Complete Genome Assembly of Pantoea stewartii subsp. stewartii DC283, a Corn Pathogen.
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ABSTRACT:
SUBMITTER: Duong DA
PROVIDER: S-EPMC5454200 | biostudies-literature | 2017 Jun
REPOSITORIES: biostudies-literature
Publications
Complete Genome Assembly of <i>Pantoea stewartii</i> subsp. <i>stewartii</i> DC283, a Corn Pathogen.
Duong Duy An DA Stevens Ann M AM Jensen Roderick V RV
Genome announcements 20170601 22
The phytopathogen <i>Pantoea stewartii</i> subsp. <i>stewartii</i> DC283 causes Stewart's wilt disease in corn after transmission from the corn flea beetle insect vector. Here, we report that the complete annotated genome of <i>P. stewartii</i> DC283 has been fully assembled into one circular chromosome, 10 circular plasmids, and one linear phage. ...[more]
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Project description:Pantoea stewartii subsp. stewartii is a bacterial phytopathogen that causes Stewart’s wilt disease in corn. It uses quorum sensing to regulate expression of some genes involved in virulence in a cell density-dependent manner as the bacterial population grows from small numbers at the initial infection site in the leaf apoplast to high cell numbers in the xylem where it forms a biofilm. There are also other genes important for pathogenesis not under quorum-sensing control such as a Type III secretion system. The purpose of this study was to compare gene expression during a high-density in planta infection versus a low-density pre-inoculum liquid culture and in a high-density culture grown on agar medium to identify genes specifically expressed in planta that may also be important for colonization and/or virulence. RNA was purified from each sample type to determine the transcriptome via RNA-Seq using Illumina sequencing of cDNA. Fold gene expression changes in the high-density in planta data set in comparison to the two in vitro grown samples were determined and a list of the most differentially expressed genes was generated to elucidate genes important for plant association. Quantitative reverse transcription PCR (qRT-PCR) was used to validate expression patterns for a select subset of genes. Analysis of the transcriptome data via gene ontology revealed that bacterial transporters and systems active under low oxygen tensions appear to play a critical role for P. stewartii as it colonizes and causes wilt disease in corn plants.
2017-04-01 | GSE87520 | GEO
Project description:Pantoea stewartii subsp. stewartii is a bacterial phytopathogen that causes Stewart's wilt disease in corn. It uses quorum sensing to regulate expression of some genes involved in virulence in a cell density-dependent manner as the bacterial population grows from small numbers at the initial infection site in the leaf apoplast to high cell numbers in the xylem where it forms a biofilm. There are also other genes important for pathogenesis not under quorum-sensing control such as a Type III secretion system. The purpose of this study was to compare gene expression during an in planta infection versus either a pre-inoculum in vitro liquid culture or an in vitro agar plate culture to identify genes specifically expressed in planta that may also be important for colonization and/or virulence. RNA was purified from each sample type to determine the transcriptome via RNA-Seq using Illumina sequencing of cDNA. Fold gene expression changes in the in planta data set in comparison to the two in vitro grown samples were determined and a list of the most differentially expressed genes was generated to elucidate genes important for plant association. Quantitative reverse transcription PCR (qRT-PCR) was used to validate expression patterns for a select subset of genes. Analysis of the transcriptome data via gene ontology revealed that bacterial transporters and systems important for oxidation reduction processes appear to play a critical role for P. stewartii as it colonizes and causes wilt disease in corn plants.
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Project description:Pantoea stewartii subsp. stewartii is the causative agent of Stewart's wilt, a bacterial disease transmitted by the corn flea beetle mainly to sweet corn (Zea mays). In many countries, it is classified as a quarantine organism and must be differentiated from other yellow enteric bacteria frequently occurring with corn. We have created novel primers from the pstS-glmS region of P. stewartii for use in conventional PCR (cPCR) and quantitative PCR (qPCR). To facilitate rapid diagnosis, we applied matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis. Using whole-cell protein extracts, profiles were generated with a Bruker microflex machine, and the bacteria classified. P. stewartii strains were clearly distinguished from strains of Pantoea agglomerans, Pantoea dispersa, and Pantoea ananatis. Dendrogram analysis of the protein profiles confirmed the score values and showed the formation of separate clades for each species. The identification achieved by MALDI-TOF MS analysis agrees with the diagnosis by specific PCR primers. The combination of both methods allows a rapid and simple identification of the corn pathogen. P. stewartii subsp. stewartii and P. stewartii subsp. indologenes are highly related and can be distinguished not only by virulence assays and indole tests but also by a characteristic pattern in the nucleotide sequence of recA.
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Project description:Following a request from the European Commission, the EFSA Panel on Plant Health performed a risk assessment of the entry of Pantoea stewartii subsp. stewartii on maize seed imported by the EU from the USA. This pest is a Gram-negative bacterium which causes Stewart's vascular wilt and leaf blight of maize (including sweet corn), a disease responsible for serious crop losses throughout the world. The following scenarios were considered: scenario A0 (current practice), scenario A1 (US request for modification of EU conditions for derogation), and scenario A2 (EU conditions for derogation). Results from the quantitative seed pathway model presented here show that, despite the low rates of plant-to-seed and seed-to-seedling transmission that have been reported in the literature for Stewart's wilt, given the amount of traded seed, and in the case of voluntary (i.e. not mandatory) inspections of seed production fields at the origin (i.e. scenario A0), the frequency of introducing the disease is in the order of magnitude of some hundred introductions per year (median number). The EU conditions for derogation would lead to a decrease in the likelihood of entry compared to scenarios A0 (about 10,000 times fewer introductions) and A1 (about 2,000 times fewer introductions). This protective effect is mainly due to the requirement that only genotypes resistant to Stewart's wilt are traded, with the additional field inspection (two instead of one per season) providing additional reassurance. The Panel also concluded that seed lot inspections, as currently carried out (e.g. with a sample of 400 seeds) are not likely to lead to a relevant reduction in the level of infected imported maize seed, given the low prevalence of Stewart's wilt at the origin. If, however, there is aggregation in infection among consignments, inspection would work towards identifying the highly infected consignments. Recently, outbreaks of Stewart's wilt have occurred in Italy (Emilia Romagna, Friuli, Lombardy and Veneto). A review is provided of the available information to assess the possible role of seed imports in these outbreaks.
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