Asparagus racemosus is known to be a very important species due to its vital application in vario... more Asparagus racemosus is known to be a very important species due to its vital application in various diseases. Shatavari is the popular name of A. racemosus. In Ayurveda, shatavari has been documented for its curative and preventive use in aging, with improved mental function and increased longevity, along with supplementing vigor and vitality to the body. A. racemosus has also been used in dyspepsia, nervous disorders, inflammation, tumors, hepatopathy, and neuropathy. There are reports regarding the pharmacological activities of extracts of A. racemosus that include antioxidant, anti-cancerous, anti-diarrheal, immunomodulatory, anti-ulcer, and anti-diabetic activities. Steroidal saponins (shatavarinI–X) are the main active components of shatavari root extract with pharmacological activity. In the same category, shatavarinIV has been classified as glycosides of sarsasapogenin. Along with shatavarin, there are a few other active components that have been identified and characterized, such as quercetin, rutin, and immunoside. Not only the roots, flowers, fruits, as well as leaves also possess many of these pharmacologically active compounds such as shataverins, diosgenin, and quercetin-3 glucuronide. In this chapter, we will try to collect the state of the art of phytochemicals discovered from shatavari with their biological application that will be beneficial for utilization in the development of specialty/functional foods.
Globally, medicinal plant conservation is among the most pressing issues. To satisfy the demand f... more Globally, medicinal plant conservation is among the most pressing issues. To satisfy the demand for medicinal plants, natural resources are being depleted at an exponential rate. A rich legacy of spirituality along with culture underpins the nation’s strategies, plans for conservation, sustainable use, and equitable resource stewardship. Asparagus racemosus (Shatavari) has been described as the “queen of herbs” for its medicinal properties. The multitude of uses of A. racemosus in conjunction with its constant, high demand, resulted in fluctuated and insufficient supply, which makes it a critical resource for conservation. The conservation of Shatavari can be achieved by using ex situ techniques such as the establishment of gene banks, field banks, seed banks, in vitro plant tissue banks, cryopreservation, vitrification, artificial propagation of plants for reintroduction into the wild, as well as the implementation of nurseries and home gardens. This chapter will provide detailed information about the advantages, disadvantages, preparations for ex situ tools (conventional and biotechnological), as well as the role of key institutions involved in ex situ conservation. A series of themed issues are intended to provide a window into the current research concerning the conservation of plants by ex situ means.
Echinacea purpurea (L.) Moench, a member of the Asteraceae (Compositae) family is an important an... more Echinacea purpurea (L.) Moench, a member of the Asteraceae (Compositae) family is an important and well-known medicinal plant. The plant is used in chemoprevention and chemotherapy for infectious disorders of the upper and lower respiratory tracts. Toothaches, gut pain, snake bites, skin problems, epilepsy, chronic arthritis, and cancer have all been treated with this species in the past. For instance, research has demonstrated the plant’s ability to cause anti-anxiety, anti-depression, cytotoxicity, and anti-mutagenic effects. Echinacea has immense decorative potential in addition to its potential medical benefits. Alkamides, caffeic acid derivatives, polysaccharides, and glycoproteins are some of the plant’s secondary metabolites that have immunostimulatory properties. Echinacea’s anti-inflammatory properties are largely due to its polysaccharides. In vitro culture offers the ability to overcome a variety of problems associated with Echinacea propagation, such as bottlenecks in growth and poor seed germination, and also to meet up the increased demand. Alkamides are thought to be responsible for the immunomodulatory actions of Echinacea extracts both in vitro and in vivo.
Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol so... more Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol solubility, chemical stability and compartmentalization, and helps plants to adapt to environmental changes. The SGTs in medicinal plants are of particular interest for their role in the biosynthesis of pharmacologically active substances. WsSGTL1, a SGT isolated from Withania somnifera, was expressed and functionally characterized in transgenic tobacco plants. Transgenic WsSGTL1-Nt lines showed an adaptive mechanism through demonstrating late germination, stunted growth, yellowish-green leaves and enhanced antioxidant system. The reduced chlorophyll content and chlorophyll fluorescence with decreased photosynthetic parameters were observed in WsSGTL1-Nt plants. These changes could be due to the enhanced glycosylation by WsSGTL1, as no modulation in chlorophyll biogenesis-related genes was observed in transgenic lines as compared to wildtype (WT) plants. Enhanced accumulation of main sterols like, campesterol, stigmasterol and sitosterol in glycosylated form was observed in WsSGTL1-Nt plants. Apart from these, other secondary metabolites related to plant's antioxidant system along with activities of antioxidant enzymes (SOD, CAT; two to fourfold) were enhanced in WsSGTL1-Nt as compared to WT. WsSGTL1-Nt plants showed significant resistance towards Spodoptera litura (biotic stress) with up to 27 % reduced larval weight as well as salt stress (abiotic stress) with improved survival capacity of leaf discs. The present study demonstrates that higher glycosylation of sterols and enhanced antioxidant system caused by expression of WsSGTL1 gene confers specific functions in plants to adapt under different environmental challenges.
Genus Ocimum of Labiatae is well known in all traditional medicinal systems like Ayurveda, Unani,... more Genus Ocimum of Labiatae is well known in all traditional medicinal systems like Ayurveda, Unani, Siddha, and Homeopathy. The pharmaceutical activities of different species of Ocimum attributed to all plant parts. Roots are the most significant vital organ of the plant, as they absorb water and nutrients from soil and transport to aerial parts of the plants. Roots of Ocimum were found helpful with free-radical scavenging activity to improve physical and mental strength as well as to treat diabetes, malaria, and liver problems. Antibacterial activity of Ocimum roots and its main component, rosmarinic acid, is very beneficial to protect against several human pathogens, including bacteria and viruses. Being so important in every way, roots of Ocimum need healthy rhizosphere. Bacteria, fungi, nematodes, types of soil, fungicide, pesticides, salt, radioactive elements, as well as heavy metal contaminations, affect roots and overall growth of Ocimum in positive or negative ways. Each component of rhizosphere (natural, treatment or contamination) affects the roots, which highlights current ecological scenario to discover biosafe and more productive approaches. For such prestigious organ of Ocimum, development of in vitro root cultures and hairy root cultures assists to reduce the efforts and timing of the traditional cultivation process along with elimination of negative factors in rhizosphere. Different strains of Agrobacterium rhizogenes, various media compositions, as well as discrete treatments, like elicitors, on nonidentical species or cultivars of Ocimum boost the root induction, biomass, and accumulation of phytoceuticals differently. Hairy roots and in vitro roots of Ocimum accumulate higher quantity of therapeutic metabolites. These metabolites include several phenolics (like rosmarinic acid, 3hydroxybenzoic acid, m-coumaric acid, p-coumaric acid, caffeic acid, ferulic acid, vanillic acid, chicoric acid, and lithospermic acid), triterpenes (such as betulinic acid, 3-epimaslinic acid, alphitolic acid, euscaphic acids, oleanolic acid, and ursolic acid) as well as flavonoids (flavones, flavonols, and dihydroflavonols). This review highlights pharmaceutical applications of Ocimum roots, a great deal of rhizosphere components and in vitro culturing techniques to enhance biomass as well as chief phytoceuticals.
This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an... more This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a-half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l(-1) kanamycin. Polymerase chain reaction analysis of T(0) transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T(1) progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was foun...
Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol so... more Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol solubility, chemical stability and compartmentalization, and helps plants to adapt to environmental changes. The SGTs in medicinal plants are of particular interest for their role in the biosynthesis of pharmacologically active substances. WsSGTL1, a SGT isolated from Withania somnifera, was expressed and functionally characterized in transgenic tobacco plants. Transgenic WsSGTL1-Nt lines showed an adaptive mechanism through demonstrating late germination, stunted growth, yellowish-green leaves and enhanced antioxidant system. The reduced chlorophyll content and chlorophyll fluorescence with decreased photosynthetic parameters were observed in WsSGTL1-Nt plants. These changes could be due to the enhanced glycosylation by WsSGTL1, as no modulation in chlorophyll biogenesis-related genes was observed in transgenic lines as compared to wildtype (WT) plants. Enhanced accumulation of main sterols like, campesterol, stigmasterol and sitosterol in glycosylated form was observed in WsSGTL1-Nt plants. Apart from these, other secondary metabolites related to plant’s antioxidant system along with activities of antioxidant enzymes (SOD, CAT; two to fourfold) were enhanced in WsSGTL1-Nt as compared to WT. WsSGTL1-Nt plants showed significant resistance towards Spodoptera litura (biotic stress) with up to 27 % reduced larval weight as well as salt stress (abiotic stress) with improved survival capacity of leaf discs. The present study demonstrates that higher glycosylation of sterols and enhanced antioxidant system caused by expression of WsSGTL1 gene confers specific functions in plants to adapt under different environmental challenges.
This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera—an ... more This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera—an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a-half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l−1 kanamycin. Polymerase chain reaction analysis of T0 transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T1 progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was found to be 1.67%.
Background: Sterol glycosyltrnasferases (SGT) are enzymes that glycosylate sterols which play imp... more Background: Sterol glycosyltrnasferases (SGT) are enzymes that glycosylate sterols which play important role in plant
adaptation to stress and are medicinally important in plants like Withania somnifera. The present study aims to find the role
of WsSGTL1 which is a sterol glycosyltransferase from W. somnifera, in plant’s adaptation to abiotic stress.
Methodology: The WsSGTL1 gene was transformed in Arabidopsis thaliana through Agrobacterium mediated transformation,
using the binary vector pBI121, by floral dip method. The phenotypic and physiological parameters like
germination, root length, shoot weight, relative electrolyte conductivity, MDA content, SOD levels, relative electrolyte
leakage and chlorophyll measurements were compared between transgenic and wild type Arabidopsis plants under
different abiotic stresses - salt, heat and cold. Biochemical analysis was done by HPLC-TLC and radiolabelled enzyme assay.
The promoter of the WsSGTL1 gene was cloned by using Genome Walker kit (Clontech, USA) and the 3D structures were
predicted by using Discovery Studio Ver. 2.5.
Results: The WsSGTL1 transgenic plants were confirmed to be single copy by Southern and homozygous by segregation
analysis. As compared to WT, the transgenic plants showed better germination, salt tolerance, heat and cold tolerance. The
level of the transgene WsSGTL1 was elevated in heat, cold and salt stress along with other marker genes such as HSP70,
HSP90, RD29, SOS3 and LEA4-5. Biochemical analysis showed the formation of sterol glycosides and increase in enzyme
activity. When the promoter of WsSGTL1 gene was cloned from W. somnifera and sequenced, it contained stress responsive
elements. Bioinformatics analysis of the 3D structure of the WsSGTL1 protein showed functional similarity with sterol
glycosyltransferase AtSGT of A. thaliana.
Conclusions: Transformation of WsSGTL1 gene in A. thaliana conferred abiotic stress tolerance. The promoter of the gene in
W.somnifera was found to have stress responsive elements. The 3D structure showed functional similarity with sterol
glycosyltransferases.
Leaf explants of the second or third node were collected from field-grown elite Jatropha curcas t... more Leaf explants of the second or third node were collected from field-grown elite Jatropha curcas trees and incubated in Murashige and Skoog’s (Physiol Plant 15:473–497, 1962) medium supplemented with growth regulators. Direct shoot organogenesis was induced when explants were incubated in a medium containing 0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 indolebutyric acid (IBA). A maximum of seven shoot buds differentiated within 6 weeks of culture incubation. Indirect shoot organogenesis was obtained when explants were incubated in the medium supplemented with 0.5 mg l−1 BA along with 1.0 mg l−1 each of 2,4-dichlorophenoxyacetic acid (2,4-D) and indoleacetic acid (IAA). A pulse treatment of 0.5 mg l−1 thidiazurone (TDZ) and 0.1 mg l−1 IBA for 5 days was necessary for shoot organogenesis in green compact callus before subculture into 0.5 mg l−1 BA and 0.1 mg l−1 IBA containing medium. Leaf explants of J. curcas, collected from the field, contained endophytic bacterial contamination, which expressed itself after 2–3 subcultures. These bacteria were cultured and identified as Enterobacter ludwigii. After staining, these were found as gram-negative bacteria. Their sensitivity against different antibiotics has been tested by culturing them with different antibiotic stabs for 72 h. Finally, Augmentin® was found as the most effective and suitable antibiotic which not only controlled the bacteria within 2–3 subcultures but also supported the regeneration system and growth of the regenerated shoots and such cultures have been grown for a long-term of over 2 years without any contamination.
Asparagus racemosus is known to be a very important species due to its vital application in vario... more Asparagus racemosus is known to be a very important species due to its vital application in various diseases. Shatavari is the popular name of A. racemosus. In Ayurveda, shatavari has been documented for its curative and preventive use in aging, with improved mental function and increased longevity, along with supplementing vigor and vitality to the body. A. racemosus has also been used in dyspepsia, nervous disorders, inflammation, tumors, hepatopathy, and neuropathy. There are reports regarding the pharmacological activities of extracts of A. racemosus that include antioxidant, anti-cancerous, anti-diarrheal, immunomodulatory, anti-ulcer, and anti-diabetic activities. Steroidal saponins (shatavarinI–X) are the main active components of shatavari root extract with pharmacological activity. In the same category, shatavarinIV has been classified as glycosides of sarsasapogenin. Along with shatavarin, there are a few other active components that have been identified and characterized, such as quercetin, rutin, and immunoside. Not only the roots, flowers, fruits, as well as leaves also possess many of these pharmacologically active compounds such as shataverins, diosgenin, and quercetin-3 glucuronide. In this chapter, we will try to collect the state of the art of phytochemicals discovered from shatavari with their biological application that will be beneficial for utilization in the development of specialty/functional foods.
Globally, medicinal plant conservation is among the most pressing issues. To satisfy the demand f... more Globally, medicinal plant conservation is among the most pressing issues. To satisfy the demand for medicinal plants, natural resources are being depleted at an exponential rate. A rich legacy of spirituality along with culture underpins the nation’s strategies, plans for conservation, sustainable use, and equitable resource stewardship. Asparagus racemosus (Shatavari) has been described as the “queen of herbs” for its medicinal properties. The multitude of uses of A. racemosus in conjunction with its constant, high demand, resulted in fluctuated and insufficient supply, which makes it a critical resource for conservation. The conservation of Shatavari can be achieved by using ex situ techniques such as the establishment of gene banks, field banks, seed banks, in vitro plant tissue banks, cryopreservation, vitrification, artificial propagation of plants for reintroduction into the wild, as well as the implementation of nurseries and home gardens. This chapter will provide detailed information about the advantages, disadvantages, preparations for ex situ tools (conventional and biotechnological), as well as the role of key institutions involved in ex situ conservation. A series of themed issues are intended to provide a window into the current research concerning the conservation of plants by ex situ means.
Echinacea purpurea (L.) Moench, a member of the Asteraceae (Compositae) family is an important an... more Echinacea purpurea (L.) Moench, a member of the Asteraceae (Compositae) family is an important and well-known medicinal plant. The plant is used in chemoprevention and chemotherapy for infectious disorders of the upper and lower respiratory tracts. Toothaches, gut pain, snake bites, skin problems, epilepsy, chronic arthritis, and cancer have all been treated with this species in the past. For instance, research has demonstrated the plant’s ability to cause anti-anxiety, anti-depression, cytotoxicity, and anti-mutagenic effects. Echinacea has immense decorative potential in addition to its potential medical benefits. Alkamides, caffeic acid derivatives, polysaccharides, and glycoproteins are some of the plant’s secondary metabolites that have immunostimulatory properties. Echinacea’s anti-inflammatory properties are largely due to its polysaccharides. In vitro culture offers the ability to overcome a variety of problems associated with Echinacea propagation, such as bottlenecks in growth and poor seed germination, and also to meet up the increased demand. Alkamides are thought to be responsible for the immunomodulatory actions of Echinacea extracts both in vitro and in vivo.
Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol so... more Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol solubility, chemical stability and compartmentalization, and helps plants to adapt to environmental changes. The SGTs in medicinal plants are of particular interest for their role in the biosynthesis of pharmacologically active substances. WsSGTL1, a SGT isolated from Withania somnifera, was expressed and functionally characterized in transgenic tobacco plants. Transgenic WsSGTL1-Nt lines showed an adaptive mechanism through demonstrating late germination, stunted growth, yellowish-green leaves and enhanced antioxidant system. The reduced chlorophyll content and chlorophyll fluorescence with decreased photosynthetic parameters were observed in WsSGTL1-Nt plants. These changes could be due to the enhanced glycosylation by WsSGTL1, as no modulation in chlorophyll biogenesis-related genes was observed in transgenic lines as compared to wildtype (WT) plants. Enhanced accumulation of main sterols like, campesterol, stigmasterol and sitosterol in glycosylated form was observed in WsSGTL1-Nt plants. Apart from these, other secondary metabolites related to plant's antioxidant system along with activities of antioxidant enzymes (SOD, CAT; two to fourfold) were enhanced in WsSGTL1-Nt as compared to WT. WsSGTL1-Nt plants showed significant resistance towards Spodoptera litura (biotic stress) with up to 27 % reduced larval weight as well as salt stress (abiotic stress) with improved survival capacity of leaf discs. The present study demonstrates that higher glycosylation of sterols and enhanced antioxidant system caused by expression of WsSGTL1 gene confers specific functions in plants to adapt under different environmental challenges.
Genus Ocimum of Labiatae is well known in all traditional medicinal systems like Ayurveda, Unani,... more Genus Ocimum of Labiatae is well known in all traditional medicinal systems like Ayurveda, Unani, Siddha, and Homeopathy. The pharmaceutical activities of different species of Ocimum attributed to all plant parts. Roots are the most significant vital organ of the plant, as they absorb water and nutrients from soil and transport to aerial parts of the plants. Roots of Ocimum were found helpful with free-radical scavenging activity to improve physical and mental strength as well as to treat diabetes, malaria, and liver problems. Antibacterial activity of Ocimum roots and its main component, rosmarinic acid, is very beneficial to protect against several human pathogens, including bacteria and viruses. Being so important in every way, roots of Ocimum need healthy rhizosphere. Bacteria, fungi, nematodes, types of soil, fungicide, pesticides, salt, radioactive elements, as well as heavy metal contaminations, affect roots and overall growth of Ocimum in positive or negative ways. Each component of rhizosphere (natural, treatment or contamination) affects the roots, which highlights current ecological scenario to discover biosafe and more productive approaches. For such prestigious organ of Ocimum, development of in vitro root cultures and hairy root cultures assists to reduce the efforts and timing of the traditional cultivation process along with elimination of negative factors in rhizosphere. Different strains of Agrobacterium rhizogenes, various media compositions, as well as discrete treatments, like elicitors, on nonidentical species or cultivars of Ocimum boost the root induction, biomass, and accumulation of phytoceuticals differently. Hairy roots and in vitro roots of Ocimum accumulate higher quantity of therapeutic metabolites. These metabolites include several phenolics (like rosmarinic acid, 3hydroxybenzoic acid, m-coumaric acid, p-coumaric acid, caffeic acid, ferulic acid, vanillic acid, chicoric acid, and lithospermic acid), triterpenes (such as betulinic acid, 3-epimaslinic acid, alphitolic acid, euscaphic acids, oleanolic acid, and ursolic acid) as well as flavonoids (flavones, flavonols, and dihydroflavonols). This review highlights pharmaceutical applications of Ocimum roots, a great deal of rhizosphere components and in vitro culturing techniques to enhance biomass as well as chief phytoceuticals.
This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an... more This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a-half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l(-1) kanamycin. Polymerase chain reaction analysis of T(0) transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T(1) progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was foun...
Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol so... more Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol solubility, chemical stability and compartmentalization, and helps plants to adapt to environmental changes. The SGTs in medicinal plants are of particular interest for their role in the biosynthesis of pharmacologically active substances. WsSGTL1, a SGT isolated from Withania somnifera, was expressed and functionally characterized in transgenic tobacco plants. Transgenic WsSGTL1-Nt lines showed an adaptive mechanism through demonstrating late germination, stunted growth, yellowish-green leaves and enhanced antioxidant system. The reduced chlorophyll content and chlorophyll fluorescence with decreased photosynthetic parameters were observed in WsSGTL1-Nt plants. These changes could be due to the enhanced glycosylation by WsSGTL1, as no modulation in chlorophyll biogenesis-related genes was observed in transgenic lines as compared to wildtype (WT) plants. Enhanced accumulation of main sterols like, campesterol, stigmasterol and sitosterol in glycosylated form was observed in WsSGTL1-Nt plants. Apart from these, other secondary metabolites related to plant’s antioxidant system along with activities of antioxidant enzymes (SOD, CAT; two to fourfold) were enhanced in WsSGTL1-Nt as compared to WT. WsSGTL1-Nt plants showed significant resistance towards Spodoptera litura (biotic stress) with up to 27 % reduced larval weight as well as salt stress (abiotic stress) with improved survival capacity of leaf discs. The present study demonstrates that higher glycosylation of sterols and enhanced antioxidant system caused by expression of WsSGTL1 gene confers specific functions in plants to adapt under different environmental challenges.
This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera—an ... more This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera—an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a-half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l−1 kanamycin. Polymerase chain reaction analysis of T0 transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T1 progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was found to be 1.67%.
Background: Sterol glycosyltrnasferases (SGT) are enzymes that glycosylate sterols which play imp... more Background: Sterol glycosyltrnasferases (SGT) are enzymes that glycosylate sterols which play important role in plant
adaptation to stress and are medicinally important in plants like Withania somnifera. The present study aims to find the role
of WsSGTL1 which is a sterol glycosyltransferase from W. somnifera, in plant’s adaptation to abiotic stress.
Methodology: The WsSGTL1 gene was transformed in Arabidopsis thaliana through Agrobacterium mediated transformation,
using the binary vector pBI121, by floral dip method. The phenotypic and physiological parameters like
germination, root length, shoot weight, relative electrolyte conductivity, MDA content, SOD levels, relative electrolyte
leakage and chlorophyll measurements were compared between transgenic and wild type Arabidopsis plants under
different abiotic stresses - salt, heat and cold. Biochemical analysis was done by HPLC-TLC and radiolabelled enzyme assay.
The promoter of the WsSGTL1 gene was cloned by using Genome Walker kit (Clontech, USA) and the 3D structures were
predicted by using Discovery Studio Ver. 2.5.
Results: The WsSGTL1 transgenic plants were confirmed to be single copy by Southern and homozygous by segregation
analysis. As compared to WT, the transgenic plants showed better germination, salt tolerance, heat and cold tolerance. The
level of the transgene WsSGTL1 was elevated in heat, cold and salt stress along with other marker genes such as HSP70,
HSP90, RD29, SOS3 and LEA4-5. Biochemical analysis showed the formation of sterol glycosides and increase in enzyme
activity. When the promoter of WsSGTL1 gene was cloned from W. somnifera and sequenced, it contained stress responsive
elements. Bioinformatics analysis of the 3D structure of the WsSGTL1 protein showed functional similarity with sterol
glycosyltransferase AtSGT of A. thaliana.
Conclusions: Transformation of WsSGTL1 gene in A. thaliana conferred abiotic stress tolerance. The promoter of the gene in
W.somnifera was found to have stress responsive elements. The 3D structure showed functional similarity with sterol
glycosyltransferases.
Leaf explants of the second or third node were collected from field-grown elite Jatropha curcas t... more Leaf explants of the second or third node were collected from field-grown elite Jatropha curcas trees and incubated in Murashige and Skoog’s (Physiol Plant 15:473–497, 1962) medium supplemented with growth regulators. Direct shoot organogenesis was induced when explants were incubated in a medium containing 0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 indolebutyric acid (IBA). A maximum of seven shoot buds differentiated within 6 weeks of culture incubation. Indirect shoot organogenesis was obtained when explants were incubated in the medium supplemented with 0.5 mg l−1 BA along with 1.0 mg l−1 each of 2,4-dichlorophenoxyacetic acid (2,4-D) and indoleacetic acid (IAA). A pulse treatment of 0.5 mg l−1 thidiazurone (TDZ) and 0.1 mg l−1 IBA for 5 days was necessary for shoot organogenesis in green compact callus before subculture into 0.5 mg l−1 BA and 0.1 mg l−1 IBA containing medium. Leaf explants of J. curcas, collected from the field, contained endophytic bacterial contamination, which expressed itself after 2–3 subcultures. These bacteria were cultured and identified as Enterobacter ludwigii. After staining, these were found as gram-negative bacteria. Their sensitivity against different antibiotics has been tested by culturing them with different antibiotic stabs for 72 h. Finally, Augmentin® was found as the most effective and suitable antibiotic which not only controlled the bacteria within 2–3 subcultures but also supported the regeneration system and growth of the regenerated shoots and such cultures have been grown for a long-term of over 2 years without any contamination.
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Papers by Vibha Pandey
adaptation to stress and are medicinally important in plants like Withania somnifera. The present study aims to find the role
of WsSGTL1 which is a sterol glycosyltransferase from W. somnifera, in plant’s adaptation to abiotic stress.
Methodology: The WsSGTL1 gene was transformed in Arabidopsis thaliana through Agrobacterium mediated transformation,
using the binary vector pBI121, by floral dip method. The phenotypic and physiological parameters like
germination, root length, shoot weight, relative electrolyte conductivity, MDA content, SOD levels, relative electrolyte
leakage and chlorophyll measurements were compared between transgenic and wild type Arabidopsis plants under
different abiotic stresses - salt, heat and cold. Biochemical analysis was done by HPLC-TLC and radiolabelled enzyme assay.
The promoter of the WsSGTL1 gene was cloned by using Genome Walker kit (Clontech, USA) and the 3D structures were
predicted by using Discovery Studio Ver. 2.5.
Results: The WsSGTL1 transgenic plants were confirmed to be single copy by Southern and homozygous by segregation
analysis. As compared to WT, the transgenic plants showed better germination, salt tolerance, heat and cold tolerance. The
level of the transgene WsSGTL1 was elevated in heat, cold and salt stress along with other marker genes such as HSP70,
HSP90, RD29, SOS3 and LEA4-5. Biochemical analysis showed the formation of sterol glycosides and increase in enzyme
activity. When the promoter of WsSGTL1 gene was cloned from W. somnifera and sequenced, it contained stress responsive
elements. Bioinformatics analysis of the 3D structure of the WsSGTL1 protein showed functional similarity with sterol
glycosyltransferase AtSGT of A. thaliana.
Conclusions: Transformation of WsSGTL1 gene in A. thaliana conferred abiotic stress tolerance. The promoter of the gene in
W.somnifera was found to have stress responsive elements. The 3D structure showed functional similarity with sterol
glycosyltransferases.
adaptation to stress and are medicinally important in plants like Withania somnifera. The present study aims to find the role
of WsSGTL1 which is a sterol glycosyltransferase from W. somnifera, in plant’s adaptation to abiotic stress.
Methodology: The WsSGTL1 gene was transformed in Arabidopsis thaliana through Agrobacterium mediated transformation,
using the binary vector pBI121, by floral dip method. The phenotypic and physiological parameters like
germination, root length, shoot weight, relative electrolyte conductivity, MDA content, SOD levels, relative electrolyte
leakage and chlorophyll measurements were compared between transgenic and wild type Arabidopsis plants under
different abiotic stresses - salt, heat and cold. Biochemical analysis was done by HPLC-TLC and radiolabelled enzyme assay.
The promoter of the WsSGTL1 gene was cloned by using Genome Walker kit (Clontech, USA) and the 3D structures were
predicted by using Discovery Studio Ver. 2.5.
Results: The WsSGTL1 transgenic plants were confirmed to be single copy by Southern and homozygous by segregation
analysis. As compared to WT, the transgenic plants showed better germination, salt tolerance, heat and cold tolerance. The
level of the transgene WsSGTL1 was elevated in heat, cold and salt stress along with other marker genes such as HSP70,
HSP90, RD29, SOS3 and LEA4-5. Biochemical analysis showed the formation of sterol glycosides and increase in enzyme
activity. When the promoter of WsSGTL1 gene was cloned from W. somnifera and sequenced, it contained stress responsive
elements. Bioinformatics analysis of the 3D structure of the WsSGTL1 protein showed functional similarity with sterol
glycosyltransferase AtSGT of A. thaliana.
Conclusions: Transformation of WsSGTL1 gene in A. thaliana conferred abiotic stress tolerance. The promoter of the gene in
W.somnifera was found to have stress responsive elements. The 3D structure showed functional similarity with sterol
glycosyltransferases.