ABSTRACT
Saccharomyces cerevisiae CCMA 0159 is reported as a promising biocontrol agent against ochratoxin A (OTA)-producing fungi in coffee. Coffea arabica and Coffea canephora (var. Conilon or Robusta) are the most widely consumed coffee species around the world, cultivated in tropical and subtropical regions, each exhibiting distinct physicochemical and sensory characteristics. The objective of this study was to compare the growth and OTA production by Aspergillus carbonarius, A. ochraceus, and A. westerdijkiae in C. arabica and C. canephora, along with assessing the efficiency of S. cerevisiae CCMA 0159 in biocontrolling ochratoxigenic fungi in both coffee varieties. A. carbonarius exhibited a higher growth rate and OTA production in both coffee varieties, with C. canephora showing particular susceptibility. Conversely, A. ochraceus and A. westerdijkiae demonstrated lower growth and OTA production. S. cerevisiae was effective in biocontrolling the fungal isolates, inhibiting over 80 % of A. carbonarius growth in both coffee varieties. Among the mechanisms of action of the biological control agent, the production of volatile organic compounds stands out. The results of this study confirm the significant potential of S. cerevisiae CCMA 0159 as a biocontrol agent against Aspergillus for application in coffee-producing areas.
Subject(s)
Aspergillus , Coffea , Ochratoxins , Saccharomyces cerevisiae , Ochratoxins/biosynthesis , Aspergillus/growth & development , Aspergillus/metabolism , Saccharomyces cerevisiae/growth & development , Saccharomyces cerevisiae/metabolism , Coffea/microbiology , Food Contamination/prevention & control , Food Contamination/analysis , Coffee/microbiology , Biological Control Agents , Food MicrobiologyABSTRACT
Somatic embryogenesis (SE) is a clear example of cellular totipotency. The SE of the genus Coffea has become a model for in vitro propagation for woody species and for the large-scale production of disease-free plants that provide an advantage for modern agriculture. Temporary immersion systems (TIS) are in high demand for the propagation of plants. The success of this type of bioreactor is based on the alternating cycles of immersion of the plant material in the culture medium, usually a few minutes, and the permanence outside the medium of the tissues for several hours. Some bioreactors are very efficient for propagating one species but not another. The efficiency of bioreactors depends on the species, the tissue used to propagate, the species' nutritional needs, the amount of ethylene produced by the tissue, and many more. In this protocol, we show how we produce C. canephora plants that are being taken to the field.
Subject(s)
Coffea , Plant Somatic Embryogenesis Techniques , Plant Somatic Embryogenesis Techniques/methods , Coffea/growth & development , Coffea/genetics , Bioreactors , Seeds/growth & development , Culture Media/chemistryABSTRACT
Since the term proteomics was coined by Marc Wilkins in 1994, there has been an explosion in the number of articles reporting the use of the proteomics technique. As the layers of biological organization and their regulation increase, the complexity of living beings increases. Thus, we go from the genome to tissues, cells, cellular compartments, and phenotypes and the complexity of the tools used to study this complexity also increases. Unlike the genome study, in the case of the proteome, we have a more complex panorama. We have a spatial and temporal proteome. Proteomics helps to answer complex biological questions since proteins' function depends on their molecular structure, subcellular localization, and posttranslational modifications. In this protocol, we describe a methodology to extract proteins using different methods, separating proteins by electrophoresis in double-dimensional gels and analyzing the gels using specialized software that allows obtaining information on the number and abundance of the proteins from the gels.
Subject(s)
Coffea , Plant Proteins , Proteomics , Proteomics/methods , Plant Proteins/metabolism , Plant Proteins/analysis , Coffea/metabolism , Coffea/chemistry , Coffea/genetics , Proteome/analysis , Electrophoresis, Gel, Two-Dimensional/methods , SoftwareABSTRACT
Omic tools have changed the way of doing research in experimental biology. The somatic embryogenesis (SE) study has not been immune to this benefit. The transcriptomic tools have been used to compare the genes expressed during the induction of SE with the genes expressed in zygotic embryogenesis or to compare the development of the different stages embryos go through. It has also been used to compare the expression of genes during the development of calli from which SE is induced, as well as many other applications. The protocol described here is employed in our laboratory to extract RNA and generate several transcriptomes for the study of SE on Coffea canephora.
Subject(s)
Coffea , Gene Expression Profiling , Gene Expression Regulation, Plant , Plant Somatic Embryogenesis Techniques , Transcriptome , Coffea/genetics , Coffea/embryology , Coffea/growth & development , Plant Somatic Embryogenesis Techniques/methods , Gene Expression Profiling/methods , Transcriptome/genetics , Seeds/genetics , Seeds/growth & development , Gene Expression Regulation, DevelopmentalABSTRACT
Consumers are increasingly looking for healthy foods without the addition of synthetic additives. The aim of this study was to evaluate the efficiency of coffee extracts as a natural antioxidant in fresh pork sausage. Firstly, the conditions for obtaining coffee green extracts were optimized (Central Composite Rotatable Design 23, variables: extraction time, ethanol-water ratio, and sample-solvent ratio) in an ultrasound bath (70 °C). The response variables were the bioactive compounds levels and antioxidant activity. Valid models were obtained (p ≤ 0.05, R2 > 0.751), with higher bioactive content and antioxidant activity in the central point region. Extracts of Robusta and Arabica coffee green (RG and AG) and medium roast (RR and AR) obtained, and central point (10 min, an ethanol concentration of 30%, and a sample-solvent ratio of 10 g/100 mL) and optimized (14.2 min, 34.2%, and 5.8 g/100 mL) parameters were characterized. The RG presented a significantly (p ≤ 0.05) higher content of caffeine (3114.8 ± 50.0 and 3148.1 ± 13.5 mg/100 g) and 5-CQA (6417.1 ± 22.0 and 6706.4 ± 23.5 mg/100 g) in both extraction conditions, respectively. The RG and RR coffee presented the highest antioxidant activity. Two concentrations of RG and RR coffee extracts were tested in fresh pork sausage. The Robusta coffee extract presented the highest antioxidant activity in both roasted and green states. However, when applied to a meat product, the extract prepared with RG coffee showed better results, with efficiency in replacing synthetic antioxidants (content of malonaldehyde/kg of sample below 0.696 ± 0.059 in 20 days of storage), without altering the sensory attributes of the product (average scores above 7.16 ± 1.43 for all attributes evaluated). Therefore, the RG coffee extract was a suitable alternative as a natural antioxidant applied to fresh pork sausage.
ABSTRACT
The rainfall regime has a significant impact on the microclimate and mass emergence of the coffee berry borer, Hypothenemus hampei (Coleoptera: Curculionidae) (CBB). Little is known, however, about the shade tree-microclimate-CBB mass emergence interaction. The objective of the present study was to compare the effect of microclimate on the mass emergence of CBB in a full sun-exposed plot with a plot shaded by trees. The experiment was conducted on a Robusta coffee farm in southern Chiapas, Mexico. In each plot, 18 traps baited with an alcohol mixture were installed to capture flying females, collecting caught individuals every hour from 8:00 to 18:00 h. A meteorological station recorded several microclimatic variables on 13 weekly sampling dates from February to May 2022. Significantly more CBB females were captured in the shaded plot. The largest number of CBB captures was recorded between 14:00 and 16:00 h for the shade plot and between 15:00 and 17:00 h for the sun-exposed plot. The mass emergence of CBB showed a positive association with precipitation, dew point, and wind speed samples and a negative association with maximum air temperature, average relative humidity, ultraviolet radiation, wind speed, and equilibrium moisture content. Our observations show that the relationship between shade trees, microclimate, and mass emergence of CBB is complex and that its study helps us to gain deeper insight into CBB bioecology and advance control techniques against this important pest.
ABSTRACT
Coffee is one of the most consumed beverages worldwide. Espírito Santo is the largest Brazilian producer of conilon coffee, and invested in the creation of new cultivars, such as "Conquista ES8152", launched in 2019. Therefore, the present study aimed to evaluate the effects of maturation and roasting on the chemical and sensorial composition of the new conilon coffee cultivar "Conquista ES8152". The coffee was harvested containing 3 different percentages of ripe fruits: 60%, 80%, and 100%, and roasted at 3 different degrees of roasting: light, medium, and dark, to evaluate the moisture and ash content, yield of soluble extract, volatile compound profile, chlorogenic acid and caffeine content, and sensory profile. "Conquista ES8152" coffee has a moisture content between 1.38 and 2.62%; ash between 4.34 and 4.72%; and yield between 30.7 and 35.8%. Sensory scores ranged between 75 and 80 and the majority of volatile compounds belong to the pyrazine, phenol, furan, and pyrrole groups. The content of total chlorogenic acids was drastically reduced by roasting, with values between 2.40 and 9.33%, with 3-caffeoylquinic acid being the majority. Caffeine was not influenced by either maturation or roasting, with values between 2.16 and 2.41%. The volatile compounds furfural, 5-methylfurfural, and 2-ethyl-5-methylpyrazine were positively correlated with the evaluated sensory attributes and 5-methylfurfural was the only one significantly correlated with all attributes. Ethylpyrazine, furfuryl acetate, 1-furfurylpyrrole, 4-ethyl-2-methoxyphenol, and difurfuryl ether were negatively correlated. The stripping did not affect the quality and composition of this new cultivar, however, the roasting caused changes in both the chemical and sensorial profiles, appropriately indicated by the principal component analysis.
Subject(s)
Coffea , Coffee , Coffee/chemistry , Coffea/chemistry , Chemometrics , Caffeine/analysis , Chlorogenic Acid/analysisABSTRACT
Plant growth regulators (PGR) are essential for somatic embryogenesis (SE) in different species, and Coffea canephora is no exception. In our study model, previously, we have been able to elucidate the participation of various genes involved in SE by using different strategies; however, until now, we have not used a proteomic approach. This research seeks to contribute to understanding the primary cellular pathways involved in developing SE in C. canephora. The process of our model consists of two stages: (1) preconditioning in MS medium with auxin (NAA) and cytokinin (KIN), and (2) induction in Yasuda liquid medium added with cytokinin (BA). Therefore, in this study, we analyzed different days of the SE induction process using shotgun label-free proteomics. An amount of 1630 proteins was found among different sampling days of the process, of which the majority were accumulated during the induction stage. We found that some of the most enriched pathways during this process were the biosynthesis of amino acids and secondary metabolites. Eighteen proteins were found related to auxin homeostasis and two to cytokinin metabolism, such as ABC, BIG, ILR, LOG, and ARR. Ten proteins and transcription factors related to SE were also identified, like SERK1, SKP1, nuclear transcription factor Y, MADS-box, and calreticulin, and 19 related to other processes of plant development, among which the 14-3-3 and PP2A proteins stand out. This is the first report on the proteomic approach to elucidate the mechanisms that operate during the induction of SE in C. canephora. So, our findings provide the groundwork for future, more in-depth research. Data are available via ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD047172.
ABSTRACT
Coffee genotypes cultivated in the Amazonian region have been gaining increasing prominence in Brazilian plantations. This study aimed to quantify nutrient accumulation in the fruits, grains, and husks of Robusta coffee genotypes cultivated in the Brazilian Amazon and estimate genetic diversity. The experiment was conducted in Alta Floresta D'Oeste-Rondônia, Brazil. To assess nutrient accumulation, fresh fruits were collected. These were dried, processed, separated into grains and husks, and subjected to chemical analysis. Nutrient accumulation in fruits, grains, and husks, as well as the grain/husk ratio, underwent analysis of variance through the F-test (p < 0.01. For each evaluated trait, the experimental coefficient of 337 variation (CVe), genetic coefficient of variation (CVg), and genotypic determination coefficient (H2) were also estimated. Variability was observed among Robusta coffee genotypes, with VP06, AS4, and AS10 being the most dissimilar. LB080 had the lowest dry fruit weight and the lowest percentage of grains in relation to husks. ZD156 accumulated more K in the grains, while VP06 and AS10 were the genotypes that accumulated more nutrients in the husks. Nutrients N, K, Ca, and P are accumulated in larger quantities, necessitating the calibration of mineral fertilization dosages and distribution.
ABSTRACT
Somatic embryogenesis (SE) is an excellent example of mass plant propagation. Due to its genetic variability and low somaclonal variation, coffee SE has become a model for in vitro propagation of woody species, as well as for large-scale production of vigorous plants that are advantageous to modern agriculture. The success of the large-scale propagation of an embryogenic system is dependent on the development, optimization, and transfer of complementary system technologies. In this study, two successful SE systems were combined with a SETIS™ bioreactor immersion system to develop an efficient and cost-effective approach for the in vitro development of somatic embryos of Coffea spp. This study used an efficient protocol for obtaining somatic embryos, utilizing direct and indirect SE for both C. canephora and C. arabica. Embryos in the cotyledonary stage were deposited in a bioreactor to complete their stage of development from embryo to plant with minimal manipulation. Following ten weeks of cultivation in the bioreactor, complete and vigorous plants were obtained. Different parameters such as fresh weight, length, number of leaves, and root length, as well as stomatal index and relative water content, were recorded. In addition, the survival rate and ex vitro development of plantlets during acclimatization was assessed. The best substrate combination was garden soil (GS), peat moss (PM), and agrolite (A) in a 1:1:0.5 ratio, in which the bioreactor-regenerated plants showed an acclimatization rate greater than 90%. This is the first report on the use of SETIS™ bioreactors for the in vitro development of somatic embryos in Coffea spp., providing a technology that could be utilized for the commercial in vitro propagation of coffee plants. A link between research and innovation is necessary to establish means of communication that facilitate technology transfer. This protocol can serve as a basis for the generation and scaling of different species of agroeconomic importance. However, other bottlenecks in the production chains and the field must be addressed.
ABSTRACT
Worldwide coffee production is threatened by climate change, which highlights the importance of heat tolerance studies. Here we tested the hypothesis that photosynthetic heat tolerance in coffee varieties changes according to acclimation to distinct light conditions. Furthermore, we tested if heat tolerance is associated with the habitat of origin of the coffee species. We evaluated heat tolerance using chlorophyll fluorescence in varieties of Coffea arabica (Mundo Novo and Catuai Amarelo) and C. canephora (Conilon) grown in a common garden under two conditions: high (HS) and low (LS) sunlight. Leaf traits associated with leaf cooling were evaluated in plants grown in LS and HS and associations of heat tolerance with these traits were determined. The varieties tested had high photosynthetic heat tolerance, with temperatures above 54 °C leading to a 50% reduction in Fv /Fm (T50 ). The heat tolerance of each Coffea variety was unaffected by growth in distinct light conditions. Leaves of plants grown in LS were larger and had a lower fraction of the leaf area occupied by stomata (nast ). Heat tolerance was positively associated with leaf size and negatively with nast . C. canephora exhibited higher heat tolerance than C. arabica. The limited plasticity of heat tolerance in response to acclimation under distinct light conditions contradicts the prediction that plants acclimated to HS would have higher photosynthetic heat tolerance than those acclimated to LS. Our results on heat tolerance among Coffea species/varieties in HS and LS indicate the possibility of selection of varieties for better acclimation to ongoing climate changes.
Subject(s)
Coffea , Thermotolerance , Coffea/physiology , Coffee , Photosynthesis/physiology , Acclimatization/physiologyABSTRACT
Coffea canephora plant stem cells can have bioactive compounds with tissue repairing and anti-inflammatory action. This study aimed to develop a liposomal stem cell extract formulation obtained from the leaves of C. canephora (LSCECC) and to investigate its capacity to contribute to the dynamic mechanisms of tissue repair. The liposome cream was developed and characterized through the dynamic light scattering technique, atomic force microscopy, and transmission electron microscopy. The excisional full-thickness skin wound model was used and daily topically treated with the LSCECC formulation or vehicle control. On days 2, 7, 14, and 21 after wounding, five rats from each group were euthanized and the rates of wound closure and re-epithelialization were evaluated using biochemical and histological tests. LSCECC resulted in faster re-epithelialization exhibiting a significant reduction in wound area of 36.4, 42.4, and 87.5% after 7, 10, and 14 days, respectively, when compared to vehicle control. LSCECC treated wounds exhibited an increase in granular tissue and a proper inflammatory response mediated by the reduction of pro-inflammatory cytokines like TNF-α and IL-6 and an increase of IL-10. Furthermore, wounds treated with LSCECC showed an increase in the deposition and organization of collagen fibers at the wound site and improved scar tissue quality due to the increase in transforming growth factor-beta and vascular endothelial growth factor. Our data showed that LSCECC improves wound healing, the formation of extracellular matrix, modulates inflammatory response, and promotes neovascularization being consider a promising bioactive extract to promote and support healthy skin. The graphical presents the action of LSCECC in all four phases of wound healing and tissue repair. The LSCECC can reduce the inflammatory infiltrate in the inflammatory phase by decreasing the pro-inflammatory cytokines like IL-6 and TNF-α, in addition to maintaining this modulation through lesser activation and recruitment of macrophages. The LSCECC can also increase the release of IL-10, an anti-inflammatory cytokine, decreasing local edema. The increase in VEGF provides neovascularization and the supply of nutrients to newly repaired tissue. Finally, signaling via TGF-ß increases the production and organization of collagen fibers in the remodeling phase.
Subject(s)
Coffea , Interleukin-10 , Rats , Animals , Interleukin-10/metabolism , Coffea/metabolism , Cell Extracts , Tumor Necrosis Factor-alpha/metabolism , Interleukin-6/metabolism , Vascular Endothelial Growth Factor A , Liposomes/metabolism , Wound Healing/physiology , Skin/pathology , Cytokines/metabolism , Anti-Inflammatory Agents/pharmacology , Collagen/metabolismABSTRACT
ABSTRACT This work evaluated the initial performance and genetic diversity of Coffea canephora genotypes cultivated in environments at contrasting altitudes. Fourteen morphophysiological traits and seven descriptors of the genus Coffea spp. of coffee trees cultivated at altitudes of 140 m and 700 m were evaluated. The design used was Federer's augmented block in a 2 × 112 factorial scheme with six blocks. The first factor was the two environments, and the second was the 112 genotypes, with eight common treatments, being five conilon coffee clones and three arabica coffee cultivars. The data were analyzed by the method of REML/BLUP and genetic correlation method. Genetic diversity was evaluated by estimating the distance matrix, applying the Gower methodology followed by the clustering method by Tocher and UPGMA. The phenotypic means were higher in the environment at an altitude of 700 m, except for plant height, number of leaves, and canopy height (CH). Genotypic effects were significant for most traits except for leaf width, CH, unit leaf area, and total leaf area. A wide genetic diversity was verified, with distances varying from 0.037 to 0.593 for the pairs of genotypes 26 × 93 and T7 × 76, respectively. Most of the traits studied showed high genotypic correlation with the environment and expressive genetic correlation between the evaluated traits thereby demonstrating the possibility of indirect selection. There is an adaptation of conilon coffee genotypes to high altitudes and the possibility of developing a specific cultivar for the southern state of Espírito Santo.
ABSTRACT
Plant cell cultures have become a promising production platform of bioactive compounds for biomedical and cosmetic uses in the last decades. However, the success so far has been limited. The study aimed to evaluate the effectiveness of this unique biotechnology process to obtain a bioactive stem cell extract of Coffea canephora (SCECC) with antioxidant, anti-inflammatory, and regenerative properties. Total phenolic and flavonoid contents were determined in the SCECC by spectrophotometry. The chemical composition of the extracts was characterized by mass spectrometry. Antioxidant activity was evaluated using the colorimetric methods of free radical scavenging 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and the ferric reducing ability of plasma (FRAP). The anti-inflammatory activity was determined in lipopolysaccharide-stimulated RAW 264.7 macrophages through the production of superoxide anion (O2•-), nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and the activity of nuclear factor kappa B (NF-κB). Moreover, the ability of SCECC to stimulate the proliferation and migration of fibroblasts was assessed. Five compounds were tentatively identified, two flavonoids, two phenolic acids, and one sugar. High phenolic content and antioxidant activity were observed in the SCECC. SCECC promoted the proliferation and migration of fibroblasts and suppressed the pro-inflammatory mediators O2•-, NO, TNF-α, and IL-6 in a dose-dependent manner. Moreover, SCECC inhibited the NF-κB transcription factor. Therefore, we obtained evidence that the extract from C. canephora stem cells can be used as a natural agent against skin damage. Hence, it could be of interest in cosmetics for preventing skin aging.
ABSTRACT
This work evaluated the initial performance and genetic diversity of Coffea canephora genotypes cultivated in environments at contrasting altitudes. Fourteen morphophysiological traits and seven descriptors of the genus Coffea spp. of coffee trees cultivated at altitudes of 140 m and 700 m were evaluated. The design used was Federer's augmented block in a 2 × 112 factorial scheme with six blocks. The first factor was the two environments, and the second was the 112 genotypes, with eight common treatments, being five conilon coffee clones and three arabica coffee cultivars. The data were analyzed by the method of REML/BLUP and genetic correlation method. Genetic diversity was evaluated by estimating the distance matrix, applying the Gower methodology followed by the clustering method by Tocher and UPGMA. The phenotypic means were higher in the environment at an altitude of 700 m, except for plant height, number of leaves, and canopy height (CH). Genotypic effects were significant for most traits except for leaf width, CH, unit leaf area, and total leaf area. A wide genetic diversity was verified, with distances varying from 0.037 to 0.593 for the pairs of genotypes 26 × 93 and T7 × 76, respectively. Most of the traits studied showed high genotypic correlation with the environment and expressive genetic correlation between the evaluated traits thereby demonstrating the possibility of indirect selection. There is an adaptation of conilon coffee genotypes to high altitudes and the possibility of developing a specific cultivar for the southern state of Espírito Santo.
Subject(s)
Genetic Variation , Coffea/growth & development , Coffea/genetics , AltitudeABSTRACT
This study aimed to investigate the effect of spray drying (SD) and freeze-drying (FD) on the microencapsulation of green coffee extracts by using polydextrose (PD) and inulin (IN) as encapsulating agents and their physicochemical, bioactive compounds' stability, phenolic compounds' bioaccessibility after digestion, and sensory effects in unfermented dairy beverages. The extract encapsulated with IN by FD had lower moisture content, water activity, and hygroscopicity, while particles encapsulated by SD exhibited a spherical shape and the structure of the FD products was irregular. No difference was observed in phenolic compounds' bioaccessibility. Dairy beverages with added encapsulated extracts had higher total phenolic content and antioxidant activity. Microencapsulation allowed a controlled release of the bioactive compounds with an increase in the content of caffeine, chlorogenic acid, and trigonelline during storage. The dairy beverage with added extract encapsulated with IN by FD had the highest scores of acceptability regarding the overall impression and purchase intent.
Subject(s)
Coffea , Plant Extracts , Antioxidants/chemistry , Beverages , Caffeine/analysis , Chlorogenic Acid/analysis , Delayed-Action Preparations , Inulin/chemistry , Phenols/chemistry , Plant Extracts/chemistry , Dairy ProductsABSTRACT
Cytokinins (CK) are plant growth regulators involved in multiple physiological processes in plants. One less studied aspect is CK homeostasis (HM). The primary genes related to HM are involved in biosynthesis (IPT), degradation (CKX), and signaling (ARR). This paper demonstrates the effect of auxin (Aux) and CK and their cross talk in a Coffea canephora embryogenic system. The transcriptome and RT-qPCR suggest that Aux in pre-treatment represses biosynthesis, degradation, and signal CK genes. However, in the induction, there is an increase of genes implicated in the CK perception/signal, indicating perhaps, as in other species, Aux is repressing CK, and CK are inducing per se genes involved in its HM. This is reflected in the endogenous concentration of CK; pharmacology experiments helped study the effect of each plant growth regulator in our SE system. We conclude that the Aux-CK balance is crucial to directing somatic embryogenesis in C. canephora.
ABSTRACT
The coffee quality is affected by 40% pre-harvest, 40% post-harvest, and 20% export handling. Besides, future risks for the coffee industry are related with climate change and increased pathogens. Considering the importance of the aroma profile and unique flavor of Arabica coffee, most literature focuses on this variety because of the high market share; however, nowadays, Robusta coffee stands out for its increasing industrial value and resistance to drought. In this review, both species are emphasized, highlighting sensory aspects of possible new products mixed with a higher proportion of Robusta given market trends for bitter beverages. In the present work, a systematic search of peer-reviewed literature evaluates how the coffee cup quality and physicochemical characteristics of Robusta and Arabica are influenced by environmental, agronomic, and further processing factors.
ABSTRACT
The aim of this study was to characterize Meloidogyne paranaensis populations collected from infested coffee crops. Methodologies used to characterize the 11 studied populations from municipalities in Paraná and Minas Gerais States involved the morphological analysis of perineal patterns, biochemical analysis by isozyme electrophoresis, sequencing of internal transcribes spacer 1 (ITS-1) and D2/D3 ribosomal DNA (rDNA) regions, reproductive fitness, and virulence characterization in coffee genotypes. Morphological evaluations showed the existence of variation between populations, although the majority of them showed typical perineal patterns. The biochemical identification was based on α-esterase isozyme analyses and resulted in the appearance of three distinct profiles: P1 (typical), P2 (atypical), and a nondescribed profile, P2b. BLAST of the ITS-1 and D2/D3 rDNA regions indicated homology (>95%) with other sequences deposited in GenBank. For reproductive fitness and virulence characterization, 13 coffee genotypes (5 Coffea arabica and 8 C. canephora) were inoculated with 11 M. paranaensis populations. Variation in the reproductive fitness of populations was observed for cultivar Mundo Novo, a genotype without resistance genes, and variation in the virulence of populations was observed in genotypes carrying resistance genes. Three populations exhibited virulence combined with high reproductive fitness, while one showed virulence with low reproductive fitness. Some hosts were resistant to 11 populations, while one of the hosts was resistant to only one population, indicating the presence of different resistance genes. Nevertheless, no relationship was observed between the origin of population and their variations in perineal patterns, esterase profiles, phylogeny, or reproductive fitness in coffee genotypes, or between the different characterizations, although differences were observed within each characteristic.