Successful scale-upof the anticancer benzyl isothiocyanate compound production was achieved from cell suspension cultures of Salvadora persica L. Cell cultures were established from the stem internodal segments derived callus. Elicitation with two biotic elicitors; chitosan and salicylic acid, increased callus biomass and benzyl isothiocyanate accumulation, but chitosan induced the maximum accumulation, which reached about four-fold higher over the control at the concentration of 100 mg/L. Benzyl isothiocyanate production from cell suspension cultures was studied in shake-flask culture and stirred-tank bioreactor with the application of the optimum concentration of chitosan in the medium. Both methods elevate the compound production; however its yield was higherin the stirred-tank bioreactor.It achieved about eight-fold increase in the production of benzyl isothiocyanate after 24 days of incubation. This protocol can act as a roadmap for large-scale production of benzyl isothiocyanate from cell suspension cultures ofSalvadora persica by using bioreactors
Screening Of the Association of Aspergillusfungi – Macerase and Cellulase Enz...IJERA Editor
In nature, the destruction of plant wastes takes place by not a single microorganism, butby complex of microorganisms which belong to different species and genera with complex of enzymes. Using of highly active monocultures to createfungal association which produce multifunctional enzymeswith desired properties is a promising approach to create relevant and effective crop production microorganisms with beneficial properties. Among15 industrial micro my cetes the most active association was from Aspergillusniger 355 and Aspergillusawamori 1-8 which had highmacerase and cellulase enzymes activity.
“Antimicrobial activity of rhizospheric bacteria of Curcuma longa (Turmeric) ...IOSRJPBS
The present study was carried out to explore the production of antibiotics from soil microbes of medicinal Plant Curcuma longa (Turmeric). Soil samples of the Curcuma longa rhizosphere regions were collected from the different region in Akola City, Western Vidharbh region of Maharashtra. The bacterial culture from the soil samples were collected by the serial dilution and spread plate technique. The total 16 culture have been isolated from the soil samples and out of total 16 only 3 have been characterized which are potent isolates. These 3 isolates are active against the selected pathogens, E. coli, P. aeruginosa and S.aureus. These samples were labelled as S1,S2 and S3. The characterized 3 cultures were identified as Bacillus megatherium, Pseudomonas fluroscenes and Globicitella sulfidifacein by using Bergey’s manual of systemic bacteriology. The Rhizospheric bacterial crude extract of Bacillus megatherium, Pseudomonas fluorescence, Globicatella sulfidifaciens were found to be more or less active against almost all tested pathogenic strains. Hence Curcuma longa can be employed as source of natural antimicrobials that can serve as an alternative to conventional medicines.It was concluded that the best activity have been shown by the Curcuma longa rhizospheric isolates (S2) which is of Pseudomonas fluorescens against all three human pathogenic organisms (E.coli , S.aureus , P.aeroginosa ). The activity of rhizospheric isolates was showing best results against S. aureus
DOI:10.21276/ijlssr.2016.2.4.5
ABSTRACT- Small Cardamom (Elettariacardamomum L. Maton) is one of the major spice crops of India, which were
the world’s largest producer and exporter of cardamom till 1980. There has however been a reduction in production,
mainly because of Katte disease, caused by cardamom mosaic virus (CdMV) a potyvirus. Viral diseases can be managed
effectively by early diagnosis using serological methods. In the present investigation, CdMV isolates were sampled from
Mudigere, Karnataka, ultra purified, and electron micro graphed for confirmation. Polyclonal antibodies were raised
against the virus and a direct antigen coating plate Enzyme linked immunosorbent Assay (DAC-ELISA) and Dot-ELISA
(DIBA) standardized to detect the virus in diseased and tissue cultured plants. Early diagnosis in planting material will aid
in using disease free material for better yields and hence increased profit to the farmer. Key-words- Cardamom mosaic virus (CdMV), Electron microscopy, Direct antigen coating Enzyme linked
immunosorbent Assay (DAC-ELISA), Dot-ELISA (DIBA)
Loss due to diseases range from 20 to 30 %, in case of severe infection, total crop may be lost.
Estimated global loss due to insect pests in potential yields of all crops is -14%.
In India losses due to insect pests ranges from 10 to 20 %
Abiotic stresses reduce average yield of crops by upto50% (Bray EA 1997)
Annually about 42% of the crop productivity is lost due to various abiotic stress factors (Oerkeet.al.,1994).
Plant Tissue culture part II by Dr. Preeti VermaPreeti Verma
This presentation is meant only for educational purpose and includes various aspects of Plant Tissue culture in brief, including Media, Requirements, Problems in PTC, Techniques, Basic requirements for PTC LAB, Adavantages and Applications
Screening Of the Association of Aspergillusfungi – Macerase and Cellulase Enz...IJERA Editor
In nature, the destruction of plant wastes takes place by not a single microorganism, butby complex of microorganisms which belong to different species and genera with complex of enzymes. Using of highly active monocultures to createfungal association which produce multifunctional enzymeswith desired properties is a promising approach to create relevant and effective crop production microorganisms with beneficial properties. Among15 industrial micro my cetes the most active association was from Aspergillusniger 355 and Aspergillusawamori 1-8 which had highmacerase and cellulase enzymes activity.
“Antimicrobial activity of rhizospheric bacteria of Curcuma longa (Turmeric) ...IOSRJPBS
The present study was carried out to explore the production of antibiotics from soil microbes of medicinal Plant Curcuma longa (Turmeric). Soil samples of the Curcuma longa rhizosphere regions were collected from the different region in Akola City, Western Vidharbh region of Maharashtra. The bacterial culture from the soil samples were collected by the serial dilution and spread plate technique. The total 16 culture have been isolated from the soil samples and out of total 16 only 3 have been characterized which are potent isolates. These 3 isolates are active against the selected pathogens, E. coli, P. aeruginosa and S.aureus. These samples were labelled as S1,S2 and S3. The characterized 3 cultures were identified as Bacillus megatherium, Pseudomonas fluroscenes and Globicitella sulfidifacein by using Bergey’s manual of systemic bacteriology. The Rhizospheric bacterial crude extract of Bacillus megatherium, Pseudomonas fluorescence, Globicatella sulfidifaciens were found to be more or less active against almost all tested pathogenic strains. Hence Curcuma longa can be employed as source of natural antimicrobials that can serve as an alternative to conventional medicines.It was concluded that the best activity have been shown by the Curcuma longa rhizospheric isolates (S2) which is of Pseudomonas fluorescens against all three human pathogenic organisms (E.coli , S.aureus , P.aeroginosa ). The activity of rhizospheric isolates was showing best results against S. aureus
DOI:10.21276/ijlssr.2016.2.4.5
ABSTRACT- Small Cardamom (Elettariacardamomum L. Maton) is one of the major spice crops of India, which were
the world’s largest producer and exporter of cardamom till 1980. There has however been a reduction in production,
mainly because of Katte disease, caused by cardamom mosaic virus (CdMV) a potyvirus. Viral diseases can be managed
effectively by early diagnosis using serological methods. In the present investigation, CdMV isolates were sampled from
Mudigere, Karnataka, ultra purified, and electron micro graphed for confirmation. Polyclonal antibodies were raised
against the virus and a direct antigen coating plate Enzyme linked immunosorbent Assay (DAC-ELISA) and Dot-ELISA
(DIBA) standardized to detect the virus in diseased and tissue cultured plants. Early diagnosis in planting material will aid
in using disease free material for better yields and hence increased profit to the farmer. Key-words- Cardamom mosaic virus (CdMV), Electron microscopy, Direct antigen coating Enzyme linked
immunosorbent Assay (DAC-ELISA), Dot-ELISA (DIBA)
Loss due to diseases range from 20 to 30 %, in case of severe infection, total crop may be lost.
Estimated global loss due to insect pests in potential yields of all crops is -14%.
In India losses due to insect pests ranges from 10 to 20 %
Abiotic stresses reduce average yield of crops by upto50% (Bray EA 1997)
Annually about 42% of the crop productivity is lost due to various abiotic stress factors (Oerkeet.al.,1994).
Plant Tissue culture part II by Dr. Preeti VermaPreeti Verma
This presentation is meant only for educational purpose and includes various aspects of Plant Tissue culture in brief, including Media, Requirements, Problems in PTC, Techniques, Basic requirements for PTC LAB, Adavantages and Applications
DOI:10.21276/ijlssr.2016.2.4.6
ABSTRACT- Amylases are among the most important enzymes used in various industries. They represent
approximately 30% of the world enzyme production. These are of ubiquitous occurrence and hold the maximum market
share of enzyme sales. These comprise hydrolases, which hydrolyze starch to diverse products as dextrins, and
progressively smaller polymers composed of glucose units. They are highly demanded in various arrays such as food,
pharmaceuticals, textiles, detergents, etc. However, enzymes from mold and bacterial source have dominated applications
in industrial sectors while few species of yeast were studied for the amylases production. This review focuses on the
amylolytic yeasts and their enzymes and we were interested at α- amylase and pullulanase, their distribution,
structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications. Key-words- Amylolytic yeast, α-amylase, Pullulanase, Industrial application
Isolation and Screening of Soil Actinomycetes for Antimicrobial Activity OlakunleAjibola
Antimicrobial resistance in pathogens has greatly increased of late and now pose a serious public health problem globally. New antimicrobials are continuously needed to inhibit the growth of these resistant strains.
The aim of this study was to isolate and screen soil actinomycetes and evaluate their secondary metabolites for antimicrobial activities against selected pathogenic bacteria and fungi.
International Journal of Medicinal Plants and Natural Products Robin Wong
International Journal of Medicinal Plants and Natural Products (IJMPNP)
Watercress, as a Functional Food, with Protective Effects on Human Health
Against Oxidative Stress: A Review...
A Review on the Antimicrobial Activity of Sesuvium Portulacastrumijtsrd
Mangroves are unique group of vascular plants that occur in saline coastal habitats and are known to tolerate extreme environmental conditions. Some mangrove plants are used for a wide range of conditions, including bacterial, fungal, and viral diseases. The rise of antibiotic-resistant microorganisms is one of the severe problems in healthcare systems of the world, and infectious diseases are the second most serious cause of death worldwide. Therefore, new drugs have to be found in order to combat such diseases and it is essential to find new compounds that have antimicrobial properties. Medicinal-plant extracts, known to produce certain bioactive molecules which react with other organisms in the environment, are known to be less toxic to humans and are environmentally friendly due to the less pollutant released during production. Antimicrobial properties of medicinal plants are being increasingly reported worldwide. Mangroves are biochemically unique and produce a wide array of novel natural products and are considered a rich source of steroids, triterpenes, saponins, flavonoids, alkaloids, and tannins. Extracts from the leaves, stems, barks, and roots of mangrove species have shown positive results for antioxidant activity tests. G. Beulah | D. Divya"A Review on the Antimicrobial Activity of Sesuvium Portulacastrum" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-4 , June 2018, URL: http://www.ijtsrd.com/papers/ijtsrd14540.pdf http://www.ijtsrd.com/biological-science/zoology/14540/a-review-on-the-antimicrobial-activity-of-sesuvium-portulacastrum/g-beulah
The chemical compounds produced by plants are collectively referred to as phytochemicals. Biotechnologists have special interest in plant tissue culture for the large scale production of commercially important compounds. These include pharmaceuticals, flavours, fragrances, cosmetics, food additives, feed stocks and antimicrobials.
Most of these products are secondary metabolites— chemical compounds that do not participate in metabolism of plants. Thus, secondary metabolites are not directly needed by plants as they do not perform any physiological function (as is the case with primary metabolites such as amino acids, nucleic acids etc.). Although the native plants are capable of producing the secondary metabolites of commercial interest, tissue culture systems are preferred.
Current approaches toward production ofsecondary plant metabolitesshahnam azizi
In this presentation you can familiar with:
Primary metabolite vs secondary metabolite
Importance and function of secondary metabolite
Approaches for increasing secondary metabolite production in plant tissue culture
Qualitative Phytochemical Screening and In Vitro Assessment of Antioxidant an...YogeshIJTSRD
In the present study, two plant species of the family Asteraceae were selected for an evaluation of their phytochemical screening, antioxidant and anti inflammatory properties. Phytochemical alkaloids, phenols, protein, flavonoids, quinines, tannins, and terpenoids are present in the aqua methanol and aqua acetone extract s of Cirsium arvense L Cronquist and Erigeron bonariensis. The selected plant species exhibit anti inflammatory properties in both solvents. The enzymatic antioxidant property of selected plant species was evaluated by superoxide dismutase SOD , peroxidase POD , and catalase CAT . E. bonariensis shows 20.05±0.02 superoxide dismutase activity which is moreover equal to C. arvense 19.47±0.31 . POD and CAT activities of C. arvense 109.35±0.69 and 41.48±0.13 and E. bonariensis 105.91±1.53 and 39.63±0.035 respectively, the POD activity of C. arvense is slightly higher than E. bonariensis but CAT activity again higher in E. bonariensis same as SOD. Deepti Rawat | P. B. Rao "Qualitative Phytochemical Screening and In-Vitro Assessment of Antioxidant and Anti-Inflammatory Potential of Aqua Methanol and Aqua Acetone Extract of Cirsium Arvense and Erigeron Bonariensis" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd43934.pdf Paper URL: https://www.ijtsrd.com/biological-science/biotechnology/43934/qualitative-phytochemical-screening-and-invitro-assessment-of-antioxidant-and-antiinflammatory-potential-of-aqua-methanol-and-aqua-acetone-extract-of-cirsium-arvense-and-erigeron-bonariensis/deepti-rawat
Using next generation sequencing to describe epiphytic microbiota associated ...Agriculture Journal IJOEAR
Abstract— Its seems likely that agricultural management as diverse as certified organic and conventional IPM practices would impact the microbiota associated with crop surfaces differently. We sampled organic and conventionally managed apples at multiple time-points in a growing season and characterized the bacterial taxa associated with replicates of each treatment type. Surprisingly, no evidence of significant differences persisting across multiple time-points was observed. Significant differential abundance of certain taxa was documented but when it was, it was primarily associated with a single time-point making it difficult to understand if these observations resulted from an environmental or a treatment effect. Principal component analyses demonstrated that sampling time-point explained more of the differences between bacterial communities than treatment. Description of dominant shared bacterial families for both organic and conventional samples included Oxalobacteraceae, Pseudomonadaceae, Sphingomonadaceae and Enterobacteriaceae.
Exogenous application with plant growth promoting rhizobacteria (PGPR) or pro...Agriculture Journal IJOEAR
Abstract— A pot experiment was conducted to investigate the effects of plant growth promoting rhizobacteria (PGPR) like Azotobacter chrocoocum A101, Pseudomonas fluorescens, pseudomonas mendocina Palleroni 1970 and Azospirillum lipoferum N040 or proline on growth traits, photosynthetic pigments, relative water content (RWC), electrolyte leakage percent (EL%), osmoprotectants such as proline and soluble sugars, activities of antioxidant enzymes like peroxidase (POD), polyphenol oxidase (PPO) and catalase (CAT), oil percent and water use efficiency (WUE) of basil plants subjected to water stress. Plants were treated with two regimes of irrigation water, i.e., 100% of evapotranspiration (ETc) (control) and 60% of ETc and PGPR or proline. Growth traits, photosynthetic pigments, RWC, EL %, proline and soluble sugars concentrations, activities of antioxidant enzymes oil percent and water use efficiency (WUE) were significantly altered by water stress and PGPR or proline treatments. Results indicated that PGPR or proline mitigated the water stress and significantly reduced the reduction in growth traits and leaf water content as compared to non-PGPR or proline-treated water-stressed plants. Water-stressed plants treated with PGPR or proline had significant higher photosynthetic pigments, proline and soluble sugars concentrations than water-stressed plants without PGPR or proline treatments. Higher POD, PPO and CAT activities were also observed in water-stressed plants treated by PGPR or proline than water-stressed plants without PGPR or proline treatments. Furthermore, water-stressed plants treated with PGPR or proline treatments had also significant higher oil percent and WUE as compared to water-stressed plants without PGPR or proline treatments. These results are important as the potential of PGPR or proline to alleviate the harmful effects of water stress and offers an opportunity to increase the resistance of basil plants to growth under drought conditions. The protective action of PGPR was more efficient than proline.
DOI:10.21276/ijlssr.2016.2.4.6
ABSTRACT- Amylases are among the most important enzymes used in various industries. They represent
approximately 30% of the world enzyme production. These are of ubiquitous occurrence and hold the maximum market
share of enzyme sales. These comprise hydrolases, which hydrolyze starch to diverse products as dextrins, and
progressively smaller polymers composed of glucose units. They are highly demanded in various arrays such as food,
pharmaceuticals, textiles, detergents, etc. However, enzymes from mold and bacterial source have dominated applications
in industrial sectors while few species of yeast were studied for the amylases production. This review focuses on the
amylolytic yeasts and their enzymes and we were interested at α- amylase and pullulanase, their distribution,
structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications. Key-words- Amylolytic yeast, α-amylase, Pullulanase, Industrial application
Isolation and Screening of Soil Actinomycetes for Antimicrobial Activity OlakunleAjibola
Antimicrobial resistance in pathogens has greatly increased of late and now pose a serious public health problem globally. New antimicrobials are continuously needed to inhibit the growth of these resistant strains.
The aim of this study was to isolate and screen soil actinomycetes and evaluate their secondary metabolites for antimicrobial activities against selected pathogenic bacteria and fungi.
International Journal of Medicinal Plants and Natural Products Robin Wong
International Journal of Medicinal Plants and Natural Products (IJMPNP)
Watercress, as a Functional Food, with Protective Effects on Human Health
Against Oxidative Stress: A Review...
A Review on the Antimicrobial Activity of Sesuvium Portulacastrumijtsrd
Mangroves are unique group of vascular plants that occur in saline coastal habitats and are known to tolerate extreme environmental conditions. Some mangrove plants are used for a wide range of conditions, including bacterial, fungal, and viral diseases. The rise of antibiotic-resistant microorganisms is one of the severe problems in healthcare systems of the world, and infectious diseases are the second most serious cause of death worldwide. Therefore, new drugs have to be found in order to combat such diseases and it is essential to find new compounds that have antimicrobial properties. Medicinal-plant extracts, known to produce certain bioactive molecules which react with other organisms in the environment, are known to be less toxic to humans and are environmentally friendly due to the less pollutant released during production. Antimicrobial properties of medicinal plants are being increasingly reported worldwide. Mangroves are biochemically unique and produce a wide array of novel natural products and are considered a rich source of steroids, triterpenes, saponins, flavonoids, alkaloids, and tannins. Extracts from the leaves, stems, barks, and roots of mangrove species have shown positive results for antioxidant activity tests. G. Beulah | D. Divya"A Review on the Antimicrobial Activity of Sesuvium Portulacastrum" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-4 , June 2018, URL: http://www.ijtsrd.com/papers/ijtsrd14540.pdf http://www.ijtsrd.com/biological-science/zoology/14540/a-review-on-the-antimicrobial-activity-of-sesuvium-portulacastrum/g-beulah
The chemical compounds produced by plants are collectively referred to as phytochemicals. Biotechnologists have special interest in plant tissue culture for the large scale production of commercially important compounds. These include pharmaceuticals, flavours, fragrances, cosmetics, food additives, feed stocks and antimicrobials.
Most of these products are secondary metabolites— chemical compounds that do not participate in metabolism of plants. Thus, secondary metabolites are not directly needed by plants as they do not perform any physiological function (as is the case with primary metabolites such as amino acids, nucleic acids etc.). Although the native plants are capable of producing the secondary metabolites of commercial interest, tissue culture systems are preferred.
Current approaches toward production ofsecondary plant metabolitesshahnam azizi
In this presentation you can familiar with:
Primary metabolite vs secondary metabolite
Importance and function of secondary metabolite
Approaches for increasing secondary metabolite production in plant tissue culture
Qualitative Phytochemical Screening and In Vitro Assessment of Antioxidant an...YogeshIJTSRD
In the present study, two plant species of the family Asteraceae were selected for an evaluation of their phytochemical screening, antioxidant and anti inflammatory properties. Phytochemical alkaloids, phenols, protein, flavonoids, quinines, tannins, and terpenoids are present in the aqua methanol and aqua acetone extract s of Cirsium arvense L Cronquist and Erigeron bonariensis. The selected plant species exhibit anti inflammatory properties in both solvents. The enzymatic antioxidant property of selected plant species was evaluated by superoxide dismutase SOD , peroxidase POD , and catalase CAT . E. bonariensis shows 20.05±0.02 superoxide dismutase activity which is moreover equal to C. arvense 19.47±0.31 . POD and CAT activities of C. arvense 109.35±0.69 and 41.48±0.13 and E. bonariensis 105.91±1.53 and 39.63±0.035 respectively, the POD activity of C. arvense is slightly higher than E. bonariensis but CAT activity again higher in E. bonariensis same as SOD. Deepti Rawat | P. B. Rao "Qualitative Phytochemical Screening and In-Vitro Assessment of Antioxidant and Anti-Inflammatory Potential of Aqua Methanol and Aqua Acetone Extract of Cirsium Arvense and Erigeron Bonariensis" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd43934.pdf Paper URL: https://www.ijtsrd.com/biological-science/biotechnology/43934/qualitative-phytochemical-screening-and-invitro-assessment-of-antioxidant-and-antiinflammatory-potential-of-aqua-methanol-and-aqua-acetone-extract-of-cirsium-arvense-and-erigeron-bonariensis/deepti-rawat
Using next generation sequencing to describe epiphytic microbiota associated ...Agriculture Journal IJOEAR
Abstract— Its seems likely that agricultural management as diverse as certified organic and conventional IPM practices would impact the microbiota associated with crop surfaces differently. We sampled organic and conventionally managed apples at multiple time-points in a growing season and characterized the bacterial taxa associated with replicates of each treatment type. Surprisingly, no evidence of significant differences persisting across multiple time-points was observed. Significant differential abundance of certain taxa was documented but when it was, it was primarily associated with a single time-point making it difficult to understand if these observations resulted from an environmental or a treatment effect. Principal component analyses demonstrated that sampling time-point explained more of the differences between bacterial communities than treatment. Description of dominant shared bacterial families for both organic and conventional samples included Oxalobacteraceae, Pseudomonadaceae, Sphingomonadaceae and Enterobacteriaceae.
Exogenous application with plant growth promoting rhizobacteria (PGPR) or pro...Agriculture Journal IJOEAR
Abstract— A pot experiment was conducted to investigate the effects of plant growth promoting rhizobacteria (PGPR) like Azotobacter chrocoocum A101, Pseudomonas fluorescens, pseudomonas mendocina Palleroni 1970 and Azospirillum lipoferum N040 or proline on growth traits, photosynthetic pigments, relative water content (RWC), electrolyte leakage percent (EL%), osmoprotectants such as proline and soluble sugars, activities of antioxidant enzymes like peroxidase (POD), polyphenol oxidase (PPO) and catalase (CAT), oil percent and water use efficiency (WUE) of basil plants subjected to water stress. Plants were treated with two regimes of irrigation water, i.e., 100% of evapotranspiration (ETc) (control) and 60% of ETc and PGPR or proline. Growth traits, photosynthetic pigments, RWC, EL %, proline and soluble sugars concentrations, activities of antioxidant enzymes oil percent and water use efficiency (WUE) were significantly altered by water stress and PGPR or proline treatments. Results indicated that PGPR or proline mitigated the water stress and significantly reduced the reduction in growth traits and leaf water content as compared to non-PGPR or proline-treated water-stressed plants. Water-stressed plants treated with PGPR or proline had significant higher photosynthetic pigments, proline and soluble sugars concentrations than water-stressed plants without PGPR or proline treatments. Higher POD, PPO and CAT activities were also observed in water-stressed plants treated by PGPR or proline than water-stressed plants without PGPR or proline treatments. Furthermore, water-stressed plants treated with PGPR or proline treatments had also significant higher oil percent and WUE as compared to water-stressed plants without PGPR or proline treatments. These results are important as the potential of PGPR or proline to alleviate the harmful effects of water stress and offers an opportunity to increase the resistance of basil plants to growth under drought conditions. The protective action of PGPR was more efficient than proline.
Proteomic analysis of the interaction between the plant growth promoting fhiz...kys9723331
Plant growth-promoting rhizobacteria (PGPR) facilitate the plant growth and enhance their
induced systemic resistance (ISR) against a variety of environmental stresses. In this study,
we carried out integrative analyses on the proteome, transcriptome, and metabolome to investigate
Arabidopsis root and shoot responses to the well-known PGPR strain Paenibacillus
polymyxa (P. polymyxa) E681. Shoot fresh and root dry weights were increased, whereas root
length was decreased by treatment with P. polymyxa E681. 2DE approach in conjunction
with MALDI-TOF/TOF analysis revealed a total of 41 (17 spots in root, 24 spots in shoot)
that were differentially expressed in response to P. polymyxa E681. Biological process- and
molecular function-based bioinformatics analysis resulted in their classification into seven different
protein groups. Of these, 36 proteins including amino acid metabolism, antioxidant,
defense and stress response, photosynthesis, and plant hormone-related proteins were upregulated,
whereas five proteins including three carbohydrate metabolism- and one amino
acid metabolism-related, and one unknown protein were down-regulated, respectively. A good
correlation was observed between protein and transcript abundances for the 12 differentially
expressed proteins during interactions as determined by qPCR analysis. Metabolite analysis
using LC-MS/MS revealed highly increased levels of tryptophan, indole-3-acetonitrile (IAN),
indole-3-acetic acid (IAA), and camalexin in the treated plants. Arabidopsis plant inoculated
P. polymyxa E681 also showed resistance to Botrytis cinerea infection. Taken together these
results suggest that P. polymyxa E681 may promote plant growth by induced metabolism and
activation of defense-related proteins against fungal pathogen.
Medium Composition Effects on Growth Kinetic of Cordyceps militaris Cells Usi...iosrjce
Cordyceps militaris is one of the well described mushrooms and has been extensively used in
Traditional Chinese Complementary Medicine (TCCM) since many centuries. It has been cultivated naturally or
in submerged culture using different types of media. Fungal mycelia contain adenosine, cordycepin, and
polysaccharides, which are responsible for its biological activities. Cordycepin is the best-known and most
potent mushroom-derived substances possessing anticancer, antitumor and immunomodulating activities. This
mushroom characterized by very low growth rate when grown in solid agar medium for inoculum preparation
during cultivation process. Thus, there was enormous need to increase the growth rate of this type of mushroom
on solid medium to reduce the time of inoculum preparation stage. In this study, optimization of agar cultivation
medium for rapid cell growth using modified potato dextrose agar (PDA) medium culture supplemented with
specific amount of malt extract (ME) together with yeast extract (YE) was investigated. The mycelial growth
diameter was monitored during 21 days of cultivation using two series of experiments of different medium
supplements: 2, 4, 6 and 8 g of ME and 6 g of ME with 0, 2, 4 and 6 g of YE to the PDA medium. The obtained
results clearly demonstrate that the highest mycelial growth diameter of about 7.5 cm was obtained in PDA
medium supplemented with 6 g/L ME and 4 g/L. YE
Effect of plant growth promoting rhizobacterial (PGPR) inoculation on growth ...IJEAB
Plant Growth promoting rhizobacteria are a heterogeneous group of bacteria that can be found in the rhizosphere, at root surfaces and in association with roots. They benefit plants through Production of plant hormones, such as auxins, asymbiotic N2 fixation, solubilization of mineral phosphates, antagonism against phytopathogenic microorganisms by production of antibiotics, siderophroes, Chitinase and other nutrients ability to effectively colonize roots are responsible for plant growth promotion. An experiment was conducted in the field of National Institute of Agronomic Research of Meknes. Morocco. The experiment was a completely randomized design with six replicates. There were four treatments viz. T1: (control; N0 -PGPR), T2: (N0 +2027-2), T3: (N0 +2066-7) and T4: (N0+2025-1). The results indicated that a remarkable increase in root growth, namely length, the diameter of the rod and the total chlorophyll. A total of three different bacteria colonies were isolated and proceed with in vitro screening for plant growth promoting activities; phosphate solubilization, nitrogen fixation, indole acetic acid (IAA), ammonia production and antimicrobial enzymes (cellulose, chitinase and protease) activity. Among the three bacterial strains, all bacterial strains are able to produce ammonia, IAA production and nitrogen fixation activity, one strain phosphate solubilizing activity, two strain are able to produce cellulase syntheses, Protease activity and Chitinase activity.
Mass multiplication procedure for tissue culture and PTC requirementDr. Deepak Sharma
This presentation include basic Micropropagation protocol: Application and advantages of mass multiplication. Beside this the requirement of tissue culture are there (Nutrient, gelling agent, energy source, vitamins and PGRs) are also included.
Tissue culture involves the use of small pieces of plant tissue (explants) which are cultured in a nutrient medium under sterile conditions. Tissue culture is in vitro maintenance and propagation of isolated cells tissues or organs in an appropriate artificial environment.
In vitro regeneration of multiple shoots from encapsulated somatic embryos of...IOSR Journals
A protocol has been established for plant regeneration from encapsulated somatic embryos of Artemisia vulgaris L. Encapsulated somatic embryo of A. vulgaris gave better response at 2% alginate, 75mM calcium chloride hardened for 30 minutes. Morphogenic response and percentage of conversion into plantlets of encapsulated beads on storage and temperature effect were studied. Further, encapsulated somatic embryo retrieved from storage conditions was evaluated for its viability. The encapsulated embryos can be stored upto 4 months at 40C, 200C and 220C. Maximum conversion frequency of 90% was observed from encapsulated somatic embryos cultured on MS medium supplemented with GA3 (1.5mg/l), IAA (0.5mg/l) and Ascorbic acid (40mg/l) and 22oC temperature was found to be optimum irrespective of storage periods. In the present investigation, the development of multiple shoots from a single synthetic seed is a recent record.
ABSTRACT- Biosurfactant is a structurally diverse group of surface-active molecule, synthesized by microorganisms. Kocuria rosea and Pseudomonas aeruginosa strains isolated from pesticide contaminated soil, which produces biosurfactant were studied. Curd whey was used as a cheap source of growth medium for biosurfactant production. There was formation of stable emulsions of biosurfactant containing broth with vegetable oil and kerosene. These strains produced a clear zone in oil spreading test, which is an indicative of the good biosurfactant activity. Both the strains produced extra cellular biosurfactant in the culture media and showed good foam stability in the culture medium. Biosurfactant was efficiently extracted from the culture broth by acetone-HCl precipitation. The biosurfactants from the two species, namely Kocuria rosea and Pseudomonas aeruginosa were found to have no effects on germinating seedlings of Glycine max, Pisum sativum and Spinacia oleracea, when treated with 25%, 50%, 75% and 100% with the combination of curd whey in the making of 100ml volume. Curd whey as a control was taken with no surfactant. Our study suggested an efficient use in surfactant aided bioremediation in agricultural land.
Key-words- Biosurfactant, Kerosene, Emulsification, Oil spreading, Kocuria rosea, Pseudomonas aeruginosa, Glycine max, Pisum sativum, Spinacia oleracea
Bioprocess development for enhanced spore production in shake flask and pilot...iosrjce
Bacillus thuringiensis subsp. Israelensis (Bti), has proven to be a safe and effective larvicide for controlling
mosquito and black fly larvae. The effect of cultivation and bioprocess development on Bti growth and sporulation was
investigated in shake flask level and batch cultivation in the semi-industrial scale 16-L stirred tank bioreactor. For
industrial production of biocontrol microorganism, it is necessary to obtain high cell mass and spore production in a short
time with low cost cultivation media. In this study, the new composition of production media was optimized which composed
of (g L
-1
): glucose, 10; yeast extract, 30; KH2PO4, 5; K2HPO4, 5; MgSO4. 7H20, 0.005; MnSO4.H2O, 0.03; FeSO4, 0.01;
CaCl2.7 H2O, 0.05; NaH2PO4, 1.5; NH4H2PO4, 1.5. The maximal cell dry mass and spore production, Sporemax for shake
flask study were 4.26 gL-1
at 36 h and 3.29106
spore mL-1
, respectively. Furthermore, studies of the cultivation conditions
under controlled and uncontrolled pH in the 16L-bioreactor was performed. The growth of Bti under uncontrolled pH
cultivation showing decreased of glucose and total protein concentration in the media was correlated with the vegetative cell
growth and sporulation. The maximal cell dry mass and Sporemax for uncontrolled pH bioreactor were 4.14 gL-1
at 36h and
3.7106
spore mL-1
, respectively. The maximal cell dry mass and spore production, Sporemax for controlled pH bioreactor
were 3.36 gL-1
at 26 h and 3.23 106
spore mL-1
, respectively. In conclusion, batch cultivation in 16-L bioreactor with the
new optimized production medium under uncontrolled pH condition increased of the cell dry mass and number of spores up to 23 % and 47 % , respectively
Here, all information about Plant Tissue Culture
HISTORY OF PLANT TISSUE CULTURE
THE TECHNIQUE OF PLANT TISSUE CULTURE
Plantlet Regeneration and Transfer to Soil
A Classification of Tissue Culture Techniques
EMBRYO CULTURE
MERISTEM CULTURE
ANTHER OR POLLEN CULTURE
TISSUE AND CELL CULTURES
SOMATIC HYBRIDIZATION
Antimicrobial activity of Lactobacillus spp. especially (L. planetarium and L. acidophilus) against S. aureus were tested using agar-plug, agar well diffusion methods to select the best isolate that could inhibit the growth of multidrug resistance isolates. Further identification for the presence of bacteriocin was done using ELISA kit. Results showed that Lactobacillus spp isolates were bacteriocin producers with different degrees and that L. planetarium (L7) was the most efficient in bacteriocin production. Therefore, L. planetarium (L7) was selected for purification using 70% saturated ammonium sulfate and gel chromatography. The effect of purified bacteriocin was tested on 16 bacterial isolates using micro-titer plate method and well diffusion method. The results showed the ability of the bacteriocin to inhibit bacteria only at concentrations 1866U/ml (50%), 3732U/ml (100%) with a diameter of inhibition zones ranges between (11-23 mm) respectively. The anti-biofilm activity of purified bacteriocin at concentration 100% was investigated and the results showed that biofilm formation was reduced by 100% in the presence of bacteriocin.
Antibacterial and Antifungal Activities of Essential Oils of Crude Extracts o...lukeman Joseph Ade shittu
Concern about the rising prevalence of antibiotics-resistant strains of pathogenic microorganisms has been expressed in the last three decades. However, intensive studies on extracts and biologically-active compounds isolated from medicinal plants have also doubled in the last decade. As a result of paucity of knowledge and folkloric claim on the leaves effectiveness in infectious disease treatments, we aimed to determine the antimicrobial activity of essential oils and lignans present in the crude Sesame radiatum leaves extracts. Ethanolic, Methanolic and Aqueous extracts of Sesame radiatum leaves were studied for their in-vitro antimicrobial activity against both Gram positive and Gram negative micro-organisms and Yeast using Agar diffusion method. The GC-MS phytochemical screening of methanolic extract showed the presence of carboxylic acids and phenolic groups in essential oils especially some of the most potent antioxidants like Sesamol, Sesamolin and Sesamin. Both the methanolic and ethanolic extracts have broad spectrum antimicrobial effect against all the tested micro-organisms except Streptococcus pneumoniae, Candida albicans and Staphylococcus aureus respectively, while the aqueous extract exhibited no inhibitory effect on Staphylococcus aureus and Streptococcus pneumoniae except on Candida albicans. The result confirmed the folkloric claims of the antimicrobial effectiveness of locally consumed Sesame leaves extracts especially against bacterial and common skin infection in many areas of the Country (Nigeria).
Similar to Establishment of suspension cultures of Salvadora persica L. for benzyl isothiocyanate production (20)
Cytogenetic an Experimental Monitoring Test for Plant ExtractsIOSRJPBS
More than two centuries have been passed since the chromosomes have been firstly observed in plant cells by Nageli in 1842. During this long period, chromosomes have been discovered in human cells and well recognized as a source of genes locations. The effects of chemicals and environmental pollution in human health and caners became an interested field of studying diver mutagens and their role in affecting the genetic materials. Cytogenetic tests were the main tools to evaluate the effects of those mutagens on human genome and chromosomes. Many techniques have been used for these purposes including in vitro and in vivo analyzing tests using human and animal cells. The intent of this article is to review the role of cytogenetic techniques in detecting the effects of mutagens on chromosomal aberrations and the role of plant extracts in monitoring these effects
Prevalence of Rota Virus Detection by Reverse TranscriptasePolymerase Chain R...IOSRJPBS
The present study was conducted for the period from 1/6/2016 to 20/1/2017 in Baquba city. The study aimed to detection of rotavirus in stool specimens of children fewer than five age and also explore the effects of certain demographic factors on the detection rates by revers transcriptase- polymerase chain reaction. The study included 49 patients with acute diarrhea, 32 were male and 17 were female. The age range was two months to 5 years. Demographic information on the patients regarding age, sex, residence, type of feeding and source of drinking water were collected from their parents. Stool specimens were collected from each patients and. Detection of rotavirus in stool specimens was done by conventional reverse transcriptase polymerase chain reaction (RT-PCR). The results of present study showed that the overall infection rate by rotavirus among patients with acute diarrhea by RT-PCR tests was 93.88%. The highest infection rate was recorded among those >10-≤15 months of age. None of the results showed significantly difference between female and male, PCR (88% vs 96.87%). Likewise, there was insignificantly difference between urban and rural residence, PCR (95.65% vs 92.30%). The results revealed insignificantly higher infection rate among patients (those below 2 years) feed mixing (91.66%) and bottled (100%) compared to that breast feeding (77.77%) by RT-PCR. The rotavirus infection rate was insignificantly higher among patients consuming municipal water for drinking (97.22%) compared to those consuming bottled water (84.61%) by the RT-PCR. The study concluded that rotavirus was detected in high rates among children less than 5 years old with acute diarrhea in Baquba city, particularly those less than 2 year old.
Antidiabetic and Cytoprotective Effect of Ethanolic Extract of SalaciaNitida ...IOSRJPBS
Objective:Many of the available therapies for diabetes have a number of serious adverse effects; therefore the search for more effective and safer hypoglycaemic agents becomes a paramount. This research was set to investigate the antidiabetic potential and cytoprotective effect of ethanolic root extract of Salacianitida on alloxan induced hyperglyceamic rats. Method:Acute toxicity and phytochemical constituents were evaluated using standardized methods. The study lasted for a period of thirty days comprising of two phases: induction phase and treatment phase. Thirty animals were grouped into five groups of six rats each. Group 1 and 2 serve as normal and toxic control respectively while groups 3, 4, and 5 were treated with 750, 1500mg of S.nitidaand 5mg glibenclamide respectively. The BGL, food and fluid intake was monitored daily while the body weight was measured on weekly. The last day of the study after an overnight fast, the animals were loaded with glucose and the OGTT measured prior to and at 30minutes interval for two hours after which the animals were sacrificed under ether anaesthesia and the organs isolated for histological examination. Results/Conclusion:The results of the present study indicate that S.nitida possessantidiabetic potentials and cytoprotective effect noted in its significant percentage reduction in the glucose level, slight increase in body weight and restoration of the toxic organs to normal.
Anti Bacterial and Anti Oxidant Activities of Evolvulus Alsinoides LinnIOSRJPBS
: Plants are very important sources of potential useful raw materials as natural chemotherapeutic agents. Shankhpushpi is botanically termed as Evolvulus alsinoides; the extracts have exhibited antioxidant, anti-ulcer, and immunomodulatory activities. The present work is to investigate the invitro therapeutic potential activities of methanolic extract of whole plant of Evolvulus alsinoides on the antimicrobial activity against five clinical pathogenic bacterial strains viz., Staphylococcus aureus, Bacillus cereus, Escherichia coli, Klebshiella pneumonia and Salmonella typhii, using agar well diffusion assay and Anti oxidant activity using 1-1-Diphenyl- 2-picryl-hydrazyl (DPPH) radical. Extract inhibited IC50: 40.2. Total phenolic content: 28.4 mg/ml & Total flavonoid content: 20.2 μg /ml. Evolvulus alsinoides (MIC): 512.5mg/ml possess 30 to 40 % inhibition, the antibacterial activity of the herbal extracts was more pronounced on the gram- negative bacteria Klebshiella pneumonia than the gram-positive bacteria Staphylococcus aureus. The free radical scavenging activity of methanolic activity of whole plant of Evolvulus alsinoides increase in a concentration dependent manner, and posses statistically significance DPPH free radical scavenging & Anti microbial activities
A Study on Biomarkers in a Spatially Distributed Type – 2 Diabetes mellitus G...IOSRJPBS
Diabetes, a dreaded disease already ravaged a huge population worldwide, whether it is inherited through generations or invited by the people by their uncontrolled way of life is still obscure. Biomarkers assay is a boon to the diseased to opt proper prophylactic measures to curb or eradicate diseases. The elevated levels of various biomarkers related to inflammation of vital organs, liver function, kidney function and other vital parameters in our diabetic cases reveals the extent of organ(s) damage in these subjects. It is also evident that the levels of various biomarkers are similar in our diabetic subjects, irrespective of their habits or habitats.
Toxicological profile of Grewia bicolor root extractIOSRJPBS
In recent years, traditional system of medicine has become a topic of global importance. Many of the plant species that provide medicinal herbs have been scientifically evaluated for their possible medicinal, pharmacological and toxicological effects. A number of species of the genus Grewia have been used as medicinal herbs to treat several diseases in different parts of the globe. Grewia bicolor is the most famous medicinal plant among the genus Grewia. Despite the wide use of the plant in traditional medicine, so far no scientific evaluation was carried out on this plant for the preclinical toxicity profile. The present study was conducted to evaluate the safety of Grewia bicolor root extract. The plant extract proved that it is relatively safe through the acute and sub-acute studies.
Flora of Kansai Basin at Lalgarh of Paschim Medinipur District in West Bengal...IOSRJPBS
Flora is systematic enumeration of plant species in a given community of a given ecosystem. It reflects the quality as well as the health of the ecosystem because the floral community are the producers which support the growth and development of ecosystem. It reflects the inflow of nutrients and recycling processes through bio-geochemical processes. Indiscriminate use of different chemicals and articles for different purposes produced by different leading factories for the modern development of human societies causes threat to the environment and loosing the biodiversity which significantly destroying the environment rapidly. The main reason is increasing load of human population which damaging the environment and degrading the ecosystem by pollution directly or indirectly. The present paper reflects the 57 floral elements under 29 families and the community structure including eco-degradation process which could be a starting point of extension through community research by a group of workers in future. Hope that interested people will come and take that opportunity to record the problems for future study and assessment about the dwindling ecosystem for our existence at Lalgarh.
Although ATP production is very important for mitochondria, little is known about the actual rate of production and the value of “unit of currency” for mitochondrial energy transactions. “Unit of currency” for all transactions in general is named “quantum” in Physics, and no value of ATP quantum is accurately given in bibliography, other than rough estimates. This work focuses to calculating produced energy unit in mitochondrial metabolism (Quantum for ATP, or QATP for sort) and the rate of ATP production (power of “mitochondrial factory”)
High Prevalence of Vitamin D Deficiency Among Saudi Children And Adolescents ...IOSRJPBS
Background: Vitamin D inadequacy constitutes an increasingly recognized comorbidity among Saudi populations. There were limited data about its prevalence in Saudi children with type 1 diabetes mellitus (T1DM). Aim: To evaluate the prevalence of vitamin D deficiency among Saudi children and adolescents with T1DM in Albaha region, southwestern of Saudi Arabia. Methods: A cross-sectional study was conducted on 117 Saudi children and adolescents with T1DM. These patients were recruited from Albaha diabetic center, Saudi Arabia. The range of age of the participants was 1 –18 years.. All participants were subjected to history taking including, age, gender, duration of diabetes, clinical presentation at the onset of diabetes. Measurement of serum 25 hydroxy vitamin D was done in all participants. Results: There were 117 Saudi children and adolescent with T1DM, 52 (44.4%) were boys, and 65 (55.6%) were girls with a mean age 8.8 ± 3.4 and 9.2 ± 3.9 for boys and girls respectively (p=0.52). The mean serum 25-hydroxy vitamin D levels among them was 17.1± 10.1 ng/ml and the prevalence of vitamin D inadequacy was 88.9% (59.9% of subjects had vitamin D deficiency and 29 % had vitamin D insufficiency) and 11.1% had sufficient levels of 25-hydroxy vitamin D. There was a significant difference in vitamin D level among different groups of vitamin D level (p˂ 0.001). The prevalence of Vitamin D deficiency was greater in the girls with 34.2%, as compared to 25.6% among boys. Vitamin D deficiency was more common among children in the range of age 1-10 years (52.1% vs 36.8%). In vitamin D deficient group, the mean levels of vitamin D were significantly lower in patients presented with DKA as compared to those presented with hyperglycemic symptoms (p= 0.016). Conclusion: Vitamin D deficiency is common in Saudi children and adolescent with T1DM in Albaha region. This is more common in girls and in the younger age groups. Therefore, screening for vitamin D deficiency, creating awareness to increase sunlight exposure, increase the fortification of food products with vitamin D and vitamin D supplementation for these patients should be warranted to prevent future morbidity.
Development of Nimbolide from Seeds of Azadirachta Indica Extract Assisted by...IOSRJPBS
The recent studies on Nimbolide, a robust cervical anticancer agent was identified for the separation of Nimbolide from Azadirachta indica berry collected from Ranga Reddy- Dist, the seeds were initially isolated, dried and transformed into coarse powder. The seed core was defatted and concentrated using methanol in a soxhlet apparatus. The methanolic extract obtained from seeds is subjected to High Performance Liquid Chromatography [HPLC] by direct reverse phase method for isolation of Nimbolide using Molecular Docking of Nimbolide through Molecular Operatin Environment [MOE] 2008 software. This declared that Nimbolide has capability of binding 12.3% to a Glycine 131 undergoing polar interactions. Nimbolide was found to interact with Cyclin E, was redeemed from the Protein Data Bank [PDB code; 1W98]. It was observed that Nimbolide showed an satisfactory increased life span (54.32 and 65.16), decreased the viable cells (52.3±1.5 and 32.5±1.3), and tumor volume (3.2±0.03 and 2.4±0.04).
A Review on Different Benefits of MushroomIOSRJPBS
This paper reviews mushrooms. In this paper, nutritional and medicinal values, other uses of mushrooms were discussed. Mushrooms have been important in human history as food, as medicine, as legands, and in folk lore and religion. Mushrooms are basically consumed for their texture and flavor. They have recently become attractive as health - beneficent food and as sources for the development of drugs. Many higher mushrooms are known to contain a number of biologically active components that show promising antitumor and immunomodulating, cardiovascular, hepatoprotective, hypocholesterolemic, antiviral, antibacterial, antiparasitic and antidiabetic effects. Nutritional value of mushrooms lies between that of meat and vegetables. The rich source of proteins, vitamins and minerals and low in fat content (2-8%) unique chemicals constitution of mushrooms makes them low calorie food 8 and choice diet for those suffering from hypertension, arthrosclerosis, diabetes, obesity. Mushrooms play a role in the development of new biological remediation techniques and filtration technologies (e.g. using fungi to lower bacterial levels in contaminated water). From the review it was observed that developing countries should harness the potentials of mushrooms as this would boost the revenue income and healthy living. It is hoped that this paper would add to existing information on this fungus
Evaluation of plasma Creatine Kinase activity and Inorganic phosphate among S...IOSRJPBS
Background: CK and phosphorus are helpful biomarker for detection of early renalabnormality particularlyCK which has been shown to be abetter predicator of acute renal failure. Objectives: This is a descriptive cross-sectional study.Conducted in Khartoum and White Nile state from March to June 2016.The aim of this study to assess serum creatine kinase (CK) activity and inorganic phosphate concentrations in Sudanese patients with sickle cell anemia. 80 subjects were enrolled in this study they were classified into 40 with sickle cell disease (SCD) as casegroup and 40 healthy apparently as control group. CK activity and phosphorus level was measured by using Spectrophotometry method. Results: The results showed significantly higher Ck activity in patients with SCD when compared with control group with (P = 0.021), and not significant differences in mean of serum phosphorus concentration in case when compared with control with (P = 0.547). Also show serum CK activity in patients used hydroxyurea when compare with the mean of patients not used hydroxyurea it is significantly decrease with (P= 0.006)
A Clinical Study of Amoebic Dysentery and Its Homoeopathic ManagementIOSRJPBS
A Clinical Study of Amoebic Dysentery and Its Homoeopathic Management, Sri Ganganagar, Rajasthan, India. Background: To study the clinical presentation of amoebic dysentery and to study the results of Homeopathic management and treatment of amoebic dysentery. To assess the influence of miasm in the case of Amoebic Dysentery. Methodology: Thirty out-patients with clinically confirmed symptomatic cases of amoebic dysentery were entered trial and treated with Homoeopathic remedies on the basis of Homoeopathic principles with fixed criteria to know the results of treatment as recovered, improved or not improved. For the present study he patients who visited on Sriganganagar Homoeopathic Medical College, Hospital and Research Center, Sri Ganganagar, Rajsthan, India and peripheral Out Patient Department were selected on the inclusion and exclusion criteria. Result: Totally 30 patients were selected for this study. At the end of the study it was observed that, out of 30 patients, 24 patients recovered (80%), 5 case improved (16.66%) and1case is not improved (3.3%). with the help ofsuitable homoeopathic medicines. Conclusion: There were no side effects during the treatment and it can be concluded that Homoeopathic Medicines can be help the patient to take a new lease on life
Biological Efficacy of Quassia Indica (Geratn) Nooteb and Centella Asiatica (...IOSRJPBS
Antimicrobial activity of different extracts of two selected plants, Quassia indica (Geartn) Nooteb and Centella asiatica (L.) Urban were tested against selected strains of bacteria viz; Escherichia coli, Staphylococcus aureus, Pseudomonas sps, Bacillus thuringiensis, Klebsiella sps and Serratia sps using agar disc diffusion technique. Petroleum ether, acetone, methanol and water extracts of seed and leaf of Q. indica and leaf of C. asiatica showed moderate to significant antimicrobial activity. Of the four extracts tested, petroleum ether and water fractions showed more activity against all the bacterial strains. The zone of inhibition of the active fractions for the petroleum ether ranged from 8.57 to 12.63 mm and 8.53 to 14.5mm for the water fraction of Q. indica seed and leaf and C. asiatica leaf; and exhibited comparable results with widely used commercial antimicrobial agents (16 to19 mm for Kanamycin and 14 to 17mm for Cefotaxime). The results prove the efficacy of the most active fractions of the selected plant extracts to be used for developing potent antimicrobial formulations.
Comparative Testing of Antibacterial Activity of Aqueous Extract of Bergenia ...IOSRJPBS
The aim of this research was to formulate a gel comprising of the both extracts of Bergenia ligulata rhizomes and Butea monosperma flowers that would aid in wound healing by exhibiting antibacterial activity at the site of wound infection if any. As per the literature survey conducted it was found that aqueous extract of Bergenia ligulata rhizomes and ethanolic extract of Butea monosperma flowers have good potential of antibacterial activity. Hence this antibacterial activity was studied with the help of agar well-diffusion assay method, against the micro-organisms-S. aureus, MRSA, Pr. vulgaris, and E.coli . Both these extracts were obtained through Soxhlet extraction process and this process was optimized to get maximum yield of extraction. By agar gel well- diffusion assay at the concentration of about 100µg/mL both the extracts exhibited maximum zone of inhibition. This concentration was helpful in deciding the dose for topical gel formulation.
Yield Performances of Pleurotus ostreatus on Different Growth Substrates as I...IOSRJPBS
This study was carried out to investigate the effects of some vegetables (Diplazium sammatii, Moringa oleifera, and Justicia insularis) as additives on the growth parameters of Pleurotus ostreatus cultivated on two substrates; banana leaves (BL) and banana leaves/sawdust (BL/SD). The substrates were amended with these vegetables at different concentration levels (0 g, 5 g, 10 g and 15 g). Spawn running was completed in 30, 35, and 40 days in banana leaves, banana leaves/sawdust and the control respectively. Parameters assessed included area of pileus, number of fruit bodies, fresh weight, dry weight, length of stipe and girth of stipe. Higher mean values for fresh weight (21.69 g), area of pileus (42.58 cm2 ), length of stipe (5.10 g) and girth of stipe (4.30 g) were obtained from BL substrate treated to 5 g M. oleifera leaf powder. These values were significantly different from values obtained for other plant additives. Considering the number of fruit bodiesthat emerged on BL/SD substrates, the control produced more fruit bodies than the substrates with additives. The best performance in BL/SD substrates was observed when the substrate was treated to 10 g J. insularis in some of the parameters assessed. These included fresh weight (20.64 g), number of fruit bodies (8.00) and stipe girth (3.44 cm).Other parameters like area of pileus (36.08 cm2 ) and dry weight (2.39 cm), control had a better performance. While for length of stipe, 15 g M. oleifera leaf powder had the highest mean value that was significantly different (P≤0.05) from what was observed in substrates amended with other additives. It is therefore evident that these vegetables have high potential for utilization as additives or supplements for growing of Pleurotus ostreatus especially when using BL as substrates. In addition, substratesadditive concentration combination played a significant role in selective organ growth of the test mushroom.
Dermatoglification Analysis for Selection and Training of Sports TalentsIOSRJPBS
: The term sports talent can be defined as the individual who, through his inherited and acquired conditions, has a special ability for sports performance, above the average of the general population. Fingerprints are genetic markers and can function as indicators of sporting talents. The objective of the present study was to use dermatoglyphics to identify the genetic-physical profile of soccer athletes. Twenty-four athletes belonging to the Palmeiras Sports Society Athletes' Capture Center participated in this study in the Nordeste sub-category category 16. The dermatoglyphic protocol of Cummins and Midlo (1961) was used. Standard fingerprint indices were calculated: the number of drawings for the 10 fingers and the delta index (D10). We also analyzed the types of digital formulas that indicate the representation in individuals of different types of drawings. Based on the classification of Fernandes (2002), the group presented somatofunctional classification for height, velocity and explosive strength, due to the delta index of 11.54. We conclude that dermatoglyphics can be a parameter to identify the potentiality of an individual and can be incorporated into a policy of selection and formation of sports talent.
Antimicrobial Activity of Bauhinia Purpurea (L) by Minimum Inhibitory Concent...IOSRJPBS
MIC methods are widely used in the comparative testing of new agents, or when a more accurate result is required for clinical management. As there are no CLSI (formerly NCCLS) recommendations for the determination of MICs of varies bio-compounds against gram positive and gram negative organisms. The minimum inhibitory concentration is the lowest concentration (in μg/ml) of an antibiotic that inhibits the growth of a given strain of bacteria. In Diagnostic laboratories these MIC’S are used to confirm resistance and also most often used to determine antimicrobial invitro activity. In the present MIC method, one can get the information about Antibiotic agent, preparation of stock solutions, media and inoculation, conditions of incubation, ultimately reading and interpretation of results. The present study is focused to evaluate antibacterial activities of ethanolic extract and purified fractions of Bauhinia purpurea using a gram-positive and a gram-negative organisms Staphylococcus aureus , Klebsiella, E.coli and enterococcus
Evaluations of Some Biological Properties of Ethanolic Leave Extract of Costu...IOSRJPBS
Costus afer plant has been known to the traditionalist for a very long time. The locals having realized its medicinal potentials have employed it in a variety of ways. This study was designed to investigate the antimicrobial, biochemical and CNS depressant properties of ethanolic leave extract of Costus afer in albino mice and anti-inflammatory activity in albino rats. Ninety albino mice (22-29g) of both sexes were selected and divided into eighteen groups of five mice each. Four groups for analgesic activity, three groups for in-vivo clothing time, four groups for spontaneous locomotor activity, three groups for heamatological evaluation and four groups for phenobarbitone induced hypnosis. Twenty albino rats (180-200g) of both sexes were also selected and divided into four groups of five rats each and used for anti-inflammatory activity. In all, the control groups receive 0.2ml of normal saline while two dose groups of the extract was used 400 and 1200mg/kg representing low and high dose in the treatment group. Indomethacin, Phenobarbitone, Chropromazine, Indomethacine, Ciprofloxacin and Fluconazole were used as reference drug for anlagesic activity, locomotor activity, hypnosis, anti-inflammatory, antibacterial and antifungal test respectively. Qualitative phytochemical analysis of ethanolic leave extract of Costus afer was performed. The result of the qualitative phytochemical analysis revealed the presence of alkaloids, saponins, flavonoids, tannins, phenols, glycosides and terpenoids whereas anthraquinones, essential oil and phlobatannins were absent. The results obtained shows that ethanolic leave extract of Costus afer posses antibacterial activity but no antifungal activity. There was a dose dependent increase in analgesic, anti-inflammatory activities, bleeding/clotting time and CNS activity. The hematological evaluation of the leaves of costusafer shows that the extract has no noticeable effect on the HB, PCV, RBC, but decreases the WBC, platelet count, neutrophils and lymphocytes components of the blood, also the clotting/bleeding time was seen to increase dose dependently in the treated groups. In conclusion, ethanol extract of costus afer ker Gawl leaves possess antimicrobial, analgesic, anti-inflammatory and CNS depressant activity.
Hepatoprotective and Antioxidant Effects of the Flavonoid-rich Fraction of th...IOSRJPBS
The leaves of Jatropha tanjorensis are edible and used in herbal medicine in the treatment of diseases associated with oxidative stress. The present study demonstrates the antioxidative effect of the flavonoid-rich fraction of the methanol extract of Jatropha tanjorensis leaves (FRJT) against CCl4-induced hepatotoxicity in rats. Hepatoprotective and antioxidant properties of FRJT were determined by serum biochemical enzymes; alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), antioxidant enzymes (SOD, CAT and GPx), heamatological pararmeters (PCV, Hb and WBC) and histology study. The results obtained showed a significant reduction (p < 0.05) in the activities of liver marker enzymes across the pre-treated groups compared with the untreated rats. Assay of antioxidant enzymes showed that the extract significantly (p < 0.05) enhanced SOD and GPx activities whereas CAT activity was non-significantly (p ˃ 0.05) increased when compared with the untreated animals. PCV, Hb and WBC levels were significantly (p < 0.05) lower in the untreated group. However, supplementation with FRJT and Silymarin ameliorated the induced depletion of blood in the pre-treated animals. Histological examination of the liver tissue showed marked reduction in fatty degeneration across the pre-treated groups when compared with the untreated group. The results in this study indicate that FRJT exhibited varying levels of protection against CCl4-induced oxidative stress in rat models. These results also indicate that the flavonoid-rich fraction contains antioxidants, which mop up free radicals in the system and support its use in the treatment of diseases resulting from oxidative damage.
Welcome to WIPAC Monthly the magazine brought to you by the LinkedIn Group Water Industry Process Automation & Control.
In this month's edition, along with this month's industry news to celebrate the 13 years since the group was created we have articles including
A case study of the used of Advanced Process Control at the Wastewater Treatment works at Lleida in Spain
A look back on an article on smart wastewater networks in order to see how the industry has measured up in the interim around the adoption of Digital Transformation in the Water Industry.
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Final project report on grocery store management system..pdfKamal Acharya
In today’s fast-changing business environment, it’s extremely important to be able to respond to client needs in the most effective and timely manner. If your customers wish to see your business online and have instant access to your products or services.
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This document will discuss each of the underlying technologies to create and implement an e- commerce website.
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Data file handling has been effectively used in the program.
The automated cosmetic shop management system should deal with the automation of general workflow and administration process of the shop. The main processes of the system focus on customer's request where the system is able to search the most appropriate products and deliver it to the customers. It should help the employees to quickly identify the list of cosmetic product that have reached the minimum quantity and also keep a track of expired date for each cosmetic product. It should help the employees to find the rack number in which the product is placed.It is also Faster and more efficient way.
Overview of the fundamental roles in Hydropower generation and the components involved in wider Electrical Engineering.
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Author: Robbie Edward Sayers
Collaborators and co editors: Charlie Sims and Connor Healey.
(C) 2024 Robbie E. Sayers
Establishment of suspension cultures of Salvadora persica L. for benzyl isothiocyanate production
1. IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS)
e-ISSN:2278-3008, p-ISSN:2319-7676. Volume 12, Issue 1 Ver. II (Jan. - Feb.2017), PP 72-78
www.iosrjournals.org
DOI: 10.9790/3008-1201027278 www.iosrjournals.org 72 | Page
Establishment of suspension cultures of Salvadora persica L. for
benzyl isothiocyanate production
Ghada Abd El-Moneim Hegazi
Tissue Culture Unit, Department of Genetic Resources, Ecology and Dry Land Agriculture Division, Desert
Research Center, 11753 El-Matareya, 1 Mathaf El-Matareya St., Cairo, Egypt
Abstract:Successful scale-upof the anticancer benzyl isothiocyanate compound production was achieved from
cell suspension cultures of Salvadora persica L. Cell cultures were established from the stem internodal
segments derived callus. Elicitation with two biotic elicitors; chitosan and salicylic acid, increased callus
biomass and benzyl isothiocyanate accumulation, but chitosan induced the maximum accumulation, which
reached about four-fold higher over the control at the concentration of 100 mg/L. Benzyl isothiocyanate
production from cell suspension cultures was studied in shake-flask culture and stirred-tank bioreactor with the
application of the optimum concentration of chitosan in the medium. Both methods elevate the compound
production; however its yield was higherin the stirred-tank bioreactor.It achieved about eight-fold increase in
the production of benzyl isothiocyanate after 24 days of incubation. This protocol can act as a roadmap for
large-scale production of benzyl isothiocyanate from cell suspension cultures ofSalvadora persica by using
bioreactors.
Keywords:plant cell culture, chitosan, salicylic acid, shake-flask culture, stirred-tank bioreactor
I. Introduction
Plants are the primary and natural source of a huge number of secondary metabolites, which are used as
pharmaceuticals, food additives, flavors, fragrances, colors, agrochemicals and biopesticides. The production of
secondary metabolites in the plants is very low (less than 1% dry weight) and depends greatly on the
physiological and developmental stage of the plant[1]
.Plant cell and tissue cultures represent sustainable, eco-
friendly and cost-effective alternative to the extraction of the whole plant material for production of valuable
phytochemical compounds. Among other advantages, plant cell and tissue cultures lead to have a higher rate of
metabolism when compared to field grown plants[2,3]
. Furthermore, in vitrocultures offer uniform secondary
product synthesis under controlled conditions by overcoming the effect of climatic conditions and diseases in
field grown or natural plants[4]
.
Various strategies have been developed to improve the in vitro production of secondary metabolites[5]
.
For the large-scale production of valuable compounds, first of all, the establishment of callus biomass from
selected highly productive plant genotypes is required. The callus has an unlimited growth capacity and can
synthesize the same compounds as the original plant. Cell suspension cultures derived from the callus are first
maintained on a small scale and then at bioreactor level[6]
. Scale-up of plant cells in the bioreactor is the key step
towards commercial production of secondary metabolites. Stirred-tank bioreactors (mechanically agitated) have
been the most versatile bioreactor system for large-scale production of plant cells. They are highly effective for
proper mixing of cell suspensions and the break-up of air bubbles to enhance oxygenation and prevent large cell
aggregates formation[7]
.
Elicitation is one the most effective strategies for improving the in vitro production of secondary
metabolites[8]
.Elicitors can stimulate plant defense, leading to the enhanced synthesis and accumulation of
secondary metabolites[2]
.According to their origin, elicitors are divided into abiotic and biotic elicitors. Abiotic
elicitors are substances of non-biological origin, being inorganic compounds; such as salts or physical
factors[9]
.Biotic elicitors include substances of pathogenic origin (exogenous elicitors) and compounds produced
by plants after the action of the pathogen (endogenous elicitors)[3,10]
.Chitosan and salicylic acid are biotic
elicitors. Chitosan is considered as an exogenous elicitor. It is a polysaccharide and the deacetylated form of
chitin, which is the main component of the cell wall of some fungi and simulate the effects of pathogenic
microorganisms on plants to biosynthesize secondary metabolites[11]
.On the other hand, salicylic acid is an
endogenous elicitor. It is a small molecule and considered as a plant hormone with a vital role in plant defense
regulatory systems to many pathogens[12]
. It stimulates a wide range of defense responses, including the
production of plant secondary metabolites[13]
.
Salvadora persica L. (family Salvadoraceae) is an economically and medicinally important plant
belonging to the flora of Egypt[14]
. It is an evergreen tree or shrub commonly known as Miswak. Miswak is a
natural toothbrush, which has been documented to have a potent antibacterial effect[15]
. The different parts of the
plant contains many active constituents that possess a broad spectrum of biological and pharmacological effects,
2. Establishment of suspension cultures of Salvadora persica L. for benzyl isothiocyanate production
DOI: 10.9790/3008-1201027278 www.iosrjournals.org 73 | Page
such as anti-microbial, anti-viral, anti-diabetic, anti-fungal, anti-cancer, anti-ulcer, anti-plaque, anti-caries, anti-
plasmodial, oral hygienic activities, antidepressant, wound-healing, anti-gingival irritation, hypoglycemia,
antioxidant, anti-fever, anti-ulcerogenic, anti-caries and antiplatelet-agression effect, corrective, deobstruent,
liver tonic, diuretic, analgesic, anthelmintic, astringent, lithontriptic, carminative, diuretic, aphrodisiac, and
stomachic[16, 17, 18]
.
Benzylisothiocyanate (BITC) is the major active constituent of S.persica. It is one of the
isothiocyanates with the strongest anticancer properties in human cancer cells. BITC reduce risk of cancers of
the lung, esophageal, breast[19, 20]
, gastrointestinal, pancreatic and colon[21]
by blocking cancer development.
Also, it inhibits growth and triggers apoptosis of human oral cancer cells with a minimal toxicity to normal
cells[22]
.BITC efficiently inhibits several cancer promoting-cytochrome enzymes, helping to prevent
carcinogenesis[23]
. Moreover, it is the main antibacterial component in S. persica and exhibits rapid and strong
bactericidal effect against oral pathogens involved in periodontal disease[24]
. A previous study was carried out by
Hegazi et al.[25]
on the in vitro production of BITC from S. persica callus cultures generated from leaf and stem
internodal explants; determining the effect of different plant growth regulators and two amino acid precursors;
phenylalanine and cysteine, on the callus growth and BITC accumulation. It proved that the callusing response
from stem internodal explants was much higher than that of the leaf, andphenylalanineincreased the
accumulation of BITC in callus cultures.
The present research work is aimed to scale-up BITC production from S. persica cells in shake-flask
culture and stirred-tank bioreactor under similar nutrient and environmental conditions, after the investigation of
the effect of chitosan and salicylic acid as biotic elicitors on BITC accumulation.
II. Materials and Methods
1. Induction ofcallus culture
Stem internodal segmentsfrom young S.persicaplantsgrown in the Egyptain Botanical Garden (Orman)
were used as explants. Explants were taken from plants originated from seeds collected from naturally grown
plants in Gabal Elba, south Egypt. The stem intermodal segments(2-3 cm long) were initially washed
thoroughlyunder running tap water for 10 minutes and surface sterilized under complete aseptic conditions with
30% (v/v) commercial bleach [containing 5.25% sodium hypochlorite (NaOCl)] for 10 minutes, followed by
dipping in 0.1% (w/v) mercuric chloride (HgCl2) for 45 seconds. Finally, explants were thoroughly rinsed five
times with sterilized distilled water. The cutting ends of the stem internodal segments were trimmed for
culturing.
Callus culture was developed from explantsand maintained on ready-made solid Murashige and Skoog
(MS) basal medium[26]
(Caisson, USA) supplemented with 100 mg/L myo-inositol, 3% w/v sucrose and 0.5
mg/L of each of 2,4-dichlorophenoxy acetic acid (2,4-D) and Kinetin (Kin)(the optimal medium composition of
callus inductionas reported by Hegazi et al.[25]
). Chemicals added to the culture medium were supplied by Sigma
Chemical Co., USA. The pH of the medium was adjusted to 5.7-5.8 and solidified with 0.7% w/v agar. Media
(40-45 ml volumes) were dispensed into glass jars. Jars were sealed with autoclavable polypropylene caps and
autoclaved at 1.1 kg/cm2
for 15 minutes at 121°C.
Cultures were maintained under a 16-hourphotoperiod provided by cool-white fluorescent tubes (F
140t9d/38, Toshiba) at a temperature of 25±2ºC and 70±10 relative humidity.Callus cultures thus raised were
subcultured to fresh medium after four weeks.
2. Elicitation
An amount of 5 g of callus fresh weight/jar was transferred to the same medium composition in
addition to different concentrations of chitosan (100, 200 and 300 mg/L) and salicylic acid (5, 10 and 20 mg/L)
as biotic elicitors, in addition to the control treatment without elicitors. Callus was collected and the fresh and
dry weights were determined. Dry weight was determined after drying the callus at 45ºC in an oven until a
constant weight was obtained. Also, BITC concentration was recorded for each treatment. Data were
documented after four weeks of culture.
3. Establishment of suspension cultures
3.1. Shake-flask culture experiment
Suspension cultures were established from 3 g fresh friable callus in 100 ml Erlenmeyer flasks
containing 30 ml of theliquid optimized MS medium that containing0.5 mg/L of each of 2,4-D and Kin, in
addition to 100 mg/L chitosan. The pH of the medium was adjusted to 5.7-5.8 before autoclaving as mentioned
previously. Flasks were closed with cotton plugs and two layers of aluminum foil and incubated on a rotary
shaker (DAIHAN Scientific, Korea) with speed of 100 rpm at 25±2°Cundera 16-hour photoperiod in the growth
room. Cultured cells were sampled at each growth stage up to the 28th
day of incubation in 7-day intervals and
BITC concentration was determined.
3. Establishment of suspension cultures of Salvadora persica L. for benzyl isothiocyanate production
DOI: 10.9790/3008-1201027278 www.iosrjournals.org 74 | Page
3.2. Bioreactor experiment
A stirred-tank bioreactor (STB) system (Applicon, Dependable Instruments, The Netherlands) was
used in the present study; consisted of 3 L autoclavable cylindrical glass vessel. An amount of 20 g of
S.persicacellswere cultivated into the culture vessel containing 2 L (working volume) ofthe optimized medium
(MS medium supplemented with 0.5 mg/L of each of 2,4-D and Kin, in addition to 100 mg/L chitosan).The
rotation speed was kept constant at 100 rpm throughout the cultivation. The cultures were aerated through a
sintered stainless-steel sparger. The air flow rate was adjusted manually to the required level using an air flow
control system. The temperature and pH in the bioreactor were controlled at 27ºC and 5.7-5.8, respectively, by
bio-controller (ADI 1032, Applicon, Dependable Instruments, The Netherlands). Dissolved oxygen (DO) and
pH were monitored with autoclavable electrodes. Impeller vertically orientated for liquid flows away from the
motor axis was used for agitation.Samples were collected in 7-day intervals up to the 28th
day for the
quantitative analysis of BITC.
4. Analysis of benzyl isothiocyanate content
The determination of BITC was performed by high performance liquid chromatograph (HPLC) as
described by Kiddle et al.[27]
and modified by Rossetto et al.[28]
. The extraction of BITC was carried out from
callus and cells from shake-flask culture and stirred-tank bioreactor experiments according to Kiddle et al.[27]
with minor modifications. Samples were homogenized in a porcelain mortar containing 70: 30 (v:v)
MeOH:water. Extracts were transferred to stoppered Erlenmeyer flasks and conditioned in a thermostatic bath
under constant agitation. The extraction was carried out at 70ºC for 30 minutes. The extracts were filtered on
MillexTM
polyvinylidene fluoride (PVDF) membranes (0.45 μm, Millipore) prior to HPLC injection. Methanolic
extracts of samples were analyzed by HPLC for BITCconcentration using an external standard. HPLC system
(Agilent 1100 series, Waldborn, Germany) was coupledwith UV–Vis detector (G1315B) and G1322A Degasser.
Sample injectionsof 10 µl were made by an Agilent 1100 Series auto-sampler; thechromatographic separations
were performed on ZORBAX-EclipseXDB-C18 column (4.6×250 mm, particle size 5 μm). Column temperature
was maintained at 30ºC and detection wavelength was 254 nm. Optimum efficiencyofseparation was obtained
using methanol (solvent A), and theacitonitrile (solvent B) (HPLC grade).The flow rate was 1 ml/minute. The
run time for each analysis was 60 minutes, and 10 minuteswere required for column cleaning and equilibrium.
The BITC concentration of the injected samples was calculated as µg/g fresh weight of sample.
5. Data analysis
Experiments were conducted in a completely randomized designwith at least 20 replicates per
treatment for the callus culture experiment and three replicates for the cell suspension culture experiments. Data
were subjected to statistical analyses by Anova programme using Duncan’s multiple range test[29]
as modified by
Snedecor and Cochran[30]
.Values marked with different letters were considered to be statistically different at p <
0.05.
III. Results and Discussion
1. Elicitation
S. persica callus cultures derived from stem internodal segments accumulated BITC as a response to
chitosan and salicylic acid, which acted as biotic elicitors (Table 1). The two elicitors were screened for various
concentrations for high quantity of BITC accumulation. Both elicitors showed significant increase in BITC
accumulation over the control cultures with insignificant increase in the fresh weight of callus. Callus treated
with chitosan increased BITC accumulation compared with that of salicylic acid. Maximum accumulation of
BITC (4.534 µg/g fresh weight) was recorded after the treatment with chitosan at the concentration of 100
mg/L, which achieved about 3.7-fold increase in the BITC production over the control treatment (1.239 µg/g
fresh weight), and also achieved the highest fresh and dry weight of callus (Fig. 1). Despite this treatment was
the optimum, BITC production decreased above this concentration level of chitosan (200 and 400 mg/L) in the
medium. The callus treated with different concentrations of salicylic acid (5-20 mg/L) showed maximum BITC
accumulation (1.690 µg/g fresh weight) at 5 mg/L, which slightly raised the yield of BITC over the control. It
recorded an increase of 1.4 times over the control treatment. The increase in salicylic acid elicitor concentration
led to a decline in the BITC accumulation. The best dose of the elicitors with maximum accumulation of BITC
was used in further experiments (100 mg/Lchitosan).
4. Establishment of suspension cultures of Salvadora persica L. for benzyl isothiocyanate production
DOI: 10.9790/3008-1201027278 www.iosrjournals.org 75 | Page
Table 1.Effect of different concentrations of chitosan and salicylic acid on the callus weight and accumulation of
BITC in callus cultures of Salvadora persicaon MS medium supplemented with 0.5 mg/L of both of 2,4-D and
Kin.
Elicitor concentration
(mg/L)
Fresh weight(g) Dry weight (g) Conc. of BITC (µg/g fresh
weight)
Control
0 5.044a
4.521d
1.239
Chitosan
100 6.104a
5.700a
4.534
200 5.518a
5.000b
2.443
400 4.914a
4.567d
2.045
Salicylic acid
5 5.678a
4.903c
1.690
10 4.900a
4.400e
1.624
20 5.500a
5.196b
1.531
In each column, mean followed by the same subscripted letter was not significantly different (p < 0.05) according to the
Duncan’s multiple range test
Fig. 1.Callus of Salvadora persica on MS medium supplemented with 0.5 mg/L of both of2,4-D and Kin in
addition to 100 mg/L chitosan.
It is observed that BITC production is highly affected by the addition of biotic elicitors. Therefore,
elicitation offers an attractive strategy for increasing the secondary metabolite in vitro production[31]
. This study
showed that both chitosan and salicylic acid increased accumulation of BITC in callus culture of S. persicaover
the control. Chitosan and salicylic acid have been reported to enhance the in vitro production of secondary
metabolites in various plants[32]
, such a stimulatory effect may be due to the stimulation of biosynthetic
enzymes[33]
. Chitosan is the main structural component of the cell wall of plant pathogen fungi, which mimics
its effects and activates the biosynthesis of defense-related secondary metabolites in plants[8, 11, 34]
.Also, salicylic
acid has an important role in the defense against attacks by microbes and herbivores, and against abiotic
stresses, hence, it is very often used in plant in vitro cultures as an elicitor[32, 35]
. It was observed that chitosan
was better for the enhancement of the production of BITC than salicylic acid. The same observation was
reported by Palee et al.[33]
, who revealed that chitosan was better than salicylic acid for the enhancement of the
in vitro production of three stemona alkaloids from Stemona curtisii. Chitosan has been used as a strong elicitor
to improve the in vitro production of secondary metabolites of other numerous plants, for example, Ocimum
basilicum[36]
andWithania somnifera[37]
.Another significant effect of the elicitors observed in this experiment was
that the accumulation of BITC is elicitor dose dependent. The increase in BITC accumulation with elicitor
application was observed and the increase in elicitor concentration over the optimum concentration led to
decrease in the BITC content in callus cultures. Similar results were reported in taxol[38]
, ajmalicine[39]
and
psoralen[4]
production.
2. Cell suspension cultures
Cell suspension cultures offer an alternative way to traditional agriculture for industrial production of
various valuable phytochemicals. The liquid culture has many advantages compared to solid one. The contacting
surface area of cells and media is much larger, which permit cells to utilize the well mixed nutrients much
easier. Also, harmful compounds that may be formed can be effectively diluted, which prevents the inhibition of
cell growth[40]
. Fig. 2 shows the effect of elicitation with the optimum concentration of chitosan (100 mg/L) on
5. Establishment of suspension cultures of Salvadora persica L. for benzyl isothiocyanate production
DOI: 10.9790/3008-1201027278 www.iosrjournals.org 76 | Page
BITC accumulation in shake-flask culture and stirred-tank bioreactor during different durations of incubation
time. Successful scale-up of BITC accumulationwas achieved from cell suspension cultures using both methods.
Its accumulation varied from 7 to 28 days of incubation and increased with increasing in the duration of
incubation time. This result indicates that the duration of cell cultured with the elicitor is important for the
maximum BITC accumulation. This agrees with Karwasara et al.[41]
and Ahmed and Baig[4]
, who stated that the
amount of metabolite production varied with duration time of incubation with elicitors. The optimum incubation
time for BITC accumulation with chitosan was found to be 21 days for both shake-flask culture and bioreactor.
Comparing BITC accumulation in S. persica cell cultures between shake-flask and stirred-tank
bioreactor systems is shown in Fig. 2. In general, BITC yield was higher in the bioreactor than that using shake-
flask culture during all the studied durations. Using the bioreactor, BITC content was 10.078 and 11.98µg/g
fresh weight after 7 and 14 days of incubation, respectively. BITC content increased significantly after 21 days,
reaching the maximum value of 36.189 µg/g fresh weight, which achieved about 8-fold increase, then it
declined (reaching 24.459 µg/g fresh weight of only 5.4-fold increase). The BITC content was higher in the
bioreactor by about 1.5, 1.1, 2.3 and 2 times than the shake-flask culture after 7, 14, 21 and 28 days,
respectively. Only after 14 days they were more or less the same. Both gave the maximum BITC accumulation
after 21 days and declined after 28 days. The decreased BITC accumulation could be due to arrest of cell
growth, degradation or conversion of BITC to some other chemical forms or due to change in medium nutrient
composition.
Fig.2.BITC yield of Salvadora persica cell suspension cultures after different durations of incubation time in
shake-flask culture and stirred-tank bioreactor in MS medium supplemented with 0.5 mg/L of both of 2,4-D and
Kin in addition to 100 mg/L chitosan.
The different performance between shake-flask culture and the stirred-tank bioreactor could be due to
the difference in hydrodynamic conditions and gas compositions between the two systems. The large surface
area for the air medium interface in the shake-flask and the bioreactor systems (larger container) provide an
adequate amount of oxygen and nutrient mass transfer[40]
. It was reported earlier that the plant cell growth
environment in a bioreactor is different from that in Erlenmeyer flasks; changes in shear stress, oxygen supply
and gas composition are expected.Cell suspension cultures grown in bioreactors are exposed to higher
hydrodynamic stress in comparison to those in Erlenmeyer flasks that causes damage of the plant cell cultures;
include alteration in single cells and aggregate size and shape, lower cell growth and viability, and/or decrease
in secondary metabolite production[42]
. However, there are some examples confirming the results of the present
study and demonstrating that exposure to shear stress as a result of cultivation in a bioreactor enhances
secondary metabolite production. Chen and Huang[43]
reported that a high shear stress favored the production
of L-DOPA by cells of Stizolobium hasjoo in a stirred-tank bioreactor. They proposed that shear stress generated
a proton elicitation, and this might be responsible for the enhancement of L-DOPA production. In addition,
Trejo-Tapia et al.[42]
found that in the stirred-tank bioreactor, Uncaria tomentosa cells afforded higher levels of
MOA than in Erlenmeyer flasks. Also, a successful scale-up of Azadirachta indica suspension culture for
azadirachtin production was done in stirred tank bioreactor[7]
.
In general, application of the bioreactor is one of the prerequisites for industrialization of plant cell
culture. They can provide well mixed media and sterile air. There are many successful researches on cell and
tissue culture of medicinal herbs to produce useful secondary metabolites through bioreactors, which
demonstrated the industrial potential of using bioreactor in producing valuable phytochemicals[40]
.
6. Establishment of suspension cultures of Salvadora persica L. for benzyl isothiocyanate production
DOI: 10.9790/3008-1201027278 www.iosrjournals.org 77 | Page
IV. Conclusion
This is the first report on the production of BITC from cell suspension cultures of S. persica in a shake-
flask and bioreactor systems. The results of this study show that the application of the biotic elicitor;
chitosan,enhanced BITC synthesis inS. persica callus culturewhen compared to the untreated callus or that
treated with salicylic acid.Moreover, the present investigation established the feasibility of large-scale BITC
production in stirred-tank bioreactors, where S. persica cells afforded higher levels of BITC production (reached
about 8-fold after 21 days of incubation) than in Erlenmeyer flasks.The results obtained from the present study
can act as a roadmap for further studies regarding large-scale production of BITC from S. persica cell
suspension cultures by using bioreactors.
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