000 02438nab|a22003257a|4500
999 _c61734
_d61726
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003 MX-TxCIM
005 20211006073554.0
008 200423s2013||||xxk|||p|op||||00||0|eng|d
022 _a1364-5072
022 _a1365-2672 (Online)
024 8 _ahttps://doi.org/10.1111/jam.12068
040 _aMX-TxCIM
041 _aeng
100 1 _aTao, G.
_912543
245 1 0 _aSpecies‐specific real‐time PCR detection of Colletotrichum kahawae
260 _aUnited Kingdom :
_bWiley,
_c2013.
500 _aPeer review
520 _aAims. Colletotrichum kahawae is a strongly aggressive pathogen causing coffee berry disease and is specific to Arabica coffee (Coffea arabica) in Africa. In this article, we developed a real‐time PCR assay for the species‐specific diagnosis of C. kahawae by designing the primers and a TaqMan probe derived from the single nucleotide polymorphism‐rich region of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) gene. Methods and Results. DNA markers from rDNA internal transcribed spacer, actin, β‐tubulin and GAPDH genes of the ex‐type culture of C. kahawae and 10 reference strains of Colletotrichum species were analysed for intra‐ and interspecific variations. The GAPDH gene was selected to develop a species‐specific DNA marker. A TaqMan real‐time PCR assay for species‐specific detection of C. kahawae was developed, and its accuracy was tested against type strains of other phylogenetically closely related species in the C. gloeosporioides species complex, with the detection sensitivity of 80 fg μl−1 of genomic DNA. Conclusions. This real‐time PCR assay is highly specific and sensitive for the diagnosis of C. kahawae and can be applied in qualitative and quantitative tests. Significance and Impact of the Study. This protocol allows for a rapid and sensitive detection of C. kahawae and will be useful in disease management and pest detection to prevent further spread of this pathogen.
546 _aText in English
650 7 _2AGROVOC
_911096
_aCoffea arabica
650 7 _2AGROVOC
_912544
_aGlomerella cingulata
650 7 _2AGROVOC
_95807
_aDiagnosis
650 7 _aPlant diseases
_gAGROVOC
_2
_91206
650 7 _2AGROVOC
_93563
_aGenes
700 1 _912545
_aHyde, K.D.
700 0 _912546
_aLei Cai
773 0 _tJournal of Applied Microbiology
_gv. 114, no. 3, p. 828-835
_dUnited Kingdom : Wiley, 2013.
_x1365-2672
942 _cJA
_n0
_2ddc