Withanolide

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Neelam S Sangwan - One of the best experts on this subject based on the ideXlab platform.

  • rnai and homologous over expression based functional approaches reveal triterpenoid synthase gene cycloartenol synthase is involved in downstream Withanolide biosynthesis in withania somnifera
    PLOS ONE, 2016
    Co-Authors: Smrati Mishra, Bhawana Mishra, Rajender Singh Sangwan, Jyoti Singh Jadaun, Shilpi Bansal, Neelam S Sangwan
    Abstract:

    Withania somnifera Dunal, is one of the most commonly used medicinal plant in Ayurvedic and indigenous medicine traditionally owing to its therapeutic potential, because of major chemical constituents, Withanolides. Withanolide biosynthesis requires the activities of several enzymes in vivo. Cycloartenol synthase (CAS) is an important enzyme in the Withanolide biosynthetic pathway, catalyzing cyclization of 2, 3 oxidosqualene into cycloartenol. In the present study, we have cloned full-length WsCAS from Withania somnifera by homology-based PCR method. For gene function investigation, we constructed three RNAi gene-silencing constructs in backbone of RNAi vector pGSA and a full-length over-expression construct. These constructs were transformed in Agrobacterium strain GV3101 for plant transformation in W. somnifera. Molecular and metabolite analysis was performed in putative Withania transformants. The PCR and Southern blot results showed the genomic integration of these RNAi and overexpression construct(s) in Withania genome. The qRT-PCR analysis showed that the expression of WsCAS gene was considerably downregulated in stable transgenic silenced Withania lines compared with the non-transformed control and HPLC analysis showed that Withanolide content was greatly reduced in silenced lines. Transgenic plants over expressing CAS gene displayed enhanced level of CAS transcript and Withanolide content compared to non-transformed controls. This work is the first full proof report of functional validation of any metabolic pathway gene in W. somnifera at whole plant level as per our knowledge and it will be further useful to understand the regulatory role of different genes involved in the biosynthesis of Withanolides.

  • comparative transcriptome analysis of different chemotypes elucidates Withanolide biosynthesis pathway from medicinal plant withania somnifera
    Scientific Reports, 2015
    Co-Authors: Parul Gupta, Neelam S Sangwan, Ridhi Goel, Aditya Vikram Agarwal, Mehar Hasan Asif, Rajender S Sangwan, Prabodh Kumar Trivedi
    Abstract:

    Withania somnifera is one of the most valuable medicinal plants synthesizing secondary metabolites known as Withanolides. Despite pharmaceutical importance, limited information is available about the biosynthesis of Withanolides. Chemo-profiling of leaf and root tissues of Withania suggest differences in the content and/or nature of Withanolides in different chemotypes. To identify genes involved in chemotype and/or tissue-specific Withanolide biosynthesis, we established transcriptomes of leaf and root tissues of distinct chemotypes. Genes encoding enzymes for intermediate steps of terpenoid backbone biosynthesis with their alternatively spliced forms and paralogous have been identified. Analysis suggests differential expression of large number genes among leaf and root tissues of different chemotypes. Study also identified differentially expressing transcripts encoding cytochrome P450s, glycosyltransferases, methyltransferases and transcription factors which might be involved in chemodiversity in Withania. Virus induced gene silencing of the sterol ∆7-reductase (WsDWF5) involved in the synthesis of 24-methylene cholesterol, Withanolide backbone, suggests role of this enzyme in biosynthesis of Withanolides. Information generated, in this study, provides a rich resource for functional analysis of Withanolide-specific genes to elucidate chemotype- as well as tissue-specific Withanolide biosynthesis. This genomic resource will also help in development of new tools for functional genomics and breeding in Withania.

  • qualitative and quantitative variations in Withanolides and expression of some pathway genes during different stages of morphogenesis in withania somnifera dunal
    Protoplasma, 2013
    Co-Authors: Farzana Sabir, Smrati Mishra, Jyoti Singh Jadaun, Rajender S Sangwan, Neelam S Sangwan
    Abstract:

    Withania somnifera Dunal is an important and extensively studied medicinal plant; however, there is no report available that relates Withanolide content and its profile in relation to the expression of pathway genes during different morphogenic stages. In this study, Withanolide A, withaferin A, and withanone, the major Withanolides of W. somnifera, were measured in different in vitro stages during organogenesis, viz., shoot to root (direct rhizogenesis)/root to shoot (indirect via callus phase) transition vis-a-vis expression levels of key pathway genes involved in Withanolide biosynthetic pathways. The morphogenic transitions were found to be tightly linked to the pattern of accumulation of Withanolides. The high expression levels of most of the pathway genes in in vitro shoots in comparison to in vitro root and callus tissues exhibited a direct co-relation with the maximum Withanolide content (>2.7 mg/gDW). The biogenesis of withaferin A, a major constituent of the leaves, was however found to be tightly linked to shoots/green tissue. In addition, we were also able to establish an efficient regeneration system from roots for their further utilization in biotechnological applications.

  • Withanolide biosynthesis recruits both mevalonate and doxp pathways of isoprenogenesis in ashwagandha withania somnifera l dunal
    Plant Cell Reports, 2012
    Co-Authors: Neelam S Sangwan, Narayan D Chaurasiya, Laxminarain Misra, Farzana Sabir, Rajender S Sangwan
    Abstract:

    Withanolides are pharmaceutically important C28-phytochemicals produced in most prodigal amounts and diversified forms by Withania somnifera. Metabolic origin of Withanolides from triterpenoid pathway intermediates implies that isoprenogenesis could significantly govern Withanolide production. In plants, isoprenogenesis occurs via two routes: mevalonate (MVA) pathway in cytosol and non-mevalonate or DOXP/MEP pathway in plastids. We have investigated relative carbon contribution of MVA and DOXP pathways to Withanolide biosynthesis in W. somnifera. The quantitative NMR-based biosynthetic study involved tracing of 13C label from 13C1-d-glucose to withaferin A in Withanolide producing in vitro microshoot cultures of the plant. Enrichment of 13C abundance at each carbon of withaferin A from 13C1-glucose-fed cultures was monitored by normalization and integration of NMR signal intensities. The pattern of carbon position-specific 13C enrichment of withaferin A was analyzed by a retro-biosynthetic approach using a squalene-intermediated metabolic model of Withanolide (withaferin A) biosynthesis. The pattern suggested that both DOXP and MVA pathways of isoprenogenesis were significantly involved in Withanolide biosynthesis with their relative contribution on the ratio of 25:75, respectively. The results have been discussed in a new conceptual line of biosynthetic load-driven model of relative recruitment of DOXP and MVA pathways for biosynthesis of isoprenoids.

  • metabolic clustering of a core collection of indian ginseng withania somnifera dunal through dna isoenzyme polypeptide and Withanolide profile diversity
    Fitoterapia, 2009
    Co-Authors: Narayan D Chaurasiya, Rajender S Sangwan, Rakesh Tuli, Laxmi N Misra, Neelam S Sangwan
    Abstract:

    Withania somnifera is one of the most important medicinal plants of Ayurveda and finds extensive uses in Indian traditional herbal preparations. In this investigation, selected accessions of the plant were examined for diversity through RAPDs, isoenzymes, polypeptide polymorphism and Withanolide profiles. The accessions clustered together with respect to their characteristic profile of major Withanolides and represented withaferin A, withanone, Withanolide D or Withanolide A rich groups. This level of phytochemical diversity as discrete chemotypes is widest and is being first ever documented to occur in Indian population of the plant.

Trivedi, Mahendra Kumar - One of the best experts on this subject based on the ideXlab platform.

  • LC-MS, GC-MS, and NMR Spectroscopy Based Evaluation of the Energy of Consciousness Healing Treated Withania somnifera (Ashwagandha) Root Extract
    2017
    Co-Authors: Trivedi, Mahendra Kumar
    Abstract:

    Withania somnifera root extract which contains Withanolides as major active constituent and it is used traditionally for the prevention and treatment of several diseases. The objective of the current study was to investigate the impact of The Trivedi Effect® - Consciousness Energy Healing Treatment on the structural properties of the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Consciousness Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in the control and Biofield Energy Treated samples, whereas the peak area% i.e. the relative amount of the phytoconstituents at respective retention time was significantly altered. The peak area% at Rt of 5.6, 6.8, 6.9, 7.2, 7.9, 8.4, 8.5, 8.6, and 9.2 minutes of the treated sample were increased significantly in the range of 1.46% to 253.06% compared to the control sample. In the contrary, the peak area% of the treated sample at Rt of 6.4 and 8.2 minutes were significantly decreased by 12.72% and 17.35%, respectively with respect to the control sample. A total of 16 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, dihydroWithanolide D, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 621, 489, 473, 767, 471, 505, and 992 at retention times of 6.4, 6.8, 7.2, 7.9, 8.2, 8.4, and 9.1 minutes, respectively with the help of LC-MS, GC-MS and NMR data of both the control and Biofield Energy Treated samples. The mass peak intensities of the Biofield Energy Treated sample were significantly changed in the range of -81.36% to 1720.90% compared with the control sample at the same retention time. These findings suggest that The Trivedi Effect® - Consciousness Energy Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging, and other chronic infections

  • LC-MS, GC-MS, and NMR Spectroscopy Based Evaluation of the Energy of Consciousness Healing Treated Withania somnifera (Ashwagandha) Root Extract
    2017
    Co-Authors: Trivedi, Mahendra Kumar, Nayak Gopal, Branton Alice, Trivedi Dahryn, Afaganis, Ariadne Esmene, Bader, Barbara Marie, Weekes, Brian A., Dumas, Daphne Luisa, Fiedler, Denise Marie, Smith, Dennille Mellesia
    Abstract:

    Withania somnifera root extract which contains Withanolides as major active constituent and it is used traditionally for the prevention and treatment of several diseases. The objective of the current study was to investigate the impact of The Trivedi Effect® - Consciousness Energy Healing Treatment on the structural properties of the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Consciousness Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in the control and Biofield Energy Treated samples, whereas the peak area% i.e. the relative amount of the phytoconstituents at respective retention time was significantly altered. The peak area% at Rt of 5.6, 6.8, 6.9, 7.2, 7.9, 8.4, 8.5, 8.6, and 9.2 minutes of the treated sample were increased significantly in the range of 1.46% to 253.06% compared to the control sample. In the contrary, the peak area% of the treated sample at Rt of 6.4 and 8.2 minutes were significantly decreased by 12.72% and 17.35%, respectively with respect to the control sample. A total of 16 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, dihydroWithanolide D, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 621, 489, 473, 767, 471, 505, and 992 at retention times of 6.4, 6.8, 7.2, 7.9, 8.2, 8.4, and 9.1 minutes, respectively with the help of LC-MS, GC-MS and NMR data of both the control and Biofield Energy Treated samples. The mass peak intensities of the Biofield Energy Treated sample were significantly changed in the range of -81.36% to 1720.90% compared with the control sample at the same retention time. These findings suggest that The Trivedi Effect® - Consciousness Energy Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging, and other chronic infections. Source: https://www.trivedieffect.com/science/lc-ms-gc-ms-and-nmr-spectroscopy-based-evaluation-of-the-energy-of-consciousness-healing-treated-withania-somnifera-ashwagandha-root-extract http://www.sciencepublishinggroup.com/journal/paperinfo?journalid=329&doi=10.11648/j.jddmc.20170302.1

  • LC-MS, GC-MS, and NMR Spectroscopy Based Evaluation of the Energy of Consciousness Healing Treated Withania somnifera (Ashwagandha) Root Extract
    2017
    Co-Authors: Trivedi, Mahendra Kumar, Nayak Gopal, Branton Alice, Trivedi Dahryn, Afaganis, Ariadne Esmene, Bader, Barbara Marie, Weekes, Brian A., Dumas, Daphne Luisa, Fiedler, Denise Marie, Smith, Dennille Mellesia
    Abstract:

    Withania somnifera root extract which contains Withanolides as major active constituent and it is used traditionally for the prevention and treatment of several diseases. The objective of the current study was to investigate the impact of The Trivedi Effect® - Consciousness Energy Healing Treatment on the structural properties of the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Consciousness Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in the control and Biofield Energy Treated samples, whereas the peak area% i.e. the relative amount of the phytoconstituents at respective retention time was significantly altered. The peak area% at Rt of 5.6, 6.8, 6.9, 7.2, 7.9, 8.4, 8.5, 8.6, and 9.2 minutes of the treated sample were increased significantly in the range of 1.46% to 253.06% compared to the control sample. In the contrary, the peak area% of the treated sample at Rt of 6.4 and 8.2 minutes were significantly decreased by 12.72% and 17.35%, respectively with respect to the control sample. A total of 16 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, dihydroWithanolide D, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 621, 489, 473, 767, 471, 505, and 992 at retention times of 6.4, 6.8, 7.2, 7.9, 8.2, 8.4, and 9.1 minutes, respectively with the help of LC-MS, GC-MS and NMR data of both the control and Biofield Energy Treated samples. The mass peak intensities of the Biofield Energy Treated sample were significantly changed in the range of -81.36% to 1720.90% compared with the control sample at the same retention time. These findings suggest that The Trivedi Effect® - Consciousness Energy Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging, and other chronic infections. https://www.trivedieffect.com/science/lc-ms-gc-ms-and-nmr-spectroscopy-based-evaluation-of-the-energy-of-consciousness-healing-treated-withania-somnifera-ashwagandha-root-extract http://www.sciencepublishinggroup.com/journal/paperinfo?journalid=329&doi=10.11648/j.jddmc.20170302.1

  • Evaluation of the Energy of Consciousness Healing Treated Withania Somnifera (Ashwagandha) Root Extract Using LC-MS, GC-MS, and NMR Spectroscopy
    2017
    Co-Authors: Balmer, Joy Angevin, Trivedi, Mahendra Kumar, Nayak Gopal, Branton Alice, Trivedi Dahryn
    Abstract:

    Withania somnifera (Ashwagandha) root extract is very useful herbal medicine since from ancient time. The current study designed to investigate the impact of The Trivedi Effect® - Energy of Consciousness Healing Treatment on the structural properties of the ashwagandha root hydroalcoholic extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Energy of Consciousness Healing Treatment remotely by seven renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in both the control and treated samples, whereas the peak area % at respective retention time was significantly altered. The peak area% of the treated sample at Rt of 5.3, 5.5, 6.4, 6.5, 6.8, 6.9, 7.1, 7.3, 7.8, 7.9, 8.0, 8.2, 8.4, 8.6, 8.8, 9.0, 9.1, and 10.1 minutes were significantly reduced by 0.57% to 38.10% compared to the control sample. In addition, the peak area% of the treated sample at Rt of 5.7, 6.7, and 8.1 minutes were significantly increased by 16.00%, 244.44% and 19.62%, respectively compared with the control sample. A total of 21 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A and Withanolide sulfoxide, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 569, 621, 489, 473, 767, 471, 505, 992, and 455 at retention times of 6.4, 6.5, 6.8, 7.1, 7.9, 8.1, 8.4, 9.1, and 10.1 minutes with the help of GC-MS and NMR data of both the control and Biofield Energy Treated samples. There were significant changes observed in the peak intensity values at the same retention time in the Biofield Energy Treated sample in the range of -75.32% to 108.51% compared with the control sample. These findings suggest that The Trivedi Effect® - Energy of Consciousness Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections

  • Evaluation of the Energy of Consciousness Healing Treated Withania Somnifera (Ashwagandha) Root Extract Using LC-MS, GC-MS, and NMR Spectroscopy
    2017
    Co-Authors: Trivedi, Mahendra Kumar, Balmer, Joy Angevin, Nayak Gopal, Branton Alice, Trivedi Dahryn, Balmer, Alan Joseph, Anagnos Dimitrius, Kinney, Janice Patricia, Holling, Joni Marie, Duprey-reed, Lauree Ann
    Abstract:

    Withania somnifera (Ashwagandha) root extract is very useful herbal medicine since from ancient time. The current study designed to investigate the impact of The Trivedi Effect® - Energy of Consciousness Healing Treatment on the structural properties of the ashwagandha root hydroalcoholic extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Energy of Consciousness Healing Treatment remotely by seven renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in both the control and treated samples, whereas the peak area % at respective retention time was significantly altered. The peak area% of the treated sample at Rt of 5.3, 5.5, 6.4, 6.5, 6.8, 6.9, 7.1, 7.3, 7.8, 7.9, 8.0, 8.2, 8.4, 8.6, 8.8, 9.0, 9.1, and 10.1 minutes were significantly reduced by 0.57% to 38.10% compared to the control sample. In addition, the peak area% of the treated sample at Rt of 5.7, 6.7, and 8.1 minutes were significantly increased by 16.00%, 244.44% and 19.62%, respectively compared with the control sample. A total of 21 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A and Withanolide sulfoxide, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 569, 621, 489, 473, 767, 471, 505, 992, and 455 at retention times of 6.4, 6.5, 6.8, 7.1, 7.9, 8.1, 8.4, 9.1, and 10.1 minutes with the help of GC-MS and NMR data of both the control and Biofield Energy Treated samples. There were significant changes observed in the peak intensity values at the same retention time in the Biofield Energy Treated sample in the range of -75.32% to 108.51% compared with the control sample. These findings suggest that The Trivedi Effect® - Energy of Consciousness Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections. https://www.trivedieffect.com/science/evaluation-of-the-energy-of-consciousness-healing-treated-withania-somnifera-ashwagandha-root-extract-using-lc-ms-gc-ms-and-nmr-spectroscopy http://www.sciencepublishinggroup.com/journal/paperinfo?journalid=655&doi=10.11648/j.ajbls.20170502.1

Rajender S Sangwan - One of the best experts on this subject based on the ideXlab platform.

  • comparative transcriptome analysis of different chemotypes elucidates Withanolide biosynthesis pathway from medicinal plant withania somnifera
    Scientific Reports, 2015
    Co-Authors: Parul Gupta, Neelam S Sangwan, Ridhi Goel, Aditya Vikram Agarwal, Mehar Hasan Asif, Rajender S Sangwan, Prabodh Kumar Trivedi
    Abstract:

    Withania somnifera is one of the most valuable medicinal plants synthesizing secondary metabolites known as Withanolides. Despite pharmaceutical importance, limited information is available about the biosynthesis of Withanolides. Chemo-profiling of leaf and root tissues of Withania suggest differences in the content and/or nature of Withanolides in different chemotypes. To identify genes involved in chemotype and/or tissue-specific Withanolide biosynthesis, we established transcriptomes of leaf and root tissues of distinct chemotypes. Genes encoding enzymes for intermediate steps of terpenoid backbone biosynthesis with their alternatively spliced forms and paralogous have been identified. Analysis suggests differential expression of large number genes among leaf and root tissues of different chemotypes. Study also identified differentially expressing transcripts encoding cytochrome P450s, glycosyltransferases, methyltransferases and transcription factors which might be involved in chemodiversity in Withania. Virus induced gene silencing of the sterol ∆7-reductase (WsDWF5) involved in the synthesis of 24-methylene cholesterol, Withanolide backbone, suggests role of this enzyme in biosynthesis of Withanolides. Information generated, in this study, provides a rich resource for functional analysis of Withanolide-specific genes to elucidate chemotype- as well as tissue-specific Withanolide biosynthesis. This genomic resource will also help in development of new tools for functional genomics and breeding in Withania.

  • de novo assembly functional annotation and comparative analysis of withania somnifera leaf and root transcriptomes to identify putative genes involved in the Withanolides biosynthesis
    PLOS ONE, 2013
    Co-Authors: Parul Gupta, Ridhi Goel, Mehar Hasan Asif, Rajender S Sangwan, Sumya Pathak, Apeksha Srivastava, Surya Pratap Singh, Prabodh Kumar Trivedi
    Abstract:

    Withania somnifera is one of the most valuable medicinal plants used in Ayurvedic and other indigenous medicine systems due to bioactive molecules known as Withanolides. As genomic information regarding this plant is very limited, little information is available about biosynthesis of Withanolides. To facilitate the basic understanding about the Withanolide biosynthesis pathways, we performed transcriptome sequencing for Withania leaf (101L) and root (101R) which specifically synthesize withaferin A and Withanolide A, respectively. Pyrosequencing yielded 8,34,068 and 7,21,755 reads which got assembled into 89,548 and 1,14,814 unique sequences from 101L and 101R, respectively. A total of 47,885 (101L) and 54,123 (101R) could be annotated using TAIR10, NR, tomato and potato databases. Gene Ontology and KEGG analyses provided a detailed view of all the enzymes involved in Withanolide backbone synthesis. Our analysis identified members of cytochrome P450, glycosyltransferase and methyltransferase gene families with unique presence or differential expression in leaf and root and might be involved in synthesis of tissue-specific Withanolides. We also detected simple sequence repeats (SSRs) in transcriptome data for use in future genetic studies. Comprehensive sequence resource developed for Withania, in this study, will help to elucidate biosynthetic pathway for tissue-specific synthesis of secondary plant products in non-model plant organisms as well as will be helpful in developing strategies for enhanced biosynthesis of Withanolides through biotechnological approaches.

  • qualitative and quantitative variations in Withanolides and expression of some pathway genes during different stages of morphogenesis in withania somnifera dunal
    Protoplasma, 2013
    Co-Authors: Farzana Sabir, Smrati Mishra, Jyoti Singh Jadaun, Rajender S Sangwan, Neelam S Sangwan
    Abstract:

    Withania somnifera Dunal is an important and extensively studied medicinal plant; however, there is no report available that relates Withanolide content and its profile in relation to the expression of pathway genes during different morphogenic stages. In this study, Withanolide A, withaferin A, and withanone, the major Withanolides of W. somnifera, were measured in different in vitro stages during organogenesis, viz., shoot to root (direct rhizogenesis)/root to shoot (indirect via callus phase) transition vis-a-vis expression levels of key pathway genes involved in Withanolide biosynthetic pathways. The morphogenic transitions were found to be tightly linked to the pattern of accumulation of Withanolides. The high expression levels of most of the pathway genes in in vitro shoots in comparison to in vitro root and callus tissues exhibited a direct co-relation with the maximum Withanolide content (>2.7 mg/gDW). The biogenesis of withaferin A, a major constituent of the leaves, was however found to be tightly linked to shoots/green tissue. In addition, we were also able to establish an efficient regeneration system from roots for their further utilization in biotechnological applications.

  • Withanolide biosynthesis recruits both mevalonate and doxp pathways of isoprenogenesis in ashwagandha withania somnifera l dunal
    Plant Cell Reports, 2012
    Co-Authors: Neelam S Sangwan, Narayan D Chaurasiya, Laxminarain Misra, Farzana Sabir, Rajender S Sangwan
    Abstract:

    Withanolides are pharmaceutically important C28-phytochemicals produced in most prodigal amounts and diversified forms by Withania somnifera. Metabolic origin of Withanolides from triterpenoid pathway intermediates implies that isoprenogenesis could significantly govern Withanolide production. In plants, isoprenogenesis occurs via two routes: mevalonate (MVA) pathway in cytosol and non-mevalonate or DOXP/MEP pathway in plastids. We have investigated relative carbon contribution of MVA and DOXP pathways to Withanolide biosynthesis in W. somnifera. The quantitative NMR-based biosynthetic study involved tracing of 13C label from 13C1-d-glucose to withaferin A in Withanolide producing in vitro microshoot cultures of the plant. Enrichment of 13C abundance at each carbon of withaferin A from 13C1-glucose-fed cultures was monitored by normalization and integration of NMR signal intensities. The pattern of carbon position-specific 13C enrichment of withaferin A was analyzed by a retro-biosynthetic approach using a squalene-intermediated metabolic model of Withanolide (withaferin A) biosynthesis. The pattern suggested that both DOXP and MVA pathways of isoprenogenesis were significantly involved in Withanolide biosynthesis with their relative contribution on the ratio of 25:75, respectively. The results have been discussed in a new conceptual line of biosynthetic load-driven model of relative recruitment of DOXP and MVA pathways for biosynthesis of isoprenoids.

  • metabolic clustering of a core collection of indian ginseng withania somnifera dunal through dna isoenzyme polypeptide and Withanolide profile diversity
    Fitoterapia, 2009
    Co-Authors: Narayan D Chaurasiya, Rajender S Sangwan, Rakesh Tuli, Laxmi N Misra, Neelam S Sangwan
    Abstract:

    Withania somnifera is one of the most important medicinal plants of Ayurveda and finds extensive uses in Indian traditional herbal preparations. In this investigation, selected accessions of the plant were examined for diversity through RAPDs, isoenzymes, polypeptide polymorphism and Withanolide profiles. The accessions clustered together with respect to their characteristic profile of major Withanolides and represented withaferin A, withanone, Withanolide D or Withanolide A rich groups. This level of phytochemical diversity as discrete chemotypes is widest and is being first ever documented to occur in Indian population of the plant.

Javier Palazon - One of the best experts on this subject based on the ideXlab platform.

  • overexpression of the arabidopsis thaliana squalene synthase gene in withania coagulans hairy root cultures
    Biologia Plantarum, 2011
    Co-Authors: Mohammad Hossein Mirjalili, Mercedes Bonfill, R. M. Cusido, Elisabet Moyano, Javier Palazon
    Abstract:

    Squalene synthase (SS) dimerizes two molecules of farnesyl diphosphate to synthesize squalene, a shared precursor in steroid and triterpenoid biosynthesis in plants. The SS1 gene encoding SS from Arabidopsis thaliana was introduced in Withania coagulans under the control of the CaMV35S promoter together with the T-DNA of Agrobacterium rhizogenes A4. The engineered hairy roots were studied for Withanolide production and phytosterol accumulation and the results were compared with those obtained from control roots harbouring only the T-DNA from pRiA4. The increased capacity of the engineered roots for biosynthesizing phytosterols and Withanolides was strongly related with the expression level of the transgene, showing the effectiveness of overexpressing 35SS1 to increase triterpenoid biosynthesis.

  • morphology and Withanolide production of withania coagulans hairy root cultures
    Engineering in Life Sciences, 2009
    Co-Authors: Hossein M Mirjalili, R. M. Cusido, Mercedes Bonfill, Seyyed M Fakhrtabatabaei, Houshang Alizadeh, Alireza Ghassempour, Javier Palazon
    Abstract:

    Inoculation of leaf sections of Withania coagulans with Agrobacterium tumefaciens strain C58C1 (pRiA4) induced transformed roots with the capacity to produce the most important bioactive compounds of Withania species, Withanolide A and withaferin A. The hairy roots obtained showed two morphologies: callus-like roots (CR) with a high capacity to produce Withanolides and typical hairy roots (HR) with faster growth capacity and lower Withanolide accumulation. The aux1 gene of pRiA4 was detected by PCR analyses in all roots showing callus-like morphology. However, this gene was only detected in 12.5% of the roots showing typical hairy root morphology. This fact suggests a significant role of aux genes in the morphology of transformed roots. Time course studies of Withanolide production showed that Withanolide A accumulated during the first part of the culture whereas the maximum accumulation of withaferin A occurred at the end of the culture period. Some transformed root lines, such as HR112 and CR26, showed considerable potential to produce Withanolides in a scaled up bioreactor system, especially the important pharmaceutical compound Withanolide A.

  • Steroidal lactones from withania somnifera, an ancient plant for novel medicine
    Molecules, 2009
    Co-Authors: Mohammad Hossein Mirjalili, Mercedes Bonfill, R. M. Cusido, Elisabeth Moyano, Javier Palazon
    Abstract:

    Withania somnifera, commonly known as Ashwagandha, is an important medicinal plant that has been used in Ayurvedic and indigenous medicine for over 3,000 years. In view of its varied therapeutic potential, it has also been the subject of considerable modern scientific attention. The major chemical constituents of the Withania genus, the Withanolides, are a group of naturally occurring C28-steroidal lactone triterpenoids built on an intact or rearranged ergostane framework, in which C-22 and C-26 are appropriately oxidized to form a six-membered lactone ring. In recent years, numerous pharmacological investigations have been carried out into the components of W. somnifera extracts. We present here an overview of the chemical structures of triterpenoid components and their biological activity, focusing on two novel activities, tumor inhibition and antiangiogenic properties of withaferin A and the effects of Withanolide A on Alzheimer's disease. The most recent attempts in biotechnological production of Withanolides are also discussed.

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  • LC-MS, GC-MS, and NMR Spectroscopy Based Evaluation of the Energy of Consciousness Healing Treated Withania somnifera (Ashwagandha) Root Extract
    2017
    Co-Authors: Trivedi, Mahendra Kumar, Nayak Gopal, Branton Alice, Trivedi Dahryn, Afaganis, Ariadne Esmene, Bader, Barbara Marie, Weekes, Brian A., Dumas, Daphne Luisa, Fiedler, Denise Marie, Smith, Dennille Mellesia
    Abstract:

    Withania somnifera root extract which contains Withanolides as major active constituent and it is used traditionally for the prevention and treatment of several diseases. The objective of the current study was to investigate the impact of The Trivedi Effect® - Consciousness Energy Healing Treatment on the structural properties of the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Consciousness Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in the control and Biofield Energy Treated samples, whereas the peak area% i.e. the relative amount of the phytoconstituents at respective retention time was significantly altered. The peak area% at Rt of 5.6, 6.8, 6.9, 7.2, 7.9, 8.4, 8.5, 8.6, and 9.2 minutes of the treated sample were increased significantly in the range of 1.46% to 253.06% compared to the control sample. In the contrary, the peak area% of the treated sample at Rt of 6.4 and 8.2 minutes were significantly decreased by 12.72% and 17.35%, respectively with respect to the control sample. A total of 16 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, dihydroWithanolide D, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 621, 489, 473, 767, 471, 505, and 992 at retention times of 6.4, 6.8, 7.2, 7.9, 8.2, 8.4, and 9.1 minutes, respectively with the help of LC-MS, GC-MS and NMR data of both the control and Biofield Energy Treated samples. The mass peak intensities of the Biofield Energy Treated sample were significantly changed in the range of -81.36% to 1720.90% compared with the control sample at the same retention time. These findings suggest that The Trivedi Effect® - Consciousness Energy Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging, and other chronic infections. Source: https://www.trivedieffect.com/science/lc-ms-gc-ms-and-nmr-spectroscopy-based-evaluation-of-the-energy-of-consciousness-healing-treated-withania-somnifera-ashwagandha-root-extract http://www.sciencepublishinggroup.com/journal/paperinfo?journalid=329&doi=10.11648/j.jddmc.20170302.1

  • LC-MS, GC-MS, and NMR Spectroscopy Based Evaluation of the Energy of Consciousness Healing Treated Withania somnifera (Ashwagandha) Root Extract
    2017
    Co-Authors: Trivedi, Mahendra Kumar, Nayak Gopal, Branton Alice, Trivedi Dahryn, Afaganis, Ariadne Esmene, Bader, Barbara Marie, Weekes, Brian A., Dumas, Daphne Luisa, Fiedler, Denise Marie, Smith, Dennille Mellesia
    Abstract:

    Withania somnifera root extract which contains Withanolides as major active constituent and it is used traditionally for the prevention and treatment of several diseases. The objective of the current study was to investigate the impact of The Trivedi Effect® - Consciousness Energy Healing Treatment on the structural properties of the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Consciousness Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in the control and Biofield Energy Treated samples, whereas the peak area% i.e. the relative amount of the phytoconstituents at respective retention time was significantly altered. The peak area% at Rt of 5.6, 6.8, 6.9, 7.2, 7.9, 8.4, 8.5, 8.6, and 9.2 minutes of the treated sample were increased significantly in the range of 1.46% to 253.06% compared to the control sample. In the contrary, the peak area% of the treated sample at Rt of 6.4 and 8.2 minutes were significantly decreased by 12.72% and 17.35%, respectively with respect to the control sample. A total of 16 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, dihydroWithanolide D, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 621, 489, 473, 767, 471, 505, and 992 at retention times of 6.4, 6.8, 7.2, 7.9, 8.2, 8.4, and 9.1 minutes, respectively with the help of LC-MS, GC-MS and NMR data of both the control and Biofield Energy Treated samples. The mass peak intensities of the Biofield Energy Treated sample were significantly changed in the range of -81.36% to 1720.90% compared with the control sample at the same retention time. These findings suggest that The Trivedi Effect® - Consciousness Energy Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging, and other chronic infections. https://www.trivedieffect.com/science/lc-ms-gc-ms-and-nmr-spectroscopy-based-evaluation-of-the-energy-of-consciousness-healing-treated-withania-somnifera-ashwagandha-root-extract http://www.sciencepublishinggroup.com/journal/paperinfo?journalid=329&doi=10.11648/j.jddmc.20170302.1

  • Evaluation of the Energy of Consciousness Healing Treated Withania Somnifera (Ashwagandha) Root Extract Using LC-MS, GC-MS, and NMR Spectroscopy
    2017
    Co-Authors: Balmer, Joy Angevin, Trivedi, Mahendra Kumar, Nayak Gopal, Branton Alice, Trivedi Dahryn
    Abstract:

    Withania somnifera (Ashwagandha) root extract is very useful herbal medicine since from ancient time. The current study designed to investigate the impact of The Trivedi Effect® - Energy of Consciousness Healing Treatment on the structural properties of the ashwagandha root hydroalcoholic extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Energy of Consciousness Healing Treatment remotely by seven renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in both the control and treated samples, whereas the peak area % at respective retention time was significantly altered. The peak area% of the treated sample at Rt of 5.3, 5.5, 6.4, 6.5, 6.8, 6.9, 7.1, 7.3, 7.8, 7.9, 8.0, 8.2, 8.4, 8.6, 8.8, 9.0, 9.1, and 10.1 minutes were significantly reduced by 0.57% to 38.10% compared to the control sample. In addition, the peak area% of the treated sample at Rt of 5.7, 6.7, and 8.1 minutes were significantly increased by 16.00%, 244.44% and 19.62%, respectively compared with the control sample. A total of 21 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A and Withanolide sulfoxide, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 569, 621, 489, 473, 767, 471, 505, 992, and 455 at retention times of 6.4, 6.5, 6.8, 7.1, 7.9, 8.1, 8.4, 9.1, and 10.1 minutes with the help of GC-MS and NMR data of both the control and Biofield Energy Treated samples. There were significant changes observed in the peak intensity values at the same retention time in the Biofield Energy Treated sample in the range of -75.32% to 108.51% compared with the control sample. These findings suggest that The Trivedi Effect® - Energy of Consciousness Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections

  • Evaluation of the Energy of Consciousness Healing Treated Withania Somnifera (Ashwagandha) Root Extract Using LC-MS, GC-MS, and NMR Spectroscopy
    2017
    Co-Authors: Trivedi, Mahendra Kumar, Balmer, Joy Angevin, Nayak Gopal, Branton Alice, Trivedi Dahryn, Balmer, Alan Joseph, Anagnos Dimitrius, Kinney, Janice Patricia, Holling, Joni Marie, Duprey-reed, Lauree Ann
    Abstract:

    Withania somnifera (Ashwagandha) root extract is very useful herbal medicine since from ancient time. The current study designed to investigate the impact of The Trivedi Effect® - Energy of Consciousness Healing Treatment on the structural properties of the ashwagandha root hydroalcoholic extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Energy of Consciousness Healing Treatment remotely by seven renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in both the control and treated samples, whereas the peak area % at respective retention time was significantly altered. The peak area% of the treated sample at Rt of 5.3, 5.5, 6.4, 6.5, 6.8, 6.9, 7.1, 7.3, 7.8, 7.9, 8.0, 8.2, 8.4, 8.6, 8.8, 9.0, 9.1, and 10.1 minutes were significantly reduced by 0.57% to 38.10% compared to the control sample. In addition, the peak area% of the treated sample at Rt of 5.7, 6.7, and 8.1 minutes were significantly increased by 16.00%, 244.44% and 19.62%, respectively compared with the control sample. A total of 21 Withanolides such as withanoside IV, coagulin Q, viscosa lactone B, Withanolide A, withaferin A, withanone, Withanolide D, ixocarpalactone A and Withanolide sulfoxide, ixocarpalactone A, Withanolide sulfoxide, Withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 569, 621, 489, 473, 767, 471, 505, 992, and 455 at retention times of 6.4, 6.5, 6.8, 7.1, 7.9, 8.1, 8.4, 9.1, and 10.1 minutes with the help of GC-MS and NMR data of both the control and Biofield Energy Treated samples. There were significant changes observed in the peak intensity values at the same retention time in the Biofield Energy Treated sample in the range of -75.32% to 108.51% compared with the control sample. These findings suggest that The Trivedi Effect® - Energy of Consciousness Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections. https://www.trivedieffect.com/science/evaluation-of-the-energy-of-consciousness-healing-treated-withania-somnifera-ashwagandha-root-extract-using-lc-ms-gc-ms-and-nmr-spectroscopy http://www.sciencepublishinggroup.com/journal/paperinfo?journalid=655&doi=10.11648/j.ajbls.20170502.1

  • Liquid Chromatography – Mass Spectrometry (LC-MS) Analysis of Withania somnifera (Ashwagandha) Root Extract Treated with the Energy of Consciousness
    2017
    Co-Authors: Starodub, Joanne Lydia, Trivedi, Mahendra Kumar, Nayak Gopal, Branton Alice, Trivedi Dahryn
    Abstract:

    Withania somnifera (ashwagandha) root extract is very popular ancient herbal medicine. The objective of the study was to characterize and evaluate the impact of The Trivedi Effect®-Biofield Energy Healing Treatment (Energy of Consciousness) on phytoconstituents present in the ashwagandha root extract using LC-MS. Ashwagandha root extract was divided into two parts. One part was denoted as the control, while the other part was defined as The Trivedi Effect® - Biofield Energy Treated sample, which received Energy of Consciousness Healing Treatment remotely from eighteen renowned Biofield Energy Healers. The LC-MS analysis of the control and treated samples showed a very close retention time (Rt), indicated that the polarity of the phytoconstituents present in the root extract are same. The numbers of peaks observed in the total ion chromatograms were 28 and 29 in the control and treated samples, respectively. The change in the peak height% of the phytoconstituents in the treated sample was altered significantly within the range of -50.91% to 118.12% compared with the control sample. Similarly, the change in the peak area% of most of the phytoconstituents in the treated ashwagandha was significantly altered within the range of -54.95% to 66.95% compared with the control sample. An additional peak was appeared in the treated sample at Rt of 5.72 minutes, which was not found in the control sample. The LC-MS spectra indicated the presence of possible Withanolides like -hydroxy-2,3-dihydro-Withanolide F, Withanolide A, withaferine A, withanone, Withanolide D, ixocarpalactone A, Withanolide S, thioWithanolide, etc. in both the samples. The peak are percentage (%) was altered in the identified Withanolides, but Withanolide sulfoxide was increased significantly by 12.44% in the treated sample compared with the control sample. These results indicated that The Trivedi Effect® - Biofield Energy Treatment might have an impact on the intrinsic physicochemical properties of the phytoconstituents present in the ashwagandha root extract. This could be the probable cause of alteration in the peak height, peak area, and appearance of a new peak in the treated sample. As a result, the concentrations of the phytoconstituents altered in the treated sample compared with the control sample. The treated ashwagandha root extract would be helpful for designing better pharmaceutical/nutraceutical formulations which might be providing a better therapeutic response against autoimmune diseases, nervous and sexual disorders, infectious diseases, antiaging, diabetes, cancer, ulcer, immunological disorders, stress, arthritis, etc

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