María Cecilia Rodríguez

Botrytis cinerea Manipulates the Antagonistic Effects between Immune Pathways to Promote Disease Development in Tomato

Botrytis cinerea is a necrotrophic pathogen that causes grey mould disease in a broad host range, including tomato, grapes, potato, and strawberry. Here, we report that B. cinerea secretes a virulence factor that hijacks the plant’s own crosstalk network to promote disease development. Plants have evolved sophisticated mechanisms to sense and respond to pathogen attacks. Resistance against necrotrophic pathogens generally requires the activation of the jasmonic acid (JA) signaling pathway, whereas the salicylic acid (SA) signaling pathway is mainly activated against biotrophic pathogens. SA can antagonize JA signaling and vice versa. Here, we report that the necrotrophic pathogen Botrytis cinerea exploits this antagonism as a strategy to cause disease development. We show that B. cinerea produces an exopolysaccharide, which acts as an elicitor of the SA pathway. In turn, the SA pathway antagonizes the JA signaling pathway, thereby allowing the fungus to develop its disease in tomato (Solanum lycopersicum). SA-promoted disease development occurs through Nonexpressed Pathogen Related1. We also show that the JA signaling pathway required for tomato resistance against B. cinerea is mediated by the systemin elicitor. These data highlight a new strategy used by B. cinerea to overcome the plant’s defense system and to spread within the host.

Reconsolidation in humans opens up declarative memory to the entrance of new information

A consolidated memory recalled by a reminder enters a vulnerability phase (labilization), followed by a process of stabilization (reconsolidation). Several authors have suggested that the labilization of the consolidated memory makes the incorporation of new information possible. Here, we demonstrate updating in the framework of memory declarative reconsolidation in humans by giving an opportune verbal instruction. Volunteers learn an association between five cue-syllables (L1) and their respective response-syllables. Twenty-four hours later, the paired-associate verbal memory is labilized by exposing the subjects to the reminder, and then they receive the verbal Instruction of adding three new cue-response syllables (INFO) with their respective responses to the former list of five. The new information is incorporated into the single former L1-memory and both INFO and L1 are successfully retrieved on the third day. However, when the Instruction is not preceded by a proper reminder, or when the instruction omits the order of adding the INFO into the former L1-memory, we observed interference in retrieval of both the original and the new information, suggesting that they are encoded independently and coexist as separate memories.

Repeated labilization-reconsolidation processes strengthen declarative memory in humans.

The idea that memories are immutable after consolidation has been challenged. Several reports have shown that after the presentation of a specific reminder, reactivated old memories become labile and again susceptible to amnesic agents. Such vulnerability diminishes with the progress of time and implies a re-stabilization phase, usually referred to as reconsolidation. To date, the main findings describe the mechanisms associated with the labilization-reconsolidation process, but little is known about its functionality from a biological standpoint. Indeed, two functions have been proposed. One suggests that destabilization of the original memory after the reminder allows the integration of new information into the background of the original memory (memory updating), and the other suggests that the labilization-reconsolidation process strengthens the original memory (memory strengthening). We have previously reported the reconsolidation of human declarative memories, demonstrating memory updating in the framework of reconsolidation. Here we deal with the strengthening function attributed to the reconsolidation process. We triggered labilization-reconsolidation processes successively by repeated presentations of the proper reminder. Participants learned an association between five cue-syllables and their respective response-syllables. Twenty-four hours later, the paired-associate verbal memory was labilized by exposing the subjects to one, two or four reminders. The List-memory was evaluated on Day 3 showing that the memory was improved when at least a second reminder was presented in the time window of the first labilization-reconsolidation process prompted by the earlier reminder. However, the improvement effect was revealed on Day 3, only when at least two reminders were presented on Day2 and not as a consequence of only retrieval. Therefore, we propose central concepts for the reconsolidation process, emphasizing its biological role and the parametrical constrains for this function to be operative.

Agar from Gracilaria gracilis (Gracilariales, Rhodophyta) of the Patagonic coast of Argentina--content, structure and physical properties.

Milled summer thalli of Gracilaria gracilis from Argentina were sequentially extracted with water at room temperature (RTW1-3), 70 degrees C (W701-3) and 90 degrees C (W901-2). Both W701 and W901 consisted of high molecular weight polysaccharides (ca. 540,000Da), but polydispersity was higher for the major product W701 (yield, 72% of the recovered). Structural analyzes by methylation and (13)C NMR spectroscopy revealed that W701 was mainly agarose. Alkaline treatment, together with structural analyzes, indicated a negligible proportion of precursor l-galactose 6-sulfate residues in this product, while they were clearly detected in the (13)C NMR spectra of RTW2-3. The presence of floridean starch in W901 had an antagonistic effect on its gel strength, which resulted nearly three times lower than that of fraction W701. Ultrastructural observation by transmission electron microscopy showed that, after extraction with hot water, a partial loss of cell wall stratification and disorganization of the cuticle had occurred. Final cellular debris exhibited swelling in the microfibrillar component. After this first thorough study of the chemical composition and physical properties of the products of G. gracilis from Bahía Bustamante we conclude that a good quality agarose is obtained in high yield after extraction with water at 70 degrees C without the requirement of alkaline pretreatment, which usually produces degradation of the polysaccharide.

Neutral polysaccharide from Cedrela tubiflora with anticomplementary activity

A water-soluble polysaccharide extracted from leaves of the Meliaceae Cedrela tubiflora was separated into neutral and acidic polysaccharide fractions. The best anticomplementary activity was exhibited by the neutral product which was further purified by means of gelpermeation chromatography. The composition and methylation analysis of the purified product were determined.

A Parachlorella kessleri (Trebouxiophyceae, Chlorophyta) strain from an extremely acidic geothermal pond in Argentina

Juárez Á.B., Vélez C.G., Iñiguez A.R., Martínez D.E., Rodríguez M.C., Vigna M.S. and Ríos de Molina M.C. 2011. A Parachlorella kessleri (Trebouxiophyceae, Chlorophyta) strain from an extremely acidic geothermal pond in Argentina. Phycologia 50: 413–421. DOI: 10.2216/10-79.1 Acidic geothermal environments are rare and very few organisms can thrive in them. Chlorella or Chlorella-like species have been described in a variety of environments but their presence in extremely acidic waters have rarely been reported. A Chlorella-like alga (strain BAFC CA10) was isolated from Laguna Verde, a volcanic mesothermal acidic pond from northern Patagonia (Argentina). Applying a multimethod approach including microscopic, ultrastructural, chemical and phylogenetic analyses, the acidotolerant strain BAFC CA10 was identified as Parachlorella kessleri ( =  Chlorella kessleri) on the basis of comparisons with the authentic strain SAG 211-11g. Our results confirmed the initial characterization of the Laguna Verde strain as C. kessleri, but since this species was transferred to the new genus Parachlorella, strain BAFC CA10 should be designated as P. kessleri. Histochemical staining and sugar composition of the cell-wall amorphous and fibrillar matrix, together with transmission electron microscopy of the cells, showed the distinctive characters of the species: (1) glucosamine was the exclusive component of the fibrillar cell wall; (2) rhamnose, galactose, glucose and xylose together with minor quantities of arabinose, mannose and fucose were present in the cell-wall hemicellulose matrix; (3) there was a single parietal chloroplast surrounding the entire cell with a small aperture (“mantel-shaped”); (4) there was one pyrenoid in the thickening of the chloroplast surrounded by two starch granules and bisected by two thylakoids; and (5) the cell wall was an electron-transparent homogeneous structure 60–80 nm thick. Phylogenetic analyses of concatenated small-subunit and internal transcribed sequence ribosomal DNA sequences of coccoid green algae within the Chlorella and the Parachlorella clades of the Chlorellaceae confirmed that strain BAFC CA10 is unequivocally related to the P. kessleri strains. Our results suggest that P. kessleri should be added to the list of species capable of adapting to live in extremely acidic environments.

THE FIBRILLAR POLYSACCHARIDES AND THEIR LINKAGE TO ALGAENAN IN THE TRILAMINAR LAYER OF THE CELL WALL OF COELASTRUM SPHAERICUM (CHLOROPHYCEAE)

Methylation and spectroscopical analyses of DMSO‐LiCl‐solubilized fractions of the fibrillar cell wall of Coelastrum sphaericum Näg. established the presence of cellulose and β‐1,4‐linked mannan. A small proportion (2–7%) of (1→2) linkages in β‐mannan that introduced an interruption in the regular ribbon chain conformation was interpreted as a component that modulated the mechanical strength of the cell wall. The trilaminar layer fractions consisted mostly of algaenan and cellulose. Evidence for ether linkages between glucose C‐6 in the β‐1,4‐glucan and algaenan was obtained.

The resistant 'biopolymer' in cell walls of Coelastrum sphaericum

The resistant ‘polymer’ in the trilaminar sheath of Coelastrum sphaericum is a complex formed by three different types of biopolymers, a hydrophobic aliphatic network mainly composed of polymethylenic chains with amido groups, together with smaller amounts of cellulose and proteins. The composition of the hydrophobic polymer is close to that of algaenan of other trilaminar layer-containing species.

Seasonal variation in agar composition and properties from Gracilaria gracilis (Gracilariales, Rhodophyta) of the Patagonian coast of Argentina

Seasonal variation of agar from specimens of a commercially exploited population of Gracilaria gracilis (Stackhouse) Steentoft, Irvine & Farnham in the Patagonian coast of Argentina was studied. For each seasonal harvest, random samples of plants were pooled for subsequent polysaccharide extraction at different water temperatures and agar physico‐chemical properties and composition were determined.

Photoprotecting Action and Phytochemical Analysis of a Multiple Radical Scavenger Lipophilic Fraction Obtained from the Leaf of the Seagrass Thalassia testudinum

The apolar fraction F1 of Thalassia testudinum was chemically characterized by gas chromatography‐mass spectrometry, which led to the identification of 43 metabolites, all of them reported for the first time in the genus Thalassia. More than 80% of the F1 composition was constituted by aromatic metabolites including the major components 1,1‐bis(p‐tolyl)ethane (6.0%), 4,4′‐diisopropylbiphenyl (4.8%) and a 1,1‐bis(p‐tolyl)ethane isomer (4.7%). This lipophilic fraction was assayed for its antioxidant effects and skin protective action. In vitro assays showed that F1 strongly scavenged DPPH* (IC50 312.0 ± 8.0 μg mL−1), hydroxyl (IC50 23.8 ± 0.5 μg mL−1) and peroxyl radical (IC50 6.6 ± 0.3 μg mL−1), as well as superoxide anion (IC50 50.0 ± 0.7 μg mL−1). Also, F1 markedly inhibited the spontaneous lipid peroxidation (LPO) in brain homogenates (IC50 93.0 ± 6.0 μg mL−1) and the LPS‐stimulated nitrite generation on RAW624.7 macrophages (58.6 ± 3.2%, 400 μg mL−1). In agreement with these findings, its topical application at 250 and 500 μg cm−2 strikingly reduced skin damage on mice exposed to acute UVB radiation by 45% and 70%, respectively and significantly attenuated the LPO developed following the first 48 h after acute exposure to UVB irradiation, as manifested by the decreased malondialdehide level and by the increased of reduced gluthatione content. Our results suggest that F1 may contribute to skin repair by attenuating oxidative stress due to its antioxidant activity.