This work was aimed at isolating and identifying the microbiota present during the semi-dry method of coffee processing using polyphasic methods and to evaluate microbial diversity with PCR-DGGE. Samples of Coffea arabica L. were collected during different processing stages in southern Minas Gerais, Brazil. The bacterial and fungal isolates were phenotypically characterised and grouped according to the ARDRA technique, in which the 16-23S and ITS1-5.8S regions of the rDNA were sequenced for species identification. The bacterial counts varied from 3.7 to 7 log CFU g(-1). The yeast counts ranged from 3.4 to 6.9 log CFU g(-1), and the filamentous fungal population varied from 2 to 3.7 log CFU g(-1). Bacillus subtilis, Escherichia coli, Enterobacter agglomerans, Bacillus cereus and Klebsiella pneumoniae were the predominant bacteria detected during the processing of the coffee, and Pichia anomala, Torulaspora delbrueckii and Rhodotorula mucilaginosa were the dominant yeasts. All of the yeast and bacterial species detected by PCR-DGGE were isolated using culture-dependent methods, with the exception of one uncultivable bacterial species. Aspergillus was the most common genus among the filamentous fungal isolates. The use of polyphasic methods allowed a better characterization of the microbiota that is naturally present in semi-dry processed coffee.

The coffee fermentation is characterized by the presence of different microorganisms belonging to the groups of bacteria, fungi and yeast. The objectives of this work were to select pectinolytic microorganisms isolated from coffee fermentations and evaluate their performance on coffee pulp culture medium. The yeasts and bacteria isolates were evaluated for their activity of polygalacturonase (PG), pectin lyase (PL) and pectin methylesterase (PME) and metabolites production. Among 127 yeasts isolates and 189 bacterial isolates, 15 were pre-selected based on their ability to produce PL and organic compounds. These isolates were strains identified as Bacillus cereus, Bacillus megaterium, Bacillus subtilis, Candida parapsilosis, Pichia caribbica, Pichia guilliermondii and Saccharomyces cerevisiae. When cultivated in Coffee peel and pulp media in single culture or two by two mixed inocula, different behavior concerning to PME, PL and PG were found. The two principal components PC1 and PC2 accounted for 45.27 and 32.02 % of the total variance. UFLA CN727 and UFLA CN731 strains were grouped in the positive part of PC1 being characterized by 1,2-propanediol, hexanoic acid, decanoic acid, nonanoic acid and ethyl acetate. The UFLA CN448 and UFLA CN724 strains were grouped in the negative part of PC1 and were mainly characterized by guaiacol, butyric acid and citronellol. S. cerevisiae UFLACN727, P. guilliermondii UFLACN731 and C. parapsilosis UFLACN448 isolates are promising candidates to be tested in future studies as coffee starter cultures.

ResumoEste trabalho objetivou identificar a divergência genética entre acessos de cafeeiro do grupo Maragogipe, situados no Banco Ativo de Germoplasma de Café do Estado de Minas Gerais, no município de Patrocínio (MG), por meio de análises multivariadas. Foram avaliados 27 descritores morfo-agronômicos, e a distância generalizada de Mahalanobis foi usada para quantificar a divergência genética entre os acessos. Como estratégias de agrupamento, foram empregados o agrupamento de Tocher, o método hierárquico Unweighted Pair-Group Method Using Arithmetic Averages (UPGMA) e a análise de variáveis canônicas. A análise de agrupamento pelos métodos de Tocher e UPGMA separaram os acessos em três e seis grupos, respectivamente. A análise da contribuição relativa de cada característica para a dissimilaridade genética destacou as características intensidade da ondulação da folha, quantidade de inflorescência por axila foliar e produtividade média como as que mais contribuíram para a obtenção da divergência genética. Observou-se que as três primeiras variáveis canônicas explicaram mais de 90% da variância total contida no conjunto das características analisadas. A combinação entre os acessos MG0167 e MG0170 é a mais promissora em um possível cruzamento para uso imediato em programas de melhoramento.Palavras-chave: Coffea arabica, análise multivariada, melhoramento genético. Genetic divergence between coffee trees of the Maragogipe germplasm AbstractThis study aimed to identify the genetic divergence among coffee trees of the Maragogipe group in the active germplasm coffee bank in Patrocínio, Minas Gerais, Brazil, using multivariate analyzes. In the study, 27 morpho-agronomical descriptors were assessed. Mahalanobis generalized distance was applied to quantify the genetic divergence among genotypes. The following methods were used: Tocher clustering, Unweighted Pair-Group Method Using Arithmetic Averages (UPGMA) hierarchical clustering and the canonical variables analysis. The cluster analysis by Tocher method and the UPGMA separated the accessions into three and six groups, respectively. The analysis of the relative contribution of each characteristic to the genetic dissimilarity highlighted the intensity of curling leaf, inflorescence number per leaf axil and average yield as those that contributed most to the achievement of genetic divergence. It was observed that the first three canonical variables explained more than 90% of the total variance contained in the set of the analyzed characteristics. The combination between MG0167 and MG0170 accessions is the most promising for a possible crossing for immediate use in breeding programs.

Two strains of Bacillus, one from a culture collection (B. subtilis ATCC 6633) and a wild type (Bacillus sp. UFLA 817CF) isolated during coffee fermentation in the south of Minas Gerais, Brazil, were evaluated in relation to secretion of alkaline proteases. The strains were grown on nutrient broth, nutrient broth with sodium caseinate and nutrient broth with three different concentrations of cheese whey powder for 72 h. Samples were collected at 24-h intervals to evaluate the proteolytic activity, protein content and cell population. Maximum protease activity was observed after 24-h growth for both the microorganisms, a period that coincided with the end of the exponential phase. The specific activity values were, respectively, 839.8 U/mg for B. subtilis ATCC 6633 and 975.9 U/mg for Bacillus sp. UFLA 817CF. The 60% saturation presented the best results for specific protease activity in all the growth culture media tested with B. sp. UFLA 817CF. Bacillus sp. UFLA 817CF showed highest enzymatic activity at pH 9.0 and 40°C in the three culture media tested. The protease obtained from culture of the wild Bacillus strain presented stability at pH 7.0 and considerable heat stability at 40°C and 50°C, and could be an alternative for the industry to utilize cheese whey to produce proteolytic enzymes.

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Through the pruning type 'esqueletamento' it was developed a technology for the management of trees called "Zero Yield" in order to keep the size of the crop and eliminate the harvesting in a low yield season, which usually compromises the earnings of the coffee grower. With this technology, the harvest takes place every two years, always in years of high yield season. This technology is highly dependent on climate, and genotype. Given this, the objective of the study was to select genotypes of Coffea arabica L. responsive to pruning type 'esqueletamento', with high productivity and high potential for quality of drink for specialty coffee production. The experiment was deployed in the Coffee Sector at the Universidade Federal de Lavras (UFLA) in December 2005 and after six crops, it has undergone the pruning type 'esqueletamento' in August 2014. 20 genotypes were evaluated, being 18 progenies in F 5 generation, derived from the cross between cultivars of the Catuaí Group with coffees of germplasm Icatu and germplasm "Hibrido de Timor" and two commercial cultivars as witnesses. (Tupi IAC 1669-33 and Obatã IAC 1669-20). The following characteristics were evaluated: productivity (before and after pruning), vegetative vigor, the incidence of leaf rust, dull fruits, sieve No.16 and above, grain type mocha, aspect, and quality of drink. The genotypes 9 (H516-2-1-1-18-1-1), 12 (H516-2-1-1-18-1-4), 16 (H419-3-4-5-2-1-3), 18 (H419-3-4-5-2-1-5) and the cultivar Tupi IAC 1669-33 were responsive to the 'esqueletamento', with productivity after pruning higher than the maximum reached before the adoption of the pruning. All genotypes with the exception of Catucaí Amarelo 24/137 showed potential for the production of specialty coffees.

The microbial community of artisanal corn fermentation called Chicha were isolated, purified and then identified using protein profile by Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and confirmed by partial ribosomal gene sequencing. Samples from Chicha beverage were chemically characterized by gas and liquid chromatography (HPLC and GC-MS). Aerobic mesophilic bacteria (AMB) (35.8% of total of isolated microorganisms), lactic acid bacteria (LAB) (21.6%) and yeast (42.6%) were identified. Species of the genera Klebsiella, Bacillus, Staphylococcus, Micrococcus, Enterobacter, and Weissella were identified. Rhodotorula mucilaginosa, Lodderomyces elongisporus, Candida metapsilosis, and C. bohicensis were the yeasts found. The LAB isolates detected were responsible for the high concentrations of lactic acid found during the fermentation process (1.2 g L), which is directly related to the decrease in pH values (from 6.95 to 3.70). Maltose was the main carbohydrate detected during corn fermentation (7.02 g L with 36 h of fermentation). Ethanol was found in low concentrations (average 0.181 g L), making it possible to characterize the beverage as non-alcoholic. Twelve volatile compounds were identified by gas chromatography; belonging to the groups acids, alcohols aldehydes, acetate and others. MALDI-TOF was successfully used for identification of microbiota. Weissella confusa and W. cibaria were detected in the final product (after 36 h of fermentation), W. confusa is often classified as probiotic and deserve further application studies.

The benefits of mycorrhization occur with the growth of hyphae in colonized roots by promoting an increase of the contact surface which improves the initial growth due to a better absorption of water and nutrients. The objective was to evaluate the effect of arbuscular mycorrhizal fungus Rhizophagus clarus inoculation over the initial development and nutritional response of six genotypes of Coffea arabica L. Six genotypes of Coffea arabica L. were used (MGS Aranas, H29-1-8-5, Red Catuai IAC 144, IPR 100, Catigua MG2, Paraíso H 419-1) and with arbuscular mycorrhizal fungus - AMF: Rhizophagus clarus and without the application of the fungus. For the production of coffee seedlings, seeds were placed to germinate in plastic trays with sterile sand. After germination, 10 seedlings of each genotype were transferred to 0.120 dm3 polyethylene tubes with substrate. Then the inoculation of five seedlings of each genotype with the AMF R. Clarus was performed. When the seedlings with and without inoculation with the AMF presented six pairs of leaves they were transplanted to 13-liter pots containing soil (Dystrophic red-yellow latosol). The inoculation favored the initial growth of the coffee plants and its intensity varied according to the genotypes. The genotypes H 29-1-8-5, Red Catuai IAC 144 and Catigua MG 2 were the ones that presented higher shoot dry mass, root dry mass, total dry mass and accumulation of P, in relation to MGS aranãs, Paraiso H 419-1 and IPR 100, so they are the most promising to be inoculated with the arbuscular mycorrhizal fungus.