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

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

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

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

UJI EKSTRAK BAWANG PUTIH (Allium sativum) SEBAGAI FUNGISIDA ALAMI JAMUR Colletotrichum gloeosporioides PENYAKIT TANAMAN JERUK

Penelitian ini bertujuan untuk mendapatkan informasi secara kuantitatif mengenai penghambatan pertumbuhan jamur dengan fungisida alami dari ekstrak bawang putih. Penelitian ini dilaksanakan pada bulan Mei 2017. Metode yang digunakan dalam penelitian ini adalah metode eksperimen dengan ekstraksi maserasi dan cara penanaman jamurnya dengan dilusi padat. Penelitian ini terdiri dari 6 perlakuan dan 4 pengulangan yaitu konsentrasi 30%, 40%, 50%, 60%, 70% dan kontrol. Parameter yang diukur yaitu diameter koloni jamur Colletotrichum gloeosporioides. Penghambatan pertumbuhan diameter koloni jamur Colletotrichum gloeosporioides dianalisis menggunakan aplikasi pengolah data SPSS 24. Identifikasi penghambatan pertumbuhan diameter koloni jamur dilakukan di Laboratorium Pendidikan Biologi FKIP Universitas Pasundan Bandung. Hasil penelitian menunjukkan bahwa ekstrak bawang putih optimum dapat menghambat pertumbuhan diameter koloni jamur Colletotrichum gloeosporioides. Dan konsentrasi yang optimum dalam menghambat pertumbuhan jamur Colletotrichum gloeosporioides yaitu pada konsentrasi 70% dengan rata-rata 0,15 mm. Kata kunci: Ekstrak Bawang Putih, Fungisida Alami, Jamur Colletotrichum gloeosporioides

Global Antifungal Profile Optimization of Chlorophenyl Derivatives against Botrytis cinerea and Colletotrichum gloeosporioides

Twenty-two aromatic derivatives bearing a chlorine atom and a different chain in the para or meta position were prepared and evaluated for their in vitro antifungal activity against the phytopathogenic fungi Botrytis cinerea and Colletotrichum gloeosporioides. The results showed that maximum inhibition of the growth of these fungi was exhibited for enantiomers S and R of 1-(40-chlorophenyl)- 2-phenylethanol (3 and 4). Furthermore, their antifungal activity showed a clear structure-activity relationship (SAR) trend confirming the importance of the benzyl hydroxyl group in the inhibitory mechanism of the compounds studied. Additionally, a multiobjective optimization study of the global antifungal profile of chlorophenyl derivatives was conducted in order to establish a rational strategy for the filtering of new fungicide candidates from combinatorial libraries. The MOOPDESIRE methodology was used for this purpose providing reliable ranking models that can be used later

First Report of Colletotrichum gloeosporioides Causing Anthracnose of Tejocote (Crataegus gracilior) Fruits in Mexico

Crataegus, is a genus classified in family Rosaceae and includes several tree species commonly called Tejocote that are widely cultivated for their pome fruits in Mexico. During fall of 2014, 2015, and 2016, severe symptoms of anthracnose were observed on approx. 60% of tejocote (Crataegus gracilior) fruits in an orchard located in Tulancingo, Oaxaca, Mexico. Affected fruits showed sunken, prominent, dark brown to black necrotic lesions, and were exuding salmon spore masses. To isolate the fungus, small pieces from tissue adjacent to the lesions of 10 symptomatic fruits were excised and surface disinfested by immersion in a 1% sodium hypochlorite solution for 2 min, rinsed three times in sterile distilled water, placed in Petri plates containing potato dextrose agar (PDA), and incubated at 25ºC for 5 to 7 days in darkness. Mycelial plugs were excised from the edge of the actively growing fungal colony and aseptically transferred to fresh PDA medium and incubated at 25°C for 6 days. Five monoconidial cultures were obtained by transferring germinated spores to Petri plates with fresh PDA. One isolate was selected as representative for morphological and molecular identification. Colonies of pure cultures exhibited greyish-white aerial mycelium and abundant salmon-pink conidial masses. Conidia (n= 100) were subcylindrical, hyaline, straight, one-celled, with rounded ends, measuring 13.6 to 17.7 × 4.4 to 5.9 μm. Conidial appressoria were ovoid and brown to dark brown. Based on morphological characteristics, the fungus was identified within the Colletotrichum gloeosporioides species complex (Weir et al. 2012). The isolate was designated UACH-177 and deposited in the Culture Collection of Phytopathogenic Fungi at the Chapingo Autonomous University. For molecular identification, the ITS region (White et al. 1990), and fragments of (Apn2) (Rojas et al. 2010), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-tubulin 2 (TUB2) genes (Weir et al. 2012) were amplified by PCR, and sequenced. The sequences were deposited in GenBank (Accessions numbers ITS:MG821312; Apn2:MG821310; GAPDH:MG821311; and TUB2:MG821313). A phylogenetic analysis using Bayesian inference and including published ITS, Apn2, GAPDH, and TUB2 data for C. gloeosporioides and other Colletotrichum species was performed. The phylogenetic analysis showed the sequences were grouped into the clade of C. gloeosporioides. To confirm the pathogenicity of the fungus, 20 tejocote fruits were surface disinfested by immersion in a 1% sodium hypochlorite solution for 1 min, washed three times with sterile distilled water and dried on sterilized filter paper. Inoculations were performed by deposition of 10 μl of a conidial suspension (106 spores ml-1) on the fruit surface. Ten fruit were mock inoculated with distilled water as a control. All fruits were kept in a moist chamber at 25°C for 10 days. Pathogenicity test was repeated twice. Disease symptoms were observed on all inoculated fruit after 7 days, whereas control fruit did not develop symptoms. Fungal colonies were re-isolated from all symptomatic fruits and were found to be morphologically identical to the original isolate inoculated on tejocote fruits, thus fulfilling Koch´s postulates. In Mexico, García-Alvarez (1976) reported Colletotrichum sp. on fruits of Crataegus mexicana, however, that report was not supported by morphological characterization nor pathogenicity tests. To our knowledge, this is the first report of C. gloeosporioides causing anthracnose of Crataegus gracilior in Mexico and worldwide.Fil: Nieto López, Edgar Humberto. University of Nebraska; Estados UnidosFil: Everhart, Sydney. University of Nebraska; Estados UnidosFil: Ayala Escobar, Victoria. Colegio de Postgraduados; MéxicoFil: Camacho Tapia, Moises. Universidad Autónoma Chapingo; MéxicoFil: Bernardi Lima, Nelson. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; ArgentinaFil: Nieto Angel, Raúl. Universidad Autónoma Chapingo; MéxicoFil: Tovar Pedraza, Juan Manuel. Universidad Autónoma Chapingo; Méxic

Mechanism study of alachlor biodegradation by Paecilomycesmarquandii with proteomic and metabolomic methods

Alachlor is an herbicide that is widely used worldwide to protect plant crops against broadleaf weedsand annual grasses. However, due to its endocrine-disrupting activity, its application had been bannedin the European Union. As described in our earlier work, Paecilomyces marquandii is a microscopic funguscapable of alachlor removal by N-acetyl oxidation. Our current work uses proteomics and metabolomicsto gain a better understanding of alachlor biodegradation by the microscopic fungus P. marquandii.The data revealed that the addition of alachlor reduced the culture growth and glucose consump-tion rates. Moreover, the rates of glycolysis and the tricarboxylic acids (TCA) cycle increased duringthe initial stage of growth, and there was a shift toward the formation of supplementary materials(UDP-glucose/galactose) and reactive oxygen species (ROS) scavengers (ascorbate). Proteomic analy-sis revealed that the presence of xenobiotics resulted in a strong upregulation of enzymes related toenergy, sugar metabolism and ROS production. However, the unique overexpression of cyanide hydratasein alachlor-containing cultures may implicate this enzyme as the key protein involved in the alachlorbiodegradation pathway. The characterization of P. marquandii-mediated alachlor removal in terms of cellstructure and function provides a deeper insight into the strategies of microorganisms toward xenobioticbiodegradation.This study was supported by the grant of the National Science Centre, Poland (Project No. UMO-2011/01/B/NZ9/02898)

Caracterització de "Colletotrichum gloeosporioides" en oliva

La malaltia coneguda com «olives sabonoses», és una de les més temudes en olivera, degut a la incidència negativa sobre la collita. El fong causant, havia estat descrit com Gloeosporium olivarum Alm.; no obstant, moltes espècies d'aquest gènere, s'han inclòs en Colletotrichum gloeosporioides que és l'anamorf de Glomerella cingulata (Ston.) Sp. & Schr.. Von Arx, també ha inclòs Gloeosporium olivarum dins d'aquesta espècie. Els estudis realitzats en olives, donen supon a aquesta inclusió dins de Colletotrichum gloeosporioides i posen de manifest la presència de diferents tipus culturals basats en la coloració i tipus de colònia que concorden en els tipus observats per C. Gloeosporioides en altres hostes.La enfermedad conocida como «aceituna jabonosa» es una de las más temibles en el olivo por su incidencia negativa en la cosecha. El hongo causante habia sido descrito como Gloeosporium olivarum Alm. No obstante, muchas especies de este género se han incluido en Colletotrichum gloeosporioides, anamorfo de Glomerella cingulata (Ston.) Sp. & Schr. Von Arx ha incluido también Gloesporium olivarum dentro de esta especie. Los estudiós realizados en aceituna apoyan esta inclusión dentro de Colletotrichum gloeosporioides y ponen de manifiesto la presencia de distintos tipos culturales basados en la coloración y tipo de colonia que concuerdan con los tipos observados de C. Gloeosporioides en otros huéspedes

Etiologija bolesti uzrokovanih Colletotrichum vrstama na mandarini Unshiu u Hrvatskoj

During the last decade anthracnose has become a major disease of Satsuma mandarin, the most important citrus crop in Croatia. The aim of this study was to determine Colletotrichum species associated with different symptoms and to identify the origin of inoculum. From 2013 to 2016, 437 samples of plant material were collected. Colletotrichum spp. was isolated from 93% of dried twigs, 35% of dropped flowers, 89% of leaf spots, all fruit (100%) with anthracnose or calyx-end rot symptoms, 12% of fruit with post-harvest soft rot and from 40% of fruit showing spots remaining on trees after harvest. Out of 258 Colletotrichum isolates, 253 has been morphologically identified as C. gloeosporioides (Penz.) Penz. & Sacc. species complex. Twenty-seven representative isolates were selected for phylogenetic analysis. Sequencing the inter-spacer gene region of ribosomal DNA confirmed the identity of the species. Artificial inoculation of flowers led to more than 2-fold higher young fruit drop compared to control. Pathogenicity tests on green fruit induced typical anthracnose symptoms on 82% of inoculated fruit two months after inoculation. Inoculation of mature fruit caused the appearance of typical anthracnose symptoms on 87% of inoculated fruit. These results showed that C. gloeosporioides species complex is responsible for different disease types on Satsuma mandarin, and that the fungus is present throughout the year on different plant organs.Antraknoza je tijekom prošlog desetljeća postala gospodarski štetna bolest mandarine Unshiu, najvažnije vrste agruma u Hrvatskoj. Cilj rada bio je istražiti. Colletotrichum vrste povezane s različitim simptomima na mandarini te utvrditi izvor zaraze. Od 2013. do 2016. sakupljena su 437 uzorka biljnog materijala. Colletotrichum vrste izolirane su iz 93 % osušenih izbojaka, 35 % otpalih cvjetova, 89 % lisnih pjega, iz svih plodova (100 %) sa simptomima antraknoze ili truleži čaške, 12 % plodova s mekom truleži u skladištu te iz 40 % plodova s pjegama koji su ostali na stablu nakon zime. Od ukupno 258 Colletotrichum izolata, 253 je morfološki determinirano kao C. gloeosporioides (Penz.) Penz. & Sacc. kompleks. Dvadeset sedam izolata odabrano je za filogenetsku analizu. Sekvenciranjem ITS regije gena ribosomske DNA potvrđena je identifikacija vrste. Umjetna zaraza cvjetova uzrokovala je više nego dvostruko veće otpadanje mladih plodića mandarine u usporedbi s kontrolom. Testovi patogenosti na nezrelim zelenim plodovima doveli su do pojave tipičnih simptoma antraknoze na 82 % inokuliranih plodova dva mjeseca nakon umjetnih zaraza. Inokulacija zrelih ubranih plodova uzrokovala je pojavu tipičnih simptoma antraknoze na 87 % inokuliranih plodova. Rezultati istraživanja pokazali su da je C. gloeosporioides kompleks vrsta uzročnik različitih tipova bolesti na mandarini Unshiu, kao i da je patogen prisutan na različitim biljnim organima tijekom cijele godine