Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene Regulation

Bradyrhizobium japonicum mutants with altered nod gene induction characteristics were isolated by screening mutants for genistein-independent nod gene expression. Plasmid pZB32, carrying a nodY∷lacZ transcriptional gene fusion, was introduced into B. japonicum cells that had been subjected to UV mut...

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Main Authors: H. Ip, F. D'Aoust, A. A. Begum, H. Zhang, D. L. Smith, B. T. Driscoll, T. C. Charles
Format: Article
Language:English
Published: The American Phytopathological Society 2001-12-01
Series:Molecular Plant-Microbe Interactions
Online Access:https://apsjournals.apsnet.org/doi/10.1094/MPMI.2001.14.12.1404
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author H. Ip
F. D'Aoust
A. A. Begum
H. Zhang
D. L. Smith
B. T. Driscoll
T. C. Charles
author_facet H. Ip
F. D'Aoust
A. A. Begum
H. Zhang
D. L. Smith
B. T. Driscoll
T. C. Charles
author_sort H. Ip
collection DOAJ
description Bradyrhizobium japonicum mutants with altered nod gene induction characteristics were isolated by screening mutants for genistein-independent nod gene expression. Plasmid pZB32, carrying a nodY∷lacZ transcriptional gene fusion, was introduced into B. japonicum cells that had been subjected to UV mutagenesis. Ten independent transformants producing a blue color on plates containing 5-bromo-4chloro-3indolyl-β-d-galactopyranoside but lacking genistein, indicative of constitutive expression of the nodY∷lacZ reporter gene, were isolated. β-Galactosidase activity assays revealed that while all of the 10 strains were sensitive to low concentrations of genistein, none exhibited truly constitutive nodY∷lacZ expression in liquid culture. Soybean plants inoculated with three of the mutants were chlorotic and stunted, with shoot dry weights close to those of the uninoculated plants, indicating the absence of nitrogen fixation. Differences in the kinetics of nodY∷lacZ expression and lipochitin oligosaccharide Nod signal production suggested that the strains carried different mutations. Some of these strains may be useful in mitigating the low root zone temperature-associated delay in soybean nodulation at the northern extent of soybean cultivation.
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spelling doaj.art-04cd2c06014e49e4a8d13b1e149a21782022-12-22T03:04:56ZengThe American Phytopathological SocietyMolecular Plant-Microbe Interactions0894-02821943-77062001-12-0114121404141010.1094/MPMI.2001.14.12.1404Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene RegulationH. IpF. D'AoustA. A. BegumH. ZhangD. L. SmithB. T. DriscollT. C. CharlesBradyrhizobium japonicum mutants with altered nod gene induction characteristics were isolated by screening mutants for genistein-independent nod gene expression. Plasmid pZB32, carrying a nodY∷lacZ transcriptional gene fusion, was introduced into B. japonicum cells that had been subjected to UV mutagenesis. Ten independent transformants producing a blue color on plates containing 5-bromo-4chloro-3indolyl-β-d-galactopyranoside but lacking genistein, indicative of constitutive expression of the nodY∷lacZ reporter gene, were isolated. β-Galactosidase activity assays revealed that while all of the 10 strains were sensitive to low concentrations of genistein, none exhibited truly constitutive nodY∷lacZ expression in liquid culture. Soybean plants inoculated with three of the mutants were chlorotic and stunted, with shoot dry weights close to those of the uninoculated plants, indicating the absence of nitrogen fixation. Differences in the kinetics of nodY∷lacZ expression and lipochitin oligosaccharide Nod signal production suggested that the strains carried different mutations. Some of these strains may be useful in mitigating the low root zone temperature-associated delay in soybean nodulation at the northern extent of soybean cultivation.https://apsjournals.apsnet.org/doi/10.1094/MPMI.2001.14.12.1404
spellingShingle H. Ip
F. D'Aoust
A. A. Begum
H. Zhang
D. L. Smith
B. T. Driscoll
T. C. Charles
Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene Regulation
Molecular Plant-Microbe Interactions
title Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene Regulation
title_full Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene Regulation
title_fullStr Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene Regulation
title_full_unstemmed Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene Regulation
title_short Bradyrhizobium japonicum Mutants with Enhanced Sensitivity to Genistein Resulting in Altered nod Gene Regulation
title_sort bradyrhizobium japonicum mutants with enhanced sensitivity to genistein resulting in altered nod gene regulation
url https://apsjournals.apsnet.org/doi/10.1094/MPMI.2001.14.12.1404
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