Differentiation of phylogenetic lineages within the ‘Colletotrichum gloeosporioides species complex’ associated with cassava anthracnose disease by PCR-RFLP

• Published in: Tropical plant pathology Vol. 43; no. 3; pp. 194 - 201
• Main Authors: Silva, Leandro L., Pestana, Kátia N., Ferreira, Claudia F., Olveira, Saulo A. S.
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01-06-2018; Springer Nature B.V
• Subjects: Anthracnose; Biomedical and Life Sciences; Cassava; Colletotrichum gloeosporioides; Differentiation; Enzymes; Life Sciences; Original Article; Phylogenetics; Phylogeny; Plant Pathology; Polymerase chain reaction; Restriction fragment length polymorphism; Species; spp; DNA fingerprinting; analysis; DNA polymorphism
• ISSN: 1983-2052; 1982-5676; 1983-2052
• DOI: 10.1007/s40858-018-0218-0

Different species were found associated with cassava anthracnose, considered a major disease of this crop. The correct identification of the causal agent is a first step for defining appropriate control strategies, such as resistant varieties.
In silico
analyses used sequences of six genomic regions of ex-type specimens from the ‘
Colletotrichum gloeosporioides
species complex’ (
C.g.
SC) and 21 restriction enzymes to identify phylogenetic lineages. The three best combinations of region/enzymes were validated in 18
Colletotrichum
spp. isolates from cassava. Dendrograms for
in silico
PCR-RFLP for CAL, ITS and TUB2 showed considerable agreement with the phylogenetic analysis of each genomic region; however, the CAL gene presented greater discriminatory power
.
Since the band patterns from
in gel
analysis were almost the same as expected for the
in silico
analysis for the CAL region, a new approach was proposed based on the combined data from these two methodologies, allowing the differentiation of five phylogenetic lineages within the
C.g.
SC (
C. tropicale
;
C. fructicola
;
C. siamense
;
C. gloeosporioides sensu stricto
and
C. theobromicola
), and one outside of this complex (
C. cliviae
). This evaluation showed to be a reliable technique for preliminary identification of species prior to sequencing.