Generation of Bradyrhizobium japonicum Mutants with Increased N 2 O Reductase Activity by Selection after Introduction of a Mutated dnaQ Gene
We obtained two beneficial mutants of Bradyrhizobium japonicum USDA110 with increased nitrous oxide (N 2 O) reductase (N 2 OR) activity by introducing a plasmid containing a mutated B. japonicum dnaQ gene (pKQ2) and performing enrichment culture under selection pressure for N 2 O respiration. Mutati...
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Veröffentlicht in: | Applied and environmental microbiology 2008-12, Vol.74 (23), p.7258-7264 |
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creator | Itakura, Manabu Tabata, Kazufumi Eda, Shima Mitsui, Hisayuki Murakami, Kiriko Yasuda, Junichi Minamisawa, Kiwamu |
description | We obtained two beneficial mutants of
Bradyrhizobium japonicum
USDA110 with increased nitrous oxide (N
2
O) reductase (N
2
OR) activity by introducing a plasmid containing a mutated
B. japonicum dnaQ
gene (pKQ2) and performing enrichment culture under selection pressure for N
2
O respiration. Mutation of
dnaQ
, which encodes the epsilon subunit of DNA polymerase III, gives a strong mutator phenotype in
Escherichia coli
. pKQ2 introduction into
B. japonicum
USDA110 increased the frequency of occurrence of colonies spontaneously resistant to kanamycin. A series of repeated cultivations of USDA110 with and without pKQ2 was conducted in anaerobic conditions under 5% (vol/vol) or 20% (vol/vol) N
2
O atmosphere. At the 10th cultivation cycle, cell populations of USDA110(pKQ2) showed higher N
2
OR activity than the wild-type strains. Four bacterial mutants lacking pKQ2 obtained by plant passage showed 7 to 12 times the N
2
OR activity of the wild-type USDA110. Although two mutants had a weak or null
fix
phenotype for symbiotic nitrogen fixation, the remaining two (5M09 and 5M14) had the same symbiotic nitrogen fixation ability and heterotrophic growth in culture as wild-type USDA110. |
doi_str_mv | 10.1128/AEM.01850-08 |
format | Article |
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Bradyrhizobium japonicum
USDA110 with increased nitrous oxide (N
2
O) reductase (N
2
OR) activity by introducing a plasmid containing a mutated
B. japonicum dnaQ
gene (pKQ2) and performing enrichment culture under selection pressure for N
2
O respiration. Mutation of
dnaQ
, which encodes the epsilon subunit of DNA polymerase III, gives a strong mutator phenotype in
Escherichia coli
. pKQ2 introduction into
B. japonicum
USDA110 increased the frequency of occurrence of colonies spontaneously resistant to kanamycin. A series of repeated cultivations of USDA110 with and without pKQ2 was conducted in anaerobic conditions under 5% (vol/vol) or 20% (vol/vol) N
2
O atmosphere. At the 10th cultivation cycle, cell populations of USDA110(pKQ2) showed higher N
2
OR activity than the wild-type strains. Four bacterial mutants lacking pKQ2 obtained by plant passage showed 7 to 12 times the N
2
OR activity of the wild-type USDA110. Although two mutants had a weak or null
fix
phenotype for symbiotic nitrogen fixation, the remaining two (5M09 and 5M14) had the same symbiotic nitrogen fixation ability and heterotrophic growth in culture as wild-type USDA110.</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>DOI: 10.1128/AEM.01850-08</identifier><language>eng</language><ispartof>Applied and environmental microbiology, 2008-12, Vol.74 (23), p.7258-7264</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c1188-d82d82388b521c823ae4373e6db40d00f0564e04ca9ab93b8c964a287d9627743</citedby><cites>FETCH-LOGICAL-c1188-d82d82388b521c823ae4373e6db40d00f0564e04ca9ab93b8c964a287d9627743</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,3189,27929,27930</link.rule.ids></links><search><creatorcontrib>Itakura, Manabu</creatorcontrib><creatorcontrib>Tabata, Kazufumi</creatorcontrib><creatorcontrib>Eda, Shima</creatorcontrib><creatorcontrib>Mitsui, Hisayuki</creatorcontrib><creatorcontrib>Murakami, Kiriko</creatorcontrib><creatorcontrib>Yasuda, Junichi</creatorcontrib><creatorcontrib>Minamisawa, Kiwamu</creatorcontrib><title>Generation of Bradyrhizobium japonicum Mutants with Increased N 2 O Reductase Activity by Selection after Introduction of a Mutated dnaQ Gene</title><title>Applied and environmental microbiology</title><description>We obtained two beneficial mutants of
Bradyrhizobium japonicum
USDA110 with increased nitrous oxide (N
2
O) reductase (N
2
OR) activity by introducing a plasmid containing a mutated
B. japonicum dnaQ
gene (pKQ2) and performing enrichment culture under selection pressure for N
2
O respiration. Mutation of
dnaQ
, which encodes the epsilon subunit of DNA polymerase III, gives a strong mutator phenotype in
Escherichia coli
. pKQ2 introduction into
B. japonicum
USDA110 increased the frequency of occurrence of colonies spontaneously resistant to kanamycin. A series of repeated cultivations of USDA110 with and without pKQ2 was conducted in anaerobic conditions under 5% (vol/vol) or 20% (vol/vol) N
2
O atmosphere. At the 10th cultivation cycle, cell populations of USDA110(pKQ2) showed higher N
2
OR activity than the wild-type strains. Four bacterial mutants lacking pKQ2 obtained by plant passage showed 7 to 12 times the N
2
OR activity of the wild-type USDA110. Although two mutants had a weak or null
fix
phenotype for symbiotic nitrogen fixation, the remaining two (5M09 and 5M14) had the same symbiotic nitrogen fixation ability and heterotrophic growth in culture as wild-type USDA110.</description><issn>0099-2240</issn><issn>1098-5336</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><recordid>eNotkN1KAzEQhYMoWKt3PkAewK2Tn91NLmuptVAt_l0v2SRLU9rdkqTK-g6-s2ktDMxhmPnOcBC6JTAihIr78fR5BETkkIE4QwMCUmQ5Y8U5GgBImVHK4RJdhbAGAA6FGKDfmW2tV9F1Le4a_OCV6f3K_XS122_xWu261umknvdRtTHgbxdXeN5qb1WwBr9gipf4zZq9jmmAxzq6Lxd7XPf43W6sPoJVE61PV9F3h8WTlzpCY6KYVr3iwyPX6KJRm2BvTn2IPh-nH5OnbLGczSfjRaYJESIzgqZiQtQ5JTopZTkrmS1MzcEANJAX3ALXSqpaslpoWXBFRWlkQcuSsyG6--dq34XgbVPtvNsq31cEqkOUVYqyOkZZgWB_uTxnhw</recordid><startdate>200812</startdate><enddate>200812</enddate><creator>Itakura, Manabu</creator><creator>Tabata, Kazufumi</creator><creator>Eda, Shima</creator><creator>Mitsui, Hisayuki</creator><creator>Murakami, Kiriko</creator><creator>Yasuda, Junichi</creator><creator>Minamisawa, Kiwamu</creator><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>200812</creationdate><title>Generation of Bradyrhizobium japonicum Mutants with Increased N 2 O Reductase Activity by Selection after Introduction of a Mutated dnaQ Gene</title><author>Itakura, Manabu ; Tabata, Kazufumi ; Eda, Shima ; Mitsui, Hisayuki ; Murakami, Kiriko ; Yasuda, Junichi ; Minamisawa, Kiwamu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c1188-d82d82388b521c823ae4373e6db40d00f0564e04ca9ab93b8c964a287d9627743</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Itakura, Manabu</creatorcontrib><creatorcontrib>Tabata, Kazufumi</creatorcontrib><creatorcontrib>Eda, Shima</creatorcontrib><creatorcontrib>Mitsui, Hisayuki</creatorcontrib><creatorcontrib>Murakami, Kiriko</creatorcontrib><creatorcontrib>Yasuda, Junichi</creatorcontrib><creatorcontrib>Minamisawa, Kiwamu</creatorcontrib><collection>CrossRef</collection><jtitle>Applied and environmental microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Itakura, Manabu</au><au>Tabata, Kazufumi</au><au>Eda, Shima</au><au>Mitsui, Hisayuki</au><au>Murakami, Kiriko</au><au>Yasuda, Junichi</au><au>Minamisawa, Kiwamu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Generation of Bradyrhizobium japonicum Mutants with Increased N 2 O Reductase Activity by Selection after Introduction of a Mutated dnaQ Gene</atitle><jtitle>Applied and environmental microbiology</jtitle><date>2008-12</date><risdate>2008</risdate><volume>74</volume><issue>23</issue><spage>7258</spage><epage>7264</epage><pages>7258-7264</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><abstract>We obtained two beneficial mutants of
Bradyrhizobium japonicum
USDA110 with increased nitrous oxide (N
2
O) reductase (N
2
OR) activity by introducing a plasmid containing a mutated
B. japonicum dnaQ
gene (pKQ2) and performing enrichment culture under selection pressure for N
2
O respiration. Mutation of
dnaQ
, which encodes the epsilon subunit of DNA polymerase III, gives a strong mutator phenotype in
Escherichia coli
. pKQ2 introduction into
B. japonicum
USDA110 increased the frequency of occurrence of colonies spontaneously resistant to kanamycin. A series of repeated cultivations of USDA110 with and without pKQ2 was conducted in anaerobic conditions under 5% (vol/vol) or 20% (vol/vol) N
2
O atmosphere. At the 10th cultivation cycle, cell populations of USDA110(pKQ2) showed higher N
2
OR activity than the wild-type strains. Four bacterial mutants lacking pKQ2 obtained by plant passage showed 7 to 12 times the N
2
OR activity of the wild-type USDA110. Although two mutants had a weak or null
fix
phenotype for symbiotic nitrogen fixation, the remaining two (5M09 and 5M14) had the same symbiotic nitrogen fixation ability and heterotrophic growth in culture as wild-type USDA110.</abstract><doi>10.1128/AEM.01850-08</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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language | eng |
recordid | cdi_crossref_primary_10_1128_AEM_01850_08 |
source | American Society for Microbiology; PubMed Central; Alma/SFX Local Collection |
title | Generation of Bradyrhizobium japonicum Mutants with Increased N 2 O Reductase Activity by Selection after Introduction of a Mutated dnaQ Gene |
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