RESUMO

In the present investigation, we employ a novel and meticulously structured database assembled by experts, encompassing macrofungi field-collected in Brazil, featuring upwards of 13,894 photographs representing 505 distinct species. The purpose of utilizing this database is twofold: firstly, to furnish training and validation for convolutional neural networks (CNNs) with the capacity for autonomous identification of macrofungal species; secondly, to develop a sophisticated mobile application replete with an advanced user interface. This interface is specifically crafted to acquire images, and, utilizing the image recognition capabilities afforded by the trained CNN, proffer potential identifications for the macrofungal species depicted therein. Such technological advancements democratize access to the Brazilian Funga, thereby enhancing public engagement and knowledge dissemination, and also facilitating contributions from the populace to the expanding body of knowledge concerning the conservation of macrofungal species of Brazil.

Assuntos

RESUMO

Aspergillus welwitschiae causes bole rot disease in sisal (Agave sisalana and related species) which affects the production of natural fibers in Brazil, the main worldwide producer of sisal fibers. This fungus is a saprotroph with a broad host range. Previous research established A. welwitschiae as the only causative agent of bole rot in the field, but little is known about the evolution of this species and its strains. In this work, we performed a comparative genomics analysis of 40 Aspergillus strains. We show the conflicting molecular identity of this species, with one sisal-infecting strain sharing its last common ancestor with Aspergillus niger, having diverged only 833 thousand years ago. Furthermore, our analysis of positive selection reveals sites under selection in genes coding for siderophore transporters, Sodium­calcium exchangers, and Phosphatidylethanolamine-binding proteins (PEBPs). Herein, we discuss the possible impacts of these gene functions on the pathogenicity in sisal.

Assuntos

RESUMO

Sisal is a common name for different plant varieties in the genus Agave (especially Agave sisalana) used for high-quality natural leaf fiber extraction. Despite the economic value of these plants, we still lack information about the diversity of viruses (virome) in non-tequilana species from the genus Agave. In this work, by associating RNA and DNA deep sequencing we were able to identify 25 putative viral species infecting A. sisalana, A. fourcroydes, and Agave hybrid 11648, including one strain of Cowpea Mild Mottle Virus (CPMMV) and 24 elements likely representing new viruses. Phylogenetic analysis indicated they belong to at least six viral families: Alphaflexiviridae, Betaflexiviridae, Botourmiaviridae, Closteroviridae, Partitiviridae, Virgaviridae, and three distinct unclassified groups. We observed higher viral taxa richness in roots when compared to leaves and stems. Furthermore, leaves and stems are very similar diversity-wise, with a lower number of taxa and dominance of a single viral species. Finally, approximately 50% of the identified viruses were found in all Agave organs investigated, which suggests that they likely produce a systemic infection. This is the first metatranscriptomics study focused on viral identification in species from the genus Agave. Despite having analyzed symptomless individuals, we identified several viruses supposedly infecting Agave species, including organ-specific and systemic species. Surprisingly, some of these putative viruses are probably infecting microorganisms composing the plant microbiota. Altogether, our results reinforce the importance of unbiased strategies for the identification and monitoring of viruses in plant species, including those with asymptomatic phenotypes.

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The order Hypocreales (Ascomycota) is composed of ubiquitous and ecologically diverse fungi such as saprobes, biotrophs, and pathogens. Despite their phylogenetic relationship, these species exhibit high variability in biomolecules production, lifestyle, and fitness. The mitochondria play an important role in the fungal biology, providing energy to the cells and regulating diverse processes, such as immune response. In spite of its importance, the mechanisms that shape fungal mitogenomes are still poorly understood. Herein, we investigated the variability and evolution of mitogenomes and its relationship with the divergence time using the order Hypocreales as a study model. We sequenced and annotated for the first time Trichoderma harzianum mitochondrial genome (mtDNA), which was compared to other 34 mtDNAs species that were publicly available. Comparative analysis revealed a substantial structural and size variation on non-coding mtDNA regions, despite the conservation of copy number, length, and structure of protein-coding elements. Interestingly, we observed a highly significant correlation between mitogenome length, and the number and size of non-coding sequences in mitochondrial genome. Among the non-coding elements, group I and II introns and homing endonucleases genes (HEGs) were the main contributors to discrepancies in mitogenomes structure and length. Several intronic sequences displayed sequence similarity among species, and some of them are conserved even at gene position, and were present in the majority of mitogenomes, indicating its origin in a common ancestor. On the other hand, we also identified species-specific introns that advocate for the origin by different mechanisms. Investigation of mitochondrial gene transfer to the nuclear genome revealed that nuclear copies of the nad5 are the most frequent while atp8, atp9, and cox3 could not be identified in any of the nuclear genomes analyzed. Moreover, we also estimated the divergence time of each species and investigated its relationship with coding and non-coding elements as well as with the length of mitogenomes. Altogether, our results demonstrated that introns and HEGs are key elements on mitogenome shaping and its presence on fast-evolving mtDNAs could be mostly explained by its divergence time, although the intron sharing profile suggests the involvement of other mechanisms on the mitochondrial genome evolution, such as horizontal transference.

RESUMO

Aspergillus welwitschiae is a species of the Nigri section of the genus Aspergillus. In nature, it is usually a saprotroph, decomposing plant material. However, it causes the bole rot disease of Agave sisalana (sisal), a plant species used for the extraction of hard natural fibers, causing great economic loss to this culture. In this study, we isolated and sequenced one genome of A. welwitschiae (isolate CCMB 674 (Collection of Cultures of Microorganisms of Bahia)) from the stem tissues of sisal and performed in silico and wet lab experimental strategies to describe its ability to produce mycotoxins. CCMB 674 possesses 64 secondary metabolite gene clusters (SMGCs) and, under normal conditions, it produces secondary metabolism compounds that could disturb the cellular cycle of sisal or induce abnormalities in plant growth, such as malformin C. This isolate also produces a pigment that might explain the characteristic red color of the affected tissues. Additionally, this isolate is defective for the production of fumonisin B1, and, despite bearing the full cluster for the synthesis of this compound, it did not produce ochratoxin A. Altogether, these results provide new information on possible strategies used by the fungi during the sisal bole rot, helping to better understand this disease and how to control it.

Assuntos

RESUMO

Approximately 75% of the worldwide production of hard natural fibers originates from sisal, an industrial crop from arid and semiarid tropical regions. Brazil is the world's largest producer of sisal fiber, accounting for more than 40% of the worldwide production, and sisal bole rot disease has been the main phytosanitary problem of this crop. All previous studies reporting Aspergillus niger as the causal agent of the disease were based on the morphological features of fungal isolates from infected plant tissues in pure cultures. Black aspergilli are one of the most complex and difficult groups to classify and identify. Therefore, we performed an integrative analysis of this disease based on the isolation of black aspergilli from the endospheres and soils in the root zones of symptomatic adult plants, in vivo pathogenicity tests, histopathology of symptomatic plants, and molecular phylogeny and worldwide genetic variability of the causal agent. All sisal isolates were pathogenic and unequivocally produced symptoms of bole rot disease in healthy plants. In all tree-based phylogenetic methods used, a monophyletic group formed by A. welwitschiae along with all sisal isolates was retrieved. Ten A. welwitschiae haplotypes have been identified in the world, and three occur in the largest sisal-producing area. Most of the isolates are from a unique haplotype, present in only the sisal-producing region. A. welwitschiae destroyed parenchymatic and vascular cylinder cells and induced the necrosis of internal stem tissues. Therefore, sisal bole disease is probably the consequence of a saprotrophic fungus that opportunistically invades sisal plants and behaves as a typical necrotrophic pathogen.

RESUMO

This study investigated the anti-viral effects of the polyphenolic compounds Quercetin and Kaempherol on the release of HTLV-1 from the surface of MT-2 cells. Atomic force microscopy (AFM) was used to scan the surface of the MT-2 cells. MT-2 cells were fixed with 100% methanol on round glass lamina or cleaved mica and dried under UV light and laminar flow. The images were captured on a Multimode equipment monitored by a NanoScope IIId controller from Veeco Instruments Inc operated in tapping mode and equipped with phase-imaging hardware. The images demonstrated viral budding structures 131 ± 57 nm in size, indicating profuse viral budding. Interestingly, cell-free viruses and budding structures visualized on the surface of cells were less common when MT-2 was incubated with Quercetin, and no particles were seen on the surface of cells incubated with Kaempherol. In summary, these data indicate that HTLV-1 is budding constantly from the MT-2 cell surface and that polyphenolic compounds were able to reduce this viral release. Biological samples were analyzed with crude cell preparations just after cultivation in the presence of Quercetin and Kaempherol, showing that the AFM technique is a rapid and powerful tool for analysis of antiviral activity of new biological compounds.

Assuntos