Mycoviruses infecting Colletotrichum spp: A comprehensive review / Micoviroses em Colletotrichum spp: uma revisão abrangente

Abstract Colletotrichum is one of the most economically important fungal genera, which affects a wide range of hosts, specifically tropical and subtropical crops. Thus far, there have been several records of mycovirus infection in Colletotrichum spp., primarily by viruses of the Partitiviridae family. There have also been records of infections by mycoviruses of the Chrysoviridae family. Mycoviruses are (+)ssRNA and dsRNA genome viruses, which may or may not be enveloped. To date, no mycovirus with a DNA genome has been isolated from Colletotrichum spp. Typically, mycoviruses cause latent infections, although hypo- and hypervirulence have also been reported in Colletotrichum spp. In addition to its effects on pathogenic behavior, mycovirus infection can lead to important physiological changes, such as altered morphological characteristics, reduced vegetative growth, and suppressed conidia production. Therefore, research on mycoviruses infecting phytopathogenic fungi can help develop alternative methods to chemical control, which can cause irreversible damage to humans and the environment. From an agricultural perspective, mycoviruses can contribute to sustainable agriculture as biological control agents via changes in fungal physiology, ultimately resulting in the total loss of or reduction in the virulence of these pathogens.

Resumo Colletotrichum é um dos gêneros fúngicos mais importantes economicamente, afetando uma ampla gama de hospedeiros, especialmente em cultivos tropicais e subtropicais. Atualmente já existem diversos registros de infecção por micovírus em Colletotrichum spp., sendo a maioria dos já identificados classificados na família Partitiviridae. Ocorrem registros também de micovírus pertencentes à família Chrysoviridae. Compreendem vírus de genoma de (+)ssRNA e dsRNA que podem ser ou não envelopados. Ainda não foram identificados micovírus com genoma de DNA isolados de Colletotrichum. A infecção por micovírus pode ocorrer de forma latente, mas já foi observado em Colletotrichum spp. o fenômeno de hipo e hipervirulência. Além de influenciar no comportamento patogênico, a infecção pode causar mudanças fisiológicas importantes como alterações das características morfológicas, redução do crescimento vegetativo e redução na produção de conídios. O estudo com micovírus em fungos fitopatogênicos traz uma alternativa ao controle químico que é um método capaz de causar danos irreversíveis ao homem e o meio ambiente. Sob a perspectiva agrícola, os micovírus podem contribuir para agricultura sustentável como agentes de controle biológico. Isso porque obsevam-se mudanças importantes na fisiologia fúngica resultando na perda total ou redução da virulência desses patógenos.

Mycoviruses infecting Colletotrichum spp: a comprehensive review / Micoviroses em Colletotrichum spp: uma revisão abrangente

Colletotrichum is one of the most economically important fungal genera, which affects a wide range of hosts, specifically tropical and subtropical crops. Thus far, there have been several records of mycovirus infection in Colletotrichum spp., primarily by viruses of the Partitiviridae family. There have also been records of infections by mycoviruses of the Chrysoviridae family. Mycoviruses are (+)ssRNA and dsRNA genome viruses, which may or may not be enveloped. To date, no mycovirus with a DNA genome has been isolated from Colletotrichum spp. Typically, mycoviruses cause latent infections, although hypo- and hypervirulence have also been reported in Colletotrichum spp. In addition to its effects on pathogenic behavior, mycovirus infection can lead to important physiological changes, such as altered morphological characteristics, reduced vegetative growth, and suppressed conidia production. Therefore, research on mycoviruses infecting phytopathogenic fungi can help develop alternative methods to chemical control, which can cause irreversible damage to humans and the environment. From an agricultural perspective, mycoviruses can contribute to sustainable agriculture as biological control agents via changes in fungal physiology, ultimately resulting in the total loss of or reduction in the virulence of these pathogens.

Colletotrichum é um dos gêneros fúngicos mais importantes economicamente, afetando uma ampla gama de hospedeiros, especialmente em cultivos tropicais e subtropicais. Atualmente já existem diversos registros de infecção por micovírus em Colletotrichum spp., sendo a maioria dos já identificados classificados na família Partitiviridae. Ocorrem registros também de micovírus pertencentes à família Chrysoviridae. Compreendem vírus de genoma de (+)ssRNA e dsRNA que podem ser ou não envelopados. Ainda não foram identificados micovírus com genoma de DNA isolados de Colletotrichum. A infecção por micovírus pode ocorrer de forma latente, mas já foi observado em Colletotrichum spp. o fenômeno de hipo e hipervirulência. Além de influenciar no comportamento patogênico, a infecção pode causar mudanças fisiológicas importantes como alterações das características morfológicas, redução do crescimento vegetativo e redução na produção de conídios. O estudo com micovírus em fungos fitopatogênicos traz uma alternativa ao controle químico que é um método capaz de causar danos irreversíveis ao homem e o meio ambiente. Sob a perspectiva agrícola, os micovírus podem contribuir para agricultura sustentável como agentes de controle biológico. Isso porque obsevam-se mudanças importantes na fisiologia fúngica resultando na perda total ou redução da virulência desses patógenos.

Resistance to diseases of sour, sweet and wild passion fruit genotypes

The cultivation of passion fruit is important for Brazil, since the country is currently the largest producer and consumer of fruit in the world. However, the fields of passion fruit still face important problems due to the incidence and severity of diseases in the field. Thus, the present study aimed to assess resistance to bacterial and fungal diseases in 13 genotypes of sour, sweet and wild passion fruit, in field conditions in the Distrito Federal, Brazil. For this, a field experiment was installed in a randomized block design, with four replications and 13 treatments (genotypes). The characteristics of incidence, severity and degree of resistance for bacteriosis, septoriosis, scab and anthracnose diseases were evaluated in 5 fruits per plot of each genotype. Genetic parameters of the evaluated traits were also estimated. High heritability values and CVg/Cve ratio were observed for most of the evaluated characteristics. The genotypes presented mean values of incidence and severity of bacteriosis, septoriosis, scab and anthracnose different among them, and the one that presented the best results in the degree of resistance for all diseases was F1 (MAR20 # 24 x ECL7 P1 R4).

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.

Evaluation of the production of antifungal metabolites against Colletotrichum gloeosporioides in Streptomyces 5.1 by random mutagenesis

Streptomyces 5.1 is a bacterium isolated from rice soils in the south of the Tolima department (Colombia). This microorganism is characterized by its antagonistic activity against rubber tree phytopathogens like Colletotrichum gloeosporioides, the causal agent of leaf anthracnose. The antifungal activity of this Streptomyces isolate has been associated with secondary metabolites production. However, the identity of those metabolites is unknown because its purification and identification have not been possible through classic chemical studies. Therefore, aiming to contribute in the study of the secondary metabolites produced by 5.1 from a molecular approach, this research seeks to identify -preliminarily- the genomic fingerprint changes associated with the production of antifungal secondary metabolites produced by Streptomyces 5.1 through the evaluation of a mutant library of 5.1 obtained by random mutagenesis using controlled ultraviolet light exposure. The antifungal activity of obtained mutants was evaluated using Colletotrichum gloeosporioides (C1) fungus as a biosensor, isolated by the Biotechnology Institute of Universidad Nacional de Colombia. In this way, the library of mutants of 5.1, initially formed by 300 isolations, was classified into two phenotypic groups of interest: enhanced mutants (1 isolate) and null mutants (11 isolates) of secondary metabolites. The genomic changes in both groups were analyzed by obtaining the genomic profile of the isolates using Repetitive Extragenic Palindromic (Rep-PCR). The obtained profiles evidenced the presence of one additional band in the enhanced mutant, and the absence of a specific band in the non-producing mutants, both in comparison with the original strain. These bands are proposed for a future sequencing study which will define their role in the production process of metabolites with antifungal activity in Streptomyces 5.1.

Colletotrichum siamense, a Mycovirus-Carrying Endophyte, as a Biological Control Strategy for Anthracnose in Guarana Plants

Abstract Guarana, the fruit of Paullinia cupana, is known for its stimulating and medicinal properties by the Amazonian indigenous population and communities. However, it presents serious phytopathological problems, such as anthracnose disease caused by Colletotrichum spp. The objective of this study was to verify if C. siamense, a mycovirus-carrying endophytic fungus, could protect guarana seedlings, by reducing or by eliminating characteristic disease symptoms. Other physiological changes in the plant caused by the presence of this endophyte were also evaluated. The cuttings of the Cereçaporanga cultivar were dipped in a biological control suspension and planted in a specific substrate. After four months in the greenhouse, the seedlings were sprayed with a suspension of phytopathogen conidia, and a portion of these seedlings received the fungicide indicated for the crop to be compared with the control seedlings. After 28 days, the number of lesions, morphophysiological and macro characteristics, and leaf micronutrients were evaluated. The seedlings treated with C. siamense showed a lower percentage of lesions and an increased aerial part and root system compared to the other treatments. There were no significant differences between treatments regarding the percentage of macronutrients and micronutrients.

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.

Genetic diversity and antibacterial activity of endophytic fungi from Zanthoxylum nitidum / 中国中药杂志

In order to reveal the distribution and population characteristics of endophytic fungi from Zanthoxylum nitidum and the antibacterial potential,this study performed molecular identification and analyzed the genetic diversity and antibacterial activity of endophytic fungi from Z. nitidum in Guangxi. Through culture and molecular identification,35 strains,belonging to 15 genera,12 families,10 orders,4 classes,and 2 phyla,were isolated from various tissues of Z. nitidum,of which Colletotrichum and Fusarium were the dominant genera,respectively accounting for 20% of total strains. The diversity of endophytic fungi was significantly different among roots,stems,and leaves,as manifested by the significantly higher Shannon index( H') in stems( 1. 678) than in roots( 0. 882 1) and leaves( 0. 515 4). The antimicrobial activity analysis showed that 14. 28% of endophytic fungi inhibited at least one indicator pathogen. Among them,Fusarium sp. ZN-34 and Fusarium sp. ZN-26 separately demonstrated the strongest inhibitory effect on Escherichia coli and Staphylococcus aureus. In general,Fusarium sp. ZN-26 and Phialemoniopsis plurioloculosa ZN-35 were advantageous in suppressing the two bacteria owing to the broad spectrum and strong efficacy. In summary,Z. nitidum in Guangxi boasts rich endophytic fungi with the majority showing strong antibacterial activity,which can be used as candidates for the extraction and separation of basic antibacterial substances and the development of natural antibacterial agents.

A new phenolic acid ester from solid culture of Colletotrichum capsici, an endophytic fungus from Paeonia lactiflora / 中国中药杂志

A new phenolic acid ester, 4'-hydroxyphenylethyl 4,8(R)-dihydroxyphenylpropionate(1), was isolated from an endophytic fungus Colletotrichum capsici of Paeonia lactiflora roots, along with eight known phenolic derivatives, tyrosol(2), 2-(4-hydroxyphenyl) ethyl acetate(3), methyl p-hydroxyphenylacetate(4), methyl m-hydroxyphenylacetate(5), 4-(4-hydroxyphene-thoxy)-4-oxobutanoic acid(6), 4-hydroxyphenethyl methyl succinate(7), trichodenol B(8) and 4-hydroxyphenethyl 2-(4-hydroxyphenyl) acetate(9). Their structures were identified by a combination of high-resolution electrospray ionization mass spectrometry(HR-ESI-MS), nuclear magnetic resonance(NMR) spectroscopy, ultraviolet(UV) spectroscopy and electronic circular dichroism(ECD) spectroscopy. Compounds 2-9 were isolated from this fungus for the first time.

A novel pathogenic Colletotrichum species for rhizome rot disease of Polygonatum kingianum / 中国中药杂志

Rhizome rot disease is one of the main disease of planted Polygonatum kingianum. In this study, six strains of pathogenic fungus was isolated from P. kingianum samples with rhizome rot disease collected from six counties in Yunnan province. Its pathogenicity was confirmed by inoculation to healthy P. kingianum rhizome according to Koch's postulates. The colonies of the isolated fungi on potato dextrose agar(PDA) were orange with abundant crescentic conidia which were eseptate with a mean size of 19. 3-24. 9 μm×5. 2-5. 9 μm and a L/W ratio of 3. 4-4. 5. There was an oil ball in the center of the conidium. It's easy to see setae on PDA colony.The phylogenetic tree based on ITS, GAPDH, CHS-1, HIS3, ACT, and TUB2 sequences by maximum likelihood(ML) method indicated that the pathogenic fungus for P. kingianum rhizome rot disease was clustered into the clade of Colletotrichum spaethianum species complex, and was close to C. spaethianum. However, there were some differences in morphological and genetic characteristics between the pathogenic fungus and C. spaethianum. Therefore, the pathogenic fungus for rhizome rot disease of P. kingianum was identified as a new Colletotrichum species named C. kingianum. The disease spreads primarily due to the plantation of infected seedlings of P. kingianum. It is necessary to choose healthy seedlings and take rigorous disinfection measures for the disease prevention.

Emergence and epidemics of a new disease in Araguaia Valley: anthracnosis of cotton caused by Colletotrichum truncatum, varietal reaction and difficulties of chemical control / Epidemias da antracnose do algodoeiro no Vale do Araguaia causada por Colletotrichum truncatum reação de cultivares e dificuldades de controle químico

Cotton culture (Gossypium hirsutum L.) is trending in an upward expansion amidst Brazilian "cerrado". Due this growth, new pathosystems are growing in incidence on tropical fields in the region of Araguaia Valley - MT. Surveys and material collection were conducted out in production areas on two counties (Bom Jesus - MT and Canarana ­ MT), both regions represents a total amount of 50.000 hectares of cultivated area. The region also is characterized by succession areas previously sowed with soybean plants (main cover crop season). Previously surveys revealed the incidence of target spot (Corynespora cassiicola), ramularia spot (Ramularia areola) and cercosporiosis (Cercospora gossypii). Disease incidence is commonly observed on several crop management methods and cultivars. Due to a reduction in efficacy of chemichal control by fungicides spray programs, this disease is increasing and spreading in a faster rate in production areas with previous harvested soybean at the biggest soybean producer group in the world (Bom Futuro). Plants on field were selected due to differences in symptoms, and isolation methods were carried out on PDA (potato-dextrose-agar) before "in vitro" pathogenicity tests conducted on seedlings, detached leaves and bolls (growth chamber conditions of 23°C / photoperiod of 12 hours). After Colletotrichum dextructor sp. pathogenicity confirmation, bioassays were carried out with several different fungicide's groups (i.e. registered for usage on Brazilian jurisdiction). This test consisted of the employment of a solution with 500 ppm of each different fungicide/a.i. that is applied on infected bolls and detached leaves who were inoculated with the target pathogen (concentration of 104 conidia per mL). This test was followed by an infection (%) evaluation during 10 days of incubation. The incidence/prevalence index (%) was also evaluated at different parts of the plant (lower, middle, and upper canopy) on different cultivars/genotypes (130-150 days after sowing). A standard level of control by different fungicides, ranged between 0 to 100% of control. Two groups of cultivars/genotypes were separated after differences on resistance response, one with susceptible traits (FM 985 GLTP) and other with partial resistance traits. No immunity response was observed. We suggest that new efficacy tests should be carried out with combination of varietal response (resistance or tolerance) combined with other chemical fungicides for better understanding of synergism or positive interaction. Valuable information will highlight the best association for greater varietal response and yield against this necrotrophic pathogen (higher B0 ­ initial inoculum) survival during successive years of rotation main crop (cotton x soybean). Isolated spray of benzimidazoles, cupric (except cuprous oxide) and triple associations with triazoles or triazolinthione (prothioconazol) combined with strobilurins and carboxamides should be wisely administrated to manage this disease due to low efficiency (below 50%). New field and laboratory essay must be carried out to input data about resistance risk and clarify damage levels on leaves and bolls impacting yield.

O algodoeiro Gossypium hirsutum L. está em plena expansão no Brasil em área da Bahia e Mato Grosso. Em função dessa expansão novos patossistemas podem ocorrer nas áreas tropicais na região do Vale do Araguaia - MT. Foram realizados levantamentos em áreas de produção nos municípios de Bom Jesus ­ MT e Canarana ­ MT nas áreas de expansão da cultura que atingem mais de 50.000 ha. Nessa região o cultivo é realizado após o cultivo da soja em uma sucessão. São comuns nessas áreas a mancha alvo (Corynespora cassiicola), mancha de ramularia (Ramularia areola) e a Cercosporiose (Cercospora gossypii). Devido as falhas nos programas, com diferentes fungicidas de controle foram realizadas inspeções nas áreas de produção do grupo Bom futuro em diferentes cultivares e sistemas de manejo. Em função da sintomatologia diferenciada e constando-se ser um novo problema foram realizados isolamentos em BDA (batata-dextrose-ágar) e realizados testes de patogenicidade em plântulas, folhas e maçãs destacadas na temperatura de 23o Celsius e regime alternado de luz do dia por 12 horas. Após a confirmação da patogenicidade em plântulas, folhas destacadas, maçãs ou capulhos para Colletotrichum truncatum foram realizados bioensaios com todos os grupos de fungicidas registrados no Brasil. Foram usada a concentração de 500 (mg.kg-1) ppm dos ingredientes ativos, de cada formulação ou fungicida. Após a pulverização dos fungicidas foi em seguida inoculado o patógeno na concentração de 104 conídios por ml. Avaliou-se 10 dias após a incubação a porcentagem de infecção nas folhas destacadas, assim como o percentual de infecção em cada maça inoculada pelo patógeno. Confirmou-se um nível de controle pelos diferentes fungicidas variando de 0 a 100 %, tanto para folhas, como para maças destacadas. Também foi avaliado o índice de prevalência (%), considerando os três terços da planta (terço inferior, médio e superior), em diferentes genótipos em condições de campo na fase final da produção das plantas (130-150 dias). Os genótipos foram suscetíveis (FM 985 GLTP) e muitos genótipos apresentaram resistência parcial. Nenhum genótipo foi imune ao patógeno. Sugere-se a revisão dos programas e manejo com fungicidas visando alternar os melhores ingredientes ativos para o novo patógeno, associado aos genótipos como maior resistência varietal, uma vez que o patógeno pode sobreviver de um ano para o outro em sementes e restos culturais. Fungicidas benzimidazóis, cúpricos (exceto óxido cuproso), benzimidazol + triazol + estrobilurina e carboxamida + triazolintiona + estrobilurina devem ser usados com cautela no controle da doença em função da sua baixa eficiência (abaixo de 50 %), em condições de laboratório (folhas e frutos ­ maçãs destacadas).

Antifungal activity of essential oils of Myrcia ovata chemotypes and their major compounds on phytopathogenic fungi / Atividade antifúngica de óleos essenciais de quimiotipos de Myrcia ovata e seus principais compostos em fungos fitopatogênicos

This work evaluated the antifungal activity of essential oils of Myrcia ovata chemotypes (MYRO-175, MYRO-156, MYRO-154, MYRO-165, and MYRO-015) and their major compounds (linalool, geraniol, citral, and (E)-nerolidol) on the phytopathogenic fungi Fusarium pallidoroseum (which causes melon postharvest rot) and Colletotrichum musae (which causes anthracnose in banana). The essential oils were obtained by hydrodistillation and analyzed by GCMS/FID. To evaluate the antifungal activity, the essential oils and their major compounds were tested at different concentrations (0.1; 0.3; 0.4; 0.5; 0.7; 1.0; 3.0, and 5.0 mL/L). The major compounds found in the essential oils were nerolic acid, linalool, geraniol, citral, and (E)-nerolidol. The essential oils of the plants MYRO-154, MYRO-165, and MYRO-015 had the minimum inhibitory concentration (MIC) (0.3 mL/L) for F. pallidoroseum and the lowest minimum fungicidal concentration (MFC) (0.7 mL/L), for C. musae. Geraniol and citral had the lowest MFC (0.5 mL / L) for the two fungi tested. For F. pallidoroseum, the essential oils of the chemotypes were more effective than their major compounds. Conversely, the major compounds geraniol of the chemotype MYRO-156 (74.37%) and citral were more effective than their respective essential oils for C. musae. (E)-nerolidol and geraniol of the chemotype MYRO-015 (33.15%) were responsible for the antifungal activity of the essential oils of their respective chemotypes.

No presente trabalho avaliou-se a atividade antifúngica de óleos essenciais de quimiotipos de Myrcia lundiana (MYRO-175, MYRO-156, MYRO-154, MYRO-165, and MYRO-015) e seus compostos majoritários (linalol, geraniol, citral e (E)-nerolidol) sobre os fungos fitopatogênicos Fusarium pallidoroseum(causa podridão em frutos de melão) e Colletotrichum musae (causa antracnose em frutos de banana). Os óleos essenciais foram obtidos hidrodestilação e analisados por CGEM/DIC. Para avaliação da atividade antifúngica foram testados os óleos essenciais e os compostos majoritários nas concentrações: 0,1; 0,3; 0,4; 0,5; 0,7; 1,0; 3,0 e 5,0 mL/L. Os principais compostos presentes nos óleos essenciais foram o ácido nerólico, o linalol, o geraniol, o citral e o (E)-nerolidol. Os óleos essenciais das plantas MYRO-154, MYRO-165 e MYRO-015 apresentaram CIM de 0,3 mL/L e a planta MYRO-015 apresentou a menor concentração fungicida mínima (CFM) (1,0 mL/L). O geraniol e o citral foram os compostos que apresentaram o menor valor de CFM, 0,5 mL/L, frente aos dois fungos testados. O óleo essencial dos quimiotipos testados foram mais promissores que seus componentes majoritários puros, frente o F. pallidoroseum. Já para o C. musae, os componentes majoritários geraniol do quimiotipo MYRO-156 (74,37%) e o citral foram mais promissores que seus respectivos óleos essenciais. Já o (E)-nerolidol e o geraniol do quimiotipo MYRO-015 (33,15%) foram os responsáveis pela atividade antifúngica apresentada pelos óleos essenciais dos respectivos quimiotipos.

Distribución espacial de antracnosis (Colletotrichum gloeosporioides Penz) en aguacate en el Estado de México, México / Spatial distribution of anthracnose (Colletotrichum gloeosporioides Penz) in avocado in the State of Mexico, Mexico

Resumen El aguacate (Persea americana) es una especie cuyo cultivo es de gran importancia nutricional y económica para México; sin embargo, como cualquier otro cultivo, a menudo se ve afectado por plagas y enfermedades que limitan su comercialización a nivel mundial. El hongo fitopatógeno Colletotrichum gloeosporioides es el agente causal de la antracnosis en el aguacate y se manifiesta en las etapas tempranas del desarrollo del fruto, así como en poscosecha y durante el almacenamiento, en condiciones de alta humedad relativa (80%) y temperaturas desde los 20 ◦C. Las pérdidas económicas a causa de este hongo pueden ser de hasta el 20% de la producción. En el presente estudio se aplicaron métodos geoestadísticos para definir la distribución espacial de antracnosis en frutos de aguacate cultivar Hass en cuatro municipios del Estado de México, durante el periodo de enero a junio de 2017. La distribución de la antracnosis se ajustó a modelos gaussianos y exponenciales en la mayoría de los casos. Los mapas de infestación realizados mediante krigeado muestran más de un centro de agregación de la enfermedad. Este análisis permitió estimar la superficie infestada: se encontró una infestación de más del 50% en los primeros muestreos y de hasta un 98% en los muestreos de junio en todas las zonas estudiadas. © 2019 Publicado por Elsevier Espana, S.L.U. en nombre de Asociacion Argentina de Microbiologıa. Este es un art´ıculo Open Access bajo la licencia CC BY-NC-ND (http://creativecommons. org/licenses/by-nc-nd/4.0/).

Abstract Persea americana is a species of great nutritional and economic importance for Mexico, however, like any other agricultural crop, it is affected by pests and diseases that limit its worldwide commercialization. The phytopathogenic fungus Colletotrichum gloeosporioides is the causative agent of anthracnose in avocado and manifests itself in the early stages of fruit development as well as in post-harvest and storage, under conditions of high relative humidity (80%) and at temperatures from 20°C, causing losses economic up to 20% of production. Applying geostatistical methods the present study aims to define the spatial distribution of anthracnose in Hass avocado fruits in four municipalities of the State of Mexico during the period from January to June 2017. The results show that the distribution of anthracnose was adjusted to gaussian and exponential models in most, the infestation maps made through the kriging show more than one centerof aggregation of the disease, based on it the infested surface was estimated, finding an infestation of more than 50% in the first samples and up to 98% in the samplings belonging to the month of June in all the areas studied. © 2019 Published by Elsevier Espana, S.L.U. on behalf of Asociación Argentina de Microbiología. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/ licenses/by-nc-nd/4.0/).

Control of anthracnose in papaya fruits by acetylsalicylic acid and 1-methylcyclopropene / Controle da antracnose em mamão por ácido acetil salicílico e 1-metilciclopropeno

This study assessed the effect of acetylsalicylic acid (ASA) and 1-methylcyclopropene (1-MCP) to control anthracnose in papaya (cultivar Golden). Disinfested-surface fruits were inoculated with Colletotrichum gloeosporioides and then the compounds were applied. The lesion diameters and the physical-chemical properties were analyzed. Assays were carried out with ASA and 1-MCP targeting the control of anthracnose and maintenance of the fruit's physical-chemical properties. The effect of ASA (20 mM; 20 min) on reducing lesion diameter occurred when applied before inoculation. Fruits treated with 1-MCP (300ppb) for 12 h showed a smaller lesion diameter than control. For the physical-chemical analysis, fruit treated with 1-MCP (200; 300 ppb; 12h) maintained fruit firmness, delayed fruit ripening and fruit fresh weight loss.

Neste trabalho avaliou-se o efeito do ácido acetilsalicílico (ASA) e 1-metilciclopropeno (1-MCP) no controle da antracnose do mamão (cv Golden). Para isso o Colletotrichum gloeosporioides foi inoculado em frutos superfícialmente desinfestados para depois aplicar-se os compostos. O diâmetro das lesões e as características físico-químicas foram determinadas. Os ensaios foram conduzidos com ASA e 1-MCP visando o controle da antracnose e a manutenção das características físico-químicas do fruto. O efeito do AAS (20 mM; 20 min) na redução do diâmetro da lesão ocorreu quando aplicado antes da inoculação. Frutos tratados com 1-MCP (300 ppb) por 12 horas apresentaram menor diâmetro de lesão que controle. Para a análise físico-química, frutos tratados com 1-MCP (200; 300 ppb; 12h) mantiveram a firmeza, o amadurecimento tardio e a perda de massa fresca dos frutos.

Bioprospection and molecular phylogeny of culturable endophytic fungi associated with yellow passion fruit

Endophytic microorganisms live inside the plants without causing any damage to their hosts. In the agricultural field, these endophytes might be a strategy of biological control for phytopathogens. We aimed to isolate endophytic fungifrom yellowpassion fruit (Passiflora edulis) leaves, evaluating its biocontrol capacity by in vitroantagonism against phytopathogen Colletotrichum sp. CNPU378. We also carried out greenhouse experiments in bean seedlings. A high colonization frequency was obtained (89%), and the molecular identification based on DNA sequencing attested Colletotrichumas the most frequent genus and minor occurrence of Curvulariaendophytes. The endophytes tested showed different types of competitive interactions in in vitro antagonism inhibition rate ranging from 28.8 to 48.8%. There were 10 promising antagonists tested for their antagonist activity of crude extracts of secondary metabolites, in which strain PE-36 (20.8%) stood out among the other strains evaluated. In the greenhouse assay, plants inoculated only with endophyte Colletotrichumsp. PE-36 was symptomless and suggest that the endophyte strengthened the growth promotion in common bean plants, especially in the root length and number of leaves when compared to control plantsand other treatments. Despite many fungiof Colletotrichumgenus being described as causative agents of anthracnose, in this study, the plant sampled was colonized predominantly by Colletotrichumendophytes living in asymptomatic relationship. By the way,we come across a Colletotrichumsp. endophyte able to antagonize a Colletotrichumsp. pathogen

In vitro antifungal activity of Myrcia ovata essential oils and their major compounds against pathogens of citrus, sweet potato, and coconut / Atividade antifungica in vitro de óleos essenciais de myrcia ovata e seus compostos majoritários sobre patógenos de citros, batata-doce e coqueiro

Myrcia ovata, an endemic species to the Brazilian Atlantic Forest, presents antifungal properties. The phytopathogens Colletotrichum acutatum, Plenodomus destruens,and Thielaviopsis paradoxa are responsible for the diseases citrus postbloom fruit drop, sweet potato foot rot, and coconut stem bleeding, respectively. The antifungal activity of the essential oils of five M. ovata chemotypes (MYRO-159, nerolic acid chemotype; MYRO-180, nerolic acid + linalool chemotype; MYRO-388, geraniol chemotype; MYRO-157, citral + (E)-nerolidol chemotype; and MYRO-174, isopulegol + linalool chemotype), four major compounds (nerolic acid, nerolic acid + linalool, geraniol, and citral + (E)-nerolidol), and threepure compounds (citral, (E)-nerolidol, and linalool) against the fungi C. acutatum, P. destruens,and T. paradoxawere evaluated. For this, in vitro tests were conducted in a completely randomized design with three replications, testing concentrations (v/v) ranging from 0.01 to 1.0 µL.mL-1. All treatments presented toxicity at different levels to the three fungi. For C. acutatum,the essential oil from the individual MYRO-180 (nerolic acid + linalool chemotype) and its major compound showed the lowest Minimal Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) of 0.03 and 0.1 µL.mL-1, respectively. For P. destruens, the essential oil from the individual MYRO-159 (nerolic acid chemotype) presented the lowest MIC of 0.05 µL.mL-1. The nerolic acid + linalool chemotype and its major compound presented an MFC of 0.07 µL.mL-1. For T. paradoxa, the major compound citral + (E)-nerolidol stood out with the lowest MIC and MFC of 0.03 and 0.2 µL.mL-1, respectively. Linalool presented the lowest toxicity to the three tested fungi.

Myrcia ovata, uma espécie nativa de fitofisionomia de Restinga, possui atividade antifúngica. Os fitopatógenos Colletotrichum acutatum, Plenodomus destruens e Thielaviopsis paradoxa são responsáveis pelas doenças podridão floral de citros, mal-do-pé da batata doce e resinose do coqueiro, respectivamente. A atividade antifúngica de cinco quimiotipos de M. ovata (MYRO-159, quimiotipo ácido nerólico; MYRO-180, ácido nerólico + linalol; MYRO-388, quimiotipo geraniol; MYRO-157, quimiotipo citral + (E)-nerolidol; e, MYRO-174, quimiotipo isopulegol + linalol), quatro compostos majoritários (ácido nerólico, ácido nerólico + linalol, geraniol e citral + (E)-nerolidol) e três compostos isolados (citral, (E)-nerolidol e linalol) foram avaliados sobre os fungos C. acutatum, P. destruens e T. paradoxa. Testes in vitro foram conduzidos em delineamento inteiramente casualizado com três repetições e concentrações (v/v), que variaram de 0,01 a 1,0 µL.mL-1. Todos os tratamentos testados apresentaram atividade antifúngica. Para o fungo C. acutatum, o óleo essencial do indivíduo MYRO-180, de quimiotipo ácido nerólico + linalol, e seu composto majoritário apresentaram menores Concentração Mínima Inibitória (CMI) e Concentração Mínima Fungicida (CMF) de 0,03 e 0,1 µL.mL-1, respectivamente. Para o fungo P. destruens, o óleo essencial do indivíduo MYRO-159, de quimiotipo ácido nerólico, apresentou menor CMI de 0,05 µL.mL-1, e o quimiotipo ácido nerólico + linalol e seu composto majoritário apresentaram a menor CMF de 0,07 µL.mL-1. Para o fungo T. paradoxa,a combinação de citral + (E)-nerolidol destacou-se com CMI e CMF de 0,03 e 0,2 µL.mL-1, respectivamente. Linalol foi o menos tóxico sobre os três fungos testados.

Use of phosphites in postharvest to control anthracnose of yellow passionfruit / Uso de fosfitos em pós-colheita para o controle da antracnose do maracujá-amarelo

Brazil is the largest producer of yellow passionfruit (Passiflora edulis f. flavicarpa) and one of its production problems is the anthracnose (Colletotrichum spp.). The use of fungicides on control of postharvest diseases is a method that protects the fruits during storage. However, precautions must be taken due to fungicide toxicity. The restriction to the use of fungicides in post-harvest led a demand for alternative methods of disease control, and, the phosphite application is one of these methods. Therefore, this work aimed to evaluate the effects of fruit immersion in phosphite on postharvest control of anthracnose. Two tests were developed in vitro to assess the effect on the fungus: phosphite Mg2 (40%P2O5+6%Mg), Zn (40%P2O5+10%Zn), Ca1 (30%P2O5+7%Ca) and K1 (40%P2O5+20%K2O). For the in vivo tests, passionfruit (Gigante Amarelo), were wounded and inoculated (50L; 106conidia mL-1). Two tests were done with: Cu (25%P2O5+5%Cu), 2.5mL L-1; Zn, 2.5mL L-1; K1, 2.5mL L-1; Mg1 (30%P2O5+4%Mg), 3mL L-1; Ca1, 3mL L-1; Ca2 (10%P2O5+6%Ca), 4mL L-1; K2 (40%P2O5+20%K2O), 1.5mL L-1; Mg2 (40%P2O5+6%Mg), 1.5mL L-1; K3 (20%P2O5+20%K2O) 1.75 mL L-1; K4 (30%P2O5+20%K2O), 1.75mL L-1. Other two tests with phosphites Mg2, Ca1, Zn and K1 were with CaCl2 (2%) was developed. In addition, phosphites were tested at 25, 50, 100 and 200% of the dose: K2 (100%; 1.5mL L-1) and Ca1 (100%; 3 mL L-1). The phosphites Mg2, Ca1, K1 and Zn in vitro have reduced mycelial growth and fungus conidia production. The phosphites K1, K2, Ca1 and Zn were the ones that most reduced the size of the anthracnose lesion. There were no differences among treatments, concerning the physico-chemical fruit properties analyzed (% fresh mass loss, total soluble solids, pH and titratable acidity).

O Brasil é o maior produtor mundial de maracujá-amarelo (Passiflora edulis f. flavicarpa) e um dos problemas para sua produção é a antracnose (Colletotrichum spp.). O uso de fungicidas no controle de doenças pós-colheita é um método que protege os frutos durante o armazenamento, mas, precauções adicionais devem ser tomadas quanto à sua toxidade, presença de resíduos e a provável seleção de fungos resistentes. A restrição ao uso de fungicidas na pós-colheita cresceu e levou à procura de alternativas de controle, e, entre tais está à aplicação de fosfitos. Diante disso, este trabalho objetivou avaliar os efeitos da imersão de frutos em soluções de fosfitos no controle da antracnose em pós-colheita. Dois testes in vitro foram feitospara avaliar o efeito de fosfitono fungo: Mg2 (40%P2O5+6%Mg), Zn (40%P2O5+10%Zn), Ca1 (30%P2O5+7%Ca) e K1 (40%P2O5+20%K2O). In vivo, frutos de maracujá (Gigante Amarelo), foram feridos e inoculados (50l; 106conídios mL-1). Dois testes foram feitos com: Cu (25%P2O5+5%Cu), 2,5mL L-1; Zn, 2,5mL L-1; K1, 2,5mL L-1; Mg1 (30%P2O5+4%Mg), 3mL L-1; Ca1, 3mL L-1; Ca2 (10%P2O5+6%Ca), 4mL L-1; K2 (40%P2O5+20%K2O), 1,5mL L-1; Mg2 (40%P2O5+6%Mg), 1,5mL L-1; K3 (20%P2O5+20%K2O), 1,75mL L-1; K4 (30%P2O5+20%K2O), 1,75mL L-1. Outros dois testes com fosfitos foram com CaCl2 (2%) e Mg2, Ca1, Zn e K1. Ainda, dois fosfitos foram testados a 25, 50, 100 e 200% da dose: K2 (100%; 1,5mL L-1) e Ca1 (100%; 3mL L-1). Os fosfitos Mg2, Ca1, K1 e Zn in vitro reduziram o crescimento micelial e a produção de conídios do fungo. Os fosfitos K1, K2, Ca1 e Zn foram que mais reduziram o diâmetro da lesão causada pelo patógeno. Ca1 e K1 com CaCl2 reduziram o tamanho das lesões. Não houve diferenças significativas entre os tratamentos, quanto as características físico-químicas analisadas (% perda de massa fresca, teor de sólidos solúveis totais, pH e acidez titulável) dos frutos.

Atividade antifúngica in vitro e in vivo de revestimento comestível ativo contra Colletotrichum sp / In vitro e in vivo antifungal activity of active edible coating against Colletotrichum sp

A atividade antifúngica in vitro e in vivo do carvacrol disperso no revestimentos comestíveis foi avaliada contra o Colletotrichum sp.. Na elaboração do revestimento foi utilizado amido de mandioca e gelatina. A concentração de carvacrol variou de 0-0,6 g.mL-1 de solução filmogênica. O revestimento ativo mostrou-se um ótimo antifúngico no controle da proliferação do Colletotrichum sp. com concentração inibitória mínima (CIM) para o carvacrol de 3,51 μg.mL-1. O revestimento de 0,3 mg carvacrol. g-1 solução inibiu a proliferação do Colletotrichum sp. nas análises antimicrobianas in vivo realizadas em morangos. Os revestimentos ativos foram capazes de inibir o aparecimento da doença fúngica antracnose em 80 a 90 % dos morangos com dois dias de incubação de Colletotrichum sp..

Efeito combinado de carvacrol e tiabendazol contra Colletotrichum gloesporioides, Fusarium solani e Alternaria alternata / Combined effect of carvacrol and thiabendazole against Colletotrichum gloesporioides, Fusarium solani and Alternaria alternata

O objetivo deste trabalho foi determinar a atividade antimicrobiana do carvacrol e sua combinação com tiabendazol no controle de fungos patogênicos deteriorantes de frutas (Colletotrichum gloesporioides, Fusarium solani e Alternaria alternata). O carvacrol apresentou uma concentração inibitória mínima (CIM) de 282 a 563 μg mL-1 para os fungos testados. Quando avaliado em conjunto com o tiabendazol apresentou efeito aditivo contra C. gloesporioides e F. solani (FICI 0,5 e 1,0, respectivamente) e sinérgico contra a A.alternata (FICI 0,1). Houve redução da CIM do carvacrol de 50 a 88%. Este estudo mostra o potencial do uso.

Isolation and identification of endophytic fungi from Chelidonium majus and their antifungal activity / 中国中药杂志

In order to find new source of antifungal agents, eleven cultivable endophytic fungi were isolated from the roots,stems and leaves of Chelidonium majus by traditional method. Seven of them were identified as Colletotrichum(L1, L2, L3, S1, S3, S4, S5), and three of them were identified as Fusarium(R1,R2,R3) by morphological features and molecular biological technology. The antifungal activity test showed that all the tested fungi displayed some inhibitory activity against five common plant pathogens(C. gloeosporioides, Curvularia lunata, Pyricularia oryza, Alternaria alternate and A. brassicae), and their inhibition rate of some test items were over 60%. Among them, R1, S2, S3 and S4 were more potent than others. This study enriches the understanding of endophytes from Ch. majus and provides a basis for the study of new microbial fungicides.