In vitro clonal propagation of Nyctanthes arbor-tristis

Abstract

Rapid shoot multiplication of Nyctanthes arbor-tristis L. was achieved from axillary meristems on Murashige and Skoog (MS) basal medium supplemented with 1.0 -1.5 mg dm -3 6-benzylaminopurine (BA), 50 mg dm -3 adenine sulfate (Ads) and 3 % (m/v) sucrose. Inclusion of indole-3-acetic acid (IAA) in the culture medium along with BA + Ads promoted a higher rate of shoot multiplication. Maximum mean number of microshoots per explant (6.65) was achieved on the MS medium supplemented with 1.5 mg dm -3 BA, 50 mg dm -3 Ads and 0.1 mg dm -3 IAA after 4 weeks of culture. The elongated shoots rooted within 13 to 14 d on half-strength MS medium supplemented with either indole-3-butyric acid (IBA), IAA or 1-naphthaleneacetic acid (NAA) with 2 % sucrose. Maximum percentage of rooting was obtained on medium having 0.25 mg dm -3 IBA and 0.1 mg dm -3 IAA. About 70 % of the rooted plantlets survived in the greenhouse. The in vitro raised plants were grown normally in the field.Additional key words: growth regulators, medicinal plant, shoot multiplication. ⎯⎯⎯⎯Nyctanthes arbor-tristis Linn. is a valuable medicinal plant which belongs to the family Oleaceae, and is distributed in Indo-Malayan regions. It is a small tree with scented white flowers. It is a native of India occurring wild in the Sub-Himalayan region. Due to unrestricted large-scale exploitatiobn of the natural resource, coupled with limited cultivation and insufficient attempts for its replenishment, the natural stock of this species has been markedly depleted. Propagation from seed is unreliable due to poor germination and death of many young seedlings under natural conditions (Thapliyal and Naithani 1996). In vitro culture is an alternative method for conservation and propagation of this species, but no report has been published so far. Hence, this investigation aimed to develop an efficient protocol for micropropagation of N. arbor-tristis, an important medicinal plant.Elongated shoots (4 -5 cm long) were collected from field grown plants of Nyctanthes arbor-tristis L. and brought to the laboratory with cut ends dipped in distilled water. Stem without leaves were washed in 2 % (m/v) Teepol (Qualigen, Mumbai, India) and surface disinfected by 0.1 % (m/v) aqueous mercuric chloride solution for 15 min. After rinsing 4 -5 times with sterile distilled water, stems were cut into smaller segments (ca. 0.5 cm long), each with one node was used as explant source.Nodal explants were placed on semisolid Murashige and Skoog (1962; MS) mineral salts plus 555 µM myoinositol, 4.06 µM nicotinic acid, 2.43 µM pyridoxineHCl, 0.296 µM thiamine-HCl supplemented with various concentrations of cytokinins, i.e. 6-benzylaminopurine (BA: 0.0, 0.5, 1.0, 1.5 and 2.0 mg dm -3 ), kinetin (Kn: 0.0, 0.5, 1.0, 1.5 and 2.0 mg dm -3 ), adenine sulfate (Ads: 25, 50 and 100 mg dm -3 ) and auxins like indole-3-acetic acid (IAA: 0.0, 0.10, 0.25 and 0.5 mg dm -3 ) and 1-naphthalene-acetic acid (NAA: 0.0, 0.10, 0.25 and 0.5 mg dm -3 ) for shoot multiplication. The pH of the medium was adjusted to 5.8 prior...

Micropropagation and assessment of genetic stability in Celastrus paniculatus: An endangered medicinal plant

“…The medium devoid of growth regulators failed to induce the formation of shoot buds (data not shown). Similar results were obtained in Crataeva nurvala (Walia et al 2003); Nyctanthus arbor-tristis (Rout et al 2008) and Celastrus paniculatus (Lal & Singh 2010) on MS basal medium. Higher numbers of multiple shoot were recorded in all concentrations of BAP than the medium supplemented with the same concentration of Kn albeit with longer internodes.…”

“…A higher concentration of BAP (> 2.0 mg/L) in the culture medium inhibited the growth of the shoots and stimulated callusing at the basal end where as a higher concentration of NAA suppress the rate of shoot multiplication and showed stunted growth. This result is contradictory to the result obtained by Lal & Singh (2010), as they found maximum rate of multiplication in MS medium supplemented with 1.0 mg/L BAP and 0.1mg/L NAA (C. paniculatus) but supported by reports given by Gopi et al (2006), Rout et al (2008) and Murthy et al (2010) in Ocimum gratissimum, Nyctanthus arbor-tristis and Ceropegia spiralis respectively. The present findings suggest a high frequency of shoot production from axillary meristems by manipulating the concentration of growth regulators.…”

“…Indiscriminate over exploitation coupled with insufficient attempts for replenishment of wild stock have contributed to its threatened status requiring scientific efforts for conservation and commercial cultivation (De Silva & Senarath 2009;Lal & Singh 2010). Due to low seed viability, germination rate and lack of vegetative propagation methods, tissue culture technique is an alternate solution for conservation of those valuable and endangered medicinal plants (Rout et al 2008;Bantawa et al 2011;Swarna & Ravindhran 2012). Though tissue culture is recognized as one of the key areas of biotechnology for large scale propagation and conservation, but it is severely hindered due to somaclonal variations (Larkin & Scowcroft 1981).…”

In vitro propagation of Drosera intermedia in a single step

“…However, dewatering, drainage in order to create productive land and cattle grazing are serious threats to these habitats and the conservation of D. intermedia. Taking into account the declining natural populations, the development of efficient micropropagation protocols is critical as they can be used to replenish wild stocks and provide an alternative source of plant material for bioactivity assays (Chen et al 2008, Rout et al 2008. In fact, micropropagation protocols for several carnivorous species have been described lately (Jang et al 2003, Gonçalves and Romano 2005, Gonçalves et al 2008.…”

Effect of Carbon Sources and Sucrose Concentrations on Shoot Organogenesis of Aloe saponaria

“…The commercial cultivation of A. saponaria is challenging because of the low seed viability, low germination rate, limited availability of raw material with high quality, and slow vegetative growth. Tissue culture technique is an alternate method for preservation of those beneficial medicinal plants [12][13][14] . Large scale propagation and conservation of plants by tissue culture is recognized as one of the key areas in biotechnology techniques.…”