Abstract:
:When a combination of hydrogen peroxide and hypochlorite was used to surface sterilize rice seeds, a 10(2)- to 10(4)-fold decrease in CFU was observed during the first 15 h after inoculation of the rice rhizosphere organism Burkholderia vietnamiensis TVV75. This artifact could not be eliminated simply by rinsing the seeds, even thoroughly, with sterile distilled water. When growth resumed, a significant increase in the frequency of rifampin- and nalidixic acid-resistant mutants in the population was observed compared to the control without seeds. This phenomenon was a specific effect of hypochlorite; it was not observed with hydrogen peroxide alone. It was also not observed when the effect of hypochlorite was counteracted by sodium thiosulfate. We hypothesized that the hypochlorite used for disinfection reacted with the rice seed surface, forming a chlorine cover which was not removed by rinsing and generated mutagenic chloramines. We studied a set of rifampin- and nalidixic acid-resistant mutants obtained after seed surface sterilization. The corresponding rpoB and gyrA genes were amplified and sequenced to characterize the induced mutations. The mutations in five of seven nalidixic acid-resistant mutants and all of the rifampin-resistant mutants studied were found to correspond to single amino acid substitutions. Hypochlorite surface sterilization can thus be a source of artifacts when the initial bacterial colonization of a plant is studied.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Miché L,Balandreau Jdoi
10.1128/AEM.67.7.3046-3052.2001keywords:
subject
Has Abstractpub_date
2001-07-01 00:00:00pages
3046-52issue
7eissn
0099-2240issn
1098-5336journal_volume
67pub_type
杂志文章abstract::An anaerobic, cellulose-degrading, gram-negative rod and a gram-positive coccus, identified as Bacteroides succinogenes and Ruminococcus flavefaciens, respectively, were isolated from pig fecal samples. These organisms were isolated from cellulolytic most-probable-number dilutions which represented 4 or 6% of the viab...
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更新日期:2004-07-01 00:00:00
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更新日期:1991-09-01 00:00:00
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更新日期:1981-05-01 00:00:00
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更新日期:1994-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.34.6.832-836.1977
更新日期:1977-12-01 00:00:00