OPTIMIZATION OF BIOMASS CULTURE YIELD AND L-DOPA COMPOUND IN THE CALLUS CULTURE FROM COTYLEDONARY LEAVES OF MUCUNA PRURIENS

ABSTRACTObjective:  The objective of the present study was to evaluate the optimization of callus biomass culture yield and high-performance liquid chromatography (HPLC) analysis of L-DOPA compound in the callus culture from cotyledonary leaves of Mucuna pruriensMethods : M. pruriens seed explants were inoculated onto Murashige and Skoog medium supplemented with different concentrations of 2-isopentenyl adenine (2iP) and Gibberellic acid (GA3)  for germination of plants. The in vitro cotyledonary leaf and hypocotyl explants were cultured on MS basal media containing various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), 1- Naphthaleneacetic acid (NAA), 6-Benzylaminopurine (BA) and 2iP for callus induction. A standard approach of Latin square method was followed for screening of media to establish optimum culturing of callus by manipulating the concentration of auxins (2, 4-D, Indole-3-acetic acid (IAA) and NAA) and cytokinins (BA and 2iP) alone and in combinations. The harvested callus biomass was screened for a major metabolite namely L- Dopa compound using HPLC.Results: Cotyledonary leaf explants showed better callus initiation than hypocotyl explants. Maximum callus induction was observed in Murashige and Skoog (MS) medium containing 4.92µM 2iP. Further screening of callus culture was carried out on MS medium supplemented with different concentrations and combinations of 2, 4-D, NAA,  IAA, (BA)  and 2iP individually and in combinations. Optimum callus biomass of 21.63 g/L dry weight (246.31 g/L fresh weight) was developed on MS media containing 2.26µM 2, 4-D, 2.22µM BA and 4.92µM 2iP. The harvested callus biomass was subjected to extraction and purification of L- Dopa compound.Conclusion: The present study concludes that  HPLC analysis of cell biomass extracts in comparison  with extracts from seeds of mother plants of Mucuna prurienss showed main component of L- Dopa was present in sufficiently large amounts in the undifferentiated cultured cells.Keywords: Mucuna pruriens, Callus biomass, L-Dopa, HPLC analysi

STUDIES ON PHYTOCHEMICAL SCREENING, TANNINS CONTENT AND ANTIBACTERIAL ACTIVITY FROM LEAF AND CALLUS EXTRACTS OF MEMECYLON UMBELLATUM.

Objective: This study aims to determine the phytochemical screening, tannins content, and antibacterial activity from leaf and callus extracts of Memecylon umbellatum.Methods: Preliminary screening involved the qualitative methods to detect the presence of terpenoids, flavonoids, phenols, tannins, steroids, quinones, saponins, cardiac glycosides, and alkaloids. Total tannins contents were quantitatively estimated with tannic acid as standard. Different concentrations of ethanolic leaf and callus extracts were tested using the agar disc diffusion technique for the antibacterial activity against Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli.Results: The phytochemical analysis of leaf and callus extract of M. umbellatum revealed the presence of significant secondary metabolites such as tannins, saponins, quinones, cardiac glycosides, phenols, flavonoids, terpenoids, steroids, and alkaloids. The total tannins content in callus and leaf extract were found to be 11.37 mg tannic acid equivalents (TAE)/g and 7.1 mg TAE/g, respectively. The antibacterial activity of ethanolic leaf and callus extracts of M. umbellatum shown more active against B. subtilis. Both the callus and leaf extract of M. umbellatum was found to be inactiveagainst E. coli.Conclusion: It was concluded that the powerful antibacterial effect is attributed to the greater amount of tannin compounds in the ethanolic callus extracts of M. umbellatum.Â

The Role of Graduality for Referring Expression Generation in Visual Scenes

International audienceReferring Expression Generation (reg) algorithms, a core component of systems that generate text from non-linguistic data, seek to identify domain objects using natural language descriptions. While reg has often been applied to visual domains, very few approaches deal with the problem of fuzziness and gradation. This paper discusses these problems and how they can be accommodated to achieve a more realistic view of the task of referring to objects in visual scenes

IMI : global trends in myopia management attitudes and strategies in clinical practice : 2022 update

PURPOSE. Surveys in 2015 and 2019 identified a high level of eye care practitioner concern/activity about myopia, but the majority still prescribed single vision interventions to young myopes. This research aimed to provide updated information. METHODS. A self-administered, internet-based questionnaire was distributed in 13 languages, through professional bodies to eye care practitioners globally. The questions examined awareness of increasing myopia prevalence, perceived efficacy and adoption of available strategies, and reasons for not adopting specific strategies. RESULTS. Of the 3195 respondents, practitioners’ concern about the increasing frequency of pediatric myopia in their practices differed between continents (P < 0.001), being significantly higher in Asia (9.0 ± 1.5 of 10) than other continents (range 7.7–8.2; P ≤ 0.001). Overall, combination therapy was perceived by practitioners to be the most effective method of myopia control, followed by orthokeratology and pharmaceutical approaches. The least effective perceived methods were single vision distance undercorrection, spectacles and contact lenses, as well as bifocal spectacles. Practitioners rated their activity in myopia control between (6.6 ± 2.9 in South America to 7.9 ± 1.2/2.2 in Australasia and Asia). Single-vision spectacles are still the most prescribed option for progressing young myopia (32.2%), but this has decreased since 2019, and myopia control spectacles (15.2%), myopia control contact lenses (8.7%) and combination therapy (4.0%) are growing in popularity. CONCLUSIONS. More practitioners across the globe are practicing myopia control, but there are still significant differences between and within continents. Practitioners reported that embracing myopia control enhanced patient loyalty, increasing practice revenue and improving job satisfaction

In vitro Plant Regeneration from Nodal Explants of Coleus forskohlii Briq. - An Important Medicinal Plant

An efficient in vitro mass propagation and promising protocol has been successfully standardized and developed for Coleus forskohlii through direct organogenesis from nodal explants. Nodal explants cultured onto MS basal medium supplemented with 4.44 μM BAP recorded the highest response and produced 24.3 ± 0.2 shoots per explant with an average shoot length 5.6 ± 0.4 cm after 30 days of culture. The in vitro shoots recorded higher response for development of rooting on half strength MS fortified with 2.46 μM IBA which produced the best response 7.8 ± 0.6 roots per in vitro shoot with an average root length of 4.3 ± 0.1 cm after 25 days. The in vitro rooted plantlets were transferred for hardening and 90% of the plantlets survived were successfully acclimatized and established in small plastic pots. This protocol recorded to be a highly repeatable, successful and rapid technique that could be utilized for the commercial mass propagation and ex situ conservation of Coleus forskohlii. It is important to note that the morphology of the in vitro plantlets of Coleus forskohlii showed a true-to-type growth habit, both in vitro and when transferred to ex vitro growth conditions.&#x0D; Plant Tissue Cult. &amp; Biotech. 30(1): 143-148, 2020 (June)</jats:p

In vitro Plant Regeneration from Shoot tip Explants of Exacum travancoricum Beedi

Shoot tip explants of Exacum travancoricum, an endangered herb mainly known for ornamental purpose grown on MS with 4.44 μM BAP and 1.34 μM NAA showed better growth response and produced 29.3 ± 0.3 shoots per explant with an average length of 4.6 ± 0.1 cm after 35 days. Roots were induced after transfer to half strength of MS supplemented with 2.46 μM IBA produced 4.8 ± 0.62 roots with an average height of 3.6 ± 0.10 cm after 30 days. The rooted plantlets were transferred for hardening, 80 per cent of plantlets survived and resumed growth in the mixture of soil, vermiculite and farm yard manure (1 : 1 : 1). Key words: Exacum travancoricum; Shoot tip; Plant regeneration D.O.I. 10.3329/ptcb.v20i2.6889 Plant Tissue Cult. &amp; Biotech. 20(2): 113-118, 2010 (December)</jats:p

A Rapid, High Frequency Regeneration of &lt;i&gt;Justicia gendarussa&lt;/i&gt; Burm.f.

An efficient plant regeneration protocol was developed for Justicia gendarussa Burm.f. (Acanthaceae), an important medicinal shrub. Nodal segments grown on Murashige and Skoog (MS) medium containing 1.0 mg/l Benzyl adenine (BAP) with 10 % coconut milk showed better growth response and produced 10.5 ± 0.6 shoots per explant with an average length of 4.4 ± 0.3 cm after 35 days. Rooting of shoots was achieved on growth regulator free half strength MS medium produced 5.3 ± 0.25 cm roots with an average height of 4.8 ± 0.2 cm after 30 days. The rooted plantlets were transferred for hardening, 80 % of plants were successfully established in the field. Key words: Justicia gendarussa; Nodal segments; Regeneration; Coconut milk. DOI: http://dx.doi.org/10.3329/bjsir.v46i2.4388 Bangladesh J. Sci. Ind. Res. 46(2), 201-204, 2011</jats:p

STUDIES ON PHYTOCHEMICAL SCREENING, ANTIOXIDANT ACTIVITY AND EXTRACTION OF ACTIVE COMPOUND (SWERTIAMARIN) FROM LEAF EXTRACT OF ENICOSTEMMA LITTORALE.

Objective: The objective of the study was to evaluate the phytochemical constituents, total phenol, total terpenoid, antioxidant activity and HPLC analysis of swertiamarin compound from the leaf extract of Enicostemma littorale.Methods: Preliminary screening involved the qualitative methods to detect the presence of terpenoids, flavonoids, phenols, tannins, steroids, quinones, saponins, cardiac glycosides and alkaloids. Total phenol and terpenoid contents were quantitatively estimated.  Total phenolic content was estimated by folin-ciocalteau method. In vitro antioxidant activity of petroleum ether, chloroform, acetone, aqueous and ethanol extracts was evaluated by studying 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity using standard procedure. The leaf extract was screened for major metabolite namely swertiamarin compound using HPLC.Results: The phytochemical analysis of leaf extract of E. littorale revealed the presence of significant secondary metabolites such as steroids, quinones, cardiac glycosides, saponins, tannins, phenols, flavonoids, terpenoids and alkaloids. The total phenol and terpenoid content in leaf extract were found to be 16.32 mg GAE /g and 71.0 mg /g respectively. The acetone leaf extract of E. littorale had showed significant radical scavenging activity. The results of High Performance Liquid Chromatography analysis in the leaf extract of E. littorale proved the presence of the active principle namely swertiamarin.Conclusion: It can be concluded that E. littorale leaf extract can be used as a potent source of natural antioxidant and thus could prevent many free radical mediated diseases. The validated HPLC method can be used for a routine quality control analysis. Key words: Enicostemma littorale, antioxidant activity, phenols, terpenoids,                    phytochemical screening.Â

Rapid and Efficient Plant Regeneration from Nodal Explants of Artemisia annua L.

An efficient plant regeneration protocol was developed for Artemisia annua, an aromatic medicinal herb. Nodal explants inoculated on MS supplemented with 4.44 ?M BAP showed better growth response and produced 116.2 ± 0.1 micro-shoots of an average length 1.9 ± 0.3 cm after 35 days culture. The cluster of small shootlets were cultured on shoot elongation medium supplemented with 1.44 ?M GA3 and 10% coconut milk (CM) showed shoot elongation up to 4.6 ± 0.7 cm. Roots were induced after transfer to half strength MS supplemented with 2.46 ?M IBA produced 14.3 ± 0.2 roots with an average height of 4.3 ± 0.53 cm after 30 days. The rooted plantlets were transferred for hardening, 80 per cent of plantlets survived and resumed growth in the mixture of soil, vermiculite and farm yard manure (1 :  1 : 1). DOI: http://dx.doi.org/10.3329/ptcb.v22i1.11257 Plant Tissue Cult. &amp; Biotech. 22(1): 33-39, 2012 (June)</jats:p

Secondary Metabolite Production in Callus Cultures of &lt;i&gt;Stevia rebaudiana&lt;/i&gt; Bertoni

Stevia rebaudiana is and an important non-caloric sweetening herb contains diterpene glycosides need to be explored for its commercialization. The evolving commercial importance of secondary metabolites in recent years has resulted in a great demand in the Pharma industry. Callus cultures were established from nodal and leaf explants. Leaf explants showed better callus initiation than nodal explants. Maximum callus biomass was observed in MS medium supplemented with 2, 4-D 1.0 mg/l. Further screening of callus culture was carried out on Murashige and Skoog (MS) medium with different concentration and combinations of 2, 4-D, NAA, IAA, IBA, BA and KN individually and in combinations. Remarkable callus biomass of 11.6 g/l dry weight (182.3 g/l fresh weight) was observed in MS media containing 0.5 mg/l 2, 4-D, 0.5 mg/l NAA and 1.0 mg/l KN. The harvested cell biomass was subjected to extraction of active principles. In this study, cell biomass extracts were compared with extracts from leaves of mother plants of Stevia rebaudiana. HPLC analysis of these extracts showed that the main components of the active principles namely Stevioside were present in sufficiently large amounts in the undifferentiated cultured cells. Keywords: In vitro culture; Biomass; Stevia rebaudiana; Stevioside DOI: 10.3329/bjsir.v45i3.6532Bangladesh J. Sci. Ind. Res. 45(3), 243-248, 2010</jats:p