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1.
Arch Virol ; 166(8): 2321-2324, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34046760

RESUMO

The complete sequence of a new viral RNA from babaco (Vasconcellea × heilbornii) was determined. The genome consisted of 4,584 nucleotides, containing two open reading frames (ORFs 1 and 2), a 9-nt-long noncoding region (NCR) at the 5' terminus, and an unusually long (1,843 nt) NCR at the 3' terminus. The presence of a potential heptameric slippery signal located 12 nt upstream the stop codon of ORF 1 suggests a -1 ribosomal frameshift mechanism for the translation of ORF 2. Sequence comparisons of ORF 2 revealed similarity to the RNA-dependent RNA polymerase (RdRp) of several umbra- and umbra-like viruses. Phylogenetic analysis of the RdRp placed the new virus in a well-supported and cohesive clade that includes umbra-like viruses reported in papaya, citrus, opuntia, maize, and sugarcane hosts. Viruses of this clade share a most recent ancestor with the umbraviruses but have different genomic features. The creation of a new genus within the family Tombusviridae is proposed for the classification of these novel viruses.


Assuntos
Caricaceae/virologia , Tombusviridae/classificação , Sequenciamento Completo do Genoma/métodos , Composição de Bases , Tamanho do Genoma , Genoma Viral , Fases de Leitura Aberta , Filogenia , Tombusviridae/genética , Tombusviridae/isolamento & purificação
2.
Sci Rep ; 10(1): 15799, 2020 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-32978465

RESUMO

The chloroplast genome is an integral part of plant genomes in a species along with nuclear and mitochondrial genomes, contributing to adaptation, diversification, and evolution of plant lineages. In the family Caricaceae, only the Carica papaya chloroplast genome and its nuclear and mitochondrial genomes were sequenced, and no chloroplast genome-wide comparison across genera was conducted. Here, we sequenced and assembled the chloroplast genome of Vasconcellea pubescens A.DC. using Oxford Nanopore Technology. The size of the genome is 158,712 bp, smaller than 160,100 bp of the C. papaya chloroplast genome. And two structural haplotypes, LSC_IRa_SSCrc_IRb and LSC_IRa_SSC_IRb, were identified in both V. pubescens and C. papaya chloroplast genomes. The insertion-deletion mutations may play an important role in Ycf1 gene evolution in family Caricaceae. Ycf2 is the only one gene positively selected in the V. pubescens chloroplast genome. In the C. papaya chloroplast genome, there are 46 RNA editing loci with an average RNA editing efficiency of 63%. These findings will improve our understanding of the genomes of these two crops in the family Caricaceae and will contribute to crop improvement.


Assuntos
Caricaceae/classificação , Caricaceae/genética , DNA de Cloroplastos/análise , Evolução Molecular , Genes de Plantas/genética , Genoma de Cloroplastos , Filogenia , DNA de Cloroplastos/genética , Genoma de Planta
3.
Sci Rep ; 10(1): 3074, 2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-32080277

RESUMO

The proteolytic fraction (P1G10) from Vasconcellea cundinamarcensis, displays gastric protective and healing activities in different skin lesions in mice and human. In an excisional model, this fraction accelerates resolution of lesions and modulates inflammatory mediators. Based on these data, we assessed its anti-inflammatory activity in murine colitis model, induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) adopted by its physiopathological similarity with human colitis. Twenty four hours after colitis induction followed by three days of treatment, P1G10 at 0.3 and 3.0 mg/Kg induced 30% increase in body weight (p < 0.0001) and ~80% reduction in colon macroscopic damage score (p < 0.05) compared to the untreated TNBS-induced colitis group. Histological analyses showed that 0.3 mg/Kg P1G10 reduced the inflammatory profile and tissue damage (47%, p < 0.05) when it was proteolytically active. Compared to TNBS group, 0.3 mg/Kg P1G10 reduced MPO activity (80%, p < 0.01), MCP-1 (47%, p < 0.05) and TNF-α (50%, no significant) and increased IL-10 (330%, p < 0.001) levels in the supernatant of colonic tissue homogenate. P1G10 treatment also reduced COX-2 expression (60%, p < 0.05) and metalloprotease-2 activity (39%, p < 0.05) while increased globet cell density (140%, p < 0.01), that contributes to mucus layer protection in colonic tissue. Taken together, these findings suggest that low doses of active P1G10 promotes lesion resolution, at least in part by its anti-inflammatory activity, in TNBS-colitis model.


Assuntos
Anti-Inflamatórios/farmacologia , Colite/induzido quimicamente , Colite/patologia , Látex/química , Proteólise , Animais , Caricaceae/química , Colite/enzimologia , Colo/efeitos dos fármacos , Colo/patologia , Ciclo-Oxigenase 2/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Hexosaminidases/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Peroxidase/metabolismo , Ácido Trinitrobenzenossulfônico , Perda de Peso/efeitos dos fármacos
4.
Burns ; 46(4): 928-936, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-31722838

RESUMO

INTRODUCTION: Chemical ocular burns are among the most frequently eye-related injuries, which require immediate and intensive evaluation and care since they may lead to potential complications such as superinfection, corneal perforation, and blindness.Vasconcellea cundinamarcensis, a species from Caricaceae family, contains highly active proteolytic enzymes in its latex that show healing activity in animal models bearing lesions of different etiologies. METHODS: We evaluate the ocular toxicity of the proteolytic fraction from V. cundinamarcensis (P1G10) by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and Hen's Egg Test-Chorioallantoic Membrane test. The corneal healing property of P1G10 was studied by the ethanol-chemical burn in the rabbit's eyes. RESULTS: P1G10 is safe for ocular administration, except when administrated at 10µg/mL. P1G10 at 1µg/mL accelerates the corneal re-epithelization achieving complete wound closure after 72h of chemical burn. Also, P1G10 modulated the inflammatory response and controlled the arrangement of collagen fibers in the stroma, demonstrating its potential corneal healing properties. CONCLUSIONS: Our work was the first one to evaluate the ophthalmic application of P1G10. Here we demonstrated that P1G10 is suitable for ocular administration and it has a promising corneal healing activity which may emerge as a new pharmacological tool to the development of a new drug for ocular surface chemical injuries in the future.


Assuntos
Queimaduras Químicas/patologia , Caricaceae/enzimologia , Córnea/efeitos dos fármacos , Lesões da Córnea/patologia , Queimaduras Oculares/patologia , Fibroblastos/efeitos dos fármacos , Peptídeo Hidrolases/farmacologia , Reepitelização/efeitos dos fármacos , Administração Oftálmica , Animais , Queimaduras Químicas/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/efeitos dos fármacos , Córnea/citologia , Córnea/metabolismo , Córnea/patologia , Lesões da Córnea/metabolismo , Relação Dose-Resposta a Droga , Etanol/toxicidade , Queimaduras Oculares/metabolismo , Humanos , Técnicas In Vitro , Inflamação , Látex/química , Coelhos , Solventes/toxicidade , Cicatrização/efeitos dos fármacos
5.
Int J Mol Sci ; 19(10)2018 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-30241282

RESUMO

Previous studies showed that P1G10, a proteolytic fraction from Vasconcellea cundinamarcensis latex, reduced the tumor mass in animals bearing melanoma, increased in vitro DNA fragmentation and decreased cell adhesion. Here, we present some molecular and cellular events related to the antimetastatic effect induced by the CMS-2 fraction derived from P1G10 in metastatic melanoma B16-F10 and melanocyte Melan-a. Using difference gel electrophoresis and mass spectrometry, we identified four proteins overexpressed in tumor cells, all of them related to proliferation, survival, migration and cell invasion, that had their expression normalized upon treatment with CMS-2: nucleophosmin 1, heat shock protein 65, calcyclin binding protein and eukaryotic translation initiation factor 4H. In addition, some antioxidant and glycolytic enzymes show increased expression after exposure to CMS-2, along with an induction of melanogenesis (differentiation marker). The down regulation of cofilin 1, a protein involved in cell motility, may explain the inhibition of cell migration and dendritic-like outgrowth in B16-F10 and Melan-a, observed after CMS-2 treatment. Taken together, it is argued that CMS-2 modulates the expression of proteins related to metastatic development, driving the cell to a more differentiated-like state. These effects support the CMS-2 antimetastatic activity and place this fraction in the category of anticancer agent.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Caricaceae/enzimologia , Cisteína Proteases/farmacologia , Melanoma/tratamento farmacológico , Metástase Neoplásica/tratamento farmacológico , Proteínas de Plantas/farmacologia , Animais , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cisteína Proteases/uso terapêutico , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Melanoma/patologia , Camundongos , Metástase Neoplásica/patologia
6.
Adv Ther ; 35(4): 494-502, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29564750

RESUMO

INTRODUCTION: The aim of the study was to investigate the role of the proteolytic fraction from Vasconcellea cundinamarcensis, designated as P1G10, on the healing of chronic foot ulcers in neuropathic patients with diabetes 2. METHODS: Fifty patients were enrolled in a prospective, randomized, double-blind trial, to verify the efficacy and safety of a topical dressing formulated with 0.1% P1G10, intended for wound healing, versus a hydrogel (control) protocol. Upon completion of the intervention, the outcome evaluated the number of patients attaining full epithelization (100%), or at least 80% healing. Statistical analysis compared the data on each group for the significance of the differences. RESULTS: Collection of data was finished in week 16, and the results were analyzed by intention to treat. The results showed that, in the control group, 5 patients attained 100% ulcer healing, 3 patients ≥ 80% healing and 11 experienced ulcer changes ≤ 80%, and the remainder showed no changes or their wounds became worse. Meanwhile, in the P1G10 group, 11 patients experienced full healing, 4 had healing ≥ 80% and 5 had ulcer changes ≤ lower than 80%, and the remainder showed no changes or their wounds became worse. The healing incidence for the first endpoint (100% healing) showed that the P1G10 group was 2.95-fold more efficacious than the control group (CI 95%) and 2.52-fold (CI, 95%) higher than its control for the second endpoint (80% healing). CONCLUSIONS: These data support the hypothesis that topical application of the proteolytic fraction identified as P1G10 significantly enhances foot ulcer healing compared to hydrogel treatment.


Assuntos
Caricaceae , Pé Diabético/terapia , Látex/uso terapêutico , Cicatrização/efeitos dos fármacos , Doença Crônica , Diabetes Mellitus Tipo 2/complicações , Pé Diabético/etiologia , Nefropatias Diabéticas/complicações , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Estudos Prospectivos
7.
J Ethnopharmacol ; 178: 40-9, 2016 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-26657577

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Aloe vera (L.) Burm. f. (Xanthorrhoeaceae), Carica papaya L. (Caricaceae) and Mimosa pudica L. (Fabaceae) are widely used in the Cameroonian ethnoveterinary medicine as a panacea, and specifically for gastrointestinal disorders as well as an anthelmintic and antibacterial. AIM OF THE STUDY: The present study evaluated the potential toxicity of the hydroalcoholic extracts of Aloe vera leaves, Carica papaya leaves or seeds, and Mimosa pudica leaves after acute and sub-chronic administration in chicks. MATERIALS AND METHODS: For the acute toxicity test a single administration of each of the four hydroalcoholic extracts was given orally at doses ranging from 40 to 5120 mg/kg (n=5/group/sex). In the sub-chronic study, these extracts were given orally as a single administration to chicks at doses of 80, 160, 320 and 640 mg/kg/day for 42 days. The anti-angiogenic properties of these extracts (5-320 µg/mg) were investigated in the chick chorioallantoic membrane in vivo. RESULTS: In the acute toxicity test, none of the four studied hydroalcoholic extracts induced mortality or significant behavioural changes. The sub-acute treatment with the four plant extracts did not alter either the body weight gain or the food and water consumption. However, the results indicated that Aloe vera leaf extract acute treatment by oral route at doses up to 2560 mg/kg did not produce death in 50% (5/10) of chicks during 24h or 14 days of observation, but 20% (2/10) chicks died. The haematological and biochemical analyses did not show significant differences in any of the parameters examined in female or male groups, with the exception of a transient rise in white blood cell counts at high doses (640 mg/kg). Additionally, these extracts did not have the potential for anti-angiogenic effects through the inhibition of neo-angiogenesis in the chick chorioallantoic membrane in vivo. CONCLUSION: The results showed that the therapeutic use of the hydroalcoholic extracts of Aloe vera leaves, Carica papaya leaves or seeds and Mimosa pudica leaves had very low toxicity in oral acute high dose administration and no toxicity in oral sub-chronic low dose administration and indicate that the plants could be considered safe for oral medication in chicks.


Assuntos
Aloe/toxicidade , Carica/toxicidade , Caricaceae/toxicidade , Mimosa/toxicidade , Folhas de Planta/toxicidade , Sementes/toxicidade , Animais , Galinhas , Fabaceae/toxicidade , Feminino , Masculino , Extratos Vegetais/toxicidade , Testes de Toxicidade Aguda
8.
J Mol Model ; 21(8): 216, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26227064

RESUMO

Alcohol acyltransferases (AAT) catalyze the esterification reaction of alcohols and acyl-CoA into esters in fruits and flowers. Despite the high divergence between AAT enzymes, two important and conserved motifs are shared: the catalytic HxxxD motif, and the DFGWG motif. The latter is proposed to play a structural role; however, its function remains unclear. The DFGWG motif is located in loop 21 and stabilized by a hydrogen bond between residues Y52 and D381. Also, this motif is distant from the HxxxD motif, and most probably without a direct role in the substrate interaction. To evaluate the role of the DFGWG motif, in silico analysis was performed in the VpAAT1 protein. Three mutants (Y52F, D381A and D381E) were evaluated. Major changes (size and shape) in the solvent channels were found, although no differences were revealed in the entire 3D structure. Molecular dynamics simulations and docking studies described unfavorable energies for interaction of the mutant proteins with different substrates, as well as unfavored ligand orientations in the solvent channel. Additionally, we examined the contribution of different energetic parameters to the total free energy of protein-ligand complexes by the MM-GBSA method. The complexes differed mainly in their van der Waals contributions and have unfavorable electrostatic interactions. VpAAT1, Y52F and D381A mutants showed a dramatic reduction in the binding capacity to several substrates, which is related to differences in electrostatic potential on the protein surfaces, suggesting that D381 from the DFGWG motif and residue Y52 play a crucial role in maintenance of the adequate solvent channel structure required for catalysis. Graphical abstract Molecular docking, molecular dynamics (MD) simulations and MM-GBSA free energy calculations were employed to obtain quantitative estimates for the binding free energies of wild type Vasconcellea pubescens alcohol acyltransferase (VpAAT1-WT) and the protein mutants. Left VpAAT1 model structure in cartoon representation showing the solvent channel in the middle of the structure. Center, right Changes in shape and structure in the solvent channel of Y52F and D381A mutant proteins, respectively, compared to WT. The results obtained reveal that the interaction between D381 and Y52 residues is important for the maintenance of solvent channel structure.


Assuntos
Caricaceae/enzimologia , Simulação de Dinâmica Molecular , Proteínas/química , Motivos de Aminoácidos , Simulação por Computador , Mutação , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estrutura Terciária de Proteína , Proteínas/genética , Proteínas/metabolismo
9.
Genetics ; 199(2): 413-21, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25480779

RESUMO

Sex chromosomes have been studied in many plant and animal species. However, few species are suitable as models to study the evolutionary histories of sex chromosomes. We previously demonstrated that papaya (Carica papaya) (2n = 2x = 18), a fruit tree in the family Caricaceae, contains recently emerged but cytologically heteromorphic X/Y chromosomes. We have been intrigued by the possible presence and evolution of sex chromosomes in other dioecious Caricaceae species. We selected a set of 22 bacterial artificial chromosome (BAC) clones that are distributed along the papaya X/Y chromosomes. These BACs were mapped to the meiotic pachytene chromosomes of Vasconcellea parviflora (2n = 2x = 18), a species that diverged from papaya ∼27 million years ago. We demonstrate that V. parviflora contains a pair of heteromorphic X/Y chromosomes that are homologous to the papaya X/Y chromosomes. The comparative mapping results revealed that the male-specific regions of the Y chromosomes (MSYs) probably initiated near the centromere of the Y chromosomes in both species. The two MSYs, however, shared only a small chromosomal domain near the centromere in otherwise rearranged chromosomes. The V. parviflora MSY expanded toward the short arm of the chromosome, whereas the papaya MSY expanded in the opposite direction. Most BACs mapped to papaya MSY were not located in V. parviflora MSY, revealing different DNA compositions in the two MSYs. These results suggest that mutation of gene(s) in the centromeric region may have triggered sex chromosome evolution in these plant species.


Assuntos
Caricaceae/genética , Centrômero , Genes de Plantas , Diferenciação Sexual/genética , Caricaceae/classificação , Cromossomos de Plantas , Flores , Heterocromatina , Hibridização in Situ Fluorescente , Fenótipo , Filogenia , Mapeamento Físico do Cromossomo , Cromossomos Sexuais
10.
J Chem Inf Model ; 53(10): 2689-700, 2013 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-24032548

RESUMO

Aroma in Vasconcellea pubescens fruit is determined by esters, which are the products of catalysis by alcohol acyltransferase (VpAAT1). VpAAT1 protein structure displayed the conserved HxxxD motif facing the solvent channel in the center of the structure. To gain insight into the role of these catalytic residues, kinetic and site-directed mutagenesis studies were carried out in VpAAT1 protein. Based on dead-end inhibition studies, the kinetic could be described in terms of a ternary complex mechanism with the H166 residue as the catalytic base. Kinetic results showed the lowest Km value for hexanoyl-CoA. Additionally, the most favorable predicted substrate orientation was observed for hexanoyl-CoA, showing a coincidence between kinetic studies and molecular docking analysis. Substitutions H166A, D170A, D170N, and D170E were evaluated in silico. The solvent channel in all mutant structures was lost, showing large differences with the native structure. Molecular docking and molecular dynamics simulations were able to describe unfavored energies for the interaction of the mutant proteins with different alcohols and acyl-CoAs. Additionally, in vitro site-directed mutagenesis of H166 and D170 in VpAAT1 induced a loss of activity, confirming the functional role of both residues for the activity, H166 being directly involved in catalysis.


Assuntos
Acil Coenzima A/química , Aciltransferases/química , Caricaceae/enzimologia , Frutas/enzimologia , Simulação de Dinâmica Molecular , Proteínas de Plantas/química , Aciltransferases/genética , Motivos de Aminoácidos , Biocatálise , Caricaceae/química , Domínio Catalítico , Ensaios Enzimáticos , Escherichia coli/genética , Escherichia coli/metabolismo , Frutas/química , Cinética , Simulação de Acoplamento Molecular , Mutagênese Sítio-Dirigida , Proteínas de Plantas/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Termodinâmica
11.
Planta ; 236(5): 1471-84, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22790602

RESUMO

Vasconcellea quercifolia (Caricaceae) latex contains several cysteine endopeptidases with high proteolytic activity. Cysteine endopeptidases are the main active compounds used by the plant as a defense mechanism. A proteolytic preparation from V. quercifolia ("oak leaved papaya") latex was purified by cation exchange chromatography. From SDS-PAGE and blotting of the selected fractions, the N-terminal amino acid sequences of polypeptides were determined by Edman's degradation. The analysis by peptide mass fingerprinting (PMF) of the enzymes allowed their characterization and confirmed the presence of seven different cysteine proteinases in the latex of V. quercifolia. Moreover, the comparison between the tryptic maps with those deposited in databases using the MASCOT tool showed that none of the isolated proteases matched with another plant protease. Notably, a propeptidase was detected in the plant latex, which is being the first report in this sense. Furthermore, the cDNA of one of the cysteine proteases that is expressed in the latex of V. quercifolia was cloned and sequenced. The consensus sequence was aligned using the ClustalX web server, which allowed detecting a high degree of identity with cysteine proteases of the Caricaceae family and establishing the evolutionary relationship between them. We also observed a high conservation degree for those amino acid residues which are essential for the catalytic activity and tridimensional structure of the plant proteases belonging to the subfamily C1A. The PMF analysis strongly suggests that the sequence obtained corresponds to the VQ-III peptidase.


Assuntos
Caricaceae/química , Látex/metabolismo , Peptídeo Hidrolases/química , Peptídeo Hidrolases/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Caricaceae/metabolismo , Clonagem Molecular , Sequência Conservada , Cisteína Proteases/química , Cisteína Proteases/genética , Cisteína Proteases/metabolismo , Eletroforese em Gel de Poliacrilamida , Modelos Moleculares , Dados de Sequência Molecular , Peptídeo Hidrolases/isolamento & purificação , Mapeamento de Peptídeos , Filogenia , Proteínas de Plantas/análise , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Estrutura Terciária de Proteína , Proteólise
12.
Gene ; 502(1): 60-8, 2012 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-22543019

RESUMO

Cysteine proteases (CPs) from the C1 family, which are similar to papain, can be found in animals and plants, as well as some viruses and prokaryotes. These enzymes have diverse physiological functions and are thus very attractive for science and industry. Jacaratia mexicana, a member of the Caricaceae plant family, contains several CPs, the principal being mexicain, found to favorably compete against papain for many industrial applications due to its high stability and specific activity. In this study, leaves of J. mexicana were used to isolate a CP-coding gene, similar to those that code for mexicain and chymomexicain. By using rapid amplification of cDNA ends (RACE) as well as oligonucleotide design from papain-like conserved amino acids (aa), a sequence of 1404 bp consisting of a 5' terminal untranslated region (UTR) of 153 bp, a 3' terminal UTR of 131 bp, with a polyadenylation (poly(A)) signal sequence and a poly(A) tail, and an open reading frame (ORF) of 1046 bp, was obtained by overlapping three partial sequences. Two full-length cDNA sequences that encode for mexicain-like proteases were cloned from mRNA (JmCP4 and JmCP5). JmCP4 is predicted to have an ORF of 1044 bp, which codifies for polypeptides that have a 26 aa signal peptide region, a 108 aa propeptide region and a mature enzyme of 214 aa. A 969 bp fragment (JmCP5) encodes for a partial sequence of a CP gene, without the signal peptide region but with a full-length propeptide region. The sequence analysis showed that this protease presented a high similarity to other plant CPs from J. mexicana, Vasconcellea cundinamarcensis, Vasconcellea stipulata, and Carica papaya, among others, mainly at the conserved catalytic site. Obtaining the sequence of this CP gene from J. mexicana provides an alternative for production in a standard system and could be an initial step towards the commercialization of this enzyme.


Assuntos
Caricaceae/genética , Cisteína Proteases/genética , Proteínas de Plantas/genética , Precursores de Proteínas/genética , RNA de Plantas/isolamento & purificação , Motivos de Aminoácidos , Sequência de Aminoácidos , Sequência de Bases , Caricaceae/enzimologia , Domínio Catalítico , Cisteína Proteases/biossíntese , Cisteína Proteases/química , DNA Complementar/biossíntese , Estabilidade Enzimática , Isoenzimas/biossíntese , Isoenzimas/química , Isoenzimas/genética , Modelos Moleculares , Dados de Sequência Molecular , Folhas de Planta/genética , Proteínas de Plantas/biossíntese , Proteínas de Plantas/química , Precursores de Proteínas/química , RNA Mensageiro/genética , RNA Mensageiro/isolamento & purificação , RNA de Plantas/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia Estrutural de Proteína
13.
Rev Biol Trop ; 60(1): 1-10, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22458206

RESUMO

The tropical dry forest is a greatly endangered ecosystem, from which Jacaratia mexicana is a native tree. With the aim to assess the levels of genetic variation and population structure, four wild populations of J. mexicana were studied in the Sierra de Huautla Biosphere Reserve, Morelos, Mexico. For this, DNA was extracted from 159 individuals and were amplified with six random primers using the Random Amplified Polymorphic DNA (RAPD). A total of 54 bands were obtained, of which 50 (92.6%) were polymorphic. The total genetic diversity found within the four populations was 0.451 when estimated by Shannon's index. An AMOVA analysis showed that 84% of the total genetic variation was found within populations and 16% was among populations. The UPGMA dendrogram showed that all individuals from one of the populations (Huaxtla) formed one distinct genetic group, while the rest of the individuals did not cluster according to population. A Mantel test did not show an association between genetic and geographical distances among populations (r=0.893, p=0.20). A Bayesian cluster analysis performed with STRUCTURE, showed that the most probable number of genetic groups in the data was four (K=4), and confirmed the distinctness of Huaxtla population. Our results showed that important genetic differentiation among populations can occur even at this small geographic scale and this has to be considered in conservation actions for this genetic resource.


Assuntos
Caricaceae/genética , Variação Genética , Árvores/genética , Teorema de Bayes , DNA de Plantas/análise , México , Técnica de Amplificação ao Acaso de DNA Polimórfico
14.
Yi Chuan ; 34(2): 145-56, 2012 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-22382056

RESUMO

In recent years, the number of sequencing data of plant whole genome have been increasing rapidly and the whole genome sequencing has been also performed widely in woody plants. However, there are a set of obstacles in investigating the whole genome sequencing in woody plants, which include larger genome, complex genome structure, limitations of assembly, annotation, functional analysis, and restriction of the funds for scientific research. Therefore, to promote the efficiency of the whole genome sequencing in woody plants, the development and defect of this field should be analyzed. The three-generation sequencing technologies (i.e., Sanger sequencing, synthesis sequencing, and single molecule sequencing) were compared in our studies. The progress mainly focused on the whole genome sequencing in four woody plants (Populus, Grapevine, Papaya, and Apple), and the application of sequencing results also was analyzed. The future of whole genome sequencing research in woody plants, consisting of material selection, establishment of genetic map and physical map, selection of sequencing technology, bioinformatic analysis, and application of sequencing results, was discussed.


Assuntos
Genoma de Planta , Análise de Sequência de DNA/métodos , Árvores/genética , Caricaceae/genética , Rosácea/genética , Vitis/genética
15.
Rev. biol. trop ; 60(1): 1-10, Mar. 2012. ilus, graf, tab
Artigo em Inglês | LILACS | ID: lil-657760

RESUMO

The tropical dry forest is a greatly endangered ecosystem, from which Jacaratia mexicana is a native tree. With the aim to assess the levels of genetic variation and population structure, four wild populations of J. mexicana were studied in the Sierra de Huautla Biosphere Reserve, Morelos, Mexico. For this, DNA was extracted from 159 individuals and were amplified with six random primers using the Random Amplified Polymorphic DNA (RAPD). A total of 54 bands were obtained, of which 50 (92.6%) were polymorphic. The total genetic diversity found within the four populations was 0.451 when estimated by Shannon’s index. An AMOVA analysis showed that 84% of the total genetic variation was found within populations and 16% was among populations. The UPGMA dendrogram showed that all individuals from one of the populations (Huaxtla) formed one distinct genetic group, while the rest of the individuals did not cluster according to population. A Mantel test did not show an association between genetic and geographical distances among populations (r=0.893, p=0.20). A Bayesian cluster analysis performed with STRUCTURE, showed that the most probable number of genetic groups in the data was four (K=4), and confirmed the distinctness of Huaxtla population. Our results showed that important genetic differentiation among populations can occur even at this small geographic scale and this has to be considered in conservation actions for this genetic resource.


Jacaratia mexicana es un árbol nativo del bosque tropical seco, que es considerado el tipo de vegetación en mayor riesgo de desaparecer completamente. Se utilizaron polimorfismos de ADN amplificados al azar (RAPD, Random Amplified Polymorphic DNA), para evaluar los niveles de variación y estructura genética en cuatro poblaciones silvestres de J. mexicana en la Reserva de la Biósfera Sierra de Huautla (Morelos, México). Se amplificó el ADN de 159 individuos utilizando seis oligonucleótidos (“primers”) aleatorios. Se obtuvieron en total 54 bandas, de las cuales 50 (92.6%) fueron polimórficas. La diversidad genética total que se encontró en las cuatro poblaciones de J. mexicana fue de 0.451 de acuerdo con el índice de Shannon. Un análisis de varianza molecular (AMOVA) mostró que el 84% de la variación genética total se encuentra dentro de las poblaciones y el 16% entre las poblaciones. Un dendrograma construido mediante el algoritmo UPGMA mostró que los individuos de una población (Huaxtla) formaron un grupo, mientras que el resto de los individuos no se agruparon de acuerdo a su población de origen. Una prueba de Mantel no mostró una asociación entre las distancias genéticas y geográficas entre las poblaciones (r=0.893, p=0.20). Un análisis de agrupamiento Bayesiano realizado mediante STRUCTURE mostró que el número más probable de grupos genéticos es cuatro (K=4) y confirmó la diferenciación de la población Huaxtla. Nuestros resultados muestran que una considerable diferenciación genética entre poblaciones puede existir incluso a esta escala geográfica, lo cual es de interés para la conservación de este recurso genético.


Assuntos
Caricaceae/genética , Variação Genética , Árvores/genética , Teorema de Bayes , DNA de Plantas/análise , México , Técnica de Amplificação ao Acaso de DNA Polimórfico
16.
J Agric Food Chem ; 58(20): 11027-35, 2010 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-20873836

RESUMO

A new proteolytic preparation from Vasconcellea quercifolia ("oak leaved papaya") latex containing several cysteine endopeptidases with high proteolytic activity has been obtained. The specific activity of the new enzymatic preparation (VQ) was higher than that of Carica papaya latex. VQ was able to coagulate milk and to hydrolyze caseins and then could be used to produce cheeses and/or casein hydrolysates. Ion exchange chromatography of VQ allowed the isolation of a new protease, named quercifoliain I, homogeneous when analyzed by SDS-PAGE, IEF and MALDI-TOF-MS. Molecular mass was 24195 Da, and its isoelectric point was >9.3. The N-terminal sequence was determined (YPESVDWRQ). Insulin B-chain cleavage showed higher specificity than that of papain and was restricted to glycyl and alanyl residues at P1' position. The tryptic peptide mass fingerprint of quercifoliain I analyzed with the MASCOT search tool did not find a match with papain or any other plant cysteine proteases.


Assuntos
Caricaceae/enzimologia , Cisteína Endopeptidases/química , Cisteína Endopeptidases/isolamento & purificação , Látex/química , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Sequência de Aminoácidos , Biocatálise , Caricaceae/química , Caricaceae/genética , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Estabilidade Enzimática , Insulina/química , Insulina/metabolismo , Ponto Isoelétrico , Dados de Sequência Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Especificidade por Substrato
17.
Plant J ; 63(5): 801-10, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20579309

RESUMO

Carica and Vasconcellea are two closely related sister genera in the family Caricaceae, and were once classified as two sections under Carica. Sex chromosomes have been found in papaya and originated approximately 2-3 million years ago. The objectives of this study were to determine whether sex chromosomes have evolved in Vasconcellea. Six X/Y gene pairs were cloned, sequenced and analyzed from three dioecious, one trioecious and one monoecious species of Vasconcellea. The isolation of distinctive X and Y alleles in dioecious and trioecious species of Vasconcellea demonstrated that sex chromosomes have evolved in this genus. Phylogenetic analyses indicated a monophyletic relationship between the X/Y alleles of Carica and those of Vasconcellea. Distinctive clusters of X/Y alleles were documented in V. parviflora and V. pulchra for all available gene sequences, and in V. goudatinana and V. cardinamarcensis for some X/Y alleles. The X and Y alleles within each species shared most single nucleotide polymorphism haplotypes that differed from other species. Limited evidence of gene conversion was documented among the X/Y alleles of some species, but was not sufficient to cause the evolutionary patterns reported herein. The Carica and Vasconcellea sex chromosomes may have originated from the same autosomes bearing the X allelic form that still exist in the monoecious species V. monoica, and have evolved independently after the speciation event that separated Carica from Vasconcellea. Within Vasconcellea, sex chromosomes have evolved at the species level, at least for some species.


Assuntos
Carica/genética , Caricaceae/genética , Cromossomos de Plantas/genética , Recombinação Genética/genética , Alelos , Teorema de Bayes , Evolução Molecular , Filogenia , Proteínas de Plantas/classificação , Proteínas de Plantas/genética , Polimorfismo de Nucleotídeo Único , Reprodução/genética , Especificidade da Espécie
18.
Phytochemistry ; 71(5-6): 524-30, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20116077

RESUMO

The lattices of Carica candamarcensis and Carica papaya, members of the Caricaceae family, contain isoforms of cysteine proteinases that help protect these plants against injury. In a prior study, we fractionated 14 discrete proteinaceous components from C. candamarcensis, two of them displaying mitogenic activity in mammalian cells. In this study, we compared the kinetic parameters of one of the mitogenic proteinases (CMS2MS2) with one of the isoforms displaying the highest enzyme activity of this group (CMS1MS2). Both enzymes display a similar Km value with either BAPNA (Benzoyl-Arg-pNA) or PFLPNA (Pyr-Phe-Leu-pNA), but the kcat of CMS1MS2 is about 14-fold higher for BAPNA and 129-fold higher with PFLPNA. While both enzymes are inhibited by E-64 and iodoacetamide, chicken cystatin fully inhibits CMS1MS2, but scarcely affects activity of CMS2MS2. Based on the structure of these proteins and other enzymes from the Caricaceae family whose structures have been resolved, it is proposed that Arg(180) located in the cleft at the active site in CMS2MS2 is responsible for its resistance to cystatin.


Assuntos
Caricaceae/enzimologia , Cistatinas , Cisteína Proteases/química , Inibidores de Cisteína Proteinase , Mitógenos/química , Peptídeo Hidrolases/química , Proteínas de Plantas/química , Animais , Galinhas , Cisteína Proteases/metabolismo , Iodoacetamida , Mitógenos/metabolismo , Peptídeo Hidrolases/metabolismo , Proteínas de Plantas/metabolismo , Isoformas de Proteínas , Relação Estrutura-Atividade
20.
Investig. amaz ; 3(1): 12-15, ene.-jun. 2009. tab
Artigo em Espanhol | LIPECS | ID: biblio-1108991

RESUMO

La presente investigación tuvo como objetivo la pulpa de frutos de maushan (Vasconcellea weberbaueri Harms V. M. Badillo) en estado de madurez sazón y maduro para la elaboración de mermelada; para lo cual se recolectaron frutos de maushan del distrito de Soloco, región de Amazonas, a los que se midió PH, °brix y acidez; posteriormente, se realizaron seis formulaciones en relación azúcar/pulpa rara para la elaboración mermelada, evaluando la preferencia de los panelistas mediante un DBCA con seis tratamiento y 23 repeticiones; de los mejores tratamientos obtenidos se realizó el estudio de la estabilidad de las características fisicoquímicas y microbiológicas después de uno y dos meses de almacenamiento. Los frutos en los estados de madurez sazón y maduro presentaron valores de pH entre 3,52 - 3,89 y 3,95 – 4,31; °brix entre 6,92 – 7,29 y 8,61 – 8,98; acidez entre 1,90 – 2,26 y 1,33 – 1,69; humedad 85,26% y 87,87%; proteína 1,75% y 1,65%; cenizas 0,65% y 0,64% y fibra cruda 3,2% y 3% respectivamente. La mermelada con mayor aceptación por los panelistas fue aquella en la que se empleó un fruto en estado maduro y una concentración azúcar / pulpa de 50/50. En la evaluación de la estabilidad fisicoquímica se observó ligeras fluctuaciones de los °brix, pH y acidez en los dos meses de almacenamiento, el recuento de mohos, levaduras y aerobios mesófilos viables fueron inferiores a 10 ufc/g.


This study to characterize fruit pulp maushan (Vasconcellea weberbaueri Harms V. M. Badillo) ripe, ripe and ripe for the production of marmalade, for which fruits were collected maushan Soloco District, Amazon region, to pH was measured, °brix and acidity, and thereafter, six formulations in sugar/pulp for jam making, assessing the preference of the panelists by DBCA with six treatments and 23 repetitions, the best treatments is obtained conducted the study on the stability of physiochemical and microbiological characteristics after one to two months of storage. The fruits in the states of maturity and mature then presented pH values between 3,52 to 3,89 and from 3,95 to 4,31; °brix between 6,92 to 7,29 and from 8,61 to 8,98; acidity from 1,90 to 2,26 and from 1,33 to 1,69; moisture 85,26% and 87,87%; protein 1,75% and 1,65%; ash 0,65% and 0,64% and crude fiber 3,2% and 3%, respectively. The jam is more widely accepted by the panelists was that in which we employed a state ripe fruit and sugar concentration / pulp 50/50. In the evaluation of physicochemical stability was observed slight fluctuations of the °brix, pH and acidity in two months of storage, the count of molds, yeasts and aerobic plate viable were below 10cfu/g.


Assuntos
Caricaceae , Geleia de Frutas
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