Despite extensive studies in eukaryotic aldehyde dehydrogenases, functional information about the ALDH7 antiquitin-like proteins is lacking. A soybean antiquitin homologue gene, designated GmTP55, has been isolated which encodes a dehydrogenase motif-containing 55 kDa protein induced by dehydration and salt stress. GmTP55 is closely related to the stress-induced plant antiquitin-like proteins that belong to the ALDH7 family. Transgenic tobacco (Nicotiana tabacum) and Arabidopsis (Arabidopsis thaliana) plants constitutively expressing GmTP55 have been obtained in order to examine the physiological role of this enzyme under a variety of stress conditions. Ectopic expression of GmTP55 in both Arabidopsis and tobacco conferred tolerance to salinity during germination and to water deficit during plant growth. Under salt stress, the germination efficiency of both transgenic tobacco and Arabidopsis seeds was significantly higher than that of their control counterparts. Likewise, under progressive drought, the transgenic tobacco lines apparently kept the shoot turgidity to a normal level, which contrasted with the leaf wilt phenotype of control plants. The transgenic plants also exhibited an enhanced tolerance to H(2)O(2)- and paraquat-induced oxidative stress. Both GmTP55-expressing Arabidopsis and tobacco seeds germinated efficiently in medium supplemented with H(2)O(2), whereas the germination of control seeds was drastically impaired. Similarly, transgenic tobacco leaf discs treated with paraquat displayed a significant reduction in the necrotic lesions as compared with control leaves. These transgenic lines also exhibited a lower concentration of lipid peroxidation-derived reactive aldehydes under oxidative stress. These results suggest that antiquitin may be involved in adaptive responses mediated by a physiologically relevant detoxification pathway in plants.
Despite extensive studies in plant virus-host interactions, the molecular mechanisms of geminivirus movement and interactions with host components remain largely unknown. A tomato kinase protein and its soybean homolog were found to interact specifically with the nuclear shuttle protein (NSP) of Tomato golden mosaic virus (TGMV) and Tomato crinkle leaf yellows virus (TCrLYV) through yeast two-hybrid screening and in vitro protein binding assays. These proteins, designated LeNIK (Lycopersicon esculentum NSP-Interacting Kinase) and GmNIK (Glycine max NIK), belong to the LRR-RLK (leucine rich-repeat receptor-like kinase) family that is involved in plant developmental processes and/or resistance response. As such, NIK is structurally organized into characteristic domains, including a serine/threonine kinase domain with a nucleotide binding site at the C-terminal region, an internal transmembrane segment and leucine-rich repeats (LRR) at the N-terminal portion. The potential significance of the NSP-NIK interaction is discussed.
Proteins are the major constituents of muscle and are key molecules regulating the metabolic changes during conversion of muscle to meat. Brazil is one of the largest exporters of beef and most Brazilian cattle are composed by zebu (Nellore) genotype. Bos indicus beef is generally leaner and tougher than Bos taurus such as Angus. The aim of this study was to compare the muscle proteomic and phosphoproteomic profile of Angus and Nellore. Seven animals of each breed previously subjected the same growth management were confined for 84 days. Proteins were extracted from Longissimus lumborum samples collected immediately after slaughter and separated by two-dimensional electrophoresis. Pro-Q Diamond stain was used in phosphoproteomics. Proteins identification was performed using matrix assisted laser desorption/ionization time-of-flight mass spectrometry. Tropomyosin alpha-1 chain, troponin-T, myosin light chain-1 fragment, cytoplasmic malate dehydrogenase, alpha-enolase and 78 kDa glucose-regulated protein were more abundant in Nellore, while myosin light chain 3, prohibitin, mitochondrial stress-70 protein and heat shock 70 kDa protein 6 were more abundant in Angus (P<0.05). Nellore had higher phosphorylation of myosin regulatory light chain-2, alpha actin-1, triosephosphate isomerase and 14-3-3 protein epsilon. However, Angus had greater phosphorylation of phosphoglucomutase-1 and troponin-T (P<0.05). Therefore, proteins involved in contraction and muscle organization, myofilaments expressed in fast or slow-twitch fibers and heat shock proteins localized in mitochondria or sarcoplasmic reticulum and involved in cell flux of calcium and apoptosis might be associated with differences in beef quality between Angus and Nellore. Furthermore, prohibitin appears to be a potential biomarker of intramuscular fat in cattle. Additionally, differences in phosphorylation of myofilaments and glycolytic enzymes could be involved with differences in muscle contraction force, susceptibility to calpain, apoptosis and postmortem glycolysis, which might also be related to differences in beef quality among Angus and Nellore.
Colleters are secretory structures that occur in vegetative or reproductive shoot apices of many botanical families. However, in the order Myrtales, reports of colleters have considered only external morphology. We therefore evaluated apical meristems of 52 species belonging to 17 genera from seven tribes of subfamily Myrtoideae (Myrtaceae), so as to analyse the incidence and morphological types of colleters. The samples were fixed for light and scanning electron microscopy. Histochemical tests were carried out on fresh and methacrylate-embedded material. Proteins of the colleter secretions were analysed by SDS-PAGE. We have classified and described the following three new colleter types: petaloid, conic and euryform. None of the species contained all three colleter types. The petaloid colleters were present in three tribes (Syzygieae, Melaleuceae and Lophostemoneae). The conic colleters were observed in three tribes (Leptospermeae, Myrteae and Melaleuceae) and the euryform type occurred in five tribes (Leptospermeae, Syncarpieae, Myrteae, Syzygieae and Melaleuceae). In the tribe Eucalypteae, we found no evidence of colleters. The presence of mucilaginous secretion that defines colleters was confirmed by histochemical tests, and no proteins were found in the secretion. The colleters in Myrtoideae may help clarify the phylogenetic relationships of the Myrtaceae family.
Bacteriocins are ribosomally synthesized antimicrobial peptides produced by Bacteria and some Archaea. The assessment of the toxic potential of antimicrobial peptides is important in order to apply these peptides on an industrial scale. The aim of the present study was to investigate the in vitro cytotoxic and haemolytic potential of bovicin HC5, as well as to determine whether cholesterol influences bacteriocin activity on model membranes. Nisin, for which the mechanism of action is well described, was used as a reference peptide in our assays. The viability of three distinct eukaryotic cell lines treated with bovicin HC5 or nisin was analysed by using the MTT assay and cellular morphological changes were determined by light microscopy. The haemolytic potential was evaluated by using the haemoglobin liberation assay and the role of cholesterol on bacteriocin activity was examined by using model membranes composed of DOPC (1,2-dioleoylsn-glycero-3-phosphocholine) and DPoPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine). The IC 50 of bovicin HC5 and nisin against Vero cells was 65.42 and 13.48 mM, respectively. When the MTT assay was performed with MCF-7 and HepG2 cells, the IC 50 obtained for bovicin HC5 was 279.39 and 289.30 mM, respectively, while for nisin these values were 105.46 and 112.25 mM. The haemolytic activity of bovicin HC5 against eukaryotic cells was always lower than that determined for nisin. The presence of cholesterol did not influence the activity of either bacteriocin on DOPC model membranes, but nisin showed reduced carboxyfluorescein leakage in DPoPC membranes containing cholesterol. In conclusion, bovicin HC5 only exerted cytotoxic effects at concentrations that were greater than the concentration needed for its biological activity, and the presence of cholesterol did not affect its interaction with model membranes.
-The objective of this work was to verify if the induced resistance mechanism is responsible for the capacity of a phylloplane resident bacteria (Bacillus cereus), isolated from healthy tomato plants, to control several diseases of this crop. A strain of Pseudomonas syringae pv. tomato was used as the challenging pathogen. The absence of direct antibiosis of the antagonist against the pathogen, the significant increase in peroxidases activity in tomato plants exposed to the antagonist and then inoculated with the challenging pathogen, as well as the character of the protection, are evidences wich suggest that biocontrol efficiency presented by the antagonist in previous works might be due to induced systemic resistance (ISR).Index terms: Lycopersicon esculentum, Solanum esculentum, biological control. Indução de resistência sistêmica em tomateiro pela bactéria autóctone residente no filoplano Bacillus cereusResumo -O objetivo deste trabalho foi investigar se o mecanismo de indução de resistência é responsável pela capacidade de um isolado bacteriano residente no filoplano (Bacillus cereus), obtido de plantas sadias de tomateiro, em controlar diversas doenças da cultura. A fim de investigar o modo de ação envolvido no controle, foi utilizado um isolado de Pseudomonas syringae pv. tomato como patógeno desafiador. A ausência do mecanismo de antibiose contra o patógeno, o aumento significativo dos níveis de peroxidases em plantas expostas ao antagonista e submetidas à inoculação do patógeno desafiador, assim como a sistemicidade da proteção, são evidências que sugerem que o mecanismo de biocontrole em questão se refere à indução de resistência sistêmica (IRS).Termos para indexação: Lycopersicon esculentum, Solanum esculentum, controle biológico.
Bioprospection of natural compounds to be used as plant protection offers exciting possibilities in agribusiness. ÔSamaraÕ (SM) and ÔSunshineÕ (SH) cultivars of bell pepper are resistant against some plant pathogens, and leaf antimicrobial peptides could account for their defence system. In the present study, 5-7 kDa peptides were partially purified from cell wall extract (CWE) of bell pepper leaves and further resolved by ionexchange chromatography. The cationic and anionic peptide-enriched fractions from both SM-CWE and SH-CWE strongly inhibited in vitro growth of Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis. Peptide extraction and purificationSoluble extract (SE) and cell wall extract (CWE) were obtained from bell pepper leaves of SM or SH cultivars (Segura et al., 1998). Frozen leaves (3 g) were powdered and extracted with 12 ml of 100 mm Tris-HCl www.blackwell-synergy.com
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