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ABSTRACT: An efficient, simple micropropagation method was developed for Alocasia amazonica using corms in semisolid and liquid cultures. Explants were cultured onto Murashige and Skoog (MS) medium (Murashige and
Skoog, Physiol. Plant. 15:473–497, 1962) supplemented with different cytokinins (Benzyladenine [BA, 2.22–13.32μM], kinetin [2.32–13.95μM], Thidiazuron [TDZ, 0.45–4.54μM])
and cytokinin in combination with auxins [naphthalene acetic acid (NAA, 0.54–5.37μM)/indole acetic acid (IAA, 0.57–5.71μM)/indole
butyric acid (IBA, 0.49–4.9μM)]. All supplementary-induced shoot proliferation and the optimal results was on the medium
supplemented with 2.27μM TDZ, which induced 5.1 shoots per explant. Among the different concentrations of sucrose (0–120g
l−1) tested for shoot proliferation, 30g l−1 was found suitable for corm cultures of Alocasia amazonica. The optimal shoot proliferation and biomass values were with the plantlets grown at 30μmol m−2s−1 photosynthetic photon flux (PPF) and 25°C. Liquid cultures found suitable for shoot proliferation and biomass accumulation
was compared to semisolid cultures. Comparative studies of bioreactor systems [continuous immersion (with or without net)
and temporary immersion in liquid media using ebb and flood] revealed that shoot multiplication and growth were greatest with
the raft bioreactor system. Plantlets (cormlets) from the bioreactor were hydroponically cultured for 30days, and 100% of
plants were acclimatized successfully. The simple efficient method of production of plantlets (cormlets) is useful for large-scale
multiplication of this important ornamental plant.
In Vitro Cellular & Developmental Biology - Plant 04/2012; 44(1):26-32. · 1.50 Impact Factor
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ABSTRACT: The present study investigated the dynamics of nutrient utilization and various growth and physiological parameters during
in vitro proliferation of apple root stock ‘M9 EMLA’ in two different bioreactor systems, i.e. temporary and continuous immersions.
Individual shoots obtained from temporary immersion system had higher dry mass and were of better quality than those obtained
from continuous immersion. In continuous immersion bioreactor, apple shoots appeared to utilize more nutrients from liquid
culture medium than that from temporary immersion. The shoot growth was limited by the availability of phosphate and nitrogen
in continuous immersion system. The shoots produced in temporary immersion bioreactor showed higher photosynthetic rate, maximum
quantum yield of photosystem-II and slow but steady rate of nutrient absorption, indicating the occurrence of higher photomixotrophic
metabolism. The study also showed that high level of antioxidant scavenging enzymes in shoots grown in continuous immersion
system induced physiological changes to foster adaptation to stresses.
Plant Growth Regulation 04/2012; 51(1):11-19. · 1.60 Impact Factor
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ABSTRACT: The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and
combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot
regeneration rate (92 %) were obtained when 2 mg dm−3 BAP was combined with 0.5 mg dm−3 IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different
plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas
roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides
than greenhouse grown plants.
Additional key wordsDPPH-flavonoid-phenolics-polysaccharides-shoot organogenesis-withaferine
Biologia Plantarum 04/2012; 54(2):357-360. · 1.97 Impact Factor
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ABSTRACT: Compared to non-embryogenic callus, proembryonic mass, globular, and heart-shaped embryos of Eleutherococcus senticosus had higher levels of endogenous reduced glutathione (GSH). GSH content declined during the course of the embryo development
(torpedo and cotyledon). Similarly, glutathione reductase that is involved in the recycling of GSH providing a constant intracellular
level of GSH was also higher in globular and heart-shaped embryos. The transient increase in GSH contents also correlated
with the changes in measured γ-glutamylcysteine synthetase activity over the same period. The endogenous levels of oxidized
glutathione showed similar trend during development of the somatic embryos, whereas it declined in maturing somatic embryos.
A pronounced increase in glutathione-S-transferase, glutathione peroxidase, catalase, and guaiacol peroxidase activity was
observed during somatic embryo maturation. Ascorbate-glutathione cycle enzymes (ascorbate peroxidase; dehydroascorbate reductase
and monodehydroascorbate reductase) activities also induced indicated that antioxidant enzymes played an important role during
embryo development. These results suggested that the coordinated up-regulations of the antioxidant enzymes and glutathione
redox system provide protection during somatic embryo development in E. senticosus. Antioxidant responses through alterations of the glutathione redox systems, have been described in the present studies have
a significant role in somatic embryo development.
Plant Cell Tissue and Organ Culture 04/2012; 89(2):121-129. · 3.09 Impact Factor
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ABSTRACT: The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot regeneration rate (92 %) were obtained when 2 mg dm(-3) BAP was combined with 0.5 mg dm(-3) IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides than greenhouse grown plants
Biologia Plantarum. 06/2010; 54(2).
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ABSTRACT: The present study relates to different polyunsaturated fatty acids (PUFAs) which were used as elicitors to enhance biomass accumulation and ginsenoside production in Panax ginseng. Adventitious root cultures of ginseng were elicited with oleic and linolenic acid at 0, 1, 5, 10 or 50 mu mol/l concentrations respectively. Elicitors were added to the medium of adventitious roots on the 40th day of culture and roots were harvested on day 47. Cultures supplemented with oleic acid decreased root biomass and ginsenoside accumulation. Cultures supplemented with 1 mu mol/l linolenic acid enhanced ginsenoside accumulation, without the decrease of adventitious root biomass. Linolenic acid enhanced the biosynthesis of both protopanxadiols (2.95 +/- 0.048 mg/g DW) and protopanxadiols (5.66 +/- 0.043 mg/g DW) compared to that of control at (1.41 +/- 0.002 mg/g DW) and (1.58 +/- 0.006 mg/g DW) respectively. No changes in polysaccharides and phenolics content have been noticed upon elicitation with PUFAs. This is the first report on linolenic acid as an elicitor for ginsenoside accumulation in ginseng adventitious root cultures. (C) 2009 SAAB. Published by Elsevier B.V. All rights reserved
South African Journal of Botany. 04/2010; 76(2).
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ABSTRACT: The effect of the root-inoculum size and axuin concentration on growth of adventitious roots and accumulation of ginsenosides
were studied during suspension cultures of ginseng (Panax ginseng C.A. Meyer). Of the various concentrations of indole-3-butyric acid (IBA) and γ-naphthaleneacetic acid (NAA) used as supplementary
growth regulators along with Murashige and Skoog medium, 25μM IBA was found suitable for lateral root induction and growth,
as well as accumulation of ginsenosides. Inoculum size of 5gL−1 was found suitable for optimal biomass (10.5gL−1 dry biomass) and ginsenosides (5.4mgg−1 DW) accumulation. Of the various length of root inocula tested (chopped to 1–3, 4–6, 7–10mm and un-chopped), root inocula
of 7–10mm was found suitable for biomass and ginsenoside accumulation.
Acta Physiologiae Plantarum 12/2008; 31(1):219-222. · 1.64 Impact Factor
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ABSTRACT: The aim of this study was to investigate the effect of CO2 at various concentrations (1, 2.5 and 5%) on antioxidant enzymes and ginsenoside accumulation in Panax ginseng roots in 51 airlift bioreactors (working volume 41). One and 2.5% CO2 was beneficial for root biomass accumulation, but 5% CO2 decreased the biomass. Ginsenoside concentration decreased with increasing concentration of CO2. No significant difference was observed in the malondialdehyde (MDA) content and lipoxygenase (LOX) activity between respective controls and CO2 treated roots. Antioxidant enzymes such as ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), glutathione reductase (GR), catalase (CAT), guaiacol peroxidase (G-POD) including reduced ascorbate and total glutathione were induced in CO2 exposed roots which emphasized the protective role of antioxidants against CO2 induced stress. Superoxide dismutase activity (SOD) which was induced after 15 days was significantly inhibited after 45 days. Glutathione-S-transferase (GST) and glutathione peroxidase (GPX) activities also increased when the roots were subjected to 1 and 2.5% CO2 compared to the respective controls but not at 5%. A higher reduced ascorbate to oxidized (ASC/DHA) ratio in CO2 treated root indicates the plant's ability to tolerate CO2 stress. These observations suggest that an increase in antioxidant enzymes may affect a defense response to the cellular damage induced by CO2. Probably, this increase could not stop the deleterious effects of CO2 concentration on ginsenoside concentration, but reduced stress severity and thereby allowing root growth to occur. (C) 2007 Elsevier B.V. All rights reserved
Environmental and Experimental Botany. 05/2008; 63(1-3).
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ABSTRACT: The present study reports on the kinetics of nutrient utilization during in vitro flowering of Spathiphyllum in air-lift bioreactor cultures. Levels of electrical conductivity (EC), anions and cations, pH, ethylene, sugar content and photosynthetic enzymes were determined in bioreactor cultures of both flowering (FPs) and non-flowering (NFPs) plantlets over a growth period of 12 weeks. A decrease in ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity with a corresponding increase in phosphoenolpyruvate carboxylase (PEPcase) activity occurred during floral induction of Spathiphyllum in vitro. Sucrose concentration decreased significantly in FPs, while no changes in glucose, fructose and total sugars were observed in both FPs and NFPs up to 8 weeks of culture. There were significant variations in mineral nutrient utilization between FP and NFP cultures. These results provide an insight to the physiological processes involved in inflorescence formation in Spathiphyllum
Plant Growth Regulation. 03/2008; 54(2).
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Environmental and Experimental Botany. 01/2008; 64(3-00988472):279-285.
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ABSTRACT: Hormonal control of flower induction and inflorescence development in vitro was investigated in Spathiphyllum. The effects of gibberellic acid (GA(3)) and sucrose on inflorescence development were studied in plantlets regenerated in tissue culture. GA(3) was mandatory for the shift from the vegetative to the reproductive stage. The effect of sucrose concentration on inflorescence bud development was studied in plantlets cultured in MS medium supplemented with 10 mg l(-1) GA(3). Sucrose concentration at 3 or 6% induced inflorescence development in, respectively, 83-85% of the plantlets. The effect of GA(3) and sucrose on inflorescence differentiation and development were also recorded in liquid culture using air-lift bioreactor. The best response was found in the same medium which was standardized as an optimum for solid culture, but the results were better than solid culture. In order to study the relationship between glutathione (GSH) and flowering, we also measured the oxidized and reduced GSH content in leaves throughout the culture period on 2 weeks interval. The GSH accumulation was more after 4 weeks until 6 weeks in GA(3) treated plantlets. Similarly, glutathione reductase which is involved in the recycling of reduced GSH providing a constant intracellular level of GSH, was also higher in GA(3) treated plantlets. The transient increase in GSH contents also correlated with the changes in measured gamma-glutamylcysteine synthetase (gamma-ECS) activity over the same period. The antioxidant enzyme activity in GA(3) treated plantlets also suggests that the plants suffered increased oxidative stress during the period of GA(3) treatment which subsequently increases GSH synthesis through activation of gamma-ECS and this promotes flowering by increasing endogenous GSH
Plant Cell Tissue and Organ Culture. 09/2007; 90(3).
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ABSTRACT: The present study investigated the dynamics of nutrient utilization and various growth and physiological parameters during in vitro proliferation of apple root stock 'M9 EMLA' in two different bioreactor systems, i.e. temporary and continuous immersions. Individual shoots obtained from temporary immersion system had higher dry mass and were of better quality than those obtained from continuous immersion. In continuous immersion bioreactor, apple shoots appeared to utilize more nutrients from liquid culture medium than that from temporary immersion. The shoot growth was limited by the availability of phosphate and nitrogen in continuous immersion system. The shoots produced in temporary immersion bioreactor showed higher photosynthetic rate, maximum quantum yield of photosystem-II and slow but steady rate of nutrient absorption, indicating the occurrence of higher photomixotrophic metabolism. The study also showed that high level of antioxidant scavenging enzymes in shoots grown in continuous immersion system induced physiological changes to foster adaptation to stresses
Plant Growth Regulation. 01/2007; 51(1).
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ABSTRACT: A large number of micropropagated Euphorbia millii shoots from temporary immersion bioreactor showed thick broad leaves that were translucent, wrinkled and/or curled and brittle, symptoms of hyperhydricity. The environment inside bioreactor normally used in plant micropropagation is characterised by high relative humidity, poor gaseous exchange between the internal atmosphere of the bioreactor and its surrounding environment, and the accumulation of ethylene, conditions that may induce physiological disorders. A comparison of hyperhydric shoots (HS) with normal plants shows marked increase in malondialdehyde (MDA) content in HS plants. MDA, a decomposition product of polyunsaturated fatty acids hydroperoxides, has been utilized very often as a suitable biomarker for lipid peroxidation, which is an effect of oxidative damage. This hypothesis is also confirmed by the higher lipoxygenase (LOX) activity in HS plants. The potential role of antioxidant enzymes in protecting hyperhydric shoots from oxidative injury was examined by analyzing enzyme activities and isozyme profiles of hyperhydric and non-hyperhydric leaves of E. millii. Superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activity were significantly higher in hyperhydric tissue as compared to non-hyperhydric normal leaf tissue. After native polyacrylamide gel electrophoresis (PAGE) analysis, seven SOD isoenzymes were detected and the increase in SOD activity observed in hyperhydric tissue seemed to be mainly due to Mn-SOD and Cu/Zn-SOD. The activity of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) was proportionally increased in HS tissue compared to normal leaves indicating a crucial role in eliminating toxic H2O2 from plant cells. The depletion of GSH and total glutathione in spite of higher GR activities observed in HS tissue indicates that mechanism of antioxidant defense was by enhanced oxidation of GSH to GSSG by DHAR yielding ascorbate (AA). The antioxidant metabolism has been shown to be important in determining the ability of plants to survive in hyperhydric stress and the up regulation of these enzymes would help to reduce the build up of ROS. (c) 2005 Elsevier B.V. All rights reserved
Environmental and Experimental Botany. 12/2006; 58(1-3).
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ABSTRACT: The effects of paclobutrazol, light emitting diodes (LEDs) and sucrose on inflorescence development in vitro were investigated in photoperiodically day-neutral Euphorbia millii. Paclobutrazol, a growth retardant, was mandatory to the shift from the vegetative to reproductive stage. Inflorescence development in the apical bud was observed after 21-30 days of culture on MS media containing paclobutrazol. Different light qualities also influenced in vitro flowering of E. millii. Highest flowering percentage (90 %), number of inflorescences per plantlet, percentage of mature inflorescences and early flowering (days to first bloom) were observed in fluorescent light. Among the LEDs treatments, red light reduced the flowering percentage, whereas blue, red plus far-red and blue plus far-red LEDs stimulated flowering in vitro. Sucrose concentration also influenced the growth and morphogenic patterns of in vitro plantlets. Sucrose at 5% enhanced inflorescence differentiation. The results indicate that inflorescence development of E. millii plantlets in tissue culture can serve as a potential model to study the role of paclobutrazol, LEDs and sucrose in the flowering process of a day-neutral plant that does not require external signals for flower induction. Also the technique described here can be used as an efficient method for production of early, synchronized and profuse flowering of E. millii pot plants
European Journal of Horticultural Science. 12/2006; 71(6).
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ABSTRACT: The effects of carbon dioxide supply within the range of 1–5 % (along with purified air), on cell culture of Panax ginseng were investigated in a balloon type bubble bioreactor containing 4 dm3 of Murashige and Skoog (MS) medium supplemented with 7.0 mg dm−3 indolebutyric acid, 0.5 mg dm−3 kinetin and 30 g dm−3 sucrose. A 1 % CO2 supply was found beneficial for the production of cell mass; however, increasing CO2 concentration to 2.5 and 5 % decreased the biomass accumulation. CO2 enrichment was not beneficial for saponin production and 1, 2.5, and 5 % CO2 supply resulted in decrease in saponin accumulation up to 11.6, 19.5, and 50.6 %, respectively.
Biologia Plantarum 11/2006; 50(4):752-754. · 1.97 Impact Factor
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ABSTRACT: We investigated different concentrations of auxins (IAA, IBA, NAA), the strength of the MS medium, sucrose and ammonium/nitrate contents, initial medium pH, and inoculum size to determine their effects on biomass increase and the accumulation of total phenols and flavonoids in adventitious roots of Echinacea angustifolia. These roots were cultured under darkness in shake flasks for 4 weeks. IBA proved the best auxin for inducing root proliferation. Root growth was inhibited when the initial pH was maintained below 5.0 or above 6.0. Nitrate, rather than ammonium, was more necessary for root growth and phenolics accumulations. Overall, biomass increased and total phenol and flavonoid contents were maximized under the following conditions: half-strength MS medium supplemented with 2 mg L-1 IBA, 5% (w/v) sucrose, 5:25 (mM) ammonium/nitrate ratio, pH adjusted to 6.0 before autoclaving, and an inoculum size of 10 g L-1 FW. These results indicate that the type of in vitro environment strongly affects growth and the accumulation of phenolics from adventitious root cultures of E angustifolia. Such optimization is beneficial to large-scale production of biomass and secondary metabolites in that species
Journal of Plant Biology. 06/2006; 49(3).
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ABSTRACT: A method for the micropropagation of Spathiphyllum cannifolium is presented using shoot tip proliferation onto Murashige and Skoog (MS) medium supplemented with different plant growth regulator concentrations and combinations. The proliferation responses were significantly influenced by. the cytokinin type and concentrations. Supplementation of the medium with benzyladenine (BA; 4.44-13.32 mu M) increased the shoot proliferation rate significantly as compared to other treatments. When cytokinins were used with auxin (indole-3-butyric acid. IBA and naphthalene acetic acid, NAA), the number of shoots per explant increased in comparison with treatments with BA alone. The largest number of shoots, 9.3 per explant, was obtained with 13.32 mu M BA and 4.9 mu M IBA. Different MS medium strengths and Sucrose concentrations were used with the aim to stimulate in vitro shoot proliferation. Full MS medium with 30 g l(-1) sucrose was found to be suitable for shoot tip culture of Spathiphyllum. Comparative studies between gelled medium and bioreactor culture [continuous immersion (with or without net) and temporary immersion in liquid media using ebb and flood] revealed that shoot multiplication and growth were more efficient in continuous immersion (with net) bioreactor with low cytokinin-supplemented media. Plantlets from the bioreactor were cultured hydroponically for 30 d and 100% of plants were rooted and acclimatized successfully. Rapid and efficient multiplication rate in bioreactor and successful transfer to greenhouse makes this protocol Suitable for large-scale multiplication of this important foliage plant
In Vitro Cellular & Developmental Biology-Plant. 05/2006; 42(3).
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ABSTRACT: A method for the micropropagation of Spathiphyllum cannifolium is presented using shoot tip proliferation onto Murashige and Skoog (MS) medium supplemented with different plant growth
regulator concentrations and combinations. The proliferation responses were significantly influenced by the cytokinin type
and concentrations. Supplementation of the medium with benzyladenine (BA; 4.44–13.32 μM) increased the shoot proliferation rate significantly as compared to other treatments. When cytokinins were used with auxin
(indole-3-butyric acid, IBA and naphthalene acetic acid. NAA), the number of shoots per explant increased in comparison with
treatments with BA alone. The largest number of shoots, 9.3 per explant, was obtained with 13.32 μM BA and 4.9 μM IBA. Different MS medium strengths and sucrose concentrations were used with the aim to stimulate in vitro shoot proliferation. Full MS medium with 30 gl−1 sucrose was found to be suitable for shoot tip culture of Spathiphyllum. Comparative studies between gelled medium and bioreactor culture [continuous immersion (with or without net) and temporary
immersion in liquid media using ebb and flood] revealed that shoot multiplication and growth were more efficient in continuous
immersion (with net) bioreactor with low cytokinin-supplemented media. Plantlets from the bioreactor were cultured hydroponically
for 30 d and 100% of plants were rooted and acelimatized successfully. Rapid and efficient multiplication rate in bioreactor
and successful transfer to greenhouse makes this protocol suitable for large-scale multiplication of this important foliage
plant.
In Vitro Cellular & Developmental Biology - Plant 04/2006; 42(3):291-297. · 1.50 Impact Factor
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ABSTRACT: Six months old in vitro-grown Anoectochilus formosanus plantlets were transferred to ex-vitro acclimation under low irradiance, LI [60 µmol(photon) m−2 s−1], intermediate irradiance, II [180 µmol(photon) m−2 s−1], and high irradiance, HI [300 µmol(photon) m−2 s−1] for 30 d. Imposition of II led to a significant increase of chlorophyll (Chl) b content, rates of net photosynthesis (P
N) and transpiration (E), stomatal conductance (g
s), electron transfer rate (ETR), quantum yield of electron transport from water through photosystem 2 (ΦPS2), and activity of ribulose-1,5-bisphosphate carboxylase/ oxygenase (RuBPCO, EC 4.1.1.39). This indicates that Anoectochilus was better acclimated at II compared to LI treatment. On the other hand, HI acclimation led to a significant reduction of
Chl a and b, P
N, E, g
s, photochemical quenching, dark-adapted quantum efficiency of open PS2 centres (Fv/Fm), probability of an absorbed photon reaching an open PS2 reaction centre (Fv′/Fm′), ETR, ΦPS2, and energy efficiency of CO2 fixation (ΦCO2/ΦPS2). This indicates that HI treatment considerably exceeded the photo-protective capacity and Anoectochilus suffered HI induced damage to the photosynthetic apparatus. Imposition of HI significantly increased the contents of antheraxanthin
and zeaxanthin (ZEA), non-photochemical quenching, and conversion of violaxanthin to ZEA. Thus Anoectochilus modifies its system to dissipate excess excitation energy and to protect the photosynthetic machinery.
Photosynthetica 01/2006; 44(3):419-424. · 1.00 Impact Factor
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ABSTRACT: This report describes in vitro shoot induction and plant regeneration from mature nodal explants of Vitex trifolia L. on Murashige and Skoog (MS) medium fortified with benzylaminopurine (BAP), kinetin (KN), thidiazuron (TDZ), adenine (ADE),
and 2-isopentenyladenine (2-iP) (0.25 – 10.0 μM). Multiple shoots differentiated directly without callus mediation within
3 weeks when explants were cultured on medium supplemented with cytokinins. The maximum number of shoots (9 shoots per explant)
was developed on a medium supplemented with 5.0 μM BAP. Shoot cultures was established repeatedly subculturing the original
nodal explant on the same medium. Rooting of shoots was achieved on half strength MS medium supplemented with 0.5 μM naphthaleneacetic
acid (NAA). Rooted plantlets transferred to pots containing autoclaved soil and vermiculite mixture (1:1) showed 90 % survival
when transferred to outdoor.
Biologia Plantarum 01/2006; 50(2):291-294. · 1.97 Impact Factor
