CRISPR genotyping as complementary tool for epidemiological surveillance of Erwinia amylovora outbreaks

Autor: João Pedro Luz, Rafael J. Mendes, Conceição Santos, Fernando Tavares
Jazyk: angličtina
Rok vydání: 2021
Předmět:
0106 biological sciences
0301 basic medicine
Pears
Fire blight
Artificial Gene Amplification and Extension
Pathogenesis
Erwinia
Pathology and Laboratory Medicine
01 natural sciences
Synthetic Genome Editing
Polymerase Chain Reaction
Genome Engineering
Geographical locations
Disease Outbreaks
Genotype
Medicine and Health Sciences
CRISPR
Genetics
Multidisciplinary
biology
Virulence
Crispr
Eukaryota
Plants
Plant disease
Europe
Medicine
Engineering and Technology
Synthetic Biology
Research Article
Genotyping
Virulence Factors
Science
030106 microbiology
Bioengineering
Research and Analysis Methods
Fruits
03 medical and health sciences
Enterobacteriaceae
Erwinia amylovora
European Union
Molecular Biology Techniques
Molecular Biology
Plant Diseases
Bacteria
Portugal
Apples
Organisms
Outbreak
Biology and Life Sciences
Synthetic Genomics
biology.organism_classification
Genes
Bacterial
People and places
Gram-Negative Bacterial Infections
010606 plant biology & botany
Zdroj: PLoS ONE
PLoS ONE, Vol 16, Iss 4, p e0250280 (2021)
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Agência para a Sociedade do Conhecimento (UMIC)-FCT-Sociedade da Informação
instacron:RCAAP
Repositório Científico de Acesso Aberto de Portugal
Repositório Científico de Acesso Aberto de Portugal (RCAAP)
ISSN: 1932-6203
Popis: Fire blight is a destructive plant disease caused by Erwinia amylovora affecting pome fruit trees, and responsible for large yield declines, long phytosanitary confinements, and high economic losses. In Portugal, the first major fire blight outbreaks occurred in 2010 and 2011, and although later considered eradicated, the emergence of other outbreaks in recent years stressed the need to characterize the E. amylovora populations associated with these outbreaks. In this regard, CRISPR genotyping, assessment of three virulence markers, and semi-quantitative virulence bioassays, were carried out to determine the genotype, and assess the virulence of thirty-six E. amylovora isolates associated with outbreaks occurring between 2010 and 2017 and affecting apple and pear orchards located in the country central-west, known as the main producing region of pome fruits in Portugal. The data gathered reveal that 35 E. amylovora isolates belong to one of the widely-distributed CRISPR genotypes (5-24-38 / D-a-α) regardless the host species, year and region. Ea 680 was the single isolate revealing a new CRISPR genotype due to a novel CR2 spacer located closer to the leader sequence and therefore thought to be recently acquired. Regarding pathogenicity, although dot-blot hybridization assays showed the presence of key virulence factors, namely hrpL (T3SS), hrpN (T3E) and amsG from the amylovoran biosynthesis operon in all E. amylovora isolates studied, pathogenicity bioassays on immature pear slices allowed to distinguish four virulence levels, with most of the isolates revealing an intermediate to severe virulence phenotype. Regardless the clonal population structure of the E. amylovora associated to the outbreaks occurring in Portugal between 2010 and 2017, the different virulence phenotypes, suggests that E. amylovora may have been introduced at different instances into the country. This is the first study regarding E. amylovora in Portugal, and it discloses a novel CRISPR genotype for this bacterium.
Databáze: OpenAIRE
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