Several molecular tools have been used to clarify the basis of plant-bacteria interaction; however, the mechanism behind the association is still unclear. In this study, we used a proteomic approach to investigate the root proteome of Zea mays (cv. DKB240) inoculated with Herbaspirillum seropedicae strain SmR1 grown in vitro and harvested 7 days after inoculation. Eighteen differentially accumulated proteins were observed in root samples, ten of which were identified by MALDI-TOF mass spectrometry peptide mass fingerprint. Among the identified proteins, we observed three proteins present exclusively in inoculated root samples and six upregulated proteins and one downregulated protein relative to control. Differentially expressed maize proteins were identified as hypothetical protein ZEAMMB73_483204, hypothetical protein ZEAMMB73_269466, and tubulin beta-7 chain. The following were identified as H. seropedicae proteins: peroxiredoxin protein, EF-Tu elongation factor protein, cation transport ATPase, NADPH:quinone oxidoreductase, dinitrogenase reductase, and type III secretion ATP synthase. Our results presented the first evidence of type III secretion ATP synthase expression during H. seropedicae-maize root interaction.
Food authentication by quantitative polymerase chain reaction (qPCR) methods assures food quality. The aim was to evaluate three qPCR assays for DNA quantification after heat processing of common bean grains, genus-specific FAS assay for Phaseolus, species-specific LEC assay for common bean (Phaseolus vulgaris) and genetically modified (GM) event-specific FGM assay for Embrapa 5.1 event GM common bean. FAS assay showed high stability among Phaseolus genus samples. Common bean grains were heat-treated in autoclave (at 120°C for 15-60 min) and target DNA copy number decreased as processing time increased. Even with DNA degradation, qPCR assays were capable to detect low DNA quantity, and the limit of detection was 100 copy number. Mean efficiency value of FGM assay was 92% in the presence of background DNA. Background DNA did not cause any interference, and 0.39% of GM material can be detected. These qPCR assays are able to quantify common bean in processed food.
Primers and probesIn this study, we used two endogenous reference assays targeting the a-phaseolin and lectin genes of P. vulgaris common bean. The primers PvFASF/PvFASR targeting
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