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Effects of rice seed surface sterilization with hypochlorite on inoculated Burkholderia vietnamiensis.

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.

SUBMITTER: Miche L

PROVIDER: S-EPMC92979 | biostudies-literature | 2001 Jul

REPOSITORIES: biostudies-literature

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Effects of rice seed surface sterilization with hypochlorite on inoculated Burkholderia vietnamiensis.

Miché L L Balandreau J J

Applied and environmental microbiology 20010701 7

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 ...[more]

PMID: 11425720

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Project description:Methyl jasmonate (MeJA) is widely used as a model chemical to study hypersensitive responses to biotic stress impacts in plants. Elevated levels of methyl jasmonate induce jasmonate-dependent defense responses, associated with a decline in primary metabolism and enhancement of secondary metabolism of plants. However, there is no information of how stress resistance of plants, and accordingly the sensitivity to exogenous MeJA can be decreased by endophytic plant growth promoting rhizobacteria (PGPR) harboring ACC (1-aminocyclopropane-1-carboxylate) deaminase. In this study, we estimated stress alleviating potential of endophytic PGPR against MeJA-induced plant perturbations through assessing photosynthetic traits and stress volatile emissions. We used mild (5?mM) to severe (20?mM) MeJA and endophytic plant growth promoting rhizobacteria Burkholderia vietnamiensis CBMB40 and studied how MeJA and B. vietnamiensis treatments influenced temporal changes in photosynthetic characteristics and stress volatile emissions. Separate application of MeJA markedly decreased photosynthetic characteristics and increased lipoxygenase pathway (LOX) volatiles, volatile isoprenoids, saturated aldehydes, lightweight oxygenated compounds (LOC), geranyl-geranyl diphosphate pathway (GGDP) volatiles, and benzenoids. However, MeJA-treated leaves inoculated by endophytic bacteria B. vietnamiensis had substantially increased photosynthetic characteristics and decreased emissions of LOX, volatile isoprenoids and other stress volatiles compared with non-inoculated MeJA treatments, especially at later stages of recovery. In addition, analysis of leaf terpenoid contents demonstrated that several mono- and sesquiterpenes were de novo synthesized upon MeJA and B. vietnamiensis applications. This study demonstrates that foliar application of endophytic bacteria B. vietnamiensis can potentially enhance resistance to biotic stresses and contribute to the maintenance of the integrity of plant metabolic activity.

Dataset Information

Effects of rice seed surface sterilization with hypochlorite on inoculated Burkholderia vietnamiensis.

Publications

Effects of rice seed surface sterilization with hypochlorite on inoculated Burkholderia vietnamiensis.

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