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Isolation and promoter analysis of a chalcone synthase gene PtrCHS4 from Populus trichocarpa.

Identifieur interne : 002E16 ( Main/Exploration ); précédent : 002E15; suivant : 002E17

Isolation and promoter analysis of a chalcone synthase gene PtrCHS4 from Populus trichocarpa.

Auteurs : Yiming Sun [République populaire de Chine] ; Qiaoyan Tian ; Li Yuan ; Yuanzhong Jiang ; Yan Huang ; Min Sun ; Shaohu Tang ; Keming Luo

Source :

RBID : pubmed:21553109

Descripteurs français

English descriptors

Abstract

As perennial plants, Populus species are constantly exposed to environmental stresses, such as wounding and pathogen attack, which lead to production of compounds including lignin, flavonoids and phytoalexins. Chalcone synthase (CHS) is a key enzyme in the flavonoid biosynthesis pathway. In this study, a cDNA clone encoding CHS was isolated from Populus trichocarpa by reverse transcription-polymerase chain reaction (RT-PCR). The full-length cDNA, named PtrCHS4, was 1,314 bp with a 1,173 bp open reading frame that corresponded to a deduced protein of 391 amino acid residues. Multiple sequence alignments showed that PtrCHS4 shared high homology with CHS proteins from other plants. Phylogenetic analysis revealed that PtrCHS4 was most closely related to PhCHS from Petunia hybrida and NaCHS from Nicotiana attenuata. Semi-quantitative RT-PCR analysis identified that the PtrCHS4 gene was abundantly expressed in the leaves and stems, while its expression was drastically reduced in the roots. Transcript abundance of PtrCHS4 was stimulated by 2.5-fold within 24 h of wounding treatment. Promoter analysis confirmed that the PtrCHS4 promoter was capable of directing expression of the GUS reporter in both wounded and unwounded leaves of transgenic Chinese white poplar (P. tomentosa Carr.), indicating that the PtrCHS4 promoter is systemically responsive to wounding stimuli. Furthermore, promoter deletion analysis showed that the proximal 1,592 bp from the transcription start site were required for promoter activation by jasmonic acid and the -1,096 to -148 region was proved to be necessary for establishing wound-induced pattern of expression.

DOI: 10.1007/s00299-011-1075-1
PubMed: 21553109


Affiliations:


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Le document en format XML

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<term>Acyltransferases (metabolism)</term>
<term>Amino Acid Sequence (MeSH)</term>
<term>Blotting, Southern (MeSH)</term>
<term>Cloning, Molecular (MeSH)</term>
<term>Cyclopentanes (pharmacology)</term>
<term>DNA, Complementary (genetics)</term>
<term>DNA, Complementary (isolation & purification)</term>
<term>Gene Expression Regulation, Plant (MeSH)</term>
<term>Gene Fusion (MeSH)</term>
<term>Genes, Plant (MeSH)</term>
<term>Genes, Reporter (MeSH)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Oxylipins (pharmacology)</term>
<term>Phylogeny (MeSH)</term>
<term>Plant Leaves (drug effects)</term>
<term>Plant Leaves (enzymology)</term>
<term>Plant Leaves (genetics)</term>
<term>Plant Leaves (physiology)</term>
<term>Plant Proteins (genetics)</term>
<term>Plant Proteins (metabolism)</term>
<term>Plant Roots (enzymology)</term>
<term>Plant Roots (genetics)</term>
<term>Plant Stems (enzymology)</term>
<term>Plant Stems (genetics)</term>
<term>Plants, Genetically Modified (drug effects)</term>
<term>Plants, Genetically Modified (enzymology)</term>
<term>Plants, Genetically Modified (genetics)</term>
<term>Plants, Genetically Modified (physiology)</term>
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<term>Populus (enzymology)</term>
<term>Populus (genetics)</term>
<term>Populus (physiology)</term>
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<term>Reverse Transcriptase Polymerase Chain Reaction (MeSH)</term>
<term>Sequence Alignment (MeSH)</term>
<term>Sequence Analysis, Protein (methods)</term>
<term>Stress, Physiological (MeSH)</term>
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<term>Feuilles de plante (génétique)</term>
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<term>Protéines végétales (métabolisme)</term>
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<term>Racines de plante (génétique)</term>
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<term>Séquence d'acides aminés (MeSH)</term>
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<term>DNA, Complementary</term>
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<term>Tiges de plante</term>
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<term>Plant Leaves</term>
<term>Plant Roots</term>
<term>Plant Stems</term>
<term>Plants, Genetically Modified</term>
<term>Populus</term>
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<term>Plant Leaves</term>
<term>Plant Roots</term>
<term>Plant Stems</term>
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<term>Tiges de plante</term>
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<term>Acyltransferases</term>
<term>Protéines végétales</term>
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<term>Analyse de séquence de protéine</term>
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<term>Cyclopentanes</term>
<term>Oxylipines</term>
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<term>Feuilles de plante</term>
<term>Populus</term>
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<keywords scheme="MESH" qualifier="physiology" xml:lang="en">
<term>Plant Leaves</term>
<term>Plants, Genetically Modified</term>
<term>Populus</term>
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<term>Amino Acid Sequence</term>
<term>Blotting, Southern</term>
<term>Cloning, Molecular</term>
<term>Gene Expression Regulation, Plant</term>
<term>Gene Fusion</term>
<term>Genes, Plant</term>
<term>Genes, Reporter</term>
<term>Molecular Sequence Data</term>
<term>Phylogeny</term>
<term>Promoter Regions, Genetic</term>
<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>Sequence Alignment</term>
<term>Stress, Physiological</term>
<term>Transcriptional Activation</term>
<term>Transformation, Genetic</term>
<term>Transgenes</term>
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<term>Alignement de séquences</term>
<term>Clonage moléculaire</term>
<term>Données de séquences moléculaires</term>
<term>Fusion de gènes</term>
<term>Gènes de plante</term>
<term>Gènes rapporteurs</term>
<term>Phylogenèse</term>
<term>RT-PCR</term>
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<term>Stress physiologique</term>
<term>Séquence d'acides aminés</term>
<term>Technique de Southern</term>
<term>Transformation génétique</term>
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<div type="abstract" xml:lang="en">As perennial plants, Populus species are constantly exposed to environmental stresses, such as wounding and pathogen attack, which lead to production of compounds including lignin, flavonoids and phytoalexins. Chalcone synthase (CHS) is a key enzyme in the flavonoid biosynthesis pathway. In this study, a cDNA clone encoding CHS was isolated from Populus trichocarpa by reverse transcription-polymerase chain reaction (RT-PCR). The full-length cDNA, named PtrCHS4, was 1,314 bp with a 1,173 bp open reading frame that corresponded to a deduced protein of 391 amino acid residues. Multiple sequence alignments showed that PtrCHS4 shared high homology with CHS proteins from other plants. Phylogenetic analysis revealed that PtrCHS4 was most closely related to PhCHS from Petunia hybrida and NaCHS from Nicotiana attenuata. Semi-quantitative RT-PCR analysis identified that the PtrCHS4 gene was abundantly expressed in the leaves and stems, while its expression was drastically reduced in the roots. Transcript abundance of PtrCHS4 was stimulated by 2.5-fold within 24 h of wounding treatment. Promoter analysis confirmed that the PtrCHS4 promoter was capable of directing expression of the GUS reporter in both wounded and unwounded leaves of transgenic Chinese white poplar (P. tomentosa Carr.), indicating that the PtrCHS4 promoter is systemically responsive to wounding stimuli. Furthermore, promoter deletion analysis showed that the proximal 1,592 bp from the transcription start site were required for promoter activation by jasmonic acid and the -1,096 to -148 region was proved to be necessary for establishing wound-induced pattern of expression.</div>
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<AbstractText>As perennial plants, Populus species are constantly exposed to environmental stresses, such as wounding and pathogen attack, which lead to production of compounds including lignin, flavonoids and phytoalexins. Chalcone synthase (CHS) is a key enzyme in the flavonoid biosynthesis pathway. In this study, a cDNA clone encoding CHS was isolated from Populus trichocarpa by reverse transcription-polymerase chain reaction (RT-PCR). The full-length cDNA, named PtrCHS4, was 1,314 bp with a 1,173 bp open reading frame that corresponded to a deduced protein of 391 amino acid residues. Multiple sequence alignments showed that PtrCHS4 shared high homology with CHS proteins from other plants. Phylogenetic analysis revealed that PtrCHS4 was most closely related to PhCHS from Petunia hybrida and NaCHS from Nicotiana attenuata. Semi-quantitative RT-PCR analysis identified that the PtrCHS4 gene was abundantly expressed in the leaves and stems, while its expression was drastically reduced in the roots. Transcript abundance of PtrCHS4 was stimulated by 2.5-fold within 24 h of wounding treatment. Promoter analysis confirmed that the PtrCHS4 promoter was capable of directing expression of the GUS reporter in both wounded and unwounded leaves of transgenic Chinese white poplar (P. tomentosa Carr.), indicating that the PtrCHS4 promoter is systemically responsive to wounding stimuli. Furthermore, promoter deletion analysis showed that the proximal 1,592 bp from the transcription start site were required for promoter activation by jasmonic acid and the -1,096 to -148 region was proved to be necessary for establishing wound-induced pattern of expression.</AbstractText>
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