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Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua

Artemisia annua, an annual herb used in traditional Chinese medicine, produces a wealth of monoterpenes and sesquiterpenes, including the well-known sesquiterpene lactone artemisinin, an active ingredient in the treatment for malaria. Here we report three new monoterpene synthases of A. annua. From...

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Autores principales: Ruan, Ju-Xin, Li, Jian-Xu, Fang, Xin, Wang, Ling-Jian, Hu, Wen-Li, Chen, Xiao-Ya, Yang, Chang-Qing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4861830/
https://www.ncbi.nlm.nih.gov/pubmed/27242840
http://dx.doi.org/10.3389/fpls.2016.00638
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author Ruan, Ju-Xin
Li, Jian-Xu
Fang, Xin
Wang, Ling-Jian
Hu, Wen-Li
Chen, Xiao-Ya
Yang, Chang-Qing
author_facet Ruan, Ju-Xin
Li, Jian-Xu
Fang, Xin
Wang, Ling-Jian
Hu, Wen-Li
Chen, Xiao-Ya
Yang, Chang-Qing
author_sort Ruan, Ju-Xin
collection PubMed
description Artemisia annua, an annual herb used in traditional Chinese medicine, produces a wealth of monoterpenes and sesquiterpenes, including the well-known sesquiterpene lactone artemisinin, an active ingredient in the treatment for malaria. Here we report three new monoterpene synthases of A. annua. From a glandular trichome cDNA library, monoterpene synthases of AaTPS2, AaTPS5, and AaTPS6, were isolated and characterized. The recombinant proteins of AaTPS5 and AaTPS6 produced multiple products with camphene and 1,8-cineole as major products, respectively, and AaTPS2 produced a single product, β-myrcene. Although both Mg(2+) and Mn(2+) were able to support their catalytic activities, altered product spectrum was observed in the presence of Mn(2+) for AaTPS2 and AaTPS5. Analysis of extracts of aerial tissues and root of A. annua with gas chromatography–mass spectrometry detected more than 20 monoterpenes, of which the three enzymes constituted more than 1/3 of the total. Mechanical wounding induced the expression of all three monoterpene synthase genes, and transcript levels of AaTPS5 and AaTPS6 were also elevated after treatments with phytohormones of methyl jasmonate, salicylic acid, and gibberellin, suggesting a role of these monoterpene synthases in plant–environment interactions. The three new monoterpene synthases reported here further our understanding of molecular basis of monoterpene biosynthesis and regulation in plant.
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spelling pubmed-48618302016-05-30 Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua Ruan, Ju-Xin Li, Jian-Xu Fang, Xin Wang, Ling-Jian Hu, Wen-Li Chen, Xiao-Ya Yang, Chang-Qing Front Plant Sci Plant Science Artemisia annua, an annual herb used in traditional Chinese medicine, produces a wealth of monoterpenes and sesquiterpenes, including the well-known sesquiterpene lactone artemisinin, an active ingredient in the treatment for malaria. Here we report three new monoterpene synthases of A. annua. From a glandular trichome cDNA library, monoterpene synthases of AaTPS2, AaTPS5, and AaTPS6, were isolated and characterized. The recombinant proteins of AaTPS5 and AaTPS6 produced multiple products with camphene and 1,8-cineole as major products, respectively, and AaTPS2 produced a single product, β-myrcene. Although both Mg(2+) and Mn(2+) were able to support their catalytic activities, altered product spectrum was observed in the presence of Mn(2+) for AaTPS2 and AaTPS5. Analysis of extracts of aerial tissues and root of A. annua with gas chromatography–mass spectrometry detected more than 20 monoterpenes, of which the three enzymes constituted more than 1/3 of the total. Mechanical wounding induced the expression of all three monoterpene synthase genes, and transcript levels of AaTPS5 and AaTPS6 were also elevated after treatments with phytohormones of methyl jasmonate, salicylic acid, and gibberellin, suggesting a role of these monoterpene synthases in plant–environment interactions. The three new monoterpene synthases reported here further our understanding of molecular basis of monoterpene biosynthesis and regulation in plant. Frontiers Media S.A. 2016-05-10 /pmc/articles/PMC4861830/ /pubmed/27242840 http://dx.doi.org/10.3389/fpls.2016.00638 Text en Copyright © 2016 Ruan, Li, Fang, Wang, Hu, Chen and Yang. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Ruan, Ju-Xin
Li, Jian-Xu
Fang, Xin
Wang, Ling-Jian
Hu, Wen-Li
Chen, Xiao-Ya
Yang, Chang-Qing
Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua
title Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua
title_full Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua
title_fullStr Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua
title_full_unstemmed Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua
title_short Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua
title_sort isolation and characterization of three new monoterpene synthases from artemisia annua
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4861830/
https://www.ncbi.nlm.nih.gov/pubmed/27242840
http://dx.doi.org/10.3389/fpls.2016.00638
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