vishnevetsky et al 2003) and to inhibit tissue browning (Mohamed-Yasseen 1994;Mohamed-Yasseen et al 1995). For an extensive review on the effects of AC in tissue culture, see e.g.…”