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Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis
Squalene epoxidase (SQE) is a crucial regulatory enzyme for the biosynthesis of several important classes of compounds including sterols and triterpenoids. The present paper identified and characterised five SQE genes (GgSQE1 to GgSQE5) from Glycyrrhiza glabra through transcriptome data mining and h...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Vienna
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7904238/ https://www.ncbi.nlm.nih.gov/pubmed/33629144 http://dx.doi.org/10.1007/s00709-021-01616-2 |
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author | Manzoor, Malik Muzafar Goyal, Pooja Pandotra, Pankaj Dar, Mohd Saleem Dar, Mohd Jamal Misra, Prashant Gupta, Ajai P. Vishwakarma, Ram A. Ahuja, Ashok Dhar, Manoj K. Gupta, Suphla |
author_facet | Manzoor, Malik Muzafar Goyal, Pooja Pandotra, Pankaj Dar, Mohd Saleem Dar, Mohd Jamal Misra, Prashant Gupta, Ajai P. Vishwakarma, Ram A. Ahuja, Ashok Dhar, Manoj K. Gupta, Suphla |
author_sort | Manzoor, Malik Muzafar |
collection | PubMed |
description | Squalene epoxidase (SQE) is a crucial regulatory enzyme for the biosynthesis of several important classes of compounds including sterols and triterpenoids. The present paper identified and characterised five SQE genes (GgSQE1 to GgSQE5) from Glycyrrhiza glabra through transcriptome data mining and homology-based cloning, for the first time. The phylogenetic analysis implied their functional divergence. The ORF corresponding to one of the five SQEs, namely, GgSQE1, was cloned and studied for its function in a heterologous system, following transient and stable expressions. The transient expression followed by GgSQE1 encoding protein purification suggested approximately 58.0-kDa protein following the predicted molecular mass of the deduced protein. The gene expression profiling based on qRT-PCR indicated its highest expression (6.4-folds) in the 10-month-old roots. Furthermore, ABA (12.4-folds) and GA(3) (2.47) treatments upregulated the expression of GgSQE1 in the shoots after 10 and 12 hours, respectively, which was also reflected in glycyrrhizin accumulation. The inductive effects of ABA and GA(3) over GgSQE1 expression were also confirmed through functional analysis of GgSQE1 promoters using GUS fusion construct. Stable constitutive expression of GgSQE1 in Nicotiana tabacum modulated the sterol contents. The study could pave the way for understanding the metabolic flux regulation concerning biosynthesis of related sterols and triterpenoids. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00709-021-01616-2. |
format | Online Article Text |
id | pubmed-7904238 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Vienna |
record_format | MEDLINE/PubMed |
spelling | pubmed-79042382021-02-25 Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis Manzoor, Malik Muzafar Goyal, Pooja Pandotra, Pankaj Dar, Mohd Saleem Dar, Mohd Jamal Misra, Prashant Gupta, Ajai P. Vishwakarma, Ram A. Ahuja, Ashok Dhar, Manoj K. Gupta, Suphla Protoplasma Original Article Squalene epoxidase (SQE) is a crucial regulatory enzyme for the biosynthesis of several important classes of compounds including sterols and triterpenoids. The present paper identified and characterised five SQE genes (GgSQE1 to GgSQE5) from Glycyrrhiza glabra through transcriptome data mining and homology-based cloning, for the first time. The phylogenetic analysis implied their functional divergence. The ORF corresponding to one of the five SQEs, namely, GgSQE1, was cloned and studied for its function in a heterologous system, following transient and stable expressions. The transient expression followed by GgSQE1 encoding protein purification suggested approximately 58.0-kDa protein following the predicted molecular mass of the deduced protein. The gene expression profiling based on qRT-PCR indicated its highest expression (6.4-folds) in the 10-month-old roots. Furthermore, ABA (12.4-folds) and GA(3) (2.47) treatments upregulated the expression of GgSQE1 in the shoots after 10 and 12 hours, respectively, which was also reflected in glycyrrhizin accumulation. The inductive effects of ABA and GA(3) over GgSQE1 expression were also confirmed through functional analysis of GgSQE1 promoters using GUS fusion construct. Stable constitutive expression of GgSQE1 in Nicotiana tabacum modulated the sterol contents. The study could pave the way for understanding the metabolic flux regulation concerning biosynthesis of related sterols and triterpenoids. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00709-021-01616-2. Springer Vienna 2021-02-24 2021 /pmc/articles/PMC7904238/ /pubmed/33629144 http://dx.doi.org/10.1007/s00709-021-01616-2 Text en © The Author(s), under exclusive licence to Springer-Verlag GmbH, AT part of Springer Nature 2021 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Original Article Manzoor, Malik Muzafar Goyal, Pooja Pandotra, Pankaj Dar, Mohd Saleem Dar, Mohd Jamal Misra, Prashant Gupta, Ajai P. Vishwakarma, Ram A. Ahuja, Ashok Dhar, Manoj K. Gupta, Suphla Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis |
title | Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis |
title_full | Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis |
title_fullStr | Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis |
title_full_unstemmed | Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis |
title_short | Transcriptome-wide identification of squalene epoxidase genes from Glycyrrhiza glabra L.: expression analysis and heterologous expression of GgSQE1 suggest important role in terpenoid biosynthesis |
title_sort | transcriptome-wide identification of squalene epoxidase genes from glycyrrhiza glabra l.: expression analysis and heterologous expression of ggsqe1 suggest important role in terpenoid biosynthesis |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7904238/ https://www.ncbi.nlm.nih.gov/pubmed/33629144 http://dx.doi.org/10.1007/s00709-021-01616-2 |
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