Moniliophthora Roreri Research Articles

Adaptive evolution of Moniliophthora PR-1 proteins towards its pathogenic lifestyle

BackgroundPlant pathogenesis related-1 (PR-1) proteins belong to the CAP superfamily and have been characterized as markers of induced defense against pathogens. Moniliophthora perniciosa and Moniliophthora roreri are hemibiotrophic fungi that respectively cause the witches’ broom disease and frosty pod rot in Theobroma cacao. Interestingly, a large number of plant PR-1-like genes are present in the genomes of both species and many are up-regulated during the biotrophic interaction. In this study, we investigated the evolution of PR-1 proteins from 22 genomes of Moniliophthora isolates and 16 other Agaricales species, performing genomic investigation, phylogenetic reconstruction, positive selection search and gene expression analysis.ResultsPhylogenetic analysis revealed conserved PR-1 genes (PR-1a, b, d, j), shared by many Agaricales saprotrophic species, that have diversified in new PR-1 genes putatively related to pathogenicity in Moniliophthora (PR-1f, g, h, i), as well as in recent specialization cases within M. perniciosa biotypes (PR-1c, k, l) and M. roreri (PR-1n). PR-1 families in Moniliophthora with higher evolutionary rates exhibit induced expression in the biotrophic interaction and positive selection clues, supporting the hypothesis that these proteins accumulated adaptive changes in response to host–pathogen arms race. Furthermore, although previous work showed that MpPR-1 can detoxify plant antifungal compounds in yeast, we found that in the presence of eugenol M. perniciosa differentially expresses only MpPR-1e, k, d, of which two are not linked to pathogenicity, suggesting that detoxification might not be the main function of most MpPR-1.ConclusionsBased on analyses of genomic and expression data, we provided evidence that the evolution of PR-1 in Moniliophthora was adaptive and potentially related to the emergence of the parasitic lifestyle in this genus. Additionally, we also discuss how fungal PR-1 proteins could have adapted from basal conserved functions to possible roles in fungal pathogenesis.

SNP markers associated with resistance to frosty pod and black pod rot diseases in an F1 population of Theobroma cacao L.

Economically, cacao (Theobroma cacao L.) is a major tropical commodity for the Americas; however, severe losses due to Moniliophthora roreri (Cif. and Par.), which causes frosty pod rot (FPR), and Phytophthora spp., which causes black pod rot (BPR), have reduced cacao production in the Americas. The objectives of this study are to (i) re-confirm the QTL using different marker set; (ii) discover new QTL associated with FPR and BPR resistance using SNP markers; and (iii) find genes in the candidate QTL regions. At CATIE in Turrialba, Costa Rica, an F1 mapping population of cacao was obtained by crossing “POUND 7,” a clone moderately susceptible to FPR and resistant to BPR, with “UF 273,” resistant to FPR and highly susceptible to BPR. A total of 179 F1 progeny were fingerprinted with 5149 SNP markers and a dense linkage map composed of 10 linkage groups was developed using 2910 polymorphic SNP markers. Also segregating F1 trees were screened for resistance to FPR and BPR diseases. Seven QTL previously reported on chromosomes 2, 7, and 8 for FPR resistance and on chromosomes 4, 8, and 10 for BPR resistance were confirmed. Additionally, eight QTL were identified for FPR resistance (chromosomes 4, 9, and 10) and BPR resistance (chromosome 2). The expression of genes commonly associated with plant defense and disease resistance that are located within the identified QTL was confirmed.

Genotype–environment interactions shape leaf functional traits of cacao in agroforests

The outbreak of frosty (Moniliophthora roreri) and black pod (Phytophthora palmivora) in Central American cacao agroforests throughout the 1980s led to the breeding of Theobroma cacao clones resistant to these diseases. However, while clonal disease resistance has been well-documented, clonal adaptability to contrasting pedoclimates remains less understood. Plant functional responses to environmental constraints can be assessed by evaluating intraspecific trait variation (ITV), but trait-based approaches have rarely been implemented in genotype assessments. The objective of our study was to determine cacao clone ITV in contrasting environments. Here, we used bivariate, multivariate, and functional trait space analyses to quantify ITV in nine leaf functional traits, among six disease-resistant cacao clones, growing in two clonal gardens with distinct environmental characteristics (a “mild dry season” with near-optimal cacao growing conditions and a “harsh dry season” site with sub-optimal conditions). All leaf traits varied among the six clones, though trait differences among clones differed depending on the site: physiological trait differences among clones were larger at the “mild dry season” site, chemical trait differences were wider at the “harsh dry season” site, and morphological trait differences among clones were similar at both sites. Multivariate and hypervolume trait space analyses revealed greater site influence on trait values of the least productive clones, while the most productive clones were characterized by higher specific leaf area, independent of the site. To our knowledge, our paper is among the first to show evidence that functional traits provide new insights into genotype × environment interactions and clonal selection in agroforestry systems and support in favor of applying functional trait-based research to plant breeding paradigms.

Quinoa saponins, biological controller against Cercospora coffeicola and Moniliophthora roreri

An extract rich in saponins obtained from quinoa residues showed in vitro anti-fungal activity against the coffee phytopathogen Cercospora coffeicola, with an antifungal index of inhibition (AI) of 22.39 ± 4.5% at 120 mg/mL, and against the cocoa phytopathogen Moniliophthora roreri, with an AI of 55.8 ± 1.6% at 60 mg/mL. The results showed a potential of this saponin-extract for the biological control of these phytopathogens.

Pathogenicity of Moniliophthora roreri isolates from selected morphology groups in harvested cacao pods and in vitro sensitivity to compost tea.

Moniliopthora roreri is the frosty pod rot disease (FPD) and one of the most devastating cacao pathogens worldwide. However, M. roreri pathogenicity on harvested cacao pods and sensitivity to compost tea have not been fully described. Monosporic cultures of M. roreri from different morphology groups were obtained. The isolates’ pathogenicity was tested by inoculation onto harvested cacao pods, and symptoms were evaluated at 3-day intervals during 16 days before estimating the area under the disease progress curve (AUDPC). The sensitivity of M. roreri to compost tea was evaluated on potato dextrose agar (PDA) amended with 1 to 5 % compost tea. All morphology groups could infect harvested cacao pods during the 16 days with a disease severity index abode 75 %. Compost tea completely inhibited the growth of M. roreri when used at 4.5 % or higher. Results suggest a shortened biotrophic phase during the infection in harvested pods and a medium to high sensitivity of M. roreri to compost tea.

Biosurfactants synthesized by endophytic Bacillus strains as control of Moniliophthora perniciosa and Moniliophthora roreri

ABSTRACT: Ecuador is responsible for up to 70 % of the world’s production of fine flavor cocoa. Although fungal diseases have reduced cocoa production, eco–friendly alternatives have been implemented to control the spread of phytopathogens. This study aimed to explore the potential of endophytic strains isolated from fine flavor Nacional cocoa trees to synthetize compounds with antimicrobial and surface active properties. It was determined that under in vitro conditions, seven strains (DS03, DS07, DS18, DS23, DS31, DS34 and DS50) exhibited antifungal activity that inhibited the mycelial growth of Moniliophthora roreri and Moniliophthora perniciosa. The strains were identified based on their morphological (pattern forming Bacilli) and genetic characteristics using the 16S SSU rRNA gene sequences. Furthermore, we determined that the endophytic strains contained genes that encoded the lipopeptides Fengycin (fenA, fenB, fenC, fenE), Iturin (ituB, ituC, ituD), and Bacyllomicin D (bamC, bamD). The production of biosurfactant compounds was determined by a reduction in surface tension in the mineral medium. Additionally, the biosurfactant crude extract (BCE) was tested for its inhibitory activity against M. perniciosa and M. roreri. The minimal inhibitory concentration (MIC) was determined to be 0.07 and 0.035 mg mL−1, respectively, in both cases in a fungistatic action mode. The swelling granulation and fragmentation of both phytopathogens’ hyphae were observed using the BCE treatment. These results suggested the biocontrol capacity of the endophytic strains, which under in vitro conditions, have the ability to synthesize antifungal and surface active compounds that might be used as an ecological alternative in cocoa plants to prevent infection by phytopathogens.

EFECTO ANTIFÚNGICO in vitro DE EXTRACTOS METANÓLICOS DE Capsicum spp. EN Moniliophthora roreri

Uno de los principales problemas fitosanitarios en la producción de cacao (Theobroma cacao L.) en Latinoamérica y en México es la moniliasis, causada por Moniliophthora roreri. Este fitopatógeno causa más del 75% de pérdidas en la producción anual. Los metabolitos secundarios del género Capsicum tienen efectos antifúngicos, por lo que se pueden usar para el control de M. roreri. El objetivo de este experimento fue evaluar el efecto in vitro de extractos metanólicos (EM) de tres especies de Capsicum sobre M. roreri. Los EM extraídos de frutos inmaduros de C. chinense, C. frutescens, y dos morfotipos de C. annuum var. glabriusculum se probaron a dos concentraciones: 1000 y 2500 mg L-1, y se evaluaron en la inhibición del crecimiento micelial, la esporulación y la viabilidad de conidios producidos de M. roreri. Ambas dosis (tratamientos) se establecieron bajo un diseño experimental completamente al azar con cinco repeticiones; cada caja de Petri fue una unidad experimental, los datos se analizaron con ANDEVA y las medias se compararon con la prueba de Tukey (p≤0.05). Moniliophthora roreri fue sensible a los EM de las tres especies. Los EM de C. annuum var. glabriusculum en sus morfotipos amashito (AM) y garbanzo (GA) a 2500 mg L-1 inhibieron el crecimiento micelial hasta en 95%. Los EM de C. annuum var. glabriusculum AM y GA a 2500 mg L-1 inhibieron totalmente la producción de conidios en ambas concentraciones. Los EM de C. chinense y C. annuum var. glabriusculum GA inhibieron hasta 35% la viabilidad de conidios de M. roreri. La sensibilidad in vitro de M. roreri a EM de C. annuum var. glabriusculum AM y GA permite sugerir el efecto de los extractos en el campo de cultivo.

Assessment of the Potential of Trichoderma spp. Strains Native to Bagua (Amazonas, Peru) in the Biocontrol of Frosty Pod Rot (Moniliophthora roreri)

The use of native Trichoderma strains has been proposed as a sustainable alternative to control cocoa diseases. The aim of this study was to assess indigenous Trichoderma strains from Bagua Province, Peru, with reference to their antagonistic characteristics in vitro and their potential for in vitro biocontrol against frosty pod rot (FPR) disease. A total of 199 strains were assessed for in vitro mycoparasitism, antibiosis, and potential antagonism. The effect of four strains was evaluated in vitro using epidemiological variables, yield, and efficacy at two sites (Copallín and La Peca). Significant differences (p < 0.05) were reported for all variables evaluated in vitro and in vitro. Mycoparasitism ranged from 32% to 100%, antibiosis from 33.36% to 57.92%, and potential antagonism from 42.36% to 78.64%. All strains were found to affect the in vitro-assessed parameters in addition to enhancing the productive yield. The efficiency ranged from 38.99% to 71.9% in Copallín, and 45.88% to 51.16% in La Peca. The CP24-6 strain showed the highest potential for biocontrol under field conditions when considering its effect on both sites.

Indole alkaloid derivatives as building blocks of natural products from Bacillus thuringiensis and Bacillus velezensis and their antibacterial and antifungal activity study.

Six known indole alkaloid derivatives have been isolated for the first time from Bacillus thuringiensis and Bacillus velezensis strains, all of them as building blocks for the synthesis of larger natural products. Their structure was elucidated by a complete spectroscopy. Their biological activities were tested against some Gram-positive and Gram-negative bacteria and three phytopathogenic fungi which cause diseases in important crops, such as Moniliophthora roreri, the causal agent of cacao disease. The results indicated that some compounds had modest antibacterial activity; however, some of them had strong antifungal activity against the probed fungi. This antifungal activity of these compounds has not been reported.

Genome-Wide Association Study Reveals Novel Candidate Genes Associated with Productivity and Disease Resistance to Moniliophthora spp. in Cacao (Theobroma cacao L.)

Cacao (Theobroma cacao L.), the source of chocolate, is one of the most important commodity products worldwide that helps improve the economic livelihood of farmers. Diseases like frosty pod rot caused by Moniliophthora roreri and witches’ broom caused by Moniliophthora perniciosa limit the cacao productivity, this can be solved by using resistant varieties. In the current study, we sequenced 229 cacao accessions using genotyping-by-sequencing to examine the genetic diversity and population structure employing 9,003 and 8,131 single nucleotide polymorphisms recovered by mapping against two cacao genomes (Criollo B97-61/B2 v2 and Matina 1-6 v1.1). In the phenotypic evaluation, three promising accessions for productivity and 10 with good tolerance to the frosty pod rot and witches’ broom diseases were found. A genome-wide association study was performed on 102 accessions, discovering two genes associated with productivity and seven to disease resistance. The results enriched the knowledge of the genetic regions associated with important cacao traits that can have significant implications for conservation and breeding strategies like marker-assisted selection.

A disease and production index (DPI) for selection of cacao (Theobroma cacao) clones highly productive and tolerant to pod rot diseases

AbstractFrosty pod rot (FPR) (Moniliophthora roreri), along with black pod rot (Phytophthora species) and witches’ broom disease (Moniliophthora perniciosa) constitute the main phytosanitary problems limiting cacao (Theobroma cacao) production causing severe yield losses. One of the main sought after methods of pod rot management is the selection of tolerant cacao genotypes. Typically, the selection is carried out through the quantification of the percentage of diseased pods (PDP). However, PDP does not consider the relative productivity, or production potential (PT) of the genotype. Production potential can vary among cacao genotypes. Consequently, genotypes with similar PT can have similar or vastly different disease tolerance levels as measured by PDP. The disease and production index (DPI) was developed to integrate a genotype's tolerance to M. roreri and other diseases as measured by PDP, with its PT. Here, we evaluated the number of healthy pods, number of diseased pods, and weight of fresh seed for 29 clones grown in replicated five‐tree plots over 4 years. The data obtained was used to calculate PDP and DPI for each clone for three different disease combinations: frosty pod rot alone, pod rots other than frosty pod rot, and the combination of all pod rots. Multivariate analysis verified that DPI discriminated between clones based on productivity and disease tolerance. Surprisingly, there was a close ranking of clones between resistance to FPR and resistance to all other pod rots. The DPI can be used in breeding programmes focused on the selection of high yielding disease‐tolerant cacao genotypes.

Dimethyl Labeling-Based Quantitative Proteomics of Recalcitrant Cocoa Pod Tissue.

Dimethyl labeling is a type of stable-isotope labeling suitable for creating isotopic variants of peptides and thus be utilized for quantitative proteomics experiments. Labeling is achieved through a reductive amination/alkylation reaction using the low-cost reagents formaldehyde and cyanoborohydride, resulting in dimethylation of free amine groups of Lys and N-termini. Availability of isotopomeric forms of these reagents allows for the generation of up to six different isotopic variants. Here we describe the application of dimethylation to create two isotopic variants, light and heavy, differing in 4Da, to label the total tryptic digest peptides of cocoa pod extracted from healthy pods from cultivars susceptible and resistant to the fungal disease called "frosty pod" caused by Moniliophthora roreri.

Factors Affecting the Dynamics of Frosty Pod Rot in the Main Cocoa Areas of Santander State, Colombia.

Frosty pod rot (FPR) caused by Moniliophthora roreri is the primary disease affecting cacao production in the major producing countries of the Americas and is one of the major threats to cacao worldwide. The incidence of FPR on clones with different levels of resistance was investigated in four localities of Santander State, Colombia, between July 2013 and May 2015. Dynamics of diseased pods were modeled using boosted regression trees, a machine learning technique that allows regressions to be performed without prior statistical assumptions. The results suggested that FPR epidemics varied according to plot location, clone, weeks of observation, and total pods produced. Dynamics in the phenology of pods had an effect on the epidemics, and this dynamic could partially explain the difference in resistance among clones. Although not total, partial resistance of ICS 95 was confirmed. An important wilt effect was observed, particularly in the resistant clones; consequently, differences in harvested pods were not significant among clones. Pod stripping remains a good practice for the management of the disease and this practice could also have an effect on the pod dynamics and wilt phenomenon.

Regional Evaluation of Fungal Pathogen Incidence in Colombian Cocoa Crops

The production of cocoa (Theobroma cacao L.) in Colombia has a significant environmental and socioeconomic importance as a promissory crop in the post-conflict process. The department of Norte de Santander has cocoa crops that are dramatically affected by fungal pathogens causing important losses during harvest and post-harvest. Therefore, the current study focused on the determination of the incidence of diseases caused by phytopathogenic fungi in cocoa crops, and the identification of primary phytopathogenic fungi found in biological material from different farms of the region. The study was conducted in four municipalities of the department, by sampling fruits infected with frosty pod rot (FPR) and black pod rot (BPR) that presented in situ incidence ranging from 0.37 to 21.58% and from 1.75 to 35.59%, respectively. The studied hybrid materials, together with clone TSH 65, were found to be the most susceptible, while the remaining clones were more tolerant, especially CCN 51, IMC 67, and ICS95. Fifteen strains were isolated using in vitro assays and then morphologically characterized both in solid media and by microscopy. Nine of them corresponded to the pathogen Moniliophthora roreri, and other six to Phytophthora palmivora. The isolated agents showed in vitro morphological variability, as well as the ability to adapt to different environments when growing in situ.

Redesign of a New Manganese Peroxidase Highly Expressed in Pichia pastoris towards a Lignin-Degrading Versatile Peroxidase.

Manganese peroxidases, lignin peroxidases, and versatile peroxidases secreted by white rot fungi are supposed to play an essential role in lignin degradation. Thus, these enzymes have attracted significant attention as potential biocatalysts. Versatile peroxidases are the most interesting ones, since they comprise activities of manganese and lignin peroxidases. Herein, we demonstrate how the properties of a new manganese peroxidase from Moniliophthora roreri, designated MrMnP1, were shifted towards those of a versatile peroxidase. MrMnP1 was cloned in Pichia pastoris X-33 and highly expressed in a fed-batch fermentation, yielding 132 mg L-1 of active enzyme. To extend the substrate spectrum of MrMnP1, a catalytically active tryptophan present in lignin and versatile peroxidases was first introduced. Additionally, the role of five amino acids at positions adjacent to the catalytic tryptophan was elucidated through their replacement by those found in a versatile peroxidase from Pleurotus eryngii. The resulting mutants demonstrated new activities towards high-redox-potential substrates, such as lignin dimers, veratryl alcohol, and the azo dye Reactive Black 5.

Hongos endófitos foliares como candidatos a biocontroladores contra <i>Moniliophthora</i> spp. de <i>Theobroma cacao</i> (Malvaceae) en Ecuador

La adaptabilidad de los hongos endófitos a sus hospedantes, los beneficios ecológicos que le brinda y los diversos mecanismos antagónicos contra plagas que poseen los convierten en una alternativa para el control biológico de enfermedades. Se determinó el potencial de 17 cepas de hongos endofíticos foliares (FEF) obtenidas de tejido sano de Theobroma cacao como candidatas para el control biológico de Moniliophthora roreri (MR) y M. perniciosa (MP). Se evaluaron: i) el micoparasitismo de los FEF frente a colonias de Moniliophthora spp., ii) la acción de los metabolitos crudos de los FEF en el crecimiento, y iii) la habilidad para recolonizar hojas sanas del hospedante mediante ensayos de hojas sueltas. Tres cepas de Lasiodiplodia theobromae fueron las más promisorias: Ec098, Ec151 and Ec157. Estas cepas inhibieron el crecimiento de MR y MP, tanto en el enfrentamiento de las colonias como mediante sus metabolitos y, adicionalmente, recolonizaron el hospedante entre el 80-100 % de las veces. Otras cepas mostraron valores destacados en un indicador, y no deseables en otros. Por ejemplo, la Ec035 (L. theobromae) mostró los niveles más altos de micoparasitismo contra ambos patógenos en la interacción de las colonias, y el segundo mejor por sus metabolitos, pero no pudo reinfectar el hospedante. La cepa Ec059 (Xylaria feejeensis) reinfectó 100 %, pero no mostró los atributos deseados de antagonismo. Por su parte, los metabolitos de Ec107 (Colletotrichum gloeosporioides s.l.) inhibieron a MR en un 60 %, pero también estimularon el crecimiento de MP. Ninguna cepa logró todas las características deseables para un agente de control biológico.

Genome sequence and effectorome of Moniliophthora perniciosa and Moniliophthora roreri subpopulations

BackgroundThe hemibiotrophic pathogens Moniliophthora perniciosa (witches’ broom disease) and Moniliophthora roreri (frosty pod rot disease) are among the most important pathogens of cacao. Moniliophthora perniciosa has a broad host range and infects a variety of meristematic tissues in cacao plants, whereas M. roreri infects only pods of Theobroma and Herrania genera. Comparative pathogenomics of these fungi is essential to understand Moniliophthora infection strategies, therefore the detection and in silico functional characterization of effector candidates are important steps to gain insight on their pathogenicity.ResultsCandidate secreted effector proteins repertoire were predicted using the genomes of five representative isolates of M. perniciosa subpopulations (three from cacao and two from solanaceous hosts), and one representative isolate of M. roreri from Peru. Many putative effectors candidates were identified in M. perniciosa: 157 and 134 in cacao isolates from Bahia, Brazil; 109 in cacao isolate from Ecuador, 92 and 80 in wild solanaceous isolates from Minas Gerais (Lobeira) and Bahia (Caiçara), Brazil; respectively. Moniliophthora roreri showed the highest number of effector candidates, a total of 243. A set of eight core effectors were shared among all Moniliophthora isolates, while others were shared either between the wild solanaceous isolates or among cacao isolates. Mostly, candidate effectors of M. perniciosa were shared among the isolates, whereas in M. roreri nearly 50% were exclusive to the specie. In addition, a large number of cell wall-degrading enzymes characteristic of hemibiotrophic fungi were found. From these, we highlighted the proteins involved in cell wall modification, an enzymatic arsenal that allows the plant pathogens to inhabit environments with oxidative stress, which promotes degradation of plant compounds and facilitates infection.ConclusionsThe present work reports six genomes and provides a database of the putative effectorome of Moniliophthora, a first step towards the understanding of the functional basis of fungal pathogenicity.

Prediction of Cacao (Theobroma cacao) Resistance to Moniliophthora spp. Diseases via Genome-Wide Association Analysis and Genomic Selection.

Cacao (Theobroma cacao) is a globally important crop, and its yield is severely restricted by disease. Two of the most damaging diseases, witches’ broom disease (WBD) and frosty pod rot disease (FPRD), are caused by a pair of related fungi: Moniliophthora perniciosa and Moniliophthora roreri, respectively. Resistant cultivars are the most effective long-term strategy to address Moniliophthora diseases, but efficiently generating resistant and productive new cultivars will require robust methods for screening germplasm before field testing. Marker-assisted selection (MAS) and genomic selection (GS) provide two potential avenues for predicting the performance of new genotypes, potentially increasing the selection gain per unit time. To test the effectiveness of these two approaches, we performed a genome-wide association study (GWAS) and GS on three related populations of cacao in Ecuador genotyped with a 15K single nucleotide polymorphism (SNP) microarray for three measures of WBD infection (vegetative broom, cushion broom, and chirimoya pod), one of FPRD (monilia pod) and two productivity traits (total fresh weight of pods and % healthy pods produced). GWAS yielded several SNPs associated with disease resistance in each population, but none were significantly correlated with the same trait in other populations. Genomic selection, using one population as a training set to estimate the phenotypes of the remaining two (composed of different families), varied among traits, from a mean prediction accuracy of 0.46 (vegetative broom) to 0.15 (monilia pod), and varied between training populations. Simulations demonstrated that selecting seedlings using GWAS markers alone generates no improvement over selecting at random, but that GS improves the selection process significantly. Our results suggest that the GWAS markers discovered here are not sufficiently predictive across diverse germplasm to be useful for MAS, but that using all markers in a GS framework holds substantial promise in accelerating disease-resistance in cacao.

Morphological variants of Moniliophthora roreri on artificial media and the biotroph/necrotroph shift

Moniliophthora roreri (Mr) causes frosty pod rot of Theobroma cacao in a hemibiotrophic association. The Mr biotroph-like phase has not been studied in culture. Mr spores (isolates Co12, Co52, and B3) were germinated on high (V8) and low (BPMM) nutrients with different media hardness (0.5% to 3% agarose). Germination was high on V8 media. Hardness affected germination on BPMM. Most colonies on V8 were slow-growing, failing to sporulate. Colony morphology depended on the isolate. On BPMM, exaggerated mycelia formed of limited length with enlarged cells. On agarose, rapidly expanding sporulating necrotrophic colonies formed rarely. Co12 and B3 spores were germinated on V8 and BPMM with low melting point (LMP) agarose. Slow-growing colonies of B3 on BPMM were unstable on LMP agarose, often forming slow-growing/rapidly expanding hybrids. Slow-growing colonies are hypothesized to represent the biotrophic phase. One nucleus was common in Mr cells, other than spores. Binucleate cells were occasionally observed in aged cells of slow-growing mycelia. Co52 cells often had more than two nuclei per cell after germination. Mr mycelia cells typically carry a single nucleus, being considered haploid. Biotroph- and necrotroph-like mycelia displayed differential gene expression but results were inconsistent with published in vivo results and require further study.

Degree of domestication influences susceptibility of Theobroma cacao to frosty pod rot: a severe disease devastating Mexican cacao

Background: The three -main cacao ( Theobroma cacao ) varieties cultivated in México are: Criollo, Forastero, and Trinitario. Each variety has a different fruit setting time and has been subjected to several selection stages. The aim of this study was to determine the susceptibility of the three cacao varieties to Frosty pod rot caused by Moniliophthora roreri . We also analyzed the physicochemical properties of the pericarp of the three varieties and its domestication age.Question: Is there a relationship between the degree of domestication of the cacao variety and its susceptibility to Frosty pod rot?Studied species: Theobroma cacao fruits.Study site: A cacao plantation in Chiapas, Mexico.Methods: The content of moisture, lignin, phenols, peroxidase activity and hardness of the pericarp were analyzed in the middle zone of healthy fruits at 4, 8 and 12 weeks of development. Number of healthy and infected fruits were recorded every week.Results: The highest susceptibility to Frosty pod rot was found in the immature stage of fruits from the Criollo variety. The disease was mostly found in the mid-zone of the fruit. The Criollo variety was the most susceptible. This variety showed the highest moisture values and the lowest values of the other measured parameters.Conclusions: The finest cacao is obtained from the Criollo variety, the one with the highest degree of domestication, and also the most susceptible to frosty pod rot. We suggest using material from wild Criollo populations and from the Forastero and Trinitario varieties in future breeding and selection programs.