A species-specific polymerase chain reaction assay for rapid and sensitive detection of Colletotrichum capsici.
Colletotrichum capsici is an important fungal species that causes anthracnose in many genera of plants causing severe economic losses worldwide. A primer set was designed based on the sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a conventional PCR assay. The primer set (CcapF/CcapR) amplified a single product of 394 bp with DNA extracted from 20 Mexican isolates of C. capsici. The specificity of primers was confirmed by the absence of amplified product with DNA of four other Colletotrichum species and eleven different fungal genera. This primer set is capable of amplifying only C. capsici from different contaminated tissues or fungal structures, thereby facilitating rapid diagnoses as there is no need to isolate and cultivate the fungus in order to identify it. The sensitivity of detection with this PCR method was 10 pg of genomic DNA from the pathogen. This is the first report of a C. capsici-specific primer set. It allows rapid pathogen detection and provides growers with a powerful tool for a rational selection of fungicides to control anthracnose in different crops and in the post-harvest stage.
Laboratorio GeMBio, Centro de Investigación Científica de Yucatán A.C, Calle 43 # 130, Col. Chuburná de Hidalgo, Mérida, Yucatán, 97200, México.
C
C
Torres-Calzada
R
R
Tapia-Tussell
A
A
Quijano-Ramayo
R
R
Martin-Mex
R
R
Rojas-Herrera
I
I
Higuera-Ciapara
D
D
Perez-Brito
D001483:Base Sequence; D029441:Carica / Q000382:microbiology; D020231:Colletotrichum / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D008800:Mexico; D025202:Molecular Diagnostic Techniques; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D017931:DNA Primers; D004271:DNA, Fungal
Methods Mol Biol. 2000;132:365-86
10547847
Mol Biotechnol. 2008 Nov;40(3):293-8
18670909
Phytopathology. 1997 May;87(5):516-21
18945106
FEMS Microbiol Ecol. 2005 Jan 1;51(2):257-64
16329874
FEMS Microbiol Lett. 2000 Aug 1;189(1):97-101
10913873
Mol Biotechnol. 2005 Oct;31(2):137-9
16170214
Appl Environ Microbiol. 1996 Mar;62(3):1014-20
8975596
J Mol Biol. 1990 Oct 5;215(3):403-10
2231712
Mol Biotechnol. 2006 May;33(1):67-70
16691008
Phytopathology. 2007 Feb;97(2):132-44
18944368