Enzyme Production by Induratia spp. Isolated from Coffee Plants in Brazil

Abstract Endophytic fungi belonging to the genus Muscodor now transferred to Induratia are known producers of bioactive volatile organic compounds (VOCs) with many industrial applications. However, the members of this genus have rarely been reported to produce non-volatile metabolites including enzyme. Enzymes of the endophytes are degraders of the polysaccharides available in the host plants and the knowledge of enzyme production by Induratia spp. may provide insights into their possible biotechnological applications. The aim of this study was to evaluate the activity of amylase, cellulase, lipase, pectinase, phytase, protease, endo β-1,4 glucanase and exo β-1,4 glucanase enzymes produced by fungi of the species Induratia coffeana, Induratia yucatanensis and Induratia sp. isolated from organic coffee plants. All Induratia spp. were able to produce the extracellular enzymes cellulase, pectinase, protease, and phytase. Eight fungi were able to produce lipase and four produced amylase. The specific activity of endo β-1, 4 glucanase and exo β-1,4 glucanase enzymes were detected for 9 and 8 endophytic fungi, respectively. This work demonstrated for the first time, the array of enzymes produced by Induratia spp. isolated from Coffea arabica in organic systems in Brazil.

Ochratoxigenic fungi associated with green coffee beans (Coffea arabica L.) in conventional and organic cultivation in Brazil

The genera Aspergillus comprises species that produce mycotoxins such as aflatoxins, ochratoxins and patulin. These are cosmopolitan species, natural contaminants of agricultural products. In coffee grains, the most important Aspergillus species in terms of the risk of presenting mycotoxins belong to the genera Aspergillus Section Circumdati and Section Nigri. The purpose of this study was to assess the occurrence of isolated ochratoxigenic fungi of coffee grains from organic and conventional cultivation from the South of Minas Gerais, Brazil, as well as to evaluate which farming system presents higher contamination risk by ochratoxin A (OTA) produced by fungi. Thirty samples of coffee grains (Coffea arabica L.) were analysed, being 20 of them of conventional coffee grains and 10 of them organic. The microbiological analysis was done with the Direct Plating Technique in a Dichloran Rose Bengal Chloramphenicol Agar (DRBC) media. The identification was done based on the macro and micro morphological characteristics and on the toxigenic potential with the Plug Agar technique. From the 30 samples analysed, 480 filamentous fungi of the genera Aspergillus of the Circumdati and Nigri Sections were isolated. The ochratoxigenic species identified were: Aspergillus auricoumus, A. ochraceus, A. ostianus, A. niger and A. niger Aggregate. The most frequent species which produces ochratoxin A among the isolated ones was A. ochraceus, corresponding to 89.55%. There was no significant difference regarding the presence of ochratoxigenic A. ochreceus between the conventional and organic cultivation systems, which suggests that the contamination risk is similar for both cultivation systems.

Efeito do silício na intensidade da cercosporiose e na nutrição mineral de mudas de cafeeiro / Effect of silicon on the intensity of brown eye spot and on the mineral nutrition of coffee seedlings

O presente trabalho objetivou estudar o efeito do silício na intensidade da cercosporiose e na nutrição mineral de mudas de cafeeiro. No experimento 1, testou-se seis doses de ácido silícico (0, 0,5; 1; 2; 4 e 6 g kg­1 de solo) em mudas da cultivar Catuaí Vermelho IAC 99 inoculadas com o fungo Cercospora coffeicola. No experimento 2, foram realizadas microanálises de raios-X para a avaliação de nutrientes presentes nas folhas das mudas de cafeeiro das cultivares Topázio MG1190 e Icatu Precoce IAC 3282, inoculadas e não inoculadas com C. coffeicola, com e sem silicato de cálcio (1 g kg-1 de solo). Com o aumento das doses de ácido silícico observou-se redução na área abaixo da curva de progresso do número de lesões por folha (AACPNLF), redução linear nos teores foliares de magnésio e fósforo e aumento nos teores de enxofre e cobre. Os teores foliares de boro apresentaram comportamento quadrático, diminuindo com o aumento das doses de ácido silícico e aumentando a partir da dose de 4 g kg-1 de solo. Em microanálise de raio X, mudas de cafeeiro com cercosporiose apresentam menores picos de potássio e cálcio, independente da cultivar utilizada.

Our objective was to verify the effect of silicon on the intensity of brown eye spot and on the mineral nutrition of coffee seedlings. In the first experiment, 6 doses of silicic acid (0, 0.5, 1, 2, 4 and 6 g kg-1 soil) were tested using a complete randomized block design with 4 replicates with 6 coffee seedlings cultivar Catuaí Vermelho IAC 99 inoculated with the fungus Cercospora coffeicola. In the second experiment, X-ray microanalysis in a scanning electron microscope was performed on 2 coffee cultivars (Topazio MG1190 and Icatu Precoce IAC 3282), inoculated and non-inoculated with C. coffeicola, treated and untreated with calcium silicate (1 g kg-1 of soil). With the increase of the silicic acid doses, there was observed a reduction in the area under the disease progress curve of the number of lesions per leaf (AUPCNLL), coupled with a linear reduction in the foliar contents of magnesium and phosphorus as well as an increase in the contents of sulfur and copper. The foliar contents of boron presented a quadratic behavior, decreasing with the increase of silicic acid and increasing with the dose of 4 g kg-1 of soil. In X-ray microanalysis, coffee seedlings with brown eye spot presented lower peaks of potassium and calcium, regardless of the cultivar used.

Diversidade de fungos filamentosos associados a Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae) e suas galerias em frutos de Coffea canephora (Pierre) / Diversity of filamentous fungi associated with Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae) and its galleries in berries of Coffea canephora (Pierre)

Amostragens de campo foram realizadas em Ouro Preto d'Oeste (10°45'S e 62°15'W) para avaliar a micobiota associada a Hypothenemus hampei Ferrari [cutícula, aparelho bucal, protórax (micângia), tubo digestivo e fezes] e suas galerias em frutos de Coffea canephora Pierre. Dez gêneros (201 isolados) estavam diretamente relacionados com o inseto, enquanto cinco gêneros (20 isolados) estavam relacionados com galerias do inseto nos frutos. Todos os gêneros identificados associados aos insetos estavam também presentes nas suas galerias, o que indica que o broqueamento pode significar um modo ativo de inoculação de fungos por H. hampei. Na broca, a diversidade de gêneros de fungos foi maior no aparelho bucal e protórax e menor nas fezes. Fusarium, Penicillium e Geotrichum, com abundâncias de 55,7, 24,3 e 10,8 por cento, respectivamente, foram os gêneros dominantes. Nas galerias Fusarium, Geotrichum, Trichoderma e Aspergillus com abundância de 33,3; 29,6; 18,5 e 14,8 por cento, respectivamente, foram os gêneros dominantes. A presença de Fusarium no inseto e suas galerias reforça indicações anteriores de uma interação íntima entre H. hampei-Fusarium. A presença de Aspergillus e Penicillium enfatiza a possibilidade de dispersão da ocratoxina pela broca. Este trabalho fornece o primeiro registro da micobiota associada a H. hampei em C. canephora. Entre os gêneros identificados, Cephalosporium, Geotrichum e Oidiodendrum foram registrados pela primeira vez em associação com o inseto e suas galerias em C. canephora.

Field sampling was carried out in Ouro Preto d'Oeste Rondônia (10°45'S and 62°15'W) to evaluate the mycobiota associated with Hypothenemus hampei Ferrari [cuticle, mouth, prothorax (mycangia), gut and feces] and its galleries on berries of Coffea canephora Pierre. Ten genera (201 isolates) were directly related with the insect while five genera (20 isolates) were related with galleries on berries. All the genera identified in the insects were also present in their galleries, what indicates that boring may be an active way of fungi inoculation by H. hampei. The fungi genera were more diverse in the mouth and prothorax of borers, and lower in feces. Fusarium, Penicillium and Geotrichum, with abundance of 55.7, 24.3 and 10.8 percent, respectively, were dominant genera. In the galleries Fusarium, Geotrichum, Trichoderma and Aspergillus with abundance of 33.3, 29.6, 18.5 and 14.8 percent, respectively, were dominant genera. The overall presence of Fusarium in coffee berry borer and its galleries) reinforces previous indications of a close interaction between H. hampei-Fusarium. The presence of Aspergillus and Penicillium emphasizes the possibility of "ochratoxin dispersion" by the borer. This work provides the first record of the mycobiota associated with H. hampei in C. canephora. Among the identified genera, Cephalosporium, Geotrichum and Oidiodendrum were recorded for the first time in association with H. hampei and its galleries in C. canephora.