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Witches’ broom disease (WBD) affects cocoa trees (Theobroma cacao L.) and is caused by the fungus Moniliophthora perniciosa that grows in the apoplast in its biotrophic phase and later progresses into the tissues, causing serious losses in the production of cocoa beans. Therefore, the apoplast of T. cacao can provide important defense responses during the interaction with M. perniciosa. In this work, the protein profile of the apoplast of the T. cacao genotypes Catongo, susceptible to WBD, and CCN-51, resistant one, was evaluated. The leaves of T. cacao were collected from asymptomatic plants grown in a greenhouse (GH) and from green witches’ brooms grown under field (FD) conditions for extraction of apoplastic washing fluid (AWF). AWF was used in proteomic and enzymatic analysis. A total of 14 proteins were identified in Catongo GH and six in Catongo FD, with two proteins being common, one up-accumulated, and one down-accumulated. In CCN-51, 19 proteins were identified in the GH condition and 13 in FD, with seven proteins being common, one up-accumulated, and six down-accumulated. Most proteins are related to defense and stress in both genotypes, with emphasis on pathogenesis-related proteins (PR): PR-2 (β-1,3-glucanases), PR-3 and PR-4 (chitinases), PR-5 (thaumatine), PR-9 (peroxidases), and PR-14 (lipid transfer proteins). Furthermore, proteins from microorganisms were detected in the AWF. The enzymatic activities of PR-3 showed a significant increase (p < 0.05) in Catongo GH and PR-2 activity (p < 0.01) in CCN-51 FD. The protein profile of the T. cacao apoplastome offers insight into the defense dynamics that occur in the interaction with the fungus M. perniciosa and offers new insights in exploring future WBD control strategies.

In plants, serpins are a superfamily of serine and cysteine protease inhibitors involved in stress and defense mechanisms, with potential for controlling agricultural pests, making them important biotechnological tools. The objective of this study was to characterize a serpin from Theobroma cacao, called TcSERPIN, to identify its endogenous targets and determine its function and biotechnological potential. TcSERPIN has 390 amino acid residues and shows conservation of the main active site, RCL. Cis-elements related to light, stress, hormones, anaerobic induction, cell cycle regulation and defense have been identified in the gene’s regulatory region. TcSERPIN transcripts are accumulated in different tissues of Theobroma cacao. Furthermore, in plants infected with Moniliophtora perniciosa and Phytophthora palmivora, the expression of TcSERPIN was positively regulated. The protein spectrum, rTcSERPIN, reveals a typical β-sheet pattern and is thermostable at pH 8, but loses its structure with temperature increases above 66°C at pH 7. At the molar ratios of 0.65 and 0.49, rTcSERPIN inhibited 55 and 28% of the activity of papain from Carica papaya and trypsin from Sus scrofa, respectively. The protease trap containing immobilized rTcSERPIN captured endogenous defense proteins from cocoa extracts that are related to metabolic pathways, stress and defense. The evaluation of the biotechnological potential against geohelminth larvae showed that rTcSERPIN and rTcCYS4 (Theobroma cacao cystatin 4) reduced the movement of larvae after 24 hours. The results of this work show that TcSERPIN has ideal biochemical characteristics for biotechnological applications, as well as potential for studies of resistance to phytopathogens of agricultural crops.

Abstract Background Tear film (TF) helps maintain and protect ocular function against damage to the ocular surface. Proteins are one of its main constituents, whose expression pattern can be used as a biomarker of ocular changes and systemic diseases. The aim of this study was to evaluate the expression of proteins in the TF of domestic cats before and after infection with Toxoplasma gondii, in the phases of acute infection and chronicity. Twelve healthy cats received orally homogenized brain matter obtained from mice inoculated with T. gondii oocysts, strain ME49. Cat feces were collected daily from the third day after infection to assess the release of oocysts. TF samples were obtained from cats, by Schirmer’s Tear Test 1, on day 0 (before infection), day 5 after infection (acute phase of infection, with maximum peak release of oocysts in feces) and on day 21 after infection (start of chronic phase, 7 days after total absence of oocyst release in feces). Tear samples were also submitted to proteomic analysis in a Q-Tof-Premier mass spectrometer. Results A total of 37 proteins with scores equal to or greater than 100 were identified on D0, followed by 36 on D5 and 42 on D21. Of these, 27 were common to D0 and D5, 33 to D0 and D21, 27 to D5 and D21, and 26 were common to the three groups, totaling 54 proteins. The most abundant proteins were lipocalin allergen Fel d, serum albumin, aldehyde dehydrogenase, lactoperoxidase and lactotransferrin. There was no significant difference in the abundance of proteins found on D0 and D5, but there was a statistical difference between D0 and D21 for ACT1_AEDAE, CERU_HUMAN and GELS_HUMAN. Regarding D5 and D21, there were significant differences for KV1_CANLF, LAC_PIG, TRFL_PIG, ACT1_AEDAE, CERU_HUMAN, GELS_HUMAN and OVOS2_HUMAN. Conclusions The main proteins identified in the TF of domestic cats are similar to those found in humans and other animal species. Most are part of the ocular surface defense system against injuries. The most expressed proteins in animals in the chronic phase of T. gondii infection are associated with the immune response to the parasite.

This study reports the virome investigation of pollinator species and other floral visitors associated with plants from the south of Bahia: Aphis aurantii, Atrichopogon sp., Dasyhelea sp., Forcipomyia taiwana, and Trigona ventralis hoozana. Studying viruses in insects associated with economically important crops is vital to understand transmission dynamics and manage viral diseases that pose as threats for global food security. Using literature mining and public RNA next-generation sequencing data deposited in the NCBI SRA database, we identified potential vectors associated with Malvaceae plant species and characterized the microbial communities resident in these insects. Bacteria and Eukarya dominated the metagenomic analyses of all taxon groups. We also found sequences showing similarity to elements from several viral families, including Bunyavirales, Chuviridae, Iflaviridae, Narnaviridae, Orthomyxoviridae, Rhabdoviridae, Totiviridae, and Xinmoviridae. Phylogenetic analyses indicated the existence of at least 16 new viruses distributed among A. aurantii (3), Atrichopogon sp. (4), Dasyhelea sp. (3), and F. taiwana (6). No novel viruses were found for T. ventralis hoozana. For F. taiwana, the available libraries also allowed us to suggest possible vertical transmission, while for A. aurantii we followed the infection profile along the insect development. Our results highlight the importance of studying the virome of insect species associated with crop pollination, as they may play a crucial role in the transmission of viruses to economically important plants, such as those of the genus Theobroma, or they will reduce the pollination process. This information may be valuable in developing strategies to mitigate the spread of viruses and protect the global industry.

Species of the genus Phytophthora cause black-pod rot, which is the disease responsible for the largest losses in cocoa production in the world. The species Phytophthora palmivora affects cacao tree cultivation in all producing countries. However, proteomic level studies of the Theobroma cacao–P. palmivora interaction are incipient. Thus, the aim of this study was to analyze this pathosystem through comparative proteomics and systems biology analyses. The proteins were extracted from leaves of T. cacao PA 150 (resistant) and SIC 23 (susceptible) clones 48 h after inoculation with P. palmivora using inoculation with sterile distilled water as controls. There were differences in the protein profile between the control and inoculated treatments of both clones. Thirty-seven distinct proteins were identified on 88 spots of the PA 150 treatments, and 39 distinct proteins were identified on 120 spots of the SIC 23 treatments. The metabolisms of ATP, carbohydrates, and nitrogen compounds had higher percentages of proteins with increased accumulation after inoculation in both clones. Systems biology analysis demonstrated that the networks contain a higher number of proteins in the clusters corresponding to processes of photosynthesis and glucose metabolism, suggesting that they are the most affected by the infection. In addition, lipoxygenase (LOX), 2-methylene-furan-3-one reductase-like, and co-chaperonin CPN20 proteins and a probable CC-NBS-LRR protein may be involved in resistance to black-pod disease caused by P. palmivora.

Diseases associated with Phytophthora cause considerable losses in cocoa production worldwide. Analyzing genes, proteins, and metabolites involved in Theobroma cacao’s interaction with Phytophthora species is essential to explaining the molecular aspects of plant defense. Through a systematic literature review, this study aims to identify reports of genes, proteins, metabolites, morphological characteristics, and molecular and physiological processes of T. cacao involved in its interaction with species of Phytophthora. After the searches, 35 papers were selected for the data extraction stage, according to pre-established inclusion and exclusion criteria. In these studies, 657 genes and 32 metabolites, among other elements (molecules and molecular processes), were found to be involved in the interaction. The integration of this information resulted in the following conclusions: the expression patterns of pattern recognition receptors (PRRs) and a possible gene-to-gene interaction participate in cocoa resistance to Phytophthora spp.; the expression pattern of genes that encode pathogenesis-related (PRs) proteins is different between resistant and susceptible genotypes; phenolic compounds play an important role in preformed defenses; and proline accumulation may be involved in cell wall integrity. Only one proteomics study of T. cacao-Phytophthora spp. was found, and some genes proposed via QTL analysis were confirmed in transcriptomic studies.

The aim of this study was to investigate the proteins found in tear film of healthy domestic cats. Schirmer tear test strips were used to collect tear samples of twelve healthy cats, which were mixed, centrifuged, and placed in a single 1.5 mL microtube that was frozen at −20°C, until analysis by two-dimensional polyacrylamide gel and mass spectrometry associated with high-performance liquid chromatography. The resulting spectra were analyzed and compared with the Swiss-Prot search tool. Forty peptides were detected in the analyzed protein fragments of 90 spots, with 16 proteins identified. Of these, the authors confirmed what has been already found in other studies: lactotransferrin, serum albumin, allergenic lipocalins, and neutrophil gelatinase-associated lipocalin. Others were considered novel in tear film samples of all species: cyclin-dependent protein kinase, serine/arginine repetitive matrix protein, apelin receptor, secretory protein related to C1q/TNF, Wee1, α-1,4 glucan phosphorylase, and WD repeat domain 1. The network was divided into 11 clusters, and a biological function was assigned. Most of the proteins have functions in the defense and maintenance of feline ocular surface homeostasis. Serum albumin is a bottleneck protein, with a high betweenness value. This paper is a pioneer in reporting, in-depth, the tear film proteome of domestic cats.

Os neutrófilos são a primeira linha de defesa contra os microrganismos, sendo assim, uma célula muito importante na imunidade inata. Objetivou-se com este estudo caracterizar o proteoma de neutrófilo de Felis catus. Para isso, os granulócitos foram obtidos de sangue de gatos, purificados e utilizados 1x107 de células para identificação das proteínas.Foram identificadas 102 proteínas de neutrófilos de gatos, sendo classificadas conforme a abundância. A proteoglicano-3 associada a alguns processos biológicos como ativação dos neutrófilos e resposta imune, foi a proteína mais abundante. Dentre os processos, nas rotas metabólicas a maior quantidade de proteínas identificadas está envolvida na glicólise, que foram nove proteínas. Verificou-se também as proteínas envolvidas em função molecular, no componente celular e nos processos biológicos. Espera-se que a identificação das proteínas nesse estudo auxilie as futuras pesquisas elucidando algumas particularidades da espécie contribuindo com possíveis marcadores ou fármacos auxiliando no diagnóstico rápido e não invasivo de doenças que acometem em particular os felinos.