Dynamic complex formation between HD-GYP, GGDEF and PilZ domain proteins regulates motility in Xanthomonas campestris

RpfG is a member of a class of wide spread bacterial two-component regulators with an HD-GYP cyclic di-GMP phosphodiesterase domain. In the plant pathogen Xanthomonas campestris, RpfG together with the sensor kinase RpfC regulates multiple factors as a response to the cell-to-cell Diffusible Signalling Factor (DSF). A dynamic physical interaction of RpfG with two diguanylate cyclase (GGDEF) domain proteins controls motility. Here we show that, contrary to expectation, regulation of motility by the GGDEF domain proteins does not depend upon their cyclic di-GMP synthetic activity. Furthermore we show that the complex of RpfG and GGDEF domain proteins recruits a specific PilZ domain adaptor protein, and this complex then interacts with the pilus motor proteins PilU and PiIT. The results support a model in which DSF signalling influences motility through the highly regulated dynamic interaction of proteins that affect pilus action. A specific motif that we identify to be required for HD-GYP domain interaction is conserved in a number of GGDEF domain proteins, suggesting that regulation via interdomain interactions is of broad relevance.

Analysis of three Xanthomonas axonopodis pv. citri effector proteins in pathogenicity and their interactions with host plant proteins

Xanthomonas axonopodis pv. citri, the bacterium responsible for citrus canker, uses effector proteins secreted by a type III protein secretion system to colonize its hosts. Among the putative effector proteins identified for this bacterium, we focused on the analysis of the roles of AvrXacE1, AvrXacE2 and Xac3090 in pathogenicity and their interactions with host plant proteins. Bacterial deletion mutants in avrXacE1, avrXacE2 and xac3090 were constructed and evaluated in pathogenicity assays. The avrXacE1 and avrXacE2 mutants presented lesions with larger necrotic areas relative to the wild-type strain when infiltrated in citrus leaves. Yeast two-hybrid studies were used to identify several plant proteins likely to interact with AvrXacE1, AvrXacE2 and Xac3090. We also assessed the localization of these effector proteins fused to green fluorescent protein in the plant cell, and observed that they co-localized to the subcellular spaces in which the plant proteins with which they interacted were predicted to be confined. Our results suggest that, although AvrXacE1 localizes to the plant cell nucleus, where it interacts with transcription factors and DNA-binding proteins, AvrXacE2 appears to be involved in lesion-stimulating disease 1-mediated cell death, and Xac3090 is directed to the chloroplast where its function remains to be clarified.

Germination and seedling morphology of four South American Smilax (Smilacaceae)

Species of Smilax,, also known as greenbrier, are widely distributed in Brazil and their commercial trades are carried out by the extractivism of native species. We the aim to provide information about the germination and development of seedlings in four Smilax species, different experiments were developed under controlled conditions. We evaluated two germination treatments: temperature (30 degrees C and 20-30 degrees C) and light (presence/absence), and for few cases the tetrazolium treatment was applied. A different treatment response was observed among the studied species. Light had a significant influence in S. brasiliensis, with the highest germination rates at 20-30 C in dark conditions. S. campestris showed significant differences among temperature treatments, but not to light; while S. cissoides showed high germination rates (66-78%), independently of treatment. However, S. polyantha had low germination rates (19-24%). After one year, the expanded leaves showed different characteristics among the studied species. Leaves of S. brasiliensis were ovate, coriaceous, three main veins and prickle-like structures only on the midrib on abaxial face. S. campestris leaves were oblong, coriaceous and prickle-like structures were located at the leaf midrib and margin. S. cissoides had ovate-elliptic, membranaceous leaves, with three main veins with prickle-like structures on the abaxial face. S. polyantha leaves showed ovate-elliptic. coriaceous leaves, with three main veins, translucent secondary veins and no prickle-like structures. A seedling identification key was elaborated based on morphological characteristics. Rev. Biol. Trop. 60 (1): 495-504. Epub 2012 March 01.

Analysis of the Organic Hydroperoxide Response of Chromobacterium violaceum Reveals That OhrR Is a Cys-Based Redox Sensor Regulated by Thioredoxin

Organic hydroperoxides are oxidants generated during bacterial-host interactions. Here, we demonstrate that the peroxidase OhrA and its negative regulator OhrR comprise a major pathway for sensing and detoxifying organic hydroperoxides in the opportunistic pathogen Chromobacterium violaceum. Initially, we found that an ohrA mutant was hypersensitive to organic hydroperoxides and that it displayed a low efficiency for decomposing these molecules. Expression of ohrA and ohrR was specifically induced by organic hydroperoxides. These genes were expressed as monocistronic transcripts and also as a bicistronic ohrR-ohrA mRNA, generating the abundantly detected ohrA mRNA and the barely detected ohrR transcript. The bicistronic transcript appears to be processed. OhrR repressed both the ohrA and ohrR genes by binding directly to inverted repeat sequences within their promoters in a redox-dependent manner. Site-directed mutagenesis of each of the four OhrR cysteine residues indicated that the conserved Cys21 is critical to organic hydroperoxide sensing, whereas Cys126 is required for disulfide bond formation. Taken together, these phenotypic, genetic and biochemical data indicate that the response of C. violaceum to organic hydroperoxides is mediated by OhrA and OhrR. Finally, we demonstrated that oxidized OhrR, inactivated by intermolecular disulfide bond formation, is specifically regenerated via thiol-disulfide exchange by thioredoxin (but not other thiol reducing agents such as glutaredoxin, glutathione and lipoamide), providing a physiological reducing system for this thiol-based redox switch.

Guatteria (Annonaceae) da Floresta Atlântica brasileira

Guatteria Ruiz et Pav. é o maior gênero de Annonaceae, com 307 espécies de distribuição neotropical. Caracteres reprodutivos constantes e vegetativos variáveis dificultam a caracterização das espécies. Como resultado do presente estudo são reconhecidas 15 espécies do gênero na Floresta Atlântica, Guatteria australis, G. campestris, G. candolleana, G. emarginata, G. ferruginea, G. latifolia, G. macropus, G. oligocarpa, G. pogonopus, G. pohliana, G. sellowiana, G. schomburgkiana, G. stenocarpa, G. tomentosa e G. villosissima. Oito nomes são sinonimizados e dois lectótipos são indicados. Chave, descrições, comentários taxonômicos, informações sobre fenologia, distribuição geográfica, hábitats de ocorrência, status de conservação e ilustrações são apresentados.