RESUMO
Aquaporins are an ancient family of membrane channel proteins present in all eukaryotes and most prokaryotes, and apart from water, allow the transport of neutral solutes and organic compounds through the pore. These proteins are essential role differentially expressed during ripening in Fragaria vesca fruits. Fv PIP2-1a is intensively expressed in fruits, inclusive several other proteins member are differentially expressed in fruit but also in other plant tissues. Phylogenetic analysis shows that Fv PIP2-1a grouped with other Fragaria proteins and far apart from other F. vesca PIP proteins. A structural model for Fv PIP2-1a protein was built by comparative modelling methodology, which was validated and refined by molecular dynamics simulation. Fv PIP2-1a structure consists of 6 transmembrane regions and two NPA domains. The mobilization of water was analyzed by molecular docking simulations in wildtype and two mutants. Interestingly, the mutant FvPIP2-1a_H214G allowed the prediction of an increment in the flux of water molecules. On contrary, structural analysis predicted that H214E mutation blocked passage of water associated to constriction of the pore.
Assuntos
Aquaporinas , Fragaria , Aquaporinas/genética , Aquaporinas/metabolismo , Fragaria/genética , Fragaria/metabolismo , Simulação de Acoplamento Molecular , Filogenia , Proteínas de Plantas/química , Água/metabolismoRESUMO
Objetivo: O consumo de alimentos contaminados por microrganismos patogênicos pode levar a quadros de doenças transmitidas por alimentos. O presente estudo objetivou realizar uma análise microbiológica de morangos frescos (Fragaria vesca) comercializados em Goiânia, Goiás, pesquisando a presença de Escherichia coli e Salmonella spp., em consonância com a RDC nº 12, de 02 de janeiro de 2001. Métodos: Foram analisadas 51 amostras de morangos frescos in natura no Laboratório Clínico da PUC Goiás, sendo 25 amostras oriundas de supermercados e 26 de comércio livre/rua, entre agosto/2019 e dezembro/2019, em diversos bairros das regiões Norte, Noroeste, Sul e Central do município de Goiânia, Goiás. Foram feitas culturas bacterianas nos ágares Eosin Methylene Blue e Salmonella-Shigella, e identificados os isolados por provas bioquímicas clássicas. Como controle de qualidade, a cada bateria de experimentos, duas cepas bacterianas conhecidas, uma Salmonella spp. (Programa Nacional de Controle de Qualidade/PNCQ) e uma E. coli ATCC 25923, foram utilizadas. Resultados: Das 51 amostras analisadas, 5,9% (3/51) apresentaram positividade para o crescimento de E. coli, sendo estas amostras adquiridas no comércio livre/rua. Foi observada ausência de Salmonella spp. Conclusão: A grande maioria (94,1%) das amostras analisadas nesta pesquisa negativaram em relação à presença de E. coli e 100% negativaram para a presença de Salmonella spp., demonstrando que independente de adquiridos em comércio livre/rua ou em supermercados, os morangos encontravam-se microbiologicamente seguros para o consumo, de acordo com a legislação brasileira vigente.
Objective: The consumption of food contaminated by pathogenic microorganisms can cause Foodborne Diseases. The present study aimed to perform a microbiological analysis of fresh strawberries (Fragaria vesca) marketed in Goiânia, Goiás, researching the presence of Escherichia coli and Salmonella spp., in line with RDC No. 12, of January 2, 2001. Methods: They have analyzed 51 samples of fresh strawberries in natura at the Clinical Laboratory of PUC Goiás, 25 samples from supermarkets and 26 from free trade/street, between August/2019 and December/2019, in several neighborhoods in the North, Northwest, South, and Central regions county of Goiânia, Goiás. Bacterial cultures cultivated on Eosin Methylene Blue and Salmonella-Shigella agar, and identification of the isolates by standard biochemical tests. As a quality control, in each battery of experiments, two known bacterial strains, one Salmonella spp. (National Quality Control Program/PNCQ) and an E. coli ATCC 25923 was used. Results: Of the 51 samples analyzed, 5.9% (3/51) were positive for the growth of E. coli, these samples were acquired in free/street commerce. Absence of Salmonella spp. Conclusion: The majority (94.1%) of the samples analyzed in this research were negative concerning the presence of E. coli, and 100% were negative for the presence of Salmonella spp., demonstrating that independent of being acquired in free trade/street or supermarkets, the strawberries analyzed were microbiologically safe for consumption, following the current Brazilian legislation.
Assuntos
Microbiologia de Alimentos , Fragaria , Escherichia coli , ColiformesRESUMO
Abstract To discover and isolate a glyphosate-resistant gene from Fragaria vesca through gene mining. An open reading frame (ORF) of 1563 bp encoding EPSPSwas amplified from Fragaria vesca (FvEPSPS). FvEPSPS (Genebank: XP004306932.1) encodes a polypeptide of 520 amino acids and it has hightly homologous with EPSPS from other plants. qRT-PCR analysis showed that the FvEPSPS was expressed extensively in all tissues including leaves, roots and stems, with higher expression in leaves. Furthermore, transgenic Arabidopsis Thaliana exhibited 10 mM glyphosate to resistance. Therefore, this research offers a new glyphosate-resistant gene for development of transgenic crops.
Assuntos
Plantas Geneticamente Modificadas , Arabidopsis , Fragaria , Herbicidas/efeitos adversosRESUMO
The phytohormone jasmonoyl-isoleucine (JA-Ile) regulates fundamental plant processes. Fragaria vesca, the woodland strawberry, is a model plant for the Rosaceae family, in which the JA-Ile perception is poorly understood at the molecular level. JA-Ile promotes binding of JAZ repressor to COI1 protein in Arabidopsis to activate jasmonate (JA)-dependent responses. The aim of this work was to understand the molecular basis of the interaction between the F. vesca COI1 (FvCOI1) and JAZ1 (FvJAZ1) promoted by JA-Ile using a computational approach. Multiple sequence alignments and phylogenetic analyses of amino acid sequences were performed for FvCOI1, FvJAZ1 and their ortholog sequences. 3D structures for FvCOI1 and FvJAZ1 proteins were built by methods of homology modeling, using AtCOI1-JA-Ile-AtJAZ1 as template and then they were further refined and validated by molecular dynamics (MD) simulation. A molecular docking approach along with MDS analysis were used to gain insights into the interaction between a putative degron-like sequence present in FvJAZ1 with the FvCOI1-JA-Ile complex. FvCOI1 and FvJAZ1 showed high and moderate sequence identity, respectively, with the corresponding ortholog proteins from other plant species including apple, grape, tomato and Arabidopsis. Moreover, the FvJAZ1 has a variant C-terminal IPMQRK sequence instead of the canonical LPIARR degron sequence located in the Jas domain of AtJAZ1. The MD simulation results showed that the FvCOI1-JA-Ile-FvJAZ1 complex was stable, and the IPMQRK peptide of FvJAZ1 directly interacted with FvCOI1 and JA-Ile. The present research provides novel insight into the molecular interactions among key JA-signaling components in the model plant F. vesca, being few examples of characterized JA-Ile receptors at a structural level in plants.
Assuntos
Ciclopentanos/química , Fragaria/genética , Isoleucina/análogos & derivados , Reguladores de Crescimento de Plantas/química , Reguladores de Crescimento de Plantas/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Sequência de Aminoácidos , Ciclopentanos/metabolismo , Fragaria/metabolismo , Isoleucina/química , Isoleucina/genética , Isoleucina/metabolismo , Ligantes , Modelos Moleculares , Conformação Molecular , Complexos Multiproteicos/química , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Ligação Proteica , Domínios e Motivos de Interação entre Proteínas , Análise de Sequência de DNA , Relação Estrutura-AtividadeRESUMO
BACKGROUND: Fragaria vesca or 'woodland strawberry' has emerged as an attractive model for the study of ripening of non-climacteric fruit. It has several advantages, such as its small genome and its diploidy. The recent availability of the complete sequence of its genome opens the possibility for further analysis and its use as a reference species. Fruit softening is a physiological event and involves many biochemical changes that take place at the final stages of fruit development; among them, the remodeling of cell walls by the action of a set of enzymes. Xyloglucan endotransglycosylase/hydrolase (XTH) is a cell wall-associated enzyme, which is encoded by a multigene family. Its action modifies the structure of xyloglucans, a diverse group of polysaccharides that crosslink with cellulose microfibrills, affecting therefore the functional structure of the cell wall. The aim of this work is to identify the XTH-encoding genes present in F. vesca and to determine its transcription level in ripening fruit. RESULTS: The search resulted in identification of 26 XTH-encoding genes named as FvXTHs. Genetic structure and phylogenetic analyses were performed allowing the classification of FvXTH genes into three phylogenetic groups: 17 in group I/II, 2 in group IIIA and 4 in group IIIB. Two sequences were included into the ancestral group. Through a comparative analysis, characteristic structural protein domains were found in FvXTH protein sequences. In complement, expression analyses of FvXTHs by qPCR were performed in fruit at different developmental and ripening stages, as well as, in other tissues. The results showed a diverse expression pattern of FvXTHs in several tissues, although most of them are highly expressed in roots. Their expression patterns are not related to their respective phylogenetic groups. In addition, most FvXTHs are expressed in ripe fruit, and interestingly, some of them (FvXTH 18 and 20, belonging to phylogenic group I/II, and FvXTH 25 and 26 to group IIIB) display an increasing expression pattern as the fruit ripens. CONCLUSION: A discrete group of FvXTHs (18, 20, 25 and 26) increases their expression during softening of F. vesca fruit, and could take part in cell wall remodeling required for softening in collaboration with other cell wall degrading enzymes.
Assuntos
Fragaria/enzimologia , Fragaria/genética , Frutas/crescimento & desenvolvimento , Frutas/genética , Perfilação da Expressão Gênica , Genômica , Glicosiltransferases/genética , Sequência de Aminoácidos , Fragaria/crescimento & desenvolvimento , Glicosiltransferases/química , Filogenia , Alinhamento de SequênciaRESUMO
The two-spotted spider mite, Tetranychus urticae Koch, is considered one of the main pests of horticultural and other crops in Brazil, in areas where a considerable volume of acaricides is annually used for its control. The objective of this work was to test the technical viability of using phytoseiids Amblyseius idaeus (Denmark & Muma) and Phytoseudus macropilis (Banks), commonly found in Brazil, to control T .urticae in cucumber and strawberry crops in Jaguaríúna,SP. A randomized complete block design was used, with 4 replicates and 4 treatments for cucumber (T1-T4) and 5 treatments for strawberry crops (T1-T5): T1, control; T2, weekly sprays of malathion for exclusion of native predators; T3, periodical releases of A.idaeus; T4, periodical releases of P. macropilis; T5, weekly sprays of avermectin for exclusion of native phytophagous and predatory mites. Only A. idaeus was successfully established on cucumber, significantly reducing T. urticae population. Both released phytoseiid species were established on strawberry and reduced significantly the population of T. urticae.
O ácaro rajado, Tetranychus urticae Koch, é considerado uma das principais pragas de hortaliças e várias outras culturas no Brasil, em áreas onde um considerável volume de acaricidas é anualmente empregado no seu controle. O objetivo deste trabalho foi testar a viabilidade técnica do emprego dos fitoseídeos Amblyseius idaeus (Denmark & Muma) e Phytoseudus macropilis (Banks), comum ente encontrados no Brasil, no controle de T. urticae em pepino e morangueiro em Jaguaríúna,SP. Utilizou-se o delineamento experimental em blocos casualizados, com 4 repetições e 4 tratamentos (T1-T4) para o pepino e 5 tratamentos para o morangueiro (T1-T5): T1, testemunha; T2, pulverizações semanais de Malation para a exclusão de predadores nativos; T3, liberações periódicas de A.idaeus; T4, liberações periódicas de P.macropilis; T5, pulverizações semanais de avermectina para a exclusão de ácaros fitófagos e predadores nativos. Apenas A.idaeus se estabeleceu na cultura de pepino, reduzindo significativamente a população de T. urticae. Ambas espécies de predadores se estabeleceram na cultura do morango e reduziram significativamente a população de T.urticae.
RESUMO
The two-spotted spider mite, Tetranychus urticae Koch, is considered one of the main pests of horticultural and other crops in Brazil, in areas where a considerable volume of acaricides is annually used for its control. The objective of this work was to test the technical viability of using phytoseiids Amblyseius idaeus (Denmark & Muma) and Phytoseudus macropilis (Banks), commonly found in Brazil, to control T .urticae in cucumber and strawberry crops in Jaguaríúna,SP. A randomized complete block design was used, with 4 replicates and 4 treatments for cucumber (T1-T4) and 5 treatments for strawberry crops (T1-T5): T1, control; T2, weekly sprays of malathion for exclusion of native predators; T3, periodical releases of A.idaeus; T4, periodical releases of P. macropilis; T5, weekly sprays of avermectin for exclusion of native phytophagous and predatory mites. Only A. idaeus was successfully established on cucumber, significantly reducing T. urticae population. Both released phytoseiid species were established on strawberry and reduced significantly the population of T. urticae.
O ácaro rajado, Tetranychus urticae Koch, é considerado uma das principais pragas de hortaliças e várias outras culturas no Brasil, em áreas onde um considerável volume de acaricidas é anualmente empregado no seu controle. O objetivo deste trabalho foi testar a viabilidade técnica do emprego dos fitoseídeos Amblyseius idaeus (Denmark & Muma) e Phytoseudus macropilis (Banks), comum ente encontrados no Brasil, no controle de T. urticae em pepino e morangueiro em Jaguaríúna,SP. Utilizou-se o delineamento experimental em blocos casualizados, com 4 repetições e 4 tratamentos (T1-T4) para o pepino e 5 tratamentos para o morangueiro (T1-T5): T1, testemunha; T2, pulverizações semanais de Malation para a exclusão de predadores nativos; T3, liberações periódicas de A.idaeus; T4, liberações periódicas de P.macropilis; T5, pulverizações semanais de avermectina para a exclusão de ácaros fitófagos e predadores nativos. Apenas A.idaeus se estabeleceu na cultura de pepino, reduzindo significativamente a população de T. urticae. Ambas espécies de predadores se estabeleceram na cultura do morango e reduziram significativamente a população de T.urticae.