Hipersensibilidade e necrose sistêmica em Nicotiana benthamiana transformada com o gene de resistência Sw-5 de tomateiro

O gene Sw-5 do tomateiro confere resistência a várias espécies de tospovírus e codifica uma proteína contendo domínios de ligação a nucleotídeos e repetições ricas em leucina. Tomateiros com Sw-5 exibem reações necróticas nas folhas inoculadas com tospovírus. Estas reações e a estrutura da proteína Sw-5 indicam que a resistência ocorre por meio do reconhecimento do patógeno e desencadeamento da resposta de hipersensibilidade. A capacidade de Sw-5 de conferir resistência a tospovírus em tabaco selvagem (Nicotiana benthamiana Domin.) foi avaliada em plantas transgênicas. Uma construção com a seqüência aberta de leitura de Sw-5 e sua região 3 não-traduzida sob controle do promotor 35S do CaMV foi utilizada para transformação de N. benthamiana via Agrobacterium tumefaciens. Plantas de progênies R1 foram inoculadas com um isolado de tospovírus e avaliadas quanto à ocorrência de reação de hipersensibilidade e resistência à infecção sistêmica. em uma progênie com segregação 3:1 (resistente:suscetível), foi selecionada uma planta homozigota e sua progênie avaliada quanto ao espectro da resistência a tospovírus. Plantas com o transgene exibiram resposta de hipersensibilidade 48 h após a inoculação, sendo resistentes à infecção sistêmica. O fenótipo da resistência foi dependente do isolado viral e um isolado de Tomato chlorotic spot virus (TCSV) causou necrose sistêmica em todas as plantas inoculadas, enquanto que isolados de Groundnut ringspot virus (GRSV) e um isolado relacionado a Chrysanthemum stem necrosis virus (CSNV) ficaram restritos ao sítio de infecção. Comparações do espectro da resistência obtido neste trabalho com aquele observado em outros membros da família Solanaceae indicam que as vias de transdução de sinais e as respostas de defesa ativadas por Sw-5 são conservadas dentro desta família e polimorfismos genéticos nas vias de transdução de sinais ou em componentes das respostas de defesa podem resultar em diferentes níveis de resistência.

PR gene families of citrus: their organ specific-biotic and abiotic inducible expression profiles based on ESTs approach

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Phytophthora parasitica transcriptome, a new concept in the understanding of the citrus gummosis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

NEST REUSE AND COMMUNAL NESTING IN MICROTHURGE-CORUMBAE (HYMENOPTERA, MEGACHILIDAE), WITH SPECIAL REFERENCE TO NEST DEFENSE

Observations on the nesting activities of Microthurge corumbae, carried out at the University Campus of Ribeirao Preto, São Paulo, Brazil, from 1977 to 1981, indicated that 61.9% of nests were re-used by succeeding generations. Re-use by one generation was more frequent than by two generations, and re-use by a third was observed only once. Nests were re-used by one or several females. Single females were more frequently in the first re-use. In these cases nest re-use did not differ essentially from the solitary foundation of a new nest, except for the adoption of a pre-existing nest without excavation. In multifemale nests, analysis of relative age (wing wear), ovarian and spermathecal conditions of associated females and the content of nests at excavation indicated that the social pattern in such colonies is communal. There is some evidence that the associated females are relatives. The chalcidoid wasp Leucospis was the principal nest parasite, and ants of the genus Crematogaster were nest predators. In multifemale nests, the rate of parasitism was significantly lower than in solitary nests, indicating that nest-sharing resulted in improved nest defense. on the other hand. The absence of predation on immatures of the first generation of M. corumbue in multifemale nests suggests that such nests are also more resistant to attack by predators.

Structure of a lectin from Canavalia gladiata seeds: new structural insights for old molecules

Background: Lectins are mainly described as simple carbohydrate- binding proteins. Previous studies have tried to identify other binding sites, which possible recognize plant hormones, secondary metabolites, and isolated amino acid residues. We report the crystal structure of a lectin isolated from Canavalia gladiata seeds ( CGL), describing a new binding pocket, which may be related to pathogen resistance activity in ConA- like lectins; a site where a non- protein amino- acid, aaminobutyric acid ( Abu), is bound.Results: the overall structure of native CGL and complexed with alpha- methyl- mannoside and Abu have been refined at 2.3 angstrom and 2.31 angstrom resolution, respectively. Analysis of the electron density maps of the CGL structure shows clearly the presence of Abu, which was confirmed by mass spectrometry.Conclusion: the presence of Abu in a plant lectin structure strongly indicates the ability of lectins on carrying secondary metabolites. Comparison of the amino acids composing the site with other legume lectins revealed that this site is conserved, providing an evidence of the biological relevance of this site. This new action of lectins strengthens their role in defense mechanisms in plants.

Interaction of pathogenic fungi with host cells: Molecular and cellular approaches

This review provides an overview of several molecular and cellular approaches that are likely to supply insights into the host-fungus interaction. Fungi present intra- and/or extracellular host-parasite interfaces, the parasitism phenomenon being dependent on complementary surface molecules. The entry of the pathogen into the host cell is initiated by the fungus adhering to the cell surface, which generates an uptake signal that may induce its cytoplasmatic internalization. Furthermore, microbial pathogens use a variety of their surface molecules to bind to host extracellular matrix (ECM) components to establish an effective infection. on the other hand, integrins mediate the tight adhesion of cells to the ECM at sites referred to as focal adhesions and also play a role in cell signaling. The phosphorylation process is an important mechanism of cell signaling and regulation; it has been implicated recently in defense strategies against a variety of pathogens that alter host-signaling pathways in order to facilitate their invasion and survival within host cells. The study of signal transduction pathways in virulent fungi is especially important in view of their putative role in the regulation of pathogenicity. This review discusses fungal adherence, changes in cytoskeletal organization and signal transduction in relation to host-fungus interaction. (c) 2005 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Increased production of beta-1,3 glucanase and proteins in Bipolaris sorokiniana pathosystem treated using commercial xanthan gum

Barley plants (cultivars Embrapa 127, 128 and 129) treated with xanthan gum, and with different time intervals between the administration of the inducer and the pathogen, demonstrated induction of resistance against Bipolaris sorokiniana. Induction was shown to have local and systemic action. In order to prove the resistance effect, biochemical analyses were performed to quantify proteins and the enzymatic activity of beta-1,3 glucanase. Results demonstrated that barley plants treated with the inducer, showed an increase in the concentration of proteins, as well as in the activity of the enzyme beta-1,3 glucanase, when compared with the extract from healthy plants. In infected plants, protein concentrations decreased and enzymatic activity was lower than in healthy plants. Results suggest that barley plants treated with xanthan gum developed mechanisms responsible for induced resistance, which are still unknown. The most important macromolecule in the defense mechanism was demonstrated to be PR-protein, due to its accumulation and concentration of proteins. However, it may not be the only macromolecule responsible for the resistance effect. (C) 2004 Elsevier SAS. All rights reserved.

Host organism defense by a peptide-polysaccharide extracted from the fungus Sporothrix schenckii

A peptide-polysaccharide, a peptide-rhamnomannan, was isolated from the pathogenic yeast form of the fungus Sporothrix schenckii. This substance, which may play a role in fungal virulence, was tested in an animal model of systemic disease, and depression of the immune response was observed in the animals between the 4th and 6th week of infection. Concomitantly, this compound showed mitogenic activity when challenged with normal lymphocytes and was also found to be involved in the inflammatory response. These results provide further information for the understanding of fungal implantation in tissues and of the pathogenicity of this systemic mycosis.

Phylogenetic and evolutionary aspects of Paracoccidioides brasiliensis reveal a long coexistence with animal hosts that explain several biological features of the pathogen

The habitat of the mycelial saprobic form of Paracoccidio ides brasiliensis, which produces the infectious propagula, has not been determined and has proven difficult for mycologists to describe. The fungus has been rarely isolated from the environment, the disease has a prolonged latency period and no outbreaks have been reported. These facts have precluded the adoption of preventive measures to avoid infection. The confirmation of natural infections in nine-banded armadillos (Dasypus novemcinctus) with P. brasiliensis, in high frequency and wide geographic distribution, has opened new avenues for the study and understanding of its ecology. Armadillos belong to the order Xenarthra, which has existed in South America ever since the Paleocene Era (65 million years ago), when the South American subcontinent was still a detached land, before the consolidation of what is now known as the American continent. on the other hand, strong molecular evidence suggests that P. brasiliensis and other dimorphic pathogenic fungi - such as Blastomyces dermatitidis, Coccidioides immitis and Histoplasma capsulatum - belong to the family Onygenaceae sensu Into (order Onygenales, Ascomycota), which appeared around 150 million years ago.P. brasiliensis ecology and relation to its human host are probably linked to the fungal evolutionary past, especially its long coexistence with and adaptation to animal hosts other than Homo sapiens, of earlier origin. Instead of being a blind alley, the meaning of parasitism for dimorphic pathogenic fungi should be considered as an open two-way avenue, in which the fungus may return to the environment, therefore contributing to preserve its teleomorphic (sexual) and anamorphic (asexual) forms in a defined and protected natural habitat. (c) 2006 Elsevier B.V. All rights reserved.

Copper resistance of biofilm cells of the plant pathogen Xylella fastidiosa

Xylella fastidiosa is a phytopathogen that causes diseases in different plant species. The development of disease symptoms is associated to the blockage of the xylem vessels caused by biofilm formation. In this study, we evaluated the sensitivity of biofilm and planktonic cells to copper, one of the most important antimicrobial agents used in agriculture. We measured the exopolysaccharides (EPS) content in biofilm and planktonic cells and used real-time reverse transcription polymerase chain reaction to evaluate the expression of the genes encoding proteins involved in cation/multidrug extrusion (acrA/B, mexE/czcA, and metI) and others associated with different copper resistance mechanisms (copB, cutA1, cutA2, and cutC) in the X. fastidiosa biofilm formed in two different media. We confirmed that biofilms are less susceptible to copper than planktonic cells. The amount of EPS seems to be directly related to the resistance and it varies according to the media where the cells are grown. The same was observed for gene expression. Nevertheless, some genes seem to have a greater importance in biofilm cells resistance to copper. Our results suggest a synergistic effect between diffusion barriers and other mechanisms associated with bacterial resistance in this phytopathogen. These mechanisms are important for a bacterium that is constantly under stress conditions in the host.

The genome sequence of the gram-positive sugarcane pathogen Leifsonia xyli subsp. xyli

The genome sequence of Leifsonia xyli subsp. xyli, which causes ratoon stunting disease and affects sugarcane worldwide, was determined. The single circular chromosome of Leifsonia xyli subsp. xyli CTCB07 was 2.6 Mb in length with a GC content of 68% and 2,044 predicted open reading frames. The analysis also revealed 307 predicted pseudogenes, which is more than any bacterial plant pathogen sequenced to date. Many of these pseudogenes, if functional, would likely be involved in the degradation of plant heteropolysaccharides, uptake of free sugars, and synthesis of amino acids. Although L. xyli subsp. xyli has only been identified colonizing the xylem vessels of sugarcane, the numbers of predicted regulatory genes and sugar transporters are similar to those in free-living organisms. Some of the predicted pathogenicity genes appear to have been acquired by lateral transfer and include genes for cellulase, pectinase, wilt-inducing protein, lysozyme, and desaturase. The presence of the latter may contribute to stunting, since it is likely involved in the synthesis of abscisic acid, a hormone that arrests growth. Our findings are consistent with the nutritionally fastidious behavior exhibited by L. xyli subsp. xyli and suggest an ongoing adaptation to the restricted ecological niche it inhabits.

A new interpretation of the defense glands of neotropical Ruptitermes (Isoptera, Termitidae, Apicotermitinae)

Suicidal altruism has been reported for some species of eusocial insects, in which the individual dies in defense of the society. The termites of the genus Ruptitermes are known for the suicidal behavior of the workers which liberate a sticky defensive secretion by body bursting. In the present paper it is given a new interpretation of the defense glands of Neotropical Ruptitermes based on the morphological analysis of three species collected at Rio Claro, SP, Brazil. Before the current study, the suicidal defensive behavior was attributed to the dehiscence of the salivary gland reservoirs. The defense or dehiscent glands of Neotropical Ruptitermes are pair structures rounded in shape that are independent of the salivary glands. The dehiscent glands consist of multiple secretory units that are kept together by thin connective tissue. Each secretory unit is composed of one cell generally with one peripheral nucleus and characteristic secretion. The three species studied here present some histological differences in the secretory units, probably related to the chemical composition of the secretion.

Nonthermal activation of transient receptor potential vanilloid-1 channels in abdominal viscera tonically inhibits autonomic cold-defense effectors

An involvement of the transient receptor potential vanilloid (TRPV) 1 channel in the regulation of body temperature (T b) has not been established decisively. To provide decisive evidence for such an involvement and determine its mechanisms were the aims of the present study. We synthesized a new TRPV1 antagonist, AMG0347 [(E)-N-(7-hydroxy-5,6,7,8-tetrahydronaphthalen-1- yl)-3-(2-(piperidin-1-yl)-6-(trifluoromethyl)pyridin-3-yl)acrylamide], and characterized it in vitro. We then found that this drug is the most potent TRPV1 antagonist known to increase T b of rats and mice and showed (by using knock-out mice) that the entire hyperthermic effect of AMG0347 is TRPV1 dependent. AMG0347-induced hyperthermia was brought about by one or both of the two major autonomic cold-defense effector mechanisms (tail-skin vasoconstriction and/or thermogenesis), but it did not involve warmth-seeking behavior. The magnitude of the hyperthermic response depended on neither T b nor tail-skin temperature at the time of AMG0347 administration, thus indicating that AMG0347-induced hyperthermia results from blockade of tonic TRPV1 activation by nonthermal factors. AMG0347 was no more effective in causing hyperthermia when administered into the brain (intracerebroventricularly) or spinal cord (intrathecally) than when given systemically (intravenously), which indicates a peripheral site of action. We then established that localized intra-abdominal desensitization of TRPV1 channels with intraperitoneal resiniferatoxin blocks the T b response to systemic AMG0347; the extent of desensitization was determined by using a comprehensive battery of functional tests. We conclude that tonic activation of TRPV1 channels in the abdominal viscera by yet unidentified nonthermal factors inhibits skin vasoconstriction and thermogenesis, thus having a suppressive effect on T b. Copyright © 2007 Society for Neuroscience.

Nest defense behavior against the attack of ants in colonies of pre-emergent Mischocyttarus cerberus (hymenoptera, vespidae)

The study was carried out at the UNESP Rio Claro campus (SP), where biotests consisting of simulated ant attacks were performed in colonies of Mischocyttarus cerberus. The behaviors of the wasps were recorded with a camcorder, for further analysis. This analysis was done using the Mann-Whitney U test and the Principal Component Analysis. In the pre-emergence development stage, colonies with a single foundress defend the nest only after the first larvae appear. When there are only eggs in the nest, the wasp abandons the nest. Before leaving, the wasp rubs its gaster against the nest, releasing the ant repellent secretion. When the nest contains larvae or larvae and pupae, the foundress defends the colony, vibrating its wings, pumping her abdomen and biting the ant.