Recubrimiento biodegradable antifúngico a base de quitosano y aceite esencial de cítricos para la conservación de papaya (Carica papaya L.) en poscosecha / Biodegradable antifungal coating based on chitosan and citrus essential oil for the conservation of papaya (Carica papaya L.) in postharvest

RESUMEN La papaya es una fruta climatérica altamente perecedera. La antracnosis, enfermedad provocada por hongos, es una de las principales causas de pérdida poscosecha. La actividad de extractos vegetales ha permitido la inhibición del desarrollo de microorganismos, en particular se ha reportado la actividad antifúngica de aceites esenciales. El objetivo de la presente investigación fue evaluar el efecto de un recubrimiento biodegradable a base de quitosano con aceite esencial de cítricos, aplicado en etapa poscosecha, en las propiedades físicoquímicas, fisiológicas e inhibición del crecimiento del hongo Colletotrichum gloeosporioides en frutos de papaya (Carica papaya L.). El recubrimiento se preparó con quitosano, glicerol, ácido acético, aceite esencial de Citrus reticulata (0.5 %, 1.0 % y 1.5 %) y Tween® 80. La solución se aplicó con una brocha en la epidermis de los frutos de papaya y se almacenaron a temperatura ambiente (24 °C ± 2 °C) por 10 d, junto con los frutos testigo. Las variables evaluadas en los frutos fueron color, pH, sólidos solubles totales (SST), firmeza, pérdida de peso, producción de CO2, de etileno y daño por antracnosis. Se utilizó un modelo estadístico completamente al azar. Los datos se sometieron al análisis de varianza (ANOVA) y comparación de medias por la prueba de Tukey (P < 0.05). El recubrimiento aplicado retrasó el cambio de color de la epidermis de los frutos de papaya, la pérdida de peso, firmeza y SST respecto a los frutos testigo. Los cambios estuvieron asociados a la modificación de la tasa de respiración y de producción de etileno de los frutos con recubrimiento. La matriz del quitosano conteniendo aceites escenciales, usada como recubrimiento, mejoró significativamente la vida de anaquel de la papaya y disminuyó más del 80 % el daño por antracnosis.

ABSTRACT Papaya is a highly perishable climacteric fruit. Anthracnose, a disease provoked by fungus, is one of the main causes of postharvest losses. The activity of plant extracts has allowed the inhibition of the development of microorganisms; in particular, the antifungal activity of essential oils has been reported. The aim of this research was to evaluate the effect of a biodegradable antifungal coating based on chitosan with citrus essential oil in the physicochemical and physiological properties and the antifungal activity of postharvest papaya (Carica papaya L.). The coating was prepared with chitosan, glycerol, acetic acid, essential oil of Citrus reticulata (0.5 %, 1.0 % and 1.5 %) and Tween® 80. The solution was applied with a brush to the epidermis of the postharvest papaya fruits and stored at room temperature (24 °C ± 2 °C) for 10 d. The variables evaluated in the fruits were color, pH, °Brix, firmness, weight loss, production of CO2 and ethylene and anthracnose damage. An analysis of variance (ANOVA) was performed under a completely randomized design and a comparison of Tukey means (P < 0.05). The coating applied delayed the color change of the epidermis of the papaya fruits, these maintained the high firmness values with respect to the control fruits. The changes were associated with the modification of the respiration rate and ethylene production of the coated fruits. The chitosan matrix containing essential oils used as a coating significantly improved shelf life of papaya and decreased anthracnose damage by 80 %.

Quiescence as a strategic stage for the infective process of Colletotrichum species

Colletotrichum, a genus of Ascomycota fungi with a diversity of species grouped into several species complexes, or clades, is associated with diseases known as "Anthracnose". It affects significantly different tropical and subtropical fruit species. Infections occur in the field and postharvest; quiescent stages are mainly responsible for postharvest losses. An analysis of this pre-infective stage is made based on consulted papers. The infective process is modulated by the length of the quiescent period of the fungus during the vegetative or pre-productive stages of trees. Quiescent stage is determined by the host's biochemical responses and the pathogen's activity. Once the pathogen is activated, it develops an infectious necrotrophic process. Colletotrichum quiescence has been studied mainly in subtropical fruits and horticultural species, and the studies highlight the role of volatile compounds, metabolites and enzymes involved in the length and breaking of quiescence, as well as the differential responses according to the phenological stage and the genotype of the plant. Transcriptomic and proteomic analysis of the host-fungus interaction have revealed the role of genes in the occurrence and breaking of quiescence. Knowledge about the quiescence of Colletotrichum in tropical fruits is necessary to improve management efficiency. Detection and induction of quiescent infections has been studied and genomics has allowed to understand the occurrence of quiescence in the infective process; however, in crops in the tropics, such as mango, an important crop in Colombia, questions about associated species and biology of their quiescence are still unanswered.

Colletotrichum es un hongo ascomicete, con diversidad de especies agrupadas en complejos o clados y se asocia a enfermedades conocidas, como "Antracnosis" y afecta significativamente especies de frutas tropicales y subtropicales. Las infecciones ocurren en el campo y postcosecha; los estados quiescentes son los principales responsables de pérdidas en postcosecha. Se realizó un análisis de esta etapa pre-infectiva, a partir de publicaciones consultadas. La infección está modulada por la duración del período de quiescencia del hongo durante las etapas vegetativa o pre-productivas de los árboles y está determinado por las respuestas bioquímicas del hospedante y la actividad del patógeno. Una vez se activa el patógeno, se desarrolla un proceso infeccioso necrotrófico. La quiescencia de Colletotrichum se ha estudiado, principalmente, en frutas subtropicales y especies hortícolas y destacan el papel de compuestos volátiles, metabolitos y enzimas en la duración y en la pérdida de la quiescencia, así como las respuestas diferenciales, según la fenología y el genotipo. Análisis transcriptómicos y proteómicos de la interacción hospedante-hongo han revelado el papel de genes en la ocurrencia y pérdida de quiescencia. Conocer la quiescencia de Colletotrichum en frutas tropicales es necesario para hacer más eficiente el manejo de la enfermedad. Se ha estudiado la detección e inducción de infecciones quiescentes y estudios genómicos han permitido entender su ocurrencia durante la infección; sin embargo, en cultivos en el trópico, como el mango, un cultivo importante en Colombia, las preguntas sobre las especies asociadas y la biología de la quiescencia de estas, aún están sin respuesta.

Análise físico-química e atividades antifúngica e antioxidante do óleo essencial da flor de Curatella americana L / Physichochemical analysis, and antifungal and antioxidant activities of the essential oil of Curatella americana L. flower

Introdução: a espécie vegetal Curatella americana produz anualmente inflorescências com aroma adocicado rica em óleo essencial. Objetivo: avaliar as características físico-químicas, e atividades antifúngica e antioxidante do óleo essencial da flor de Curatella americana. Metodologia: as flores foram coletadas em quatro áreas de Cerrado no estado de Goiás; o rendimento de óleo essencial foi obtido através de hidrodestilação; as características físicas foram determinadas para densidade e solubilidade, a atividade antioxidante foi determinada pela redução do radical livre DPPH; a atividade antifúngica foi determinada por inibição das cepas de Candida, Sclerotinia sclerotiorum, Colletotrichum gloeosporioides e Aspergillus flavus. Resultados: o rendimento de óleo foi de 0,18%, densidade de 0,907 g mL-1, solubilidade positiva para EtOH 70%, atividade antioxidante de CI50 µL mL-1 1,95. Atividade de inibição fúngica apenas para Candida tropicalis na concentração de 8% com halo de antibiose de 10 mm. Sensibilidade discreta nas maiores concentrações de 25, 50 e 100 µL-1 para Aspergillus flavus e Sclerotinia sclerotiorum e baixa atividade de inibição para Colletotrichum gloeosporioides. Conclusão: o óleo essencial da flor de Curatella americana apresentou baixo rendimento, entretanto, alta eficiência na redução do radical livre DPPH. As atividades antifúngicas apresentaram bons resultados de inibição, entretanto, torna-se necessário a adição de outros óleos essenciais para aumento das taxas de inibição micelial.

Introduction: the plant species Curatella americana produces annual inflorescences with a sweet flavour rich in essential oil. Objective: to evaluate the physicochemical characteristics, antifungal and antioxidant activities of the essential oil of the Curatella americana flower. Methodology: the flowers were collected in four areas of Cerrado in the state of Goiás; the essential oil yield was obtained through hydrodistillation; the physical characteristics were determined for density and solubility, the antioxidant activity was determined by the reduction of the free radical DPPH; antifungal activity was determined by inhibiting the strains of Candida, Sclerotinia sclerotiorum, Colletotrichum gloeosporioides and Aspergillus flavus. Results: the oil yield was 0.18%, density 0.907 g mL-1, positive solubility for EtOH 70%, antioxidant activity of IC50 µL mL-1 1.95. Fungal inhibition activity only for Candida tropicalis at a concentration of 8% with a 10 mm antibiosis halo. Discrete sensitivity in the highest concentrations of 25, 50 and 100 µL-1 for Aspergillus flavus and Sclerotinia sclerotiorum and low inhibition activity for Colletotrichum gloeosporioides. Conclusion: The essential oil of the Curatella americana flower showed low yield, however, high efficiency in reducing DPPH free radical. Antifungal activities showed good inhibition results, however, it is necessary to add other essential oils to increase mycelial inhibition rates.

Antagonistic activity of bacteria from wild honey against Colletotrichum musae, and testing of wild honey as biopesticide spray to control banana anthracnose

Anthracnose is a foliar and fruit disease caused by Colletotrichum spp. affecting a wide range of crops. Infection occurs early followed by quiescence in fruits, such as in banana, where chemical-based pesticides are used as a dependable fungal control for many years. There is an increasing need for a safe control and as implicated in the Organic Agriculture Act of 2010 (RA 10068) in the Philippines. This scenario drove the use of alternative pest control such as the use of biologicals and natural products. In this study, seven bacteria were isolated from wild honey, produced by Apis mellifera, wherein four (BC2, BC3, BC6 and BC7) were found to be an effective antagonist against Colletotrichum musae in in vitro conditions. These bacteria were identified to belong to the genus Lactobacillus spp. (BC2, BC3, BC7) and Bacillus spp. (BC6) based on sugar utilization tests, morphological and cultural growth in PDPA. For the in vivo test, different dilutions of wild honey were used and it was found out that lower concentrations were effective as biopesticide spray to prevent anthracnose infection. Lastly, we report herewith the first isolation of bacteria with biological control potential from wild honey, and to apply the raw or natural product as biopesticide in postharvest fruits.

Phaseolin Induction on Common-Bean Cultivars and Biological Control of Colletotrichum lindemuthianum 89 Race by Baccharis trimera (Less) Dc

Abstract The objective of this work was to evaluate the fungitoxic effect of the aqueous extracts of Baccharis trimera on the mycelial growth of Colletotrichum lindemuthianum 89 race, as well as its effect on the accumulation of phaseolin in hypocotyls of different cultivars and common bean varieties. It was obtained 20% aqueous extract from plants collected in municipalities of the Western Region of Paraná. Blocks containing C. lindemuthianum mycelium were transferred to Petri dishes containing medium with the different extracts and incubated at 25 °C. The colonies diameter was measured until the 12th day. Effects of aqueous extracts on phaseolin production was evaluated in hypocotyls of Carioca, Cnpf 8104, Soberana, Tibatã, Uirapurú cultivars, as well as Rosinha and Vermelho varieties. Each one cultivar and variety hypocotyl was transferred separately to test tubes containing 500 μL of 20% aqueous extracts. Sterile water, Bion®, and UV was used as controls. The phaseolin production was measured in spectrophotometer [280 nm]. Results of the evaluation of the antifungal activity of aqueous extracts of Baccharis sp. specimens collected indicate that approximately 50% of the samples presented capacity to reduce between 74 and 92% of C. lindemuthianum growth. Cultivar Tibatã and Vermelho variety showed greater sensitivity over the applied treatments. Results of fungal filtrates and vegetal aqueous extracts presented a low capacity to induce the production of phaseolin in hypocotyls.

Variability in Colletotrichum infecting chilli plants of Andaman and Nicobar Islands, India

Aim: The aim of the present study was to understand the basic pathogen dynamics of Colletotrichum species infecting chilli crop in the Andaman and Nicobar Islands for disease management practices. Methodology: Twenty fungal isolates associated with chilli (Capsicum annuum) anthracnose disease from diverse locations of tropical Andaman, India were characterized using polyphasic approaches. Results: Upon morphological studies, out of twenty isolates, fourteen isolates were found chromogenic and eleven showed concentric ring pattern of mycelial growth. Based on conidial morphology, the isolates were grouped into two species (C. capsici and C. gloesporioides). The pathogenicity tests on three chilli varieties revealed difference in the pathogenic potential among the collected isolates. The sequence analysis using internal transcriber spacer (ITS) region revealed the presence of two other species C. siamense and C. plurivorum, in addition to C. gloesporioides and C. capsici. Interpretation: The present study revealed the presence of high level of genetic and pathogenic variability among the Colletotrichum isolates infecting chilli in Andaman Islands.

Effect of Alcoholic Extracts of Cymbopogon citratus upon the Control of Colletotrichum gloeosporioides in vitro and upon the Post-harvest Quality of Guavas

Aims: This work aimed at evaluating the effects of ethanolic and methanolic extracts of lemongrass upon the control in vitro of Colletotrichum gloeosporioides and upon the post-harvest quality of guavas “Paluma”. Methodology: We analyzed the inhibition of mycelial growth and sporulation of the pathogen at different concentrations of the extracts (8%; 5%; 3%; 1.5% and 0.5%). In the post-harvest assay, the guavas were treated by immersion in distilled water, ethanolic and methanolic extracts (1%; 0.5% and 0.25%) and stored at 25ºC ± 2ºC for eight days. We evaluated mass loss, total soluble solids, total titratable acidity, ratio, reducing and non-reducing sugars, ascorbic acid and pH and the incidence of anthracnose. Results: In the test in vitro, the pathogen growth inhibition was dose-dependent and the sporulation was completely inhibited upon higher concentrations of extract. At post-harvest, the fruits maintained their physicochemical characteristics, and the treatments were not efficient at retarding fruit ripening. Although the tested treatments inhibited the plant pathogen C. gloesporioides in vitro, they were not efficient at controlling the disease in vivo. Conclusion: The extracts showed control in vitro of C. gloeosporioides at 8%. However, the extracts were not effective at controlling the disease after harvest.

Identification and Characterizations of Pathogenic Fungal Species Associated with Symptoms of Cassava Anthracnose in Ivory Coast

Cassava anthracnose is a plant disease that affects cassava stems, petioles and fruits. The aim of this study was to analyze the diversity of cassava anthracnose symptoms in Ivory Coast and then to identify and characterize the associated fungal genera. Surveys were carried out in all agricultural zones of the country from July to November, in 2014, 2015, 2016 and 2017. Infected samples consisting of stems cut with a small number of superficial cankers (0.3%), distorted stems (25.77%), and necrotic stems and petioles (65.18%) were collected. Also, withered and dried apical buds (8.76%) were harvested. Fungal pathogens derived from samples were Colletotrichum gloeosporioides (35.08%), Fusarium sp. (27.19%) and Botrytis sp. (19.73%) genera and undetermined strains (17.98%). Genera were characterized by morphological and microscopic characteristics. Parasitic pressure increased to 80 and 100% respectively for Botrytis sp. genus and Colletotrichum gloeosporioides and Fusarium sp genera. Fungal genera have caused lesions on stem and petioles in green house with diameters sizes 46, 71 and 72 mm respectively for genera Botrytis sp, Fusarium sp and Colletotrichum gloeosporioides. Aggressiveness index of Botrytis sp. genus was 3 and 4 respectively for Colletotrichum gloeosporioides and Fusarium sp. genera. The mycoflora of cassava aerial organs alteration, linked to the symptoms of anthracnose, is composed of genera of great economic importance and scientific interest.

The potential of compounds isolated from Xylaria spp. as antifungal agents against anthracnose

ABSTRACT Anthracnose is a crop disease usually caused by fungi in the genus Colletotrichum or Gloeosporium. These are considered one of the main pathogens, causing significant economic losses, such as in peppers and guarana. The current forms of control include the use of resistant cultivars, sanitary pruning and fungicides. However, even with the use of some methods of controlling these cultures, the crops are not free of anthracnose. Additionally, excessive application of fungicides increases the resistance of pathogens to agrochemicals and cause harm to human health and the environment. In order to find natural antifungal agents against guarana anthracnose, endophytic fungi were isolated from Amazon guarana. The compounds piliformic acid and cytochalasin D were isolated by chromatographic techniques from two Xylaria spp., guided by assays with Colletotrichum gloeosporioides. The isolated compounds were identified by spectrometric techniques, as NMR and mass spectrometry. This is the first report that piliformic acid and cytochalasin D have antifungal activity against C. gloeosporioides with MIC 2.92 and 2.46 µmol mL-1 respectively. Captan and difenoconazole were included as positive controls (MIC 16.63 and 0.02 µmol mL-1, respectively). Thus, Xylaria species presented a biotechnological potential and production of different active compounds which might be promising against anthracnose disease.

Induction of resistance to Colletotrichum truncatum in lima bean / Indução de resistência à Colletotrichum truncatum em feijão-fava

Anthracnose (Colletotrichum truncatum) is one of the major diseases of the lima bean culture, found in production fields, causing decrease in productivity. The objective of this study was to evaluate the effect of abiotic and biotic inducers applied in lima bean plants to reduce anthracnose severity. Lima bean accessions were used and treated with: the abiotic inducers calcium silicate (Agrosilício Plus) and silicate clay (Rocksil), using the 3 g dose.L-1; the biotic inducer citric biomass extract (Ecolife), at the dose of 3 mL.L-1; and distilled water as control. To evaluate the resistance induction, the results of severity and degree of resistance of fava bean at 7, 11, 15, 19 and 23 days after inoculation were considered according to the scale of grades and classes of reactions. The design was a randomized block in a factorial arrangement 4 × 15 (treatments × hits) with four blocks. Resistance inducers Agrosilicon Plus, Ecolife and Rocksil have potential to be used in the management of anthracnose in fava bean. The studied accesses showed degree of resistance, being able to be explored in relation to obtaining anthracnose resistant cultivars in future breeding programs of this crop.(AU)

A antracnose é uma das principais doenças da cultura do feijão-fava (Phaseolus lunatus L.), sendo encontrada em campos de produção e com grande potencial de causar redução na produtividade. O objetivo do trabalho foi determinar o efeito de indutores abiótico e biótico aplicados em plantas de feijão-fava, a fim de diminuir a severidade da antracnose. Foram utilizados 15 acessos de feijão-fava, tratados com os indutores abióticos: silicato de cálcio (Agrosilício Plus) e argila silicatada (Rocksil), utilizando a dose de 3 g.L-1 para ambos os produtos; e o indutor biótico biomassa cítrica (Ecolife), na dose de 3 mL.L-1. Como testemunha, aplicou-se apenas água destilada. Para avaliar a indução de resistência, foram considerados os resultados de severidade e grau de resistência dos acessos de feijão-fava aos 7, 11, 15, 19 e 23 dias após a inoculação, de acordo com escala de notas e classes de reações. O delineamento utilizado foi em blocos casualizados, em arranjo fatorial simples 4 × 15 (tratamentos × acessos) com quatro blocos. Os indutores de resistência Agrosilício Plus, Ecolife e Rocksil apresentaram potencial para serem utilizados no manejo da antracnose em feijão-fava. Os acessos estudados revelaram grau de resistência, podendo ser explorados quanto à obtenção de cultivares resistentes à antracnose em futuros programas de melhoramento genético dessa cultura.(AU)

Dual Biocontrol Potential of the Entomopathogenic Fungus, Isaria javanica, for Both Aphids and Plant Fungal Pathogens

Dual biocontrol of both insects and plant pathogens has been reported for certain fungal entomopathogens, including Beauveria bassiana and Lecanicillum spp. In this study, we demonstrate, for the first time, the dual biocontrol potential of two fungal isolates identified by morphological and phylogenetic analyses as Isaria javanica. Both these isolates caused mortality in the greater wax moth, and hence can be considered entomopathogens. Spores of the isolates were also pathogenic to nymphs of the green peach aphid (Myzus persicae), with an LC₅₀ value of 10⁷ spores/mL 4 days after inoculation and an LT₅₀ of 4.2 days with a dose of 10⁸ spores/mL. In vitro antifungal assays also demonstrated a strong inhibitory effect on the growth of two fungi that are pathogenic to peppers, Colletotrichum gloeosporioides and Phytophthora capsici. These results indicate that I. javanica isolates could be used as novel biocontrol agents for the simultaneous control of aphids and fungal diseases, such as anthracnose and Phytophthora blight, in an integrated pest management framework for red pepper.

Agronomic characterization and resistance of banana genotypes to Colletotrichum musae / Caracterização agronômica e resistência genótipos de banana a Colletotrichum Musae

Banana is the most consumed fruit in the world as it is a delicious and aromatic fruit. However, its production is limited mainly due post-harvest problems such as diseases, especially anthracnose, which accelerates senescence and causes early fall. The aim of this work was to evaluate the agronomic traits and the resistance of banana genotypes (Fhia 18, Fhia 01, Thap Mae, Caipira, Prata (control) and Maçã (control)) to the fungus Colletotrichum musae. The weight of bunch (Wb), fruit length (Fl), fruit diameter (Fd), the number of fruits per bunch (Nfc), number of hands per bunch (Nhb), yield (Yd), pH, total sugars (Brix) and finger detachment were evaluated. The AUDPC (area under the disease progress curve) showed that the genotypes Fhia 01, Maçã and Prata (15.77, 15.55 and 11.60 respectively) are more susceptible to the fungus C. musae than Caipira, Fhia 18 and Thap maeo (6.66, 7.29 and 8.76 respectively). The experiment with Fhias 18 and Thap Maeo genotypes showed lower finger detachment of 18 and 15%, and Thap maeo, Fhia18 and Caipira were resistant to C musae and presented the best agronomic characteristics.

A banana é fruta mais consumida no mundo, tradicionalmente exploradas em várias regiões do mundo, pois é uma fruta de muito saborosa e aromática. Contudo sua produção é limitada principalmente por problemas pós-colheita que mais acometem o fruto, dentre elas aparecimento de doenças principalmente a antracnose que acelera a senescência e o despencamento precoce. O objetivo desde trabalho foram avaliadas as características agronômicas e a resistência dos genótipos de banana Fhia 18, Fhia 01, Thap Mae, Caipira, Prata (controle) e Maçã (Controle), ao fungo Colletotrichum musae. A avaliação das características agronômicas como; peso do cacho (PC), comprimento dos frutos (CF), diâmetro do fruto (DF), número de frutos por cacho (NFC), número de pencas por cacho (NPC), Produtividade (Pd), pH, açúcares totais (Brix) e despencamento. A AACPD Área abaixo da curva da progressão da doença nos genótipos Fhia 01, Maçã e Prata (15,77, 15,55 e 11,60 respectivamente) mostraram se mais suscetível ao fungo C. musae que os demais genótipos Caipira, Fhia 18 e Thap maeo (6,66, 7,29 e 8,76 respectivamente), enquanto que para o experimento de despencamento genótipos Fhias 18 e Thap Maeo foram os que tiveram menor despencamento 18 e 15%, e a Os genótipos Thap maeo Fhia18 e Caipira foram resistentes a C musae e tiveram as melhores características agronômicas.

Caracterización química y evaluación de la actividad antifúngica del aceite esencial foliar de Lippia alba contra Colletotrichum gloeosporioides

El objetivo de este estudio fue caracterizar y evaluar in vitro la eficiencia del aceite esencial foliar de Lippia alba contra Colletotrichum gloeosporioides. Se extrajo el aceite esencial a partir de las hojas mediante el método de hidrodestilación asistida por microondas (MWHD) y se identificaron sus metabolitos mediante una cromatografía de gases acoplada a espectrometría de masas (GC/MS). La evaluación in vitro se realizó empleando 4 concentraciones del aceite esencial (500, 1000, 3000 y 10000 ppm) diluidas en acetona; se utilizó un control positivo con Benomil 1 g/L, un control negativo con acetona y un testigo absoluto; la eficiencia del aceite se midió mediante el porcentaje de índice antifúngico (%I.A). El componente mayoritario en el aceite esencial fue el citral (34.62 a 40.03%) y que el mayor %I.A se encontró a la concentración de 10000 ppm (97.8%), muy similar a la del Benomil (100%). Lo anterior demostraría la eficacia del aceite esencial de L. alba para controlar a C. gloeosporioides, y su posible uso como fungicida biológico.

The aim of this study was to characterize and evaluate the in vitro efficiency of leaf essential oil of Lippia alba against Colletotrichum gloeosporioides. Essential oil from the leaves was extracted by the method of assisted microwave hydrodistillation (MWHD) and metabolites were identified by gas chromatography coupled to mass spectrometry (GC/ MS). The in vitro evaluation was performed using 4 concentrations of essential oil (500, 1000, 3000 and 10000 ppm) diluted in acetone; positive control with Benomyl 1 g /L, a negative control with acetone and absolute control was used; oil efficiency was measured by the percentage of antifungal index (% IA). It was found that the major component was in the essential oil citral (34.62 to 40.03%) and the highest %I.A was found to 10000 ppm concentration (97.8%) very similar to Benomyl (100%). This would demonstrate the efficacy of the L. alba essential oil to control C. gloeosporioides, and its possible use as a biological fungicide.

Characterization and Pathogenicity of New Record of Anthracnose on Various Chili Varieties Caused by Colletotrichum scovillei in Korea

The anthracnose disease caused by Colletotrichum species is well-known as a major plant pathogen that primarily causes fruit rot in pepper and reduces its marketability. Thirty-five isolates representing species of Colletotrichum were obtained from chili fruits showing anthracnose disease symptoms in Chungcheongnam-do and Chungcheongbuk-do, South Korea. These 35 isolates were characterized according to morphological characteristics and nucleotide sequence data of internal transcribed spacer, glyceraldehyde-3-phosphate-dehydrogenase, and β-tubulin. The combined dataset shows that all of these 35 isolates were identified as C. scovillei and morphological characteristics were directly correlated with the nucleotide sequence data. Notably, these isolates were recorded for the first time as the causes of anthracnose caused by C. scovillei on pepper in Korea. Forty cultivars were used to investigate the pathogenicity and to identify the possible source of resistance. The result reveals that all of chili cultivars used in this study are susceptible to C. scovillei.

Caracterización molecular de Colletotrichum gloeosporioides aislado de plantas de ñame de la Costa Atlántica Colombiana utilizando la técnica "DNA Amplification Fingerprinting (DAF)" / Molecular characterization of Colletotrichum gloeosporioides isolated from yam plants of Colombia Atlantic Coast using DNA Amplification Fingerprinting technique (DAF)

La antracnosis en ñame, causada por el hongo Colletotrichum gloeosporioides, es una enfermedad relevante que posee el potencial de destruir el 100% de la cosecha, convirtiéndose en la principal limitante fitosanitaria para el rendimiento del cultivo en el país. Esta es una situación preocupante considerando que aproximadamente 35.000 familias de pequeños y medianos agricultores de la Costa Atlántica Colombiana subsisten de este cultivo; es por esto que el objetivo de este trabajo fue caracterizar molecularmente 42 aislamientos del hongo procedentes de plantas de ñame con síntomas de la enfermedad, utilizando la técnica molecular "DNA Amplification Fingerprinting (DAF)", caracterizada por su resolución en la determinación de la variabilidad genética de diferentes organismos. Para la determinación de polimorfismos, se amplificaron 16 marcadores DAF implementando iniciadores tipo decámero, los cuales fueron visualizados por electroforesis en microchip con el equipo MCE-202 MultiNA. Se evaluó la reproducibilidad de la técnica DAF. La amplificación arrojó 391 bandas inequívocamente polimórficas en todas las muestras, el coeficiente de Dice identificó cinco grupos con 0.30% de similaridad y el índice de diversidad genética fue de 0.28; datos que reflejan un alto grado de variabilidad en la colección estudiada de C. gloeosporioides. Ésta puede deberse, al intercambio de germoplasma, a su condición heterotálica, a las mutaciones y al alto potencial de dispersión de las conidias que le permiten mantener la viabilidad bajo condiciones adversas. Por último, se encontró que DAF es una técnica reproducible, confirmando que es una metodología fiable para la caracterización molecular de hongos.

Yam anthracnose, caused by the fungus Colletotrichum gloeosporioides, is a significant disease that has the potential to destroy harvests in 100%, causing the decrease in crop yields nationally. Consequently, its situation affects approximately 35.000 families of small farmers the Atlantic Coast for whom it is the livelihoods. Due to above, the objective of this work was to perform the molecular characterize of 42 C. gloeosporioides isolates from yam plants with symptoms of the disease, using the molecular technique "DNA Amplification Fingerprinting (DAF)" useful for its resolution in determining the genetic variability of different organisms. For polymorphisms determination, were amplified 16 DAF markers by using primers decamers type, the banding patron was visualized by electrophoresis in microchip 202 MCE-MultiNA equipment also, we assessed of DAF technique reproducibility. The amplification process yielded 391 unequivocally polymorphic bands in all samples, Through Dice coefficient we identified five groups with 0.30% of similarity and a genetic diversity index of 0.28. Our data show a high degree of variability in the collection of C. gloeosporioides studied. The high variability could be due to several reasons like germplasm exchanges, the heterothallic condition of fungal isolates, a presence of mutations and the high potential for conidia dispersal that allow it to maintain its viability under adverse conditions. Finally, we found that DAF technique is reproducible, confirming that it is a reliable methodology for the fungi molecular characterization.

Óleos essenciais e vegetais no controle in vitro de Colletotrichum gloeosporioides / Essential and vegetal oils in the in vitro control of Colletotrichum gloeosporioides

RESUMO O objetivo deste trabalho foi avaliar a atividade antifúngica de óleos essenciais e vegetais no controle in vitro de Colletotrichum gloeosporioides, agente causal da antracnose em pós-colheita de frutíferas. Treze óleos essenciais foram utilizados em concentrações de 0,00%, 0,40%, 0,80%, 1,70%, 3,20%, 6,25%, 12,50%, 25,00%, 50,00% e 100,00%, e uma linhagem padrão de Colletotrichum gloeosporioides. Foram avaliadas a concentração inibitória mínima e a concentração mínima fungicida a fim de caracterizar o potencial de cada um dos óleos essenciais avaliados. Verificou-se que os óleos utilizados apresentaram atividade fungicida em diferentes concentrações, as quais variaram de 0,80% (melaleuca), 3,20%, (eucalipto), 6,25% (limão, capim limão, cravo da índia, canela e nim), 12,5% (hortelã e citronela), 25% (copaíba), 50% (coco e gengibre) e 100% (manjericão). O óleo de nim apresentou maior redução da carga microbiana em função do tempo de exposição, sendo necessários 30 minutos para anulação da contagem microbiana. O efeito antifúngico dos óleos essenciais, para controle de Colletotrichum gloeosporioides, depende da planta e da concentração empregada.

ABSTRACT This study aimed to evaluate the antifungal effect of essential and vegetal oils in the in vitro control of Colletotrichum gloeosporioides, a causal agent of anthracnose in fruit postharvest. Thirteen essential oils were used at concentrations of 0.00%, 0.40%, 0.80%, 1.70%, 3.20%, 6.25%, 12.50%, 25.00%, 50.00%, and 100.00%, and also a standard strain of Colletotrichum gloeosporioides, The minimum inhibitory concentration and minimum fungicidal concentration were assessed to characterize the potential of each of the essential oils tested. We found that used oils showed fungicidal activity at different concentrations, which varied in 0.80% (Melaleuca alternifólia), 3.20%, (Eucalyptus globulus), 6.25% (Citrus limonium, Cymbopogon citratus, Syzygium aromaticum, Cinnamomum zeylanicum, and Azadirachta indica), 12.5% (Mentha piperita and Cymbopogon winterianus), 25% (Copaifera langsdorfii), 50% (Cocos nucifera and Zingiber officinale), and 100% (Ocimum basilicum). The Azadirachta indica oil showed greater reduction of microbial load because of the exposure time, and took 30 minutes for annulment of microbial count. The antifungal effect of essential oils to control Colletotrichum gloeosporioides depends on the plant and quantity of concentration.

Purification and Characterization of Peroxidase From Anthracnose Disease Infected Papaya (Carica papaya L.).

Peroxidase enzyme was isolated and purified from the pulp of disease infected ripen papaya of local variety by 90% ammonium sulphate precipitation, chromatography on DEAEcellulose followed by hydrophobic chromatography on Phenyl Sepharose CL-4B and the purifications achieved was about 7.2 fold with 2.5% recovery. The purified enzyme was homogeneous as judged by polyacrylamide slab gel electrophoresis. The purified enzyme had a Mr of about 55,000 and 50 000 as determined by gel filtration on Sephadex G-100 and SDS-PAGE, respectively. The molecular mass of the enzyme was found to be very similar under both reducing and non-reducing conditions indicating that the enzyme contains no subunit. The enzyme has the following characteristics: pH optima at 6.0, temperature optima around 38°C, enzyme activity was found to be strongly inhibited in the presence of potassium cyanide and Fe+2 while the activity was found to be remarkably increased in the presence of ammonium sulphate. The Km value for the peroxidase obtained with pyrogallol as substrate was 0.027 mM.

Spot Anthracnose Disease Caused by Colletotrichum gloeosporioides on Tulip Tree in Korea

The tulip tree (Liriodendron chinense) has been widely cultivated in Korea as a street or garden tree for its large flowers, which have a superficial resemblance to tulips. Occurrence of anthracnose disease on the leaves of tulip trees growing on the campus of Gyeongsang National University, Jinju, Korea, has been observed. Based on mycological characteristics, pathogenicity, and internal transcribed spacer sequence, the causal fungus was identified as Colletotrichum gloeosporioides. This is the first report on anthracnose disease caused by C. gloeosporioides on tulip trees in Korea.

Molecular Analysis in the differentiation of Colletotrichum gloeosporioides isolates from the cashew and mango trees

The aim of the present work was to analyze the molecular methods in the differentiation of Colletotrichum gloeosporioides isolates obtained from the cashew and mango trees. The different molecular taxonomic methods used proved to be efficient regarding intraspecific characterization. Similarly, molecular methods also proved to be efficient in differentiation of the C. gloeosporioides isolates in relation to host specificity. In the analysis of the ITS sequence of the ribosomal DNA, all the isolates amplified with the CgInt and ITS4 primers, confirming that they pertained to C. gloeosporioides. The results from this study suggested that methods based on the pathogenicity, isozyme analysis and RAPD were effective in differentiating C. gloeosporioides isolates from the cashew and mango trees.