Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i>
In the methyl-D-erythritol-4-phosphate (MEP) pathway, 1-deoxy-D-xylose-5-phosphate synthase (DXS) is considered the key enzyme for the biosynthesis of terpenoids. In this study, <i>PmDXS </i>(MK970590) was isolated from <i>Pinus massoniana</i>. Bioinformatics analysis reveale...
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doaj-06b5eedb47d74b14846d2aff17e0d6b02021-01-16T00:06:16ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-01-012284884810.3390/ijms22020848Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i>Rong Li0Peizhen Chen1Lingzhi Zhu2Fan Wu3Yu Chen4Peihuang Zhu5Kongshu Ji6Key Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, ChinaKey Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, ChinaKey Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, ChinaKey Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, ChinaKey Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, ChinaKey Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, ChinaKey Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, ChinaIn the methyl-D-erythritol-4-phosphate (MEP) pathway, 1-deoxy-D-xylose-5-phosphate synthase (DXS) is considered the key enzyme for the biosynthesis of terpenoids. In this study, <i>PmDXS </i>(MK970590) was isolated from <i>Pinus massoniana</i>. Bioinformatics analysis revealed homology of MK970590 with DXS proteins from other species. Relative expression analysis suggested that<i> PmDXS</i> expression was higher in roots than in other plant parts, and the treatment of <i>P. massoniana</i> seedlings with mechanical injury via 15% polyethylene glycol 6000, 10 mM H<sub>2</sub>O<sub>2</sub>, 50 μM ethephon (ETH), 10 mM methyl jasmonate (MeJA), and 1 mM salicylic acid (SA) resulted in an increased expression of <i>PmDXS</i>. pET28a-<i>PmDXS</i> was expressed in <i>Escherichia coli </i>TransB (DE3) cells, and stress analysis showed that the recombinant protein was involved in resistance to NaCl and drought stresses. The subcellular localization of <i>PmDXS</i> was in the chloroplast. We also cloned a full-length 1024 bp <i>PmDXS </i>promoter. <i>GUS</i> expression was observed in <i>Nicotiana benthamiana</i> roots, stems, and leaves. <i>PmDXS </i>overexpression significantly increased carotenoid, chlorophyll a, and chlorophyll b contents and DXS enzyme activity, suggesting that <i>DXS</i> is important in isoprenoid biosynthesis. This study provides a theoretical basis for molecular breeding for terpene synthesis regulation and resistance.https://www.mdpi.com/1422-0067/22/2/848<i>Pinus massoniana</i>DXSexpression patternterpenoidprokaryotic expressionchlorophyll |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Rong Li Peizhen Chen Lingzhi Zhu Fan Wu Yu Chen Peihuang Zhu Kongshu Ji |
spellingShingle |
Rong Li Peizhen Chen Lingzhi Zhu Fan Wu Yu Chen Peihuang Zhu Kongshu Ji Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i> International Journal of Molecular Sciences <i>Pinus massoniana</i> DXS expression pattern terpenoid prokaryotic expression chlorophyll |
author_facet |
Rong Li Peizhen Chen Lingzhi Zhu Fan Wu Yu Chen Peihuang Zhu Kongshu Ji |
author_sort |
Rong Li |
title |
Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i> |
title_short |
Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i> |
title_full |
Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i> |
title_fullStr |
Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i> |
title_full_unstemmed |
Characterization and Function of the 1-Deoxy-D-xylose-5-Phosphate Synthase (DXS) Gene Related to Terpenoid Synthesis in <i>Pinus massoniana</i> |
title_sort |
characterization and function of the 1-deoxy-d-xylose-5-phosphate synthase (dxs) gene related to terpenoid synthesis in <i>pinus massoniana</i> |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-01-01 |
description |
In the methyl-D-erythritol-4-phosphate (MEP) pathway, 1-deoxy-D-xylose-5-phosphate synthase (DXS) is considered the key enzyme for the biosynthesis of terpenoids. In this study, <i>PmDXS </i>(MK970590) was isolated from <i>Pinus massoniana</i>. Bioinformatics analysis revealed homology of MK970590 with DXS proteins from other species. Relative expression analysis suggested that<i> PmDXS</i> expression was higher in roots than in other plant parts, and the treatment of <i>P. massoniana</i> seedlings with mechanical injury via 15% polyethylene glycol 6000, 10 mM H<sub>2</sub>O<sub>2</sub>, 50 μM ethephon (ETH), 10 mM methyl jasmonate (MeJA), and 1 mM salicylic acid (SA) resulted in an increased expression of <i>PmDXS</i>. pET28a-<i>PmDXS</i> was expressed in <i>Escherichia coli </i>TransB (DE3) cells, and stress analysis showed that the recombinant protein was involved in resistance to NaCl and drought stresses. The subcellular localization of <i>PmDXS</i> was in the chloroplast. We also cloned a full-length 1024 bp <i>PmDXS </i>promoter. <i>GUS</i> expression was observed in <i>Nicotiana benthamiana</i> roots, stems, and leaves. <i>PmDXS </i>overexpression significantly increased carotenoid, chlorophyll a, and chlorophyll b contents and DXS enzyme activity, suggesting that <i>DXS</i> is important in isoprenoid biosynthesis. This study provides a theoretical basis for molecular breeding for terpene synthesis regulation and resistance. |
topic |
<i>Pinus massoniana</i> DXS expression pattern terpenoid prokaryotic expression chlorophyll |
url |
https://www.mdpi.com/1422-0067/22/2/848 |
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