Micropropagation of Red Kino Tree (Pterocarpus marsupium Roxb.): A Medicinally Important Plant (original) (raw)
Related papers
This paper describes multiple shoot regeneration of Pterocarpus santalinus L.f, an endangered tree endemic to the Deccan region which is commercially and medicinally most valuable for its heartwood on the international market. Seed germination under in vivo (in field conditions) and in vitro conditions were 60% and 100%, respectively. The nodal segments from in vitro regenerated shoots and from mature trees proliferated into multiple shoots (mean = 6.5) on Murashige and Skoog (MS) medium supplemented with 6-benzyl amino purine (BAP), kinetin (KN) and thidiazuron (3.0 mg L-1) individually. The seed explants (embryonic axis along with the cotyledons), when cultured on MS medium containing a combination of KN and BAP (1.0 + 2.0 mg L-1) formed 19-20 multiple shoots. MS basal medium was found suitable for rooting (70-80%). On transfer to a glasshouse 20% (3 plantlets/explant) survived.
Micropropagation of Adult Tree of Pterocarpus Marsupium Roxb . Using Nodal Explants
2015
Attempts were made for in vitro propagation of Pterocarpus marsupium Roxb., belonging to family Fabaceae, an economically important multipurpose tree. The tree is scared with noval antidiabetic properties. The tree shows poor seed germination capacity (30%) due to hard seed coat and conventional vegetative regeneration methods are a complete failure. Therefore, the propagation of this tree by tissue culture techniques is an urgent need and well justified. Nodal segments containing axillary bud from 10 years old tree of P. marsupium were evaluated for axillary shoot proliferation on Murashige and Skoog’s (MS) basal medium fortified with BAP (6–benzylaminopurine) and kinetin (Kn) singly or in combinations with auxins at different concentrations. The best shoot proliferation was obtained with 13.95 μM Kn + additives (568 μM Ascorbic acid, 260 μM Citric acid, 605 μM Ammonium sulphate and 217 μM Adenine sulphate) in MS medium where 64.44% of the axillary buds responded with development o...
Plant Cell, Tissue and Organ Culture (PCTOC), 2020
Pterocarpus marsupium Roxb., which belongs to the Fabaceae family, is a promising herbal drug producing forest tree widely known as the 'Indian Kino'. An effective regeneration protocol using in vitro seedlings of P. marsupium under the gibberellic acid and thidiazuron regimen is described in the present study. Gibberellic acid was also tested to have an effect on in vitro rhizogenesis from microshoots. Combination of gibberellic acid and thidiazuron in culture media improved the germination percentage, multiplication and subsequent elongation of shoots. Murashige and Skoog medium containing 0.50 µM gibberellic acid with 0.50 µM thidiazuron was found to be the most effective treatment for producing the highest number of shoots per seedling (23.0) with an average shoot length 5.14 cm in 85% cultures, after 8 weeks. For in vitro rhizogenesis, 100 μM of indole 3-butyric acid was treated for 5 days in basal ends of isolated microshoots. Thereafter, pretreated microshoot was transferred to gibberellic acid (0.50 µM) supplemented MS medium. This treatment was found to be the most effective for in vitro root formation, where 10.54 roots with an average root length of 4.60 cm per microshoot in 80% cultures were recorded, after 4 weeks. During acclimatization photosynthetic traits and their attributes such as net photosynthetic rate, stomatal conductance, intercellular CO 2 concentration, and Chlorophyll content were analyzed and these attributes were found to be fluctuating initially for 35 days, thereafter, an increasing trend was recorded up to 84 days of acclimatization. The well acclimatized plantlets were shifted to fields where they grew normally without any morphological changes with an 86.7% of survival. Genetic integrity in micropropagated plants was validated by DNA-based ISSR primers. These primers were amplified in monomorphic banding pattern suggesting a high-level of genetic uniformity in micropropagated plants. Overall, an effective and high throughput plant regeneration method has been developed for obtaining true-to-type regenerants of P. marsupium. Key message The present study revealed the high throughput regeneration of P. marsupium through in vitro seedlings method which may be an alternative tool for mass propagation and germplasm conservation of woody trees directly from seed, thus reducing an explant preparation and recovery of seedlings capable of multiple shoot formation under the regimes of gibberellic acid and thidiazuron. Both the growth regulators are involved in the improvement of plantlet regeneration processes as well as maintenance of physiological stresses during in vitro cultures. Communicated by Ranjith Pathirana.
Micropropagation of red kino tree (Pterocarpus marsupium Roxb.)
An efficient in vitro propagation protocol for Liparis elliptica (Rees) Lindl. using transverse thin cell layers (TCLs) was established. The initiation of protocorm-like bodies (PLBs) and the regeneration of shoot buds from PLB TCLs significantly relied on the concentration of 24-epi brassinolide (24-epiBL)-supplemented Mitra et al. basal medium. The highest percentage of PLB-TCL explants (93.0%) producing PLBs (71.0 ± 2.1) was recorded on 4.0 µM 24-epiBL in a period of 12 weeks. The cultures were maintained for 6-12 weeks for the initiation of PLBs or proliferating shoot buds. After nearly 12 weeks, small bud-like structures formed healthy shoots. The highest number of shoots with well developed roots was developed on 10.74 µM NAA-supplemented basal medium. This successful protocol will allow for the mass multiplication of L. elliptica, fulfilling the timely demand for clonal plantlets. This is the first ever report of in vitro culture for this epiphytic orchid.
Rapid in vitro multiplication of Melia azedarach L. (a multipurpose woody tree
Acta Physiologiae Plantarum, 2009
An efficient regeneration protocol for rapid multiplication of Melia azedarach, an economically as well as medicinally important timber-yielding tree, was developed. Nearly 90% of the culture exhibited axillary bud sprouting and multiple shoot formation from nodal segments derived from 20-year-old candidate plus tree on Murashige and Skoog (MS) medium supplemented with 5 μM 6-benzyladenine (BA). The highest shoot regeneration frequency (92%), maximum number of multiple shoots (19.7 ± 0.31) as well as shoot length (4.9 ± 0.08 cm) was induced from nodal explants on MS medium amended with 5.0 μM BA, 0.5 μM indole-3-acetic acid (IAA) and 30 μM adenine sulfate (AdS). Addition of 250 mg l−1 ammonium sulphate, (NH4)2SO4, and 100 mg l−1 K2SO4, prevented defoliation and tip burning without affecting the number of shoots. The explant harvest period also influenced the bud break and shoot sprouting from nodal segments. Repeated subculturing of nodal explants on fresh MS medium containing lower concentration of BA (2.5 μM) along with IAA (0.5 μM), AdS (30 μM) and additives was found most suitable growth regulator regime for achieving 1.2-fold increase in shoot multiplication rate. The percentage of shoot multiplication as well as the number of shoots per node remained the same during first three subculture passages, afterwards a decline was recorded. About 90% of the in vitro regenerated shoots were successfully rooted ex vitro by giving a pulse treatment of 250 μM indole-3-butyric acid for 15 min, followed by their transfer to thermocol cups containing soilrite. The raised plantlets were successfully acclimatized first under culture room conditions, then to green house with 85% survival rate.
A rapid in vitro propagation of red sanders (Pterocarpus santalinus L.) using shoot tip explants
Acta Physiologiae Plantarum, 2011
An efficient protocol has been developed for the in vitro propagation of Pterocarpus santalinus L. using shoot tip explants which is a valuable woody medicinal plant. Various parts of this plant are pharmaceutically used for the treatment of different diseases. Multiple shoots were induced from shoot tip explants derived from 20 days old in vivo germinated seedlings on 1:1 ratio of sand and soil after treating with gibberellic acid (GA 3). The highest frequency for shoot regeneration (83.3%) with maximum number of shoot buds (11) per explant was obtained on Murashige and Skoog (MS) medium supplemented with 1.0 mg/l of 6-benzylaminopurine (BAP) along with 0.1 mg/l of thidiazuron (TDZ) after 45 days of culture. A proliferating shoot culture was established by repeatedly subculturing the original shoot tip explants on fresh medium after each harvest of the newly formed shoots. Sixty percent of the shoots produced roots were transferred to rooting medium containing MS salts and 0.1 mg/l indole-3butyric acid (IBA) after 30 days. About 73.33% of the in vitro raised plantlets were established successfully in earthen pots. Random amplified polymorphic DNA (RAPD)-based DNA fingerprinting profiles were generated for the first time using shoot tip explants of this species and confirmed that there was no genetic variability. This protocol might be helpful for the mass multiplication of P. santalinus in the future. Keywords Apical meristem Á RAPD fingerprinting Á MS medium Á Multiple shoots Á Pterocarpus santalinus Abbreviations BAP 6-benzylaminopurine GA 3 Gibberellic acid IAA Indole-3-acetic acid IBA Indole-3-butyric acid KN Kinetin MS Murashige and Skoog NAA a-Naphthalene acetic acid TDZ Thidiazuron Communicated by E. Lojkowska.
Tissue Culture, In Vitro Organogenesis and Regeneration of
Introduction: Plantago lanceolata is one the most important species of Plantago genus and has valuable medicinal secondary metabolites. Materials and Methods: The effect of different factors on germination of P. lanceolata seeds was studied and leaf and root explants of in vitro growth seedling were cultured on Murashige and Skoog (MS) medium supplemented with combination of 6-benzylaminopurine (BAP) or thidiazuron (TDZ) (0, 0.5, 1, 1.5, 2 and 3 mg/L) and auxins: α-naphthaleneacetic acid (NAA) (0, 0.2, 0.5, 1 mg/L), indole-3-acetic acid (IAA) (0.1, 0.5, 1, 1.5 mg/L) or indole-3-butyric acid (0.5, 1, 1.5, 2 mg/L) (IBA) at different concentrations. Results: The results showed that cold pre-treatment, daylight and 1/8 MS salt concentration are more suitable for high germination. The best shoot organogenesis rate (95%) in leaf explants was observed in 1:1 mg/L, and 1.5:1 mg/L TDZ: IBA. The highest percentage of shoot organogenesis (100%) was observed in most of the plant growth regulator (PGR) treatments in root explants. About 58.67 and 60 shoot numbers obtained with 2 mg/L TDZ in leaf and 1:1.5 mg/L BAP: IBA in root explants, respectively. Conclusions: It can be suggested that the best shoot organogenesis and proliferation medium is MS basal medium containing cytokinin TDZ and auxin IBA in comparison with other hormone compounds on leaf and root explants. The result behind this fact is high callus induction and regeneration potential of explants in all concentrations.
Adventitious shoot regeneration and rooting of Prunus serotina in vitro cultures
2006
A complete regeneration protocol was developed for Prunus serotina Ehrh., an important hardwood species for timber and sawlog production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 µM 6-benzylaminopurine (BA), 0.49 µM indole-3-butyric acid (IBA), and 0.29 µM gibberellic acid (GA 3 ). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 µM thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 µM naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 µM BA in combination with 0, 0.54, 1.07, or 5.37 µM NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a signifi cant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 µM TDZ plus 0.54 µM NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 µM TDZ plus 1.07 µM NAA. The highest rooting (27%) of adventitious shoots and number of roots per shoot (2.3 ± 0.
Shoot Proliferation and Multiplication from Nodes of Andrographis Paniculata
International Research Journal of Pharmacy, 2015
Advanced biotechnological researches and exploitation of pharmacological potential of Andrographis paniculata Nees (Acanthaceae) require spontaneous and reliable in vitro propagation protocol. The purpose of the present study was to develop a reliable methodology for the high frequency direct shoot regeneration from nodal explants. Efforts were made for the in vitro shoot regeneration by collecting explants by two approaches: in vitro raised seeds and mother plants from greenhouse. The regeneration was carried out on the Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP), kinetin (Kn) and 2isopentenyl adenine (2iP) at concentrations of 0.5-2.5 mg l-1 and BAP (2.0 mg l-1) in combination with other cytokinins like Kn (0.5-2.0 mg l-1) and auxins like 1-naphthalene acetic acid (NAA) and indole-3-acetic acid (IAA) at concentrations of 0.1-0.5 mg l-1 by using nodal explants. Among all the combinations tried, MS medium supplemented with BAP (2.0 mg l-1) and NAA (0.5 mg l-1) gave the maximum of 28.0 of shoots per explant. For the induction of roots, regenerated shoots were cultured on MS medium supplemented with NAA, IAA and indole-3-butyric acid (IBA) at concentrations of 0.5-3.0 mg l-1. 100% rooting was observed on transferring the cultures to full strength MS medium supplemented with IBA (3.0 mg l-1).