Showing papers on "Murashige and Skoog medium published in 2007"
TL;DR: In vitro and in vivo dried leaf extract showed similar antimicrobial activity, and commercial manufacture of active constituents from these improved elite lines of S. rebaudiana would be useful and profitable.
Abstract: A procedure has been outlined for plant regeneration and antimicrobial screening of a medicinal herb, Stevia rebaudiana Bertoni, through in vitro culture of nodal segments with axillary buds. Murashige and Skoog (MS) medium supplemented with 2.0 mg/L N6-benzyl amino purine and 1.13 mg/L indole-3-acetic acid in combination were found to be most effective in inducing bud break and growth, and in initiating multiple shoot proliferation at the rate of 39 microshoots per nodal explant after 30 d of culture. By repeated subculturing a high-frequency multiplication rate was established for production of elite lines of S. rebaudiana.Elongated shoots were transferred to rooting medium. MS medium supplemented with 2.0 mg/L indole-3-butyric acid was found to be best for rooting. In vitro and in vivo grown leaf extracts in different solvent system were screened for potential antimicrobial activity against medically important bacterial and fungal strains by agar well diffusion method. The chloroform and methanol extract exhibited a concentration dependent antibacterial and antifungal inhibition. Both in vitro and in vivo dried leaf extract showed similar antimicrobial activity. Therefore, commercial manufacture of active constituents from these improved elite lines would be useful and profitable.
Key words: Regeneration, antimicrobial, Stevia rebaudiana, medicinal plant
152 citations
TL;DR: The whole process required 12–16 weeks of culture for completion of all steps of plant regeneration, and this protocol of somatic embryogenesis in Jatropha curcas may be an ideal system for future transgenic research.
Abstract: Jatropha curcas L. is one potential source of non-edible biofuel-producing energy crop. Its importance also lies in its medicinal properties. The species is primarily propagated through heterozygous seeds, and thus the seed oil content varies from 4 to 40%. Moreover, due to its perennial nature, seed setting requires 2 to 3 years time. The seed viability and rate of germination are low, and quality seed screening is another laborious task; thus, seed propagation alone cannot provide quality planting material for sustainable use. Somatic embryogenesis, a powerful tool of plant biotechnology for faster and quality plant production has been successfully applied to regenerate plants in Jatropha curcas for the first time. Embryogenic calli were obtained from leaf explants on MS basal medium supplemented with only 9.3 μM Kn. Induction of globular somatic embryos from 58% of the cultures was achieved on MS medium with different concentrations of 2.3–4.6 μM Kn and 0.5–4.9 μM IBA; 2.3 μM Kn and 1.0 μM IBA proved to be the most effective combination for somatic embryo induction in Jatropha curcas. Addition of 13.6 μM adenine sulphate stimulated the process of development of somatic embryos. Mature somatic embryos were converted to plantlets on half strength MS basal medium with 90% survival rate in the field condition. The whole process required 12–16 weeks of culture for completion of all steps of plant regeneration. This protocol of somatic embryogenesis in Jatropha curcas may be an ideal system for future transgenic research.
138 citations
TL;DR: Adventitious roots of Echinacea purpurea were cultured in airlift bioreactors using Murashige and Skoog medium with 2 mg indole butyric acid l−1 and 50 g sucrose l−2 for the production of chichoric acid, chlorogenic acid and caftaric acid.
Abstract: Adventitious roots of Echinacea purpurea were cultured in airlift bioreactors (20 l, 500 l balloon-type, bubble bioreactors and 1,000 l drum-type bubble bioreactor) using Murashige and Skoog (MS) medium with 2 mg indole butyric acid l−1 and 50 g sucrose l−1 for the production of chichoric acid, chlorogenic acid and caftaric acid. In the 20 l bioreactor (containing 14 l MS medium) a maximum yield of 11 g dry biomass l−1 was achieved after 60 days. However, the amount of total phenolics (57 mg g−1 DW), flavonoids (34 mg g−1 DW) and caffeic acid derivatives (38 mg g−1 DW) were highest after 50 days. Based on these studies, pilot-scale cultures were established and 3.6 kg and 5.1 kg dry biomass were achieved in the 500 l and 1,000 l bioreactors, respectively. The accumulation of 5 mg chlorogenic acid g−1 DW, 22 mg chichoric acid g−1 DW and 4 mg caftaric acids g−1 DW were achieved with adventitious roots grown in 1,000 l bioreactors.
99 citations
TL;DR: The described method can be successfully employed for large-scale multiplication and long-term in vitro conservation of T. indica and shows any immediate detectable phenotypic variation.
Abstract: An efficient protocol is described for the rapid in vitro multiplication of an endangered medicinal plant, Tylophora indica (Burm. f.) Merrill, via enhanced axillary bud proliferation from nodal explants collected from young shoots of a two-year-old plant. The physiological effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) thidiazuron (TDZ), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA)], ascorbic acid (AA), different strengths of Murashige and Skoog (MS) medium and various pH levels on in vitro morphogenesis were investigated. The highest number (8.6 ± 0.71) of shoots and the maximum average shoot length (5.2 ± 0.31 cm) were recorded on MS medium supplemented with 2.5 μM BA, 0.5 μM NAA and 100 mg/l AA at pH 5.8. Rooting was best achieved on half-strength MS medium augmented with 0.5 μM IBA. The plantlets regenerated in vitro with well-developed shoot and roots were successfully established in pots containing garden soil and grown in a greenhouse with a 90% survival rate. The regenerated plants did not show any immediate detectable phenotypic variation. The described method can be successfully employed for large-scale multiplication and long-term in vitro conservation of T. indica.
96 citations
01 Jan 2007
TL;DR: For rooting different concentrations of IBA, NAA and IAA 11 were used and highest rooting percentage was recorded on MS medium with 0.1 mg l IAA.
Abstract: Shootlets were regenerated from nodal explants of Stevia rebaudiana Bertoni through axillary shoot proliferation. The induction of multiple shoots from nodal segments was the highest in MS medium supplemented with 1.5 mg l BA + 0.5 mg l Kn. For rooting different concentrations of IBA, NAA and IAA 11 were used and highest rooting percentage (97.66%) was recorded on MS medium with 0.1 mg l IAA. The 1
95 citations
TL;DR: For the first time, the phosphomannose isomerase (PMI, EC 5.3.1.8)/mannose-based “positive” selection system has been used to obtain genetically engineered sugarcane plants and expression of manA in the transgenic plants was evaluated.
Abstract: For the first time, the phosphomannose isomerase (PMI, EC 5.3.1.8)/mannose-based “positive” selection system has been used to obtain genetically engineered sugarcane (Saccharum spp. hybrid var. CP72-2086) plants. Transgenic lines of sugarcane were obtained following biolistic transformation of embryogenic callus with an untranslatable sugarcane mosaic virus (SCMV) strain E coat protein (CP) gene and the Escherichia coli PMI gene manA, as the selectable marker gene. Postbombardment, transgenic callus was selectively proliferated on modified MS medium containing 13.6 μM 2,4-D, 20 g l−1 sucrose and 3 g l−1 mannose. Plant regeneration was obtained on MS basal medium with 2.5 μM TDZ under similar selection conditions, and the regenerants rooted on MS basal medium with 19.7 μM IBA, 20 g l−1 sucrose, and 1.5 g l−1 mannose. An increase in mannose concentration from permissive (1.5 g l−1) to selective (3 g l−1) conditions after 3 weeks improved the overall transformation efficiency by reducing the number of selection escapes. Thirty-four vigorously growing putative transgenic plants were successfully transplanted into the greenhouse. PCR and Southern blot analyses showed that 19 plants were manA-positive and 15 plants were CP-positive, while 13 independent transgenics contained both transgenes. Expression of manA in the transgenic plants was evaluated using a chlorophenol red assay and enzymatic analysis.
92 citations
TL;DR: An efficient protocol was developed for micropropagation of an economically important timber-yielding multipurpose tree, Pterocarpus marsupium Roxb, and mother tissue was repeatedly subcultured on fresh shoot induction medium after each harvest of newly formed shoots to enhance the number of shoots per explant.
Abstract: An efficient protocol was developed for micropropagation of an economically important timber-yielding multipurpose tree, Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes (CNs) derived from 18-d-old axenic seedlings on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.1–10 μM). The highest shoot regeneration frequency (90%) and maximum number (15.2 ± 0.20) of shoots per explant was recorded on MS medium amended with 0.4 μM TDZ. Continuous presence of TDZ inhibited shoot elongation. In the primary medium, TDZ-initiated cultures were transferred to the secondary medium supplemented with another cytokinin, 6-benzyladenine (BA), for shoot growth and elongation. Maximum (90%) shoot elongation with an average shoot length of 5.4 ± 0.06 cm was observed at 5 μM BA. To further enhance the number of shoots per explant, mother tissue was repeatedly subcultured on fresh shoot induction medium after each harvest of newly formed shoots. Thus, by adopting this strategy, an average of 44 shoots per explant could be obtained. About 65% of in vitro regenerated shoots produced a maximum number (4.4 ± 0.2) of roots per shoot by a two-step culture procedure employing pulse treatment and subsequent transfer of treated shoots to a low concentration of 0.2 μM indole-3-butyric acid along with phloroglucinol (3.96 μM). The in vitro-raised plantlets were successfully acclimatized first under culture room conditions, then to greenhouse with 70% survival rate.
91 citations
TL;DR: An efficient, rapid and reproducible plant regeneration protocol was successfully developed for Cassia angustifolia using nodal explants excised from 14-day-old aseptic seedlings using 6-benzyladenine and thidiazuron as supplements to Murashige and Skoog medium.
Abstract: An efficient, rapid and reproducible plant regeneration protocol was successfully developed for Cassia angustifolia using nodal explants excised from 14-day-old aseptic seedlings. Of the two cytokinins, 6-benzyladenine (BA) and thidiazuron (TDZ) evaluated as supplements to Murashige and Skoog (MS) medium, TDZ at an optimal concentration of 5.0 μM was effective in inducing multiple shoots. The highest rate of shoot multiplication was achieved on MS medium supplemented with 5.0 μM TDZ and 1.0 μM indole-3-acetic acid (IAA) at pH 5.8. The regenerated shoots when subcultured on hormone free MS medium considerably increased the rate of shoot multiplication and shoot length by end of fourth subculture passage. Rooting was achieved on the isolated shoots using MS medium with 60 μM indole -3- butyric acid (IBA) and 1% activated charcoal for 1 week and subsequently transferring the shootlets to half strength MS liquid media without IBA and activated charcoal. The in vitro raised plantlets with well-developed shoot and roots were successfully established in earthen pots containing garden soil and grown in greenhouse.
82 citations
TL;DR: Under elevated stress condition (medium supplemented with 250mM NaCl), selected calli derived regenerants (S1 plants) exhibited significantly higher SOD and APX activities over both PC and NC, and could be considered as NaCl-tolerant line showing all positive adaptive features towards the salinity stress.
79 citations
TL;DR: Adopting this procedure, positive transformed plants could be obtained within 2–3 months from mature seeds germination to transplant to soil, and more than 1,000 transgenic plants with several engineered constructs encoding different genes of interest were produced in the lab in the past two years.
Abstract: A high-frequency and simple procedure for Agrobacterium tumefaciens-mediated genetic transformation of the medicinal plant Salvia miltiorrhiza was developed. Leaf discs were pre-cultured on MS medium supplemented with 6.6 μmol l−1 BAP and 0.5 μmol l−1 NAA for one day, then co-cultured with A. tumefaciens strain EHA105 harboring the plasmid pCAMBIA 2301 for three days on the same medium. Regenerated buds were obtained on selection medium (co-culture medium supplemented with 60 mg l−1 kanamycin and 200 mg l−1 cefotaxime) after two cycles’ culture of 10 days each and then transferred to fresh MS medium with 60 mg l−1 kanamycin for rooting. Fifteen days later, the rooted plantlets were obtained and then successfully transplanted to soil. The transgenic nature of the regenerated plants was confirmed by PCR, Southern hybridization analysis and GUS histochemical assay. Averagely, 1.1 independent verified transgenics per explant plated were obtained through this protocol. Adopting this procedure, positive transformed plants could be obtained within 2–3 months from mature seeds germination to transplant to soil, and more than 1,000 transgenic plants with several engineered constructs encoding different genes of interest were produced in our lab in the past two years.
78 citations
TL;DR: A protocol for plant regeneration by somatic embryogenesis was developed in guava cv by using immature zygotic embryo explants and over 90% of rooted shoots survived acclimatization.
TL;DR: It is concluded that WPM supplemented with 1mg/L NAA yields higher phenolic content as well as higher antioxidant activity, and rosmarinic acid (RA) was determined to be the primary compound.
TL;DR: An efficient in vitro propagation protocol for Dendrobium candidum Wall ex Lindl using transverse thin cell layer (tTCL) culture system was established, and it was revealed that the shoots originated from the stem vascular bundles.
Abstract: An efficient in vitro propagation protocol for Dendrobium candidum Wall ex Lindl. using transverse thin cell layer (tTCL) culture system was established. The frequency of shoot regeneration and the number of adventitious buds produced from the regenerated shoots significantly relied on the concentration of plant growth regulators, and the position and orientation of the explant. Murashige and Skoog (MS) medium with half-strength macronutrients and 2% sucrose, supplemented with 1.2 mg l−1 naphthaleneacetic acid (NAA) and 1.2 mg l−1 6-benzyladenine (6-BA), was optimal for shoot regeneration. Upon this medium, the youngest explant inoculated in the upright orientation exhibited a high frequency of shoot regeneration (92%), and the highest number of adventitious buds (an average of 24.5) per explant. Rooting of shoots and adventitious buds was achieved on MS medium with half-strength macronutrients and 2% sucrose with 1.0 mg l−1 NAA and 1.0 mg l−1 indole-3-acetic acid (IAA). Plantlets were transplanted into vermiculite with a 95% survival rate in a greenhouse. Ontogenetic studies revealed that the shoots originated from the stem vascular bundles.
TL;DR: Data suggested the mutants of sweet potato induced with ethylmethanesulphonate were more salt tolerant than control plants.
Abstract: Salt tolerant cultivars of sweet potato (Ipomoea batatas L.) can be obtained from induced mutation. The objective of the present study was to induce mutation for salt tolerance using ethylmethanesulphonate (EMS) in calli of sweet potato, followed by cell line selection and subsequent plant regeneration. Calli initiated from leaf explants were treated with 0.5% EMS for 0, 1, 1.5, 2, 2.5 and 3 h, followed by rinsing with sterile distilled water for four times. Preliminary experiments showed that 200 mM NaCl could be used as selection pressure. Salt tolerant calli were sub-cultured on medium supplemented with 200 mM NaCl for selection of mutant cell lines and this process repeated 5 times (20 days each). The selected calli were transferred onto somatic embryo formation medium, which was Murashige and Skoog (MS) medium supplemented with 4 mg l−1 abscisic acid (ABA), 10 mg l−1 gibberellic acid (GA). After 15 days, somatic embryos were transferred onto MS medium supplemented with 0.05 mg l−1 ABA, 0.2 mg l−1 zeatin (ZT) for regeneration. Plants designated as ML1, ML2 and ML3 were regenerated from the somatic embryos formed by calli treated with 0.5% EMS for 2 and 2.5 h. After propagation, salt tolerance of these mutants was investigated. Data suggested the mutants were more salt tolerant than control plants.
TL;DR: A method was developed to initiate multiple shoots from mature nodal explants of Sida cordifolia Linn and Regenerated shoots were successfully rooted on half-strength MS medium supplemented with 2.0 mg l−1 indole-3-butyric acid and 3% (w/v) sucrose.
Abstract: A method was developed to initiate multiple shoots from mature nodal explants of Sida cordifolia Linn. High frequency of regeneration was achieved on Murashige and Skoog (MS) medium supplemented with 2.0 mg l−1 6-benzylaminopurine, 0.5 mg l−1 α-naphthalene acidic acid, 1.0 mg l−1 adenine sulfate, and 10% (v/v) coconut milk. Multiple shoots were initiated within 21 d and the above media was capable of inducing the formation of more than 20 shoots from each explant. Regenerated shoots were successfully rooted on half-strength MS medium supplemented with 2.0 mg l−1 indole-3-butyric acid and 3% (w/v) sucrose. Rooted plantlets were established in soil. The regenerated plantlets showed no morphological differences from the parent material. This protocol could be useful for germplasm conservation, cultivation, and genetic improvement of S. cordifolia.
TL;DR: The results revealed that genotype is a key factor influencing the in vitro gynogenesis in squash, and about 65% of the regenerants were haploid and 35% were double haploid.
TL;DR: Histological analyses showed that cell proliferation arose from subepidermal cells adjacent to vascular bundles, resulting in primary callus formed by a meristematic zone from which somatic embryos arose, and prompted an efficient regenerative protocol based on somatic embryogenesis.
Abstract: The factors affecting the induction and development of somatic embryos and plantlet acclimatization of peach palm (Bactris gasipaes Kunth) were evaluated to establish an efficient regenerative protocol based on somatic embryogenesis. Mature zygotic embryos were cultured in Murashige and Skoog (MS) medium supplemented with 0–40 μM of picloram (4-amino-3,5,6-trichloropicolinic acid) and 0 or 5 μM of 2-isopentyladenine (6-dimethylaminopurine) (2-iP). After 5 mo. in culture embryogenic callus arose from primary calli. Picloram (10 μM) was effective in inducing embryogenic calli in 9.8% of the explants. The use of 1 μM of AgNO3 enhanced embryogenic competence. Embryogenic calli showed an organized structure, a globular aspect, and were white to yellowish in color. Histological analyses showed that cell proliferation arose from subepidermal cells adjacent to vascular bundles, resulting in primary callus formed by a meristematic zone from which somatic embryos arose. Protein profile analyses revealed two high molecular mass bands in these embryogenic calli, but not in other tissues. Embryogenic calli were transferred to a culture medium containing 40 μM of 2,4-dichlorophenoxyacetic acid, 10 μM of 2-iP, plus 1 g l−1 of glutamine, hydrolyzed 0.5 g l−1 casein, and activated 1.5 g l−1 of charcoal. Morphogenetic responses achieved in this medium were the development of somatic embryos, rooting, and loss of embryogenic capacity. Somatic embryos were converted to plantlets on MS medium plus 24.6 μM of 2-iP and 0.44 μM of naphthalene acetic acid. Plantlets were maintained in MS medium with activated charcoal (1.5 g l−1) until they were 6 cm tall, and then acclimatized. After 16 wk, 84.2 ± 6.4% survival was observed.
TL;DR: In vitro flowering system to overcome problems associated with flower growth and development as well as fruit and seed production in vitro is described.
TL;DR: Findings indicate that hairy root cultures of buckwheat culture could be a valuable alternative approach for rutin production, and supplementation with auxins increased the growth rate, but had no significant effect on r Rutin production by H8.
Abstract: Buckwheat (Fagopyrum esculentum Moench.) is a potentially important source of rutin, a natural flavonoid with antihyperglycemic, antihypertensive, and antioxidative properties. To examine in vitro production of rutin, we established a hairy root culture of buckwheat by infecting leaf explants with Agrobacterium rhizogenes R1000, and tested the growth conditions and rutin production rates of these cultures. Ten hairy root clones were established; their growth and rutin production rates ranged from 233 to 312 (mg dry wt per 30 mL flask, and 0.8 to 1.2 (mg/g dry wt), respectively. Clone H8, which had high growth and rutin production rates (312 mg dry wt per 30 mL flask and 1.2 mg/g dry wt, respectively), was selected for further experiments. H8 showed maximal growth and rutin content at 30 days in culture in MS medium. Of four tested culture media, half-strength MS medium was found to induce the highest levels of growth (378 mg dry wt per 30 mL flask) and rutin production (1.4 mg/g dry wt) by clone H8. In contrast, supplementation with auxins (0.1-1 mg/l IAA, IBA and NAA) increased the growth rate, but had no significant effect on rutin production by H8. Collectively, these findings indicate that hairy root cultures of buckwheat culture could be a valuable alternative approach for rutin production.
TL;DR: High frequency plantlet regeneration was achieved in cotyledonary nodes of Aegle marmelos, and in vitro responded shoots were transferred to root induction medium consisting of half-strength MS supplemented with auxins IAA, indole-3-butyric acid (IBA), or α-naphthalene acetic acid (NAA).
Abstract: High frequency plantlet regeneration was achieved in cotyledonary nodes of Aegle marmelos Cotyledonary nodes from 1 mo old in vitro grown seedlings of A marmelos were cultured on Murashige and Skoog (MS) medium supplemented with benzyl adenine (BA) (0–88 μM), kinetin (KIN) (0–94 μM), and indole-3-acetic acid (IAA) (0–114 μM) either alone or in combinations The highest regenerative response was observed on medium containing 66 μM BA + 114 μM IAA where approximately 866% of the cultures responded with an average shoot numbers of 4875 per explant in 7-wk time Cultures maintained on KIN-supplemented medium showed very poor response In vitro responded shoots were transferred to root induction medium consisting of half-strength MS supplemented with auxins IAA, indole-3-butyric acid (IBA), or α-naphthalene acetic acid (NAA) Rooting was best in medium supplemented with 147 μM IBA Rooted plantlets were acclimatized and transferred to the field with 80% survival rate
TL;DR: The effect of copper sulphate on differentiation and elongation of shoot buds from cotyledonary explants of Capsicum annuum L. cv X-235 was investigated and the number of shoots formed per explant was increased two fold as compared to those formed on control.
Abstract: The effect of copper sulphate on differentiation and elongation of shoot buds from cotyledonary explants of Capsicum annuum L. cv X-235 was investigated. Shoot buds were induced on medium supplemented with 22.2 μM BAP and 14.7 μM PAA. Elongation of shoot buds was obtained on MS medium containing 13.3 μM BAP + 0.58 μM GA3. Both shoot induction and elongation media were supplemented with different levels of CuSO4 (0–5 μM). The levels of CuSO4 in the induction as well as elongation medium highly influenced the shoot bud formation and their subsequent elongation. Highest number of shoot buds per explant was obtained when the concentration of CuSO4 was increased 30 times to the normal MS level. Shoot buds formation frequency i.e., the number of shoots formed per explant was increased two fold as compared to those formed on control. Elongation both in terms of percentage and length of shoots was better than that on control. Healthy elongated shoots were rooted on MS medium supplemented with 5.7 μM IAA. Rooted plantlets were transferred to field conditions.
TL;DR: A system of multiple shoot regeneration from rice shoot apical meristem and the gus gene was found to be integrated into the genome of the T0 transformant plantlets and increased transient GUS expression in three-day co-cultivated seedlings.
Abstract: Rice (Oryza sativa ssp. indica) is an important economic crop in many countries. Although a variety of conventional methods have been developed to improve this plant, manipulation by genetic engineering is still complicated. We have established a system of multiple shoot regeneration from rice shoot apical meristem. By use of MS medium containing 4 mg L(-1) thidiazuron (TDZ) multiple shoots were successfully developed directly from the meristem without an intervening callus stage. All rice cultivars tested responded well on the medium and regenerated to plantlets that were readily transferred to soil within 5-8 weeks. The tissue culture system was suitable for Agrobacterium-mediated transformation and different factors affecting transformation efficiency were investigated. Agrobacterium strain EHA105 containing the plasmid pCAMBIA1301 was used. The lowest concentration of hygromycin B in combined with either 250 mg L(-1) carbenicillin or 250 mg L(-1) cefotaxime to kill the rice shoot apical meristem was 50 mg L(-1) and carbenicillin was more effective than cefotaxime. Two-hundred micromolar acetosyringone had no effect on the efficiency of transient expression. Sonication of rice shoot apical meristem for 10 s during bacterial immersion increased transient GUS expression in three-day co-cultivated seedlings. The gus gene was found to be integrated into the genome of the T(0) transformant plantlets.
TL;DR: This paper describes an efficient in vitro micropropagation of Artemisia vulgaris using shoot tip and nodal explants and the survival rate of plantlets varied under acclimatization, the first report on plant regeneration via organogenesis of A. vulgaris.
Abstract: This paper describes an efficient in vitro micropropagation of Artemisia vulgaris using shoot tip and nodal explants. Among the various growth regulators tested, MS medium and B5 vitamins supplemented with BA (4.44 μM) and KN (2.32 μM) combination was found to yield a better response than BA (4.44–13.32 μM) or KN (0.46–13.92 μM) alone in the medium. BA and KN combinations produced a maximum of 23.3 shoots per explant with 99.8% shooting frequency. Multiple shoots raised were elongated on MS medium containing 0.44 μM BA and 1.44 μM GA3. Rooting was highest (98.2%) on MS medium containing 8.56 μM IAA. Rooted plantlets were successfully transferred to plastic cups containing autoclaved garden soil, farmyard soil and sand (2:1:1) for hardening. After 65 days, the plantlets were transferred to Botanical Evaluation Garden and maintained. The survival rate of plantlets varied under acclimatization. Plants looked healthy with no visually detectable phenotypic variations. This is the first report on plant regeneration via organogenesis of A. vulgaris.
TL;DR: Dioscorea bulbifera L. containing the pharmaceutically important compound, diosgenin, was regenerated in vitro through nodal segments on supplemented Murashige and Skoog medium through random amplified polymorphic DNA (RAPD) analysis showed genetic fidelity of regenerants.
Abstract: Dioscorea bulbifera L. containing the pharmaceutically important compound, diosgenin, was regenerated in vitro through nodal segments on supplemented Murashige and Skoog medium (MS). Diosgenin was at 12 mg g−1dry wt in 12-week-old plantlets raised on MS with various growth hormones. Random amplified polymorphic DNA (RAPD) analysis showed genetic fidelity of regenerants. Encapsulation of shoot tips in 3% (w/v) calcium alginate for storage and germplasm exchange was achieved.
TL;DR: The performance of micropropagated plants were evaluated on the basis of some physiological and biochemical parameters and compared with the in vivo-grown plants of the same age and the sodium dodecyl sulphate polyacrylamide gel electrophoresis protein profile was same between regenerated and naturally growing shoots.
Abstract: An efficient protocol has been developed for high-frequency shoot regeneration and plant establishment of Clitoria ternatea - a potential medicinal legume. Adventitious shoots were regenerated from young excised root segments of aseptic seedlings on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzyladenine (BA), kinetin, α-naphthalene acetic acid (NAA) or 2,4-dichlorophenoxy acetic acid (2,4-D) either singly or in various combinations. The highest frequency (100%) of shoot regeneration and maximum number (16.4 ± 0.24) of shoots per explant was obtained on MS medium supplemented with 20 μ m BA and 2.0 μ m NAA. Organogenic calli were produced on a medium containing 2,4-D (10 or 20 μ m) and BA (5.0 μ m). The calli were differentiated into multiple shoots on MS medium supplemented with 2.5-10 μ m BA and 2.0 μ m NAA. The microshoots were rooted on half-strength MS medium supplemented with 5.0 μ m indole-3-butyric acid and transplanted successfully in field conditions. After 12 months of transfer to ex vitro conditions, the performance of micropropagated plants were evaluated on the basis of some physiological and biochemical parameters and compared with the in vivo-grown plants of the same age. The sodium dodecyl sulphate polyacrylamide gel electrophoresis protein profile was same between regenerated and naturally growing shoots. Total soluble protein in aerial part as well as in seeds of in vitro-regenerated and in vivo-grown plants was almost the same. The mitotic study showed normal chromosomal movement and numbers (2x = 16).
TL;DR: The effects of sucrose, plant growth regulators, MS (Murashige and Skoog), and ½MS salt media formulations were investigated for the development of shoot cultures, microtuber induction, and plantlet regeneration in Dioscorea nipponica.
Abstract: The effects of sucrose, plant growth regulators, MS (Murashige and Skoog), and ½MS salt media formulations were investigated for the development of shoot cultures, microtuber induction, and plantlet regeneration in Dioscorea nipponica. The cytokinin N-benzyladenine (BA) in the range of 0.5–2.0 mg/l showed strong enhancing effects on microtuber induction only when used in conjunction with the auxin alpha-naphthalene acetic acid (NAA), with the effect that NAA increased from 0.5 to 2.0 mg/l. Murashige and Skoog salt media supplemented with sucrose at 3% (w/v) gave the highest frequencies of shoot induction (86%) when BA was present at 2.0 mg/l and NAA at 1.0 mg/l. Sucrose at 7% (w/v) was the single most significant medium constituent for microtuber growth. The heaviest microtubers were formed on media containing 1.0 mg/l BA and 2.0 mg/l (0.073 g), especially with 7% sucrose (3.46 g). With media containing ½MS, 2% sucrose, and 0.1% (w/v) activated charcoal, the percentage of rooting was maximal when supplemented with 1.0 mg/l BA and 0.5 mg/l NAA for the in vitro produced shoots (95%) and BA and NAA both at 0.5 mg/l for the microtubers (100%). When removed from culture flasks and transferred into sterilized soil in a greenhouse, most of the hardened plantlets survived (over 91% after 1 week), and they were suitable for field planting after 1 month.
Journal Article•
TL;DR: It is suggested that artimisinin production can be enhanced with the manipulation of medium by different hormones and amino acids in the callus cultured on sterile filter paper bridge.
Abstract: Studies were conducted on the effect of amino acids and growth regulators on the production of artemisinin, an antimalarial compound, in the callus of Artemisia absinthium. Callus was initiated on solid MS medium supplemented with BAP and NAA from leaf explant. For the production of artemisinin, the callus was proliferated on sterile filter paper bridge in MS medium supplemented with different concentrations of plant growth regulators and amino acids. Estimation of artemisinin contents showed that leaves contained 223 μg/g artemisinin while no artemisinin was observed in the stem extract. Callus culture initiated from leaf explant on MS medium without any growth regulator, failed to show the presence of artemisinin. The amount of artemisinin in the callus culture was influenced with the addition of different growth regulators and amino acids to the medium; 3.1 μg/g artemisinin was present in the callus cultured on MS medium to which valine (12.5 mg/l) was added. Addition of cystine (12.5 mg/l) to the medium resulted in 2.8 μg/g artemisinin production. The amount of artemisinin in the callus cultures was 3.05μg/g and 1.95 μg/g when BAP (2 mg/l) and NAA (2 mg/l), respectively were present in the medium. Addition of other growth regulators and amino acids resulted in nominal or no artemisinin production. Present study suggest that artimisinin production can be enhanced with the manipulation of medium by different hormones and amino acids in the callus cultured on sterile filter paper bridge.
TL;DR: An inventory of differentially expressed proteins and genes in rice seedling grown on Murashige and Skoog medium with and without jasmonic acid was developed, providing a new insight into the role of JA in Rice seedling growth and development.
Abstract: Two global approaches were applied to develop an inventory of differentially expressed proteins and genes in rice (cv. Nipponbare) seedling grown on Murashige and Skoog medium with and without jasmonic acid (JA). JA significantly reduced the growth of shoot, root, leaf, and leaf sheath depending on JA concentration (1, 2, 5, 10, 25, and 50 μM) as compared with control. Almost 50% growth inhibition of seedling was observed with 5 μM JA. Shoots and roots of seedlings grown on 5 μM JA for 7 days were then used for proteomics and transcriptomics analyses. Two-dimensional gel electrophoresis revealed 66 and 68 differentially expressed protein spots in shoot and root, respectively, compared to their respective controls. Tandem mass spectrometry analysis of these proteins identified 52 (shoot) and 56 (root) nonredundant proteins, belonging to 10 functional categories. Proteins involved in photosynthesis (44%), cellular respiratory (11%), and protein modification and chaperone (11%) were highly represented in sho...
01 Mar 2007
TL;DR: A successful protocol for the in vitro propagation of Maktoom cv.
Abstract: This study aimed to determine the best combinations of plant growth regulators and other conditions in order to achieve organogenesis and multiplication directly from shoot tips of date palm (Phoenix dactylifera L.) var. Maktoom without callus formation so as to avoid any possibility of undesirable genetic variability. Results revealed that MS modified medium supplemented with 2.0 mg/L 2ip, 1.0 mg/L BA, 1.0 mg/L NAA and 1.0 mg/L NOA was the best for bud formation from shoot tip after 16 weeks (6.2 bud per explant). Subculturing the formed buds on liquid agitated multiplication medium supplemented with 4.0 mg/L 2ip, 2 mg/L BA, 1.0 mg/L NAA and 1.0 mg/L NOA gave the optimum average of buds number (12.6 buds). In elongation stage MS medium with 0.5 mg/L GA3 and 0.1 mg/L NAA enhanced plantlet length to 5.3 cm. Optimum rooting percentage 90% was achieved when shoots were transferred to a medium with 1.0 mg/L NAA. The average root number after 8 weeks was 5.4 with 9.0 cm length. Rooted shoots (plantlets) were transplanted in small pots containing a mixture of peatmoss and perlite (2:1) and placed in plastic tunnels or in a greenhouse. The survival percentage was 85% after 3 months when the plants were transferred to bigger pots. These results define a successful protocol for the in vitro propagation of Maktoom cv. date palm.
Journal Article•
TL;DR: Hypocotyl explants were more responsive than cotyledon explants in terms of callus induction and subsequent plant regeneration and Initiation of multiple shoots through organogenesis from the calli was histologically proven.
Abstract: Efficient plant regeneration via organogenesis was established for 2 summer squash (Cucurbita pepo L.) cultivars, viz. Bulum and Rumbo, using hypocotyl and cotyledon derived calli. Seeds were surface sterilized in 0.1% HgCl2 for 5 min, and germinated in vitro in plant growth regulator free MS media. The maximum morphogenic callus induction rate (86%) was observed from a hypocotyl explant by culturing in MS medium supplemented with 2.5 mg l-1 2,4-D. Calli size and fresh weight increased substantially through subculturing. The highest percentage of shoot regeneration (85%) and highest mean number of shoots (6.89) per culture were obtained with 0.5 mg l-1 thidiazuron. Initiation of multiple shoots through organogenesis from the calli was histologically proven. Hypocotyl explants were more responsive than cotyledon explants in terms of callus induction and subsequent plant regeneration. Regenerated shoots were rooted in MS medium supplemented with 1.0 mg l-1 IBA. About 70% of regenerated plantlets survived and showed new branch development under ex vitro conditions.