Full TGIF Record # 310028
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DOI:10.1094/PDIS-10-18-1727-RE
Web URL(s):https://apsjournals.apsnet.org/doi/10.1094/PDIS-10-18-1727-RE
    Last checked: 01/15/2020
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https://apsjournals.apsnet.org/doi/pdf/10.1094/PDIS-10-18-1727-RE
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Publication Type:
i
Refereed
Author(s):Martinez, J. Francisco Iturralde; Flores, Francisco J.; Koch, Alma R.; Garz贸n, Carla D.; Walker, Nathan R.
Author Affiliation:Martinez and Koch: Departamento de Ciencias de la Vida y la Agricultura, Universidad de las Fuerzas Armadas ESPE, Sangolqu铆, Ecuador; Flores: Departamento de Ciencias de la Vida y la Agricultura, Universidad de las Fuerzas Armadas ESPE, Sangolqu铆, Ecuador and Centro de Investigaci贸n de Alimentos, CIAL, Facultad de Ciencias de la Ingenier铆a e Industrias, Universidad Technol贸gica Equinoccial, Quito, Ecuador; Garz贸n and Walker: Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK
Title:Multiplex end-point PCR for the detection of three species of Ophioshaerella causing spring dead spot of bermudagrass
Section:Research
Other records with the "Research" Section
Source:Plant Disease. Vol. 103, No. 8, August 2019, p. 2010-2014.
Publishing Information:[Washington, D.C.]: Bureau of Plant Industry, U.S. Dept. of Agriculture
# of Pages:5
Related Web URL:https://apsjournals.apsnet.org/doi/abs/10.1094/PDIS-10-18-1727-RE
    Last checked: 01/15/2020
    Notes: Abstract only
Keywords:TIC Keywords: Cynodon; DNA extraction; Disease identification; Genetic analysis; Multiplex polymerase chain reaction; Ophiosphaerella; Spring dead spot
Author-Supplied Keywords: Fungi; Pathogen; Detection; Turf
Abstract/Contents:"A multiplex end-point polymerase chain reaction (PCR) assay was developed for identifying the three-fungal species in the genus Ophiosphaerella that cause spring dead spot (SDS), a devastating disease of bermudagrass. These fungi are difficult to identify by morphology because they seldom produce pseudothecia. To achieve species-specific diagnosis, three pairs of primers were designed to identify fungal isolates and detect the pathogen in infected roots. The internal transcribed spacer region, the translation elongation factor 1-伪, and the RNA polymerase II second-largest subunit were selected as targets and served as templates for the design of each primer pair. To achieve uniform melting temperatures, three to five random nucleotide extensions (flaps) were added to the 5' terminus of some of the designed specific primers. Temperature cycling conditions and PCR components were standardized to optimize specificity and sensitivity of the multiplex reaction. Primers were tested in multiplex on DNA extracted from axenic fungal cultures and from field-collected infected and uninfected roots. A distinct amplicon was produced for each Ophiosphaerella sp. tested. The DNA from Ophiosphaerella close relatives and other common bermudagrass pathogens did not amplify during the multiplex assay. Metagenomic DNA from infected bermudagrass produced species-specific amplicons while DNA extracted from noninfected roots did not. This multiplex end-point PCR approach is a sensitive and specific molecular technique that allows for correct identification of SDS-associated Ophiosphaerella spp. from field-collected roots."
Language:English
References:28
Note:Pictures, b/w
Tables
ASA/CSSA/SSSA Citation (Crop Science-Like - may be incomplete):
Martinez, J. F. I., F. J. Flores, A. R. Koch, C. D. Garz贸n, and N. R. Walker. 2019. Multiplex end-point PCR for the detection of three species of Ophioshaerella causing spring dead spot of bermudagrass. Plant Disease. 103(8):p. 2010-2014.
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DOI: 10.1094/PDIS-10-18-1727-RE
Web URL(s):
https://apsjournals.apsnet.org/doi/10.1094/PDIS-10-18-1727-RE
    Last checked: 01/15/2020
    Access conditions: Item is within a limited-access website
https://apsjournals.apsnet.org/doi/pdf/10.1094/PDIS-10-18-1727-RE
    Last checked: 01/15/2020
    Requires: PDF Reader
    Access conditions: Item is within a limited-access website
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