Identification of Colletotrichum species associated with anthracnose disease of coffee in Vietnam

Colletotrichum gloeosporioides, C. acutatum, C. capsici and C. boninense associated with anthracnose disease on coffee (Coffea spp.) in Vietnam were identified based on morphology and DNA analysis. Phylogenetic analysis of DNA sequences from the internal transcribed spacer region of nuclear rDNA and a portion of mitochondrial small subunit rRNA were concordant and allowed good separation of the taxa. We found several Colletotrichum isolates of unknown species and their taxonomic position remains unresolved. The majority of Vietnamese isolates belonged to C. gloeosporioides and they grouped together with the coffee berry disease (CBD) fungus, C. kahawae. However, C. kahawae could be distinguished from the Vietnamese C. gloeosporioides isolates based on ammonium tartrate utilization, growth rate and pathogenictity. C. gloeosporioides isolates were more pathogenic on detached green berries than isolates of the other species, i.e. C. acutatum, C capsici and C. boninense. Some of the C. gloeosporioides isolates produced slightly sunken lesion on green berries resembling CBD symptoms but it did not destroy the bean. We did not find any evidence of the presence of C. kahawae in Vietnam

Colletotrichum species with curved conidia from herbaceous hosts

Colletotrichum (Glomerellaceae, Sordariomycetes) species with dark setae and curved conidia are known as anthracnose pathogens of a number of economically important hosts and are often identified as C. dematium. Colletotrichum dematium has been synonymised with many species, including the type of the genus, C. lineola. Since there is no living strain of the original material of either species available, we re-collected C. lineola from the original location to serve as an epitype of that name, and chose an appropriate epitype specimen and associated strain of C. dematium from the CBS collection. A multilocus molecular phylogenetic analysis (ITS, ACT, Tub2, CHS-1, GAPDH, HIS3) of 97 isolates of C. lineola, C. dematium and other Colletotrichum species with curved conidia from herbaceous hosts resulted in 20 clades, with 12 clades containing strains that had previously been identified as C. dematium. The epitype strains of C. lineola and C. dematium reside in two closely related clades. Other clades represent four previously undescribed species, C. anthrisci, C. liriopes, C. rusci and C. verruculosum, isolated respectively from Anthriscus in the Netherlands, Liriope in Mexico, Ruscus in Italy and Crotalaria in Zimbabwe. The new combinations C. spaethianum and C. tofieldiae are made. Colletotrichum truncatum is epitypified, as well as C. circinans, C. curcumae and C. fructi. Three further unidentified Colletotrichum taxa were detected in the phylogenetic analysis, which may require description after further research. Each species is comprehensively described and illustrate

Molecular diversity of anthracnose pathogen populations associated with UK strawberry production suggests multiple introductions of three different Colletotrichum species.

Fragaria × ananassa (common name: strawberry) is a globally cultivated hybrid species belonging to Rosaceae family. Colletotrichum acutatum sensu lato (s.l.) is considered to be the second most economically important pathogen worldwide affecting strawberries. A collection of 148 Colletotrichum spp. isolates including 67 C. acutatum s.l. isolates associated with the phytosanitary history of UK strawberry production were used to characterize multi-locus genetic variation of this pathogen in the UK, relative to additional reference isolates that represent a worldwide sampling of the diversity of the fungus. The evidence indicates that three different species C. nymphaeae, C. godetiae and C. fioriniae are associated with strawberry production in the UK, which correspond to previously designated genetic groups A2, A4 and A3, respectively. Among these species, 12 distinct haplotypes were identified suggesting multiple introductions into the country. A subset of isolates was also used to compare aggressiveness in causing disease on strawberry plants and fruits. Isolates belonging to C. nymphaeae, C. godetiae and C. fioriniae representative of the UK anthracnose pathogen populations showed variation in their aggressiveness. Among the three species, C. nymphaeae and C. fioriniae appeared to be more aggressive compared to C. godetiae. This study highlights the genetic and pathogenic heterogeneity of the C. acutatum s.l. populations introduced into the UK linked to strawberry production

The distinctive population structure of Colletotrichum species associated with olive anthracnose in the Algarve region of Portugal reflects a host–pathogen diversity hot spot

Anthracnose (Colletotrichum spp.) is an important disease of olive fruits. Diversity and biogeographic relationships of the olive anthracnose pathogens in the Algarve (Portugal) were investigated, along with host association patterns and disease levels during 2004–2007, to test the hypothesis that this region is a host–pathogen diversity hot spot. Diverse Colletotrichum acutatum and Colletotrichum gloeosporioides populations were identified based on rRNA-internal transcribed spacer and partial β-tubulin 2 gene sequences of 95 isolates. Spatial and temporal variations in the occurrence of the eight genetic entities of the pathogens were linked to olive biogeography. Disease occurrence patterns suggest that C. acutatum populations are more stable pathogens, while C. gloeosporioides populations appear to be more influenced by favourable conditions. Three unique C. acutatum populations were identified, but none of the eight populations were dominant, with the most frequent type representing only 27%. Thus, the population structure of olive anthracnose pathogens in the Algarve is distinct from other parts of Portugal and other world locations, where only one or two genetic entities are dominant. This pattern and level of genetic diversity in a restricted area, where oleaster (wild olive tree), ancient landraces and modern cultivars of olive occur in close proximity, suggests the Algarve as a centre of diversity of the anthracnose pathogens and corroborates recent work suggesting western Mediterranean as an important centre of olive diversity and domestication

Draft genome sequence of Colletotrichum acutatum sensu lato (Colletotrichum fioriniae)

In addition to its economic impact, Colletotrichum acutatum sensu lato is an interesting model for molecular investigations due to the diversity of host-determined specialization and reproductive lifestyles within the species complex. The pathogen Colletotrichum fioriniae forms part of this species complex and causes anthracnose in a wide range of crops and wild plants worldwide. Some members of this species have also been reported to be entomopathogenic. Here, we report the draft genome sequence of a heterothallic reference isolate of C. fioriniae (strain PJ7). This sequence provides a range of new resources that serve as a useful platform for further research in the field

A Fungal Effector With Host Nuclear Localization and DNA-Binding Properties Is Required for Maize Anthracnose Development

Plant pathogens have the capacity to manipulate the host immune system through the secretion of effectors. We identified 27 putative effector proteins encoded in the genome of the maize anthracnose pathogen Colletotrichum graminicola that are likely to target the host’s nucleus, as they simultaneously contain sequence signatures for secretion and nuclear localization. We functionally characterized one protein, identified as CgEP1. This protein is synthesized during the early stages of disease development and is necessary for anthracnose development in maize leaves, stems, and roots. Genetic, molecular, and biochemical studies confirmed that this effector targets the host’s nucleus and defines a novel class of double-stranded DNA-binding protein. We show that CgEP1 arose from a gene duplication in an ancestor of a lineage of monocot-infecting Colletotrichum spp. and has undergone an intense evolution process, with evidence for episodes of positive selection. We detected CgEP1 homologs in several species of a grass-infecting lineage of Colletotrichum spp., suggesting that its function may be conserved across a large number of anthracnose pathogens. Our results demonstrate that effectors targeted to the host nucleus may be key elements for disease development and aid in the understanding of the genetic basis of anthracnose development in maize plants.Fil: Vargas, Walter Alberto. Universidad de Salamanca; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sanz Martín, José M.. Universidad de Salamanca; EspañaFil: Rech, Gabriel E.. Universidad de Salamanca; EspañaFil: Armijos Jaramillo, Vinicio D.. Universidad de Salamanca; EspañaFil: Rivera Rodriguez, Lina Patricia. Universidad de Salamanca; EspañaFil: Echeverria, María de Las Mercedes. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; ArgentinaFil: Díaz Mínguez, José M.. Universidad de Salamanca; EspañaFil: Thon, Michael R.. Universidad de Salamanca; EspañaFil: Sukno, Serenella A.. Universidad de Salamanca; Españ

Relevamiento de la producción de lacasa en aislamientos de Colletotrichum (Ascomycota), agente causal de antracnosis de la Soja

Colletotrichum truncatum es el hongo patógeno más comúnmente asociado con la antracnosis de soja. Aunque la enzima ligninolítica lacasa se relaciona con la patogenicidad de un amplio rango de hongos fitopatógenos, su rol biológico en la interacción Colletotrichum -soja aún se desconoce. La extensión de la infección en la Argentina , nos ha llevado a examinar la producción de lacasa en cepas aisladas de plantas enfermas de soja de diferentes regiones de nuestro país. Se evaluó la producción in vitro de lacasa en diez cepas (ocho de ellas identificadas como C. truncatum ). Sólo seis, todas correspondientes a C. truncatum , produjeron lacasa cuando fueron cultivadas en un medio con pectina y asparagina. La máxima actividad en este medio (44 U/L) se detectó en la cepa BAFC 3102 (aislada en la Provincia del Chaco). La electroforesis en SDS PAGE de los sobrenadantes de cultivo reveló una única banda con actividad lacasa (peso molecular 67 kDa). La adición de CuSO 4 al medio incrementó hasta siete veces su producción (280 U/L en medio con glucosa), sin embargo la edad del cultivo o la composición del medio no produjeron variaciones en el patrón isoenzimático. Este es el primer reporte sobre producción de lacasa por C. truncatum.Colletotrichum truncatum is the most common pathogen fungus associated with soybean anthracnose. Although the lignin-degrading enzyme laccase has been implicated in pathogenicity of a wide range of plant pathogenic fungi, its biological role in the Colletotrichum -soybean disease system is unknown. The extent of the infection in our country led us to examine laccase production in Argentinean Colletotrichum strains isolated from diseased soybean plants from different geographic locations. Ten strains (eight of them identified as C. truncatum) , were screened for in vitro laccase production. Only six of the isolates, all of them C. truncatum , produced laccase activity when cultured on a defined medium based on pectin and asparagine as carbon and nitrogen sources, respectively. Strain BAFC 3102 (isolated from Chaco province), yielded the highest laccase titers (44 U/L) in this medium. Denaturing polyacrylamide gel electrophoresis of extracellular culture fluids revealed one band with laccase activity (mol wt 67 kDa). CuSO 4 addition to media with either glucose or pectin as carbon sources increased up to 7-fold laccase production (280 U/L in the glucose medium), but the pattern of isoenzyme was not affected by culture age or medium composition. This is the first report on laccase production by C. truncatum.Fil: Levin, Laura Noemí. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; ArgentinaFil: Ramos, Araceli Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; ArgentinaFil: Parisi, Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; ArgentinaFil: Gally, Marcela Edith. Universidad de Buenos Aires. Facultad de Agronomía. Departamento de Producción Vegetal. Cátedra de Fitopatología; Argentin

Preliminary Evaluation of a Dodder Anthracnose Fungus from China as a Mycoherbicide for Dodder Control in the US

Dodder (Cuscuta spp.) is a noxious, parasitic, annual weed throughout most of the United States. A fungus used to control it in China was imported under permit for studies with U.S. dodder species in containment. The fungus, Colletotrichum gloeosporioides, sporulated on liquid and solid media at room temperature. Conidia from 7-12 day old cultures were diluted to 3.5 to 7 X 16⁶ spores ml^-1 for host range inoculations. Germination on water agar at 24 hrs was higher at 28 than 30 or 24 C. Inoculated plants were exposed to dew periods of 12-1 4 hrs at 24 or 28 C, then transferred to growth chambers with 1 2-hr photoperiods at constant temperatures of 24, 28, and 32C. Dodder species were severely diseased but rarely killed. Symptoms were most severe on native collections of Cuscuta campestris after 4 to 5 days incubation when this species on periwinkle seedlings was inoculated with 3.5 to 7 X 10⁵ spores ml^-1 . Cuscuta cuspidata, C. pentagona, and C. campestris from a California seedlot were also tested under optimum conditions for disease. The C. campestris from California was the most susceptible. Inoculation of 16 species in eight plant families revealed no other host except sweet potato which developed a necrotic fleck. This research indicates a need for strain improvement prior to field tests