Unknown

Dataset Information

0

Single nucleotide polymorphism-based diagnostic system for crop-associated Sclerotinia species.


ABSTRACT: A molecular diagnostic system using single nucleotide polymorphisms (SNPs) was developed to identify four Sclerotinia species: S. sclerotiorum (Lib.) de Bary, S. minor Jagger, S. trifoliorum Erikss., and the undescribed species Sclerotinia species 1. DNAs of samples are hybridized with each of five 15-bp oligonucleotide probes containing an SNP site midsequence unique to each species. For additional verification, hybridizations were performed using diagnostic single nucleotide substitutions at a 17-bp sequence of the calmodulin locus. The accuracy of these procedures was compared to that of a restriction fragment length polymorphism (RFLP) method based on Southern hybridizations of EcoRI-digested genomic DNA probed with the ribosomal DNA-containing plasmid probe pMF2, previously shown to differentiate S. sclerotiorum, S. minor, and S. trifoliorum. The efficiency of the SNP-based assay as a diagnostic test was evaluated in a blind screening of 48 Sclerotinia isolates from agricultural and wild hosts. One isolate of Botrytis cinerea was used as a negative control. The SNP-based assay accurately identified 96% of Sclerotinia isolates and could be performed faster than RFLP profiling using pMF2. This method shows promise for accurate, high-throughput species identification.

SUBMITTER: Andrew M 

PROVIDER: S-EPMC2737919 | biostudies-literature | 2009 Sep

REPOSITORIES: biostudies-literature

altmetric image

Publications

Single nucleotide polymorphism-based diagnostic system for crop-associated Sclerotinia species.

Andrew Marion M   Kohn Linda M LM  

Applied and environmental microbiology 20090706 17


A molecular diagnostic system using single nucleotide polymorphisms (SNPs) was developed to identify four Sclerotinia species: S. sclerotiorum (Lib.) de Bary, S. minor Jagger, S. trifoliorum Erikss., and the undescribed species Sclerotinia species 1. DNAs of samples are hybridized with each of five 15-bp oligonucleotide probes containing an SNP site midsequence unique to each species. For additional verification, hybridizations were performed using diagnostic single nucleotide substitutions at a  ...[more]

Similar Datasets

| S-EPMC5825101 | biostudies-literature
| S-EPMC7033337 | biostudies-literature
| S-EPMC2607568 | biostudies-literature
2023-08-28 | GSE239367 | GEO
| S-EPMC8687305 | biostudies-literature
| S-EPMC3187137 | biostudies-literature
| S-EPMC179785 | biostudies-literature
| S-EPMC2686434 | biostudies-literature
| S-EPMC4486411 | biostudies-literature
2010-06-24 | E-GEOD-4227 | biostudies-arrayexpress