Direct and Callus-mediated Protocorm-like Body Induction and High Frequency Adventitious Shoot Regeneration in an Endangered Orchid -Dendrobium farmeri Paxt. (Orchidaceae (original) (raw)
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Lindl. is one of the horticulturally important orchids of Nepal due to its beautiful yellowish flower and medicinal properties. The present study was carried out for plant regeneration from shoot tip explants of D. densiflorum by tissue culture technique. The shoot tip explants of this species, obtained from in vitro grown seedlings were cultured on Murashige and Skoog (MS) media alone and MS medium supplemented with different combination and concentration of 6benzylaminopurine (BAP) and napthalenacetic acid (NAA). The maximum number of healthy shoot was observed on MS+ BAP (2 mg/l) + NAA (0.5 mg/l) (4 shoots/ culture). The shoot multiplication started after three weeks of culture.
Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids
Plant cell, tissue and organ culture, 2004
Leaf explants of Paphiopedilum phiIippinense hybrids (hybrid PH59 and PH60) directly formed adventitious shoots from wound regions within 1 month, when cultured on modified Murashige and Skoog medium (1/2strength macro-and full-strength micro-elements) free of plant growth regulator in darkness. The combinations of 2,4-dichlorophenoxyacetic acid ((2,4-D) acid (0, 4.52 and 45.25 µM) and 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ) (0, 0.45, 4.54 and 22.71 µM) were used to test their effects on direct shoot bud formation from two types of explants (1.5-cm long intact leaf explants and 0.5-cm long leaf segment explants). In hybrid PH59, 4.54 µM TDZ increased mean numbers of shoots per explant with leaf segment explants. In hybrid PH60, 4.52 µM 2,4-D plus 0.45 µM TDZ promoted direct shoot bud formation from leaf segment explants. In addition, three treatments (4.52 µM 2,4-D, 22.71 µM TDZ, 4.52 µM 2,4-D plus 4.54 µM TDZ) gave a higher response than control on mean numbers of shoots per explant with intact leaf explants. Healthy plantlets each with one to three roots were obtained from leaf-derived shoots after transfer onto a hormone-free medium for 22 months. These plantlets were acclimatized in a greenhouse and grew well with 100% survival rate.
Scientia Horticulturae, 2003
Shoot-tip explants of Dendrobium ®mbriatum Lindl. var. oculatum Hk. f. were cultured on modi®ed nutrient solution of Knudson's C, supplemented with 10% (v/v) coconut water, 0.5 mg l À1 niacin, 0.5 mg l À1 pyridoxine HCl, 0.1 mg l À1 thiamine HCl and different concentrations of N 6benzylaminopurine (BAP) and a-naphthaleneacetic acid (NAA) in factorial combinations. After 2 weeks, the explants produced a compact mass of translucent embryogenic callus. Application of both plant growth regulators (PGRs) was essential for the induction of callus. Optimum callusing was recorded in the presence of 0.5 mg l À1 NAA and 1 mg l À1 BAP (66.7%). Following transfer to the PGR-free medium, the callus further proliferated with side-by-side regeneration of protocorm-like bodies (PLBs). These PLBs readily germinated into well-rooted plantlets in the same medium. Although the presence of exogenous PGRs was essential for the induction of callus, it was possible to maintain this callus through subculturing in the PGR-free medium for more than 2 years, without any loss of embryogenic potential. The explants that failed to respond to the callus induction media, developed elongated leafy shoots that later multiplied by axillary shoot formation. The process of axillary branching was primarily dependent on BAP, but better response was obtained in the presence of additional NAA. #
Direct shoot regeneration from nodes of Phalaenopsis orchids
2004
Nodes with dormant buds from flower stalks of orchid Phalaenopsis sp. were plated on six culture media. The composition of the culture media affected the induction, regeneration, number and form of Phalaenopsis regenerants. We managed to obtain direct shoot regeneration without callus formation on all media except for medium F. Medium A, supplemented with 2 mg/l of 6-benzylaminopurine (BAP) and 0.5 mg/l of α–naphtalenacetic acid (NAA) was found to be the most appropriate of all the media used for rapid micropropagation of a large number of vegetative shoots (multiplication factor 8.35 per node) without roots 160 days after inoculation. We found medium B, supplemented with 4.41 mg/l BAP and 1 mg/l NAA less appropriate for vegetative shoot production (multiplication factor 2.08 per node) also because the regenerants ceased to elongate. Medium C, containing BAP (2 mg/l) and a lower nitrogen content showed the highest multiplication rate (0.54 per node) for generative regenerants format...
The Agriculturists
An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration. It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium. For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoniae.
The Agriculturists, 2017
An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration. It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium. For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoni...
In vitro regeกneration from protocorms in Dendrobium aqueum Lindley – An imperiled orchidา
An efficient in vitro plant regeneration protocol from protocorms of Dendrobium aqueum was developed. The uniformly developed protocorms (in vitro origin) having shoot initials were cultured on half macro strength MS medium (1/2 MS) supplemented with cytokinins (BA, 2iP, KIN and TDZ) at 1, 3, 5, 7, 10 mg l À1 , natural additives (BP and CW) at 1%, 3%, 5%, 7%, 10% and auxins (IBA, NAA, 2,4-D) at 1, 3, 5, 7, 10 mg l À1 to study their efficacy on complete plant development. A maximum of 9.4 shoots per explant were generated on 3 mg l À1 of NAA followed by 3% of BP (7.0 shoots). Shoot elongation (1.52 cm) was achieved on 1/2 MS medium fortified with NAA 7 mg l À1 followed by TDZ 7 mg l À1 (1.37 cm). Shoots cultured on 1/2 MS medium supplemented with IBA 5 mg l À1 produced an average of 8.75 roots per shoot, however the lengthiest roots (1.48 cm) were noted in NAA 7 mg l À1 . Healthy rooted plantlets successfully acclimatized in ex vitro condition. The role of complete plantlet production by natural additives could be useful for conservation and cost effective commercial production of orchids.
Walailak Journal of Science and Technology (WJST), 2021
To investigate the suitable medium for in vitro shoot regeneration and plantlet growth of Dendrobium crumenatum Sw., individual protocorm-like bodies (PLBs) (about 4 - 5 mm in diameter) of Dendrobium crumenatum Sw. derived from MS medium supplemented with 0.5 mg/L TDZ for 60 days of culture were cultured on 6 culture media; Murashige and Skoog (MS) medium, MS medium supplemented with 15 % (v/v) CW, MS medium supplemented with 15 % (v/v) CW and 0.2 % (w/v) AC, Vacin and Went (VW) medium, VW medium supplemented with 15 % (v/v) CW, VW medium supplemented with 15 % (v/v) CW and 0.2 % (w/v) AC. After 4 months of culture, MS medium containing 15 % coconut water (CW) gave the highest percentage of shooting and number of shoots per explant of 96.0 and 9.5, respectively with a significant difference from other media. The addition of 0.2 % (w/v) activated charcoal (AC) significantly increased the number of leaves and roots. PLBs developed into complete plantlets. MS medium supplemented with 1...
Plant Cell Tissue and Organ Culture, 2010
Malaxis acuminata is a terrestrial orchid that grows in shady areas of semi-evergreen to shrubby forests. It is highly valued for its medicinal properties as dried pseudo-bulbs are important ingredients of several Ayurvedic preparations. In this study, adventitious shoot buds were induced from internodal explants of M. acuminata grown on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn), and thidiazuron (TDZ). Of the three cytokinins used, TDZ at 3 mg l−1 induced the highest frequency (82%) of organogenic explants. However, all responding explants produced only a single adventitious shoot irrespective of the type and concentration of the cytokinin. Adding 0.5 mg l−1 α naphthaleneacetic acid (NAA) to the medium enhanced adventitious shoot formation. In the presence of 3 mg l−1 TDZ and 0.5 mg l−1 NAA, frequency of organogenesis was 96% with a mean number of 6.1 shoots per explant. Prolonged culture or subculture on the same medium did not promote further shoot production. However, transfer of these cultures to MS medium supplemented with 3 mg l−1 TDZ and 0.5 mg l−1 NAA and various concentrations of different polyamines (PAs), including spermine, spermidine, and putrescine, significantly increased mean shoot number per explant. The highest frequency of shoot induction (100%) and mean shoot number per explant (14.6) was observed on MS medium with 3 mg l−1 TDZ, 0.5 mg l−1 NAA, and 0.4 mM spermidine. Regenerated shoots were excised and subcultured on an elongation medium consisting of MS medium with 3 mg l−1 BA. Moreover, the highest frequency of rooting (96%) and mean number of roots per shoot (3.3) was observed on MS medium with 4 mg l−1 indole-3-butyric acid (IBA) and 1.5 mg l−1 activated charcoal (AC). Almost 90% of rooted shoots were successfully acclimatized and established ex vitro.
In vitro Propagation of orchid (Dendrobium ovatum (L.) Kraenzl.) through Somatic Embryogenesis
Plant Tissue Culture and Biotechnology
An efficient in vitro regeneration protocol through somatic embryogenesis was established from longitudinally bisected protocorm (lTCL) of an endangered orchid Dendrobium ovatum. The efficiency of EC and SEs from the protocorms significantly relied on the concentration of PGRs. MS medium supplemented with TDZ (1.0 mg/l) induced optimum of EC (31.8%) and SEs (28.1/explant). Similarly, ZEA (0.5 mg/l) induced EC (27.6%) and SEs (17.1/explant). The combined treatment of TDZ (1.0 mg/l) and NAA (0.5 mg/l) resulted in the maximum induction of EC (58.6%) and SEs (39.8/explant) in an upright incubated explant. In another combined effect of ZEA (1.0 mg/l) and NAA (0.5 mg/l) induced EC (43.8%) and SEs (24.3/explant), whereas the combination of BAP (0.5 mg/l) and NAA (0.5 mg/l) produced EC (34.4%) and SEs (16.8/explants). The induced EC and SEs where healthier on medium containing TDZ + NAA than on the medium containing ZEA+NAA and BAP+NAA. The orientation of lTCL explants seemed to interact wi...