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Overexpression of a developing xylem cDNA library in transgenic poplar generates high mutation rate specific to wood formation.

Identifieur interne : 000234 ( Main/Exploration ); précédent : 000233; suivant : 000235

Overexpression of a developing xylem cDNA library in transgenic poplar generates high mutation rate specific to wood formation.

Auteurs : James Rauschendorfer [États-Unis] ; Yordan Yordanov [États-Unis] ; Petre Dobrev [République tchèque] ; Radomira Vankova [République tchèque] ; Robert Sykes [États-Unis] ; Carsten Külheim [États-Unis] ; Victor Busov [États-Unis]

Source :

RBID : pubmed:31799778

Descripteurs français

English descriptors

Abstract

We investigated feasibility of the Full-length complementary DNA OvereXpression (FOX) system as a mutagenesis approach in poplar, using developing xylem tissue. The main goal was to assess the overall mutation rate and if the system will increase instances of mutants affected in traits linked to the xylem tissue. Indeed, we found a high mutation rate of 17.7%, whereas 80% of all mutants were significantly affected in cellulose, lignin and/or hemicellulose. Cell wall biosynthesis is a major process occurring during xylem development. Enrichment of mutants affected in cell wall composition suggests that the tissue source for the FOX library influenced the occurrence of mutants affected in a trait linked to this tissue. Additionally, we found that FLcDNAs from mutants affected in cell wall composition were homologous to genes known to be involved in cell wall biosynthesis and most recovered FLcDNAs corresponded to genes whose native expression was highest in xylem. We characterized in detail a mutant line with increased diameter. The phenotype was caused by a poplar homolog of LONELY GUY 1 (LOG1), which encodes an enzyme in cytokinin biosynthesis and significantly increased xylem proliferation. The causative role of LOG1 in the observed phenotype was further reaffirmed by elevated cytokinin concentration in the mutant and recapitulation overexpression experiment wherein multiple independent lines phenocopied the original FOX mutant. Our experiments show that the FOX approach can be efficiently used for gene discovery and molecular interrogation of traits specific to woody perennial growth and development.

DOI: 10.1111/pbi.13309
PubMed: 31799778
PubMed Central: PMC7207001


Affiliations:


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Le document en format XML

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<term>Cell Wall (genetics)</term>
<term>Cell Wall (metabolism)</term>
<term>Gene Expression Regulation, Plant (genetics)</term>
<term>Gene Library (MeSH)</term>
<term>Lignin (metabolism)</term>
<term>Mutation Rate (MeSH)</term>
<term>Plant Proteins (genetics)</term>
<term>Plant Proteins (metabolism)</term>
<term>Populus (genetics)</term>
<term>Populus (metabolism)</term>
<term>Wood (genetics)</term>
<term>Wood (metabolism)</term>
<term>Xylem (genetics)</term>
<term>Xylem (metabolism)</term>
</keywords>
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<term>Banque de gènes (MeSH)</term>
<term>Bois (génétique)</term>
<term>Bois (métabolisme)</term>
<term>Lignine (métabolisme)</term>
<term>Paroi cellulaire (génétique)</term>
<term>Paroi cellulaire (métabolisme)</term>
<term>Populus (génétique)</term>
<term>Populus (métabolisme)</term>
<term>Protéines végétales (génétique)</term>
<term>Protéines végétales (métabolisme)</term>
<term>Régulation de l'expression des gènes végétaux (génétique)</term>
<term>Taux de mutation (MeSH)</term>
<term>Xylème (génétique)</term>
<term>Xylème (métabolisme)</term>
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<term>Plant Proteins</term>
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<term>Plant Proteins</term>
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<term>Cell Wall</term>
<term>Gene Expression Regulation, Plant</term>
<term>Populus</term>
<term>Wood</term>
<term>Xylem</term>
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<term>Bois</term>
<term>Paroi cellulaire</term>
<term>Populus</term>
<term>Protéines végétales</term>
<term>Régulation de l'expression des gènes végétaux</term>
<term>Xylème</term>
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<term>Cell Wall</term>
<term>Populus</term>
<term>Wood</term>
<term>Xylem</term>
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<term>Mutation Rate</term>
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<div type="abstract" xml:lang="en">We investigated feasibility of the Full-length complementary DNA OvereXpression (FOX) system as a mutagenesis approach in poplar, using developing xylem tissue. The main goal was to assess the overall mutation rate and if the system will increase instances of mutants affected in traits linked to the xylem tissue. Indeed, we found a high mutation rate of 17.7%, whereas 80% of all mutants were significantly affected in cellulose, lignin and/or hemicellulose. Cell wall biosynthesis is a major process occurring during xylem development. Enrichment of mutants affected in cell wall composition suggests that the tissue source for the FOX library influenced the occurrence of mutants affected in a trait linked to this tissue. Additionally, we found that FLcDNAs from mutants affected in cell wall composition were homologous to genes known to be involved in cell wall biosynthesis and most recovered FLcDNAs corresponded to genes whose native expression was highest in xylem. We characterized in detail a mutant line with increased diameter. The phenotype was caused by a poplar homolog of LONELY GUY 1 (LOG1), which encodes an enzyme in cytokinin biosynthesis and significantly increased xylem proliferation. The causative role of LOG1 in the observed phenotype was further reaffirmed by elevated cytokinin concentration in the mutant and recapitulation overexpression experiment wherein multiple independent lines phenocopied the original FOX mutant. Our experiments show that the FOX approach can be efficiently used for gene discovery and molecular interrogation of traits specific to woody perennial growth and development.</div>
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<AbstractText>We investigated feasibility of the Full-length complementary DNA OvereXpression (FOX) system as a mutagenesis approach in poplar, using developing xylem tissue. The main goal was to assess the overall mutation rate and if the system will increase instances of mutants affected in traits linked to the xylem tissue. Indeed, we found a high mutation rate of 17.7%, whereas 80% of all mutants were significantly affected in cellulose, lignin and/or hemicellulose. Cell wall biosynthesis is a major process occurring during xylem development. Enrichment of mutants affected in cell wall composition suggests that the tissue source for the FOX library influenced the occurrence of mutants affected in a trait linked to this tissue. Additionally, we found that FLcDNAs from mutants affected in cell wall composition were homologous to genes known to be involved in cell wall biosynthesis and most recovered FLcDNAs corresponded to genes whose native expression was highest in xylem. We characterized in detail a mutant line with increased diameter. The phenotype was caused by a poplar homolog of LONELY GUY 1 (LOG1), which encodes an enzyme in cytokinin biosynthesis and significantly increased xylem proliferation. The causative role of LOG1 in the observed phenotype was further reaffirmed by elevated cytokinin concentration in the mutant and recapitulation overexpression experiment wherein multiple independent lines phenocopied the original FOX mutant. Our experiments show that the FOX approach can be efficiently used for gene discovery and molecular interrogation of traits specific to woody perennial growth and development.</AbstractText>
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