Genetic Differentiation of Colletotrichum gloeosporioides and C. truncatum Associated with Anthracnose Disease of Papaya (Carica papaya L.) and Bell Pepper (Capsium annuum L.) Based on ITS PCR-RFLP Fingerprinting

Abstract

Members of the genus Colletotrichum include some of the most economically important fungal pathogens in the world. Accurate diagnosis is critical to devising disease management strategies. Two species, Colletotrichum gloeosporioides and C. truncatum, are responsible for anthracnose disease in papaya (Carica papaya L.) and bell pepper (Capsicum annuum L.) in Trinidad. The ITS1-5.8S-ITS2 region of 48 Colletotrichum isolates was sequenced, and the ITS PCR products were analyzed by PCR-RFLP analysis. Restriction site polymorphisms generated from 11 restriction enzymes enabled the identification of specific enzymes that were successful in distinguishing between C. gloeosporioides and C. truncatum isolates. Species-specific restriction fragment length polymorphisms generated by the enzymes AluI, HaeIII, PvuII, RsaI, and Sau3A were used to consistently resolve C. gloeosporioides and C. truncatum isolates from papaya. AluI, ApaI, PvuII, RsaI, and SmaI reliably separated isolates of C. gloeosporioides and C. truncatum from bell pepper. PvuII, RsaI, and Sau3A were also capable of distinguishing among the C. gloeosporioides isolates from papaya based on the different restriction patterns that were obtained as a result of intra-specific variation in restriction enzyme recognition sites in the ITS1-5.8S-ITS2 rDNA region. Of all the isolates tested, C. gloeosporioides from papaya also had the highest number of PCR-RFLP haplotypes. Cluster analysis of sequence and PCR-RFLP data demonstrated that all C. gloeosporioides and C. truncatum isolates clustered separately into species-specific clades regardless of host species. Phylograms also revealed consistent topologies which suggested that the genetic distances for PCR-RFLP-generated data were comparable to that of ITS sequence data. ITS PCR-RFLP fingerprinting is a rapid and reliable method to identify and differentiate between Colletotrichum species.

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

“…However, due to the high cost of sequencing multiple regions in many fungal isolates, other techniques based on existing "in silico" polymorphisms within these regions of the genome can be used to guide decision making prior to further sequencing. PCR-RFLP analysis is among those techniques that can be used for differentiating species of Colletotrichum based on the existing polymorphisms in regions of the amplified DNA after cleavage with restriction enzymes (Tapia-Tussell et al 2008;Maharaj and Rampersad 2012). In silico tests have shown that it is possible to distinguish strains of species within the C. gloeosporioides complex based on band profiles (Ramdeen and Rampersad 2013), however, this technique was not validated as to its reliability and discriminatory power for different species/phylogenetic lineages within this complex and for the pathosystem of interest (C. gloeosporioides sensu lato versus M. esculenta).…”

Molecular and morpho-cultural characterization of Colletotrichum spp. associated with anthracnose on Capsicum spp. in northeastern Brazil

“…Colletotrichum truncatum (COUFAL0057 and COUFAL0058) has been commonly associated with Capsicum spp. in various regions of the world, such as Trinidad (Maharaj and Rampersad 2012), Australia (Ranathunge et al 2012), Thailand (Ramdial and Rampersad 2015) and China (Liu et al 2016). Moreover, the species C. capsici, a C. truncatum synonymy (Damm et al 2009) was also reported in Capsicum spp.…”

Characterization and Pathogenicity of Colletotrichum truncatum Causing Stem Anthracnose of Red‐Fleshed Dragon Fruit (Hylocereus polyrhizus) in Malaysia

“…The species has been reported to cause anthracnose on various types of beans and lentils (Ford et al. ), papaya (Maharaj and Rampersand ) as well as chilli (formerly known as C. capsici ) (Than et al. ).…”

“…Colletotrichum truncatum is pathogenic to many tropical crops and generally not host specific. The species has been reported to cause anthracnose on various types of beans and lentils (Ford et al 2004), papaya (Maharaj and Rampersand 2012) as well as chilli (formerly known as C. capsici) (Than et al 2008).…”