Katia C. Scortecci

Profiling the resting venom gland of the scorpion Tityus stigmurus through a transcriptomic survey

BackgroundThe scorpion Tityus stigmurus is widely distributed in Northeastern Brazil and known to cause severe human envenoming, inducing pain, hyposthesia, edema, erythema, paresthesia, headaches and vomiting. The present study uses a transcriptomic approach to characterize the gene expression profile from the non-stimulated venom gland of Tityus stigmurus scorpion.ResultsA cDNA library was constructed and 540 clones were sequenced and grouped into 153 clusters, with one or more ESTs (expressed sequence tags). Forty-one percent of ESTs belong to recognized toxin-coding sequences, with transcripts encoding antimicrobial toxins (AMP-like) being the most abundant, followed by alfa KTx- like, beta KTx-like, beta NaTx-like and alfa NaTx-like. Our analysis indicated that 34% of the transcripts encode “other possible venom molecules”, which correspond to anionic peptides, hypothetical secreted peptides, metalloproteinases, cystein-rich peptides and lectins. Fifteen percent of ESTs are similar to cellular transcripts. Sequences without good matches corresponded to 11%.ConclusionsThis investigation provides the first global view of gene expression of the venom gland from Tityus stigmurus under resting conditions. This approach enables characterization of a large number of venom gland component molecules, which belong either to known or non yet described types of venom peptides and proteins from the Buthidae family.

Challenges, Opportunities and Recent Advances in Sugarcane Breeding

Katia C. Scortecci1, Silvana Creste2, Tercilio Calsa Jr.3, Mauro A. Xavier2, Marcos G. A. Landell2, Antonio Figueira4 and Vagner A. Benedito5* 1Department of Cell Biology and Genetics, Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN 2Centro de Cana – Instituto Agronomico de Campinas (IAC), Ribeirao Preto, SP 3Department of Genetics, Universidade Federal de Pernambuco (UFPE), Recife, PE 4Plant Breeding Laboratory , Centro de Energia Nuclear na Agricultura (CENA), Universidade de Sao Paulo (USP), Piracicaba, SP 5Laboratory of Plant Functional Genetics, Genetics and Developmental Biology Program, Plant & Soil Sciences Division, West Virginia University (WVU), Morgantown, WV 1,2,3,4Brazil 5USA

Antioxidant and Antiproliferative Activities of Leaf Extracts from Plukenetia volubilis Linneo (Euphorbiaceae)

Plukenetia volubilis Linneo, or Sacha inca, is an oleaginous plant from the Euphorbiaceae family. The aim of this work was to perform a chemical and biological analysis of different leaf extracts from P. volubilis such as aqueous extract (AEL), methanol (MEL), ethanol (EEL), chloroform (CEL), and hexane (HEL). Thin layer chromatography analysis revealed the presence of phenolic compounds, steroids, and/or terpenoídes. Furthermore, the antioxidant activities were analyzed by in vitro assays and their effects on cell lineages by in vivo assays. The Total Antioxidant Capacity (TCA) was expressed as equivalent ascorbic acid (EEA/g) and it was observed that the extracts showed values ranging from 59.31 to 97.76 EAA/g. Furthermore, the DPPH assay values ranged from 62.8% to 88.3%. The cell viability assay showed that the extracts were able to reduce viability from cancer cells such as HeLa and A549 cells. The extracts MEL and HEL (250 µg/mL) were able to reduce the proliferation of HeLa cells up to 54.3% and 48.5%, respectively. The flow cytometer results showed that these extracts induce cell death via the apoptosis pathway. On the other hand, the extracts HEL and AEL were able to induce cell proliferation of normal fibroblast 3T3 cells.

Freshwater Plants Synthesize Sulfated Polysaccharides: Heterogalactans from Water Hyacinth (Eicchornia crassipes)

Sulfated polysaccharides (SP) are found mainly in seaweeds and animals. To date, they have only been found in six plants and all inhabit saline environments. Furthermore, there are no reports of SP in freshwater or terrestrial plants. As such, this study investigated the presence of SP in freshwaters Eichhornia crassipes, Egeria densa, Egeria naja, Cabomba caroliniana, Hydrocotyle bonariensis and Nymphaea ampla. Chemical analysis identified sulfate in N. ampla, H. bonariensis and, more specifically, E. crassipes. In addition, chemical analysis, FT-IR spectroscopy, histological analysis, scanning electron microscopy (SEM) and energy-dispersive X-ray analysis (EDXA), as well as agarose gel electrophoresis detected SP in all parts of E. crassipes, primarily in the root (epidermis and vascular bundle). Galactose, glucose and arabinose are the main monosaccharides found in the sulfated polysaccharides from E. crassipes. In activated partial thromboplastin time (APTT) test, to evaluate the intrinsic coagulation pathway, SP from the root and rhizome prolonged the coagulation time to double the baseline value, with 0.1 mg/mL and 0.15 mg/mL, respectively. However, SP from the leaf and petiole showed no anticoagulant activity. Eichornia SP demonstrated promising anticoagulant potential and have been selected for further studies on bioguided fractionation; isolation and characterization of pure polysaccharides from this species. Additionally in vivo experiments are needed and are already underway.

Phosphorus-deficiency reduces aluminium toxicity by altering uptake and metabolism of root zone carbon dioxide.

The role of phosphorus (P) status in root-zone CO(2) utilisation for organic acid synthesis during Al(3+) toxicity was assessed. Root-zone CO(2) can be incorporated into organic acids via Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31). P-deficiency and Al(3+) toxicity can induce organic acid synthesis, but it is unknown how P status affects the utilisation of PEPC-derived organic acids during Al(3+) toxicity. Two-week-old Solanum lycopersicum seedlings were transferred to hydroponic culture for 3 weeks. The hydroponic culture consisted of a standard Long Ashton nutrient solution containing either 0.1μM or 1mM P. Short-term Al(3+) toxicity was induced by a 60-min exposure to a pH-buffered solution (pH 4.5) containing 2mM CaSO(4) and 50μM AlCl(3). Al(3+) toxicity induced a decline in root respiration, adenylate concentrations and an increase in root-zone CO(2) utilisation for both P sufficient and P-deficient plants. However during Al(3+) toxicity, P deficiency enhanced the incorporation and metabolism of root-zone CO(2) via PEPC. Moreover, P deficiency led to a greater proportion of the PEPC-derived organic acids to be exuded during Al(3+) toxicity. These results indicate that P-status can influence the response to Al(3+) by inducing a greater utilisation of PEPC-derived organic acids for Al(3+) detoxification.

Biological activities of the sulfated polysaccharide from the vascular plant Halodule wrightii

A sulfated polysaccharide (SPSG) was successfully isolated from seagrass Halodule wrightii Asch., Cymodoceaceae, and its antioxidant and anticoagulant activities were investigated. The data presented here showed that the SPSG is a 11 kDa sulfated heterogalactan with a sulfatation degree of 20.63% and it also contains glucose and xylose. SPSG antioxidant activities were evaluated using several in vitro assays and the anticoagulant activity was evaluated by aPTT and PT tests. These assays suggested that the SPSG possessed remarkable antioxidant properties in different in vitro assays and an outstanding anticoagulant activity 2.5-fold higher than that of heparin Clexane® in the aPTT test. This data represents the first reported on the sulfated polysaccharide biological activities from seagrass. These results indicate that SPSG can be considered in the future as a drug utilized in treating diseases from these systems.

Identification, characterisation and molecular modelling of two AP endonucleases from base excision repair pathway in sugarcane provide insights on the early evolution of green plants

Unlike bacteria and mammals, plant DNA repair pathways are not well characterised, especially in monocots. The understanding of these processes in the plant cell is of major importance, since they may be directly involved in plant acclimation and adaptation to stressful environments. Hence, two sugarcane ESTs were identified as homologues of AP endonuclease from the base-excision repair pathway: ScARP1 and ScARP3. In order to understand their probable function and evolutionary origin, structural and phylogenetic studies were performed using bioinformatics approaches. The two predicted proteins present a considerable amino acid sequence similarity, and molecular modelling procedures indicate that both are functional, since the main structural motifs remain conserved. However, inspection of the sort signal regions on the full-length cDNAs indicated that these proteins have a distinct organelle target. Furthermore, variances in their promoter cis-element motifs were also found. Although the mRNA expression pattern was similar, there were significant differences in their expression levels. Taken together, these data raise the hypothesis that the ScARP is an example of a probable gene duplication event that occurred before monocotyledon/dicotyledon segregation, followed by a sub-functionalisation event in the Poaceae, leading to new intracellular targeting and different expression levels.

Molecular Genetic Dissection of Sugarcane Flowering under Equatorial Field Conditions

Sugarcane is a tropical crop used for sugar and biofuel production in tropical and equatorial regions of the globe. Sugarcane flowering is intrinsically induced in equatorial regions due to long-day conditions. Flower development is problematic for this crop because it halts vegetative growth, leading to a reduction of the sugar accumulated in the stalks, and significant yield loss. Notwithstanding, the identification of genes differentially expressed in contrasting cultivars can potentially reveal markers and tools to generate genotypes more suitable for expanding the geographical limits of this crop. Thus, dissecting the flowering gene expression network under field conditions is highly relevant for breeding. We report the analysis of subtractive cDNA libraries produced from shoot apical meristem of cultivars contrasting for flowering time growing in production fields under equatorial conditions. Transcripts with homology to POLYPHENOL OXIDASE (PPO), CALMODULIN (CAM), PHOSPHATIDYLCHOLINE/PHOSPHATIDYLINOSITOL-TRANSFER PROTEIN (SEC14), OBTUSIFOLIOL-14-Α-DEMETHYLASE (CYP51), 14–3-3, and the phosphotransferases SHAGGY KINASE (GSK), PROTEIN KINASE C INHIBITOR (PKCI), and SERINE/THREONINE PHOSPHATASE (PP1) were identified as differentially expressed in the subtractive libraries and further chosen for RT-qPCR validation and in silico interactome analyses. Our results suggest that ScPPO, ScSEC14 and Sc14–3-3 may act as flowering inhibitors. RT-qPCR data also revealed two 14–3-3 isoforms as potential flowering markers. Sc14–3-3 was structurally and phylogenetically characterized and its genomic architecture was analysed in two BAC clones, showing that they probably correspond to two different loci with confirmed synteny to other grass genomes. This work reveals potential novel mechanisms of flowering in grasses with implications to crop breeding.