Topological data analysis reveals core heteroblastic and ontogenetic programs embedded in leaves of grapevine (Vitaceae) and maracuyá (Passifloraceae).

Leaves are often described in language that evokes a single shape. However, embedded in that descriptor is a multitude of latent shapes arising from evolutionary, developmental, environmental, and other effects. These confounded effects manifest at distinct developmental time points and evolve at different tempos. Here, revisiting datasets comprised of thousands of leaves of vining grapevine (Vitaceae) and maracuyá (Passifloraceae) species, we apply a technique from the mathematical field of topological data analysis to comparatively visualize the structure of heteroblastic and ontogenetic effects on leaf shape in each group. Consistent with a morphologically closer relationship, members of the grapevine dataset possess strong core heteroblasty and ontogenetic programs with little deviation between species. Remarkably, we found that most members of the maracuyá family also share core heteroblasty and ontogenetic programs despite dramatic species-to-species leaf shape differences. This conservation was not initially detected using traditional analyses such as principal component analysis or linear discriminant analysis. We also identify two morphotypes of maracuyá that deviate from the core structure, suggesting the evolution of new developmental properties in this phylogenetically distinct sub-group. Our findings illustrate how topological data analysis can be used to disentangle previously confounded developmental and evolutionary effects to visualize latent shapes and hidden relationships, even ones embedded in complex, high-dimensional datasets.

Breast cancer cell growth arrest and chemopreventive effects of Passiflora edulis Sims (Passifloraceae) ethanolic leaves extract on a rat model of mammary carcinoma.

ETHNOPHARMACOLOGICAL RELEVANCE: Despite various prevention and treatment measures, the incidence and mortality due to breast cancer has been increasing globally. Passiflora edulis Sims is a plant used for the treatment of various diseases in traditional medicine, including cancers. AIM OF THE STUDY: To assess the anti-breast cancer activity of the ethanolic extract of P. edulis leaves in vitro and in vivo. MATERIAL AND METHODS: In vitro, the cell growth and proliferation were determined based on the MTT and BrdU assays. The flow cytometry was used to analyze the cell death mechanism while, cell migration, cell adhesion and chemotaxis were assayed for anti-metastatic potential. In vivo, 56 female Wistar rats aged 45-50 days (∼75 g) were exposed to 7,12-dimethylbenz(a)anthracene-DMBA except the normal group. Negative control group (DMBA) received solvent dilution throughout the study; standards groups (tamoxifen - 3.3 mg/kg BW and letrozole - 1 mg/kg BW) as well as P. edulis leaves ethanolic extract groups (50, 100 and 200 mg/kg) treated for 20 weeks. Tumor incidence, tumor burden and volume, CA 15-3 serum' level, antioxidant, inflammatory status and histopathology were assessed. RESULTS: P. edulis extract showed a significant and concentration-dependent inhibition of MCF-7 and MDA-MB 231 cells growth at 100 µg/mL. It inhibited cell proliferation and clones' formation and induced apoptosis in MDA-MB 231 cells. The migration of cell into the zone freed of cells and the number of invading cells after the 48 and 72 h were significantly diminished while, it increased their adherence to collagen and fibronectin extracellular matrix as does Doxorubicin. In vivo, all rats in the DMBA group exhibited a significant (p < 0.001) increase in tumor volume, tumor burden and grade (adenocarcinoma of SBR III) and pro-inflammatory cytokine levels (TNF-α, INF-γ, IL-6 and IL-12). P. edulis extract at all tested doses significantly inhibited the DMBA-induced increase in tumor incidence, tumor burden and grade (SBR I) as well as pro-inflammatory cytokines. Moreover, it increased enzymatic and non-enzymatic antioxidants (SOD, catalase, and GSH) and decreased MDA levels although a greater effect was observed with Tamoxifen and Letrozole. P. edulis has medium content on polyphenols, flavonoids and tannins. CONCLUSION: P. edulis has chemo-preventive effects against DMBA-induced breast cancer in rats probably through its antioxidative, anti-inflammatory and apoptosis-inducing potentials.

Hypolipidemic potential and safety profile of the ethanolic extract and flour of the peel of Passiflora Cincinnata Mast. (Passifloraceae) in mice.

Passiflora cincinnata Mast. is described as a native Caatinga species, used by nutritional and medicinal purposes, although there are still few studies and pharmacological data related to this species. This paper aims to evaluate the safety profile and hypolipidemic potential of the fruit peel of this species in mice. It was analyzed the chemical composition of ethanolic extract (EtOH-Pc) by HPLC-DAD-MS/MS, and the organic and inorganic composition of flour (MF-Pc). Also were evaluated the acute toxicity, the lipid-lowering potential of these samples, through of a pretreatment (oral: 100 and 200 mg/kg), and a single treatment with the same doses, after hyperlipidemic induction with triton WR-1339, using as animal model Swiss Mus musculus mice, besides histopathological analysis. The presence of flavonoids in the extract was confirmed, mainly C-glycosides, and antioxidant minerals and pectin, in flour. No clinical signs of toxicity or death were reported in the study. In the hyperlipidemia study model used, the analyzed substances, at all doses, notably decreased the lipid levels of TC, TG, LDL-c and VLDL-c and increase the HDL-c levels in the induced hyperlipidemic mice (p < 0.05). The results of the histopathological analysis showed that in the group only induced was identified the discrete presence of hepatic steatosis, in 2 animals at the analysis of 24 h, not being visualized in the groups treated with the substances evaluated. The results obtained in the present study suggest a hypolipidemic potential of the extract and flour, obtained from the fruit peel of Passiflora cincinnata Mast.

Seed morphoanatomy of the genus Passiflora L. (Passifloraceae) by scanning electron microscopy and light microscopy.

Morphoanatomical analysis of seeds contributes to knowledge of the development of seedlings and identification of species, as well as supporting conservation studies. The conservation of the species belonging to the Passiflora genus is crucial due to of the threats to the genetic resources of these species. Thus, the objective of this study was to morphoanatomically characterize Passiflora seeds, verify possible injuries to the tissues after cryopreservation and thus contribute to the conservation strategies of the species of this genus. Initially, seeds of Passiflora coccinea, P. edulis, P. gibertii, P. maliformis, P. morifolia, P. setacea, P. suberosa, and P. tenuifila collected from the Passion Fruit Active Germplasm Bank of the Embrapa Cassava and Fruits research unit (Embrapa Mandioca e Fruticultura) were analyzed. Then, their length, width and thickness, shape of the base and tip, and ornamentations present on the body and edge of the seeds were evaluated. The seeds of the species were placed in cryotubes and immersed in liquid nitrogen to assess possible cryoinjuries. The tegument and tissues of the seeds were examined by scanning electron microscopy. The seeds had varied biometric data, with average values of 4.63 mm for length, 3.28 mm for width, and 1.51 mm for thickness. Six ornamentation types were observed: reticulate for the species P. coccinea; finely reticulate for P. edulis; foveolate reticulate for P. gibertii and P. setacea; alveolate reticulate for P. maliformis and P. tenuifila; coarsely reticulate for P. morifolia; and falsifoveolate reticulate for P. suberosa. Some seeds suffered tegument cracks due to the freezing in liquid nitrogen, but without physiological damages to the embryo and endosperm. The cryopreservation of the seeds in the presence of the tegument significantly reduced the cryoinjuries caused to the embryo. Cryopreservation can be promising for long-term conservation of passion fruit seeds.

Nuclear variations and tapetum polyploidy related to pollen grain development in Passiflora L. (Passifloraceae).

Tapetal cells comprise an anther tissue fundamental to pollen grain development. They are associated with endoreduplication events, which culminate in polyploid and multinucleated cells, high metabolic activity, and different organelle arrangements to support all the development of the pollen grains. Passiflora species present a secretory tapetum, with diversity in the number and size of nuclei. Tapetal cells undergo numerous changes in a short period of development when compared to the plant's life span. To improve our knowledge of tapetum development, tests assessing ploidy levels, anatomy, cytochemistry, transmission electron microscopy, flow cytometry, as well as conventional and molecular cytogenetics were used in Passiflora actinia and P. elegans. The current data show striking differences in nuclear organisation during tapetal cell development, including mono to quadrinucleate cells, and ploidy levels from 2n to 32n. One of the most peculiar features was the atypical behaviour of the endoplasmic reticulum (ER), which accumulated in the cell border, similar to a 'cER', as well as large dictyosomes. This endomembrane configuration may be related to the tapetum nutritional network and secretion of compounds at the end of meiosis. Another atypical feature of the ER was the formation of an invagination to establish 'binucleated' polyploid cells. This membrane projection appears when the nuclei form two lobes, as well as when it organises a nucleoplasmic reticulum. These data demonstrate that there are important ultrastructural changes in tapetal cells, including organelle arrangements, ploidy levels, and nuclear activity, common to P. actinia and P. elegans, but different from the plant model A. thaliana.

Sequence, Structure, and Binding Site Analysis of Kirkiin in Comparison with Ricin and Other Type 2 RIPs.

Kirkiin is a new type 2 ribosome-inactivating protein (RIP) purified from the caudex of Adenia kirkii with a cytotoxicity compared to that of stenodactylin. The high toxicity of RIPs from Adenia genus plants makes them interesting tools for biotechnology and therapeutic applications, particularly in cancer therapy. The complete amino acid sequence and 3D structure prediction of kirkiin are here reported. Gene sequence analysis revealed that kirkiin is encoded by a 1572 bp open reading frame, corresponding to 524 amino acid residues, without introns. The amino acid sequence analysis showed a high degree of identity with other Adenia RIPs. The 3D structure of kirkiin preserves the overall folding of type 2 RIPs. The key amino acids of the active site, described for ricin and other RIPs, are also conserved in the kirkiin A chain. Sugar affinity studies and docking experiments revealed that both the 1α and 2γ sites of the kirkiin B chain exhibit binding activity toward lactose and D-galactose, being lower than ricin. The replacement of His246 in the kirkiin 2γ site instead of Tyr248 in ricin causes a different structure arrangement that could explain the lower sugar affinity of kirkiin with respect to ricin.

The karyotype of Adenia and the origin of the base number x = 12 in Passifloroideae (Passifloraceae).

Adenia is an Old World genus of Passifloroideae closely related to Passiflora. The two genera comprise the large majority of Passifloroideae species, although most studies are concentrated on Passiflora. Cytological analyses reveal that changes in chromosome numbers played an important role in the evolution of Passiflora, whereas in the remaining genera little is known, hindering the identification of the base number of the family. Here we analyzed the chromosome number and the 35S rDNA sites of three species of Adenia and reevaluated the base number (x) of the subfamily Passifloroideae and the family Passifloraceae, including chromosome data for Turneroideae and Malesherbioideae. The chromosome number of Adenia species seemed to be stable with 2n = 24 or 48 and one or two pairs of rDNA sites, very similar to Passiflora subgenus Astrophea, suggesting a common ancestral karyotype with x = 12. Differently, Turneroideae and Malesherbioideae present x = 7. A whole genomic duplication detected after the separation of Passifloroideae and Malesherbioideae suggests that the base number of Passifloraceae most probably was x = 7, which by dysploidy and polyploidy generated x = 12 for the subfamily Passifloroideae.

A Polylactosamine Specific Lectin from Adenia hondala Induces Apoptosis and Necrosis in Human Epithelial Colon Cancer HT-29 Cells.

BACKGROUND: Altered expression of N-glycans such as polylactosamine is observed in colon cancer. AHL, a polylactosamine specific lectin from Adenia hondala from a medicinal plant from the Passifloraceae family has been reported earlier. OBJECTIVE: The aim of the present study is to study the interaction of AHL with human colon cancer epithelial HT-29 cells and colon cancer tissues. METHODS: Cell viability was determined by MTT [3-[4, 5- dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide] assay, while cell surface binding, apoptosis by Annexin-V-PI assay and ROS production using DCFDA [2',7' - dichlorofluorescindiacetate] kit method were analysed by flowcytometry, immunohistochemistry was performed using biotinylated AHL, protein purification by affinity chromatography using asialofetuin-coupled Sepharose -4B column. RESULTS: AHL strongly binds to HT-29 cells with a Mean Fluorescence Intensity of 12.4, which could be blocked by competing glycoprotein asialofetuin. AHL inhibits HT-29 cell growth in a dose and time-dependent manner with IC50 of 2.5 µg/mL and differentially binds to human normal and cancerous tissues. AHL induces apoptosis and slight necrosis in HT-29 cells with an increase in the early apoptotic population of 25.1 and 36% for 24 h and 48 h respectively and necrotic population of 1.5 and 4.6% at 24 h and 48 h respectively as revealed by Annexin-V-PI assay. AHL induces the release of Reactive Oxygen Species in HT-29 cells in a dose-dependent manner. CONCLUSION: To the best of knowledge, this is the first report on lectin from Adenia hondala which is not a RIP with apoptotic and necrotic effects. These findings support the promising potential of AHL in cancer research.

In Vitro Phytotherapeutic Properties of Aqueous Extracted Adenia viridiflora Craib. towards Civilization Diseases.

Adenia viridiflora Craib. is an indigenous edible plant that became an endangered species due to limited consumption of the local population with unknown reproduction and growth conditions. The plant is used as a traditional herb; however, its health applications lack scientific-based evidence. A. viridiflora Craib. plant parts (old leaves and young shoots) from four areas as Kamphaeng Phet (KP), Muang Nakhon Ratchasima (MN), Pakchong Nakhon Ratchasima (PN), and Uthai Thani (UT) origins were investigated for phenolic compositions and in vitro health properties through the inhibition of key enzymes relevant to obesity (lipase), diabetes (α-glucosidase and dipeptidyl peptidase-IV), Alzheimer's disease (cholinesterases and ß-secretase), and hypertension (angiotensin-converting enzyme). Phenolics including p-coumaric acid, sinapic acid, naringenin, and apigenin were detected in old leaves and young shoots in all plant origins. Old leaves exhibited higher total phenolic contents (TPCs) and total flavonoid contents (TFCs), leading to higher enzyme inhibitory activities than young shoots. Besides, PN and MN with higher TPCs and TFCs tended to exhibit greater enzyme inhibitory activities than others. These results will be useful to promote this plant as a healthy food with valuable medicinal capacities to support its consumption and agricultural stimulation, leading to sustainable conservation of this endangered species.

Kirkiin: A New Toxic Type 2 Ribosome-Inactivating Protein from the Caudex of Adenia kirkii.

Ribosome-inactivating proteins (RIPs) are plant toxins that irreversibly damage ribosomes and other substrates, thus causing cell death. RIPs are classified in type 1 RIPs, single-chain enzymatic proteins, and type 2 RIPs, consisting of active A chains, similar to type 1 RIPs, linked to lectin B chains, which enable the rapid internalization of the toxin into the cell. For this reason, many type 2 RIPs are very cytotoxic, ricin, volkensin and stenodactylin being the most toxic ones. From the caudex of Adenia kirkii (Mast.) Engl., a new type 2 RIP, named kirkiin, was purified by affinity chromatography on acid-treated Sepharose CL-6B and gel filtration. The lectin, with molecular weight of about 58 kDa, agglutinated erythrocytes and inhibited protein synthesis in a cell-free system at very low concentrations. Moreover, kirkiin was able to depurinate mammalian and yeast ribosomes, but it showed little or no activity on other nucleotide substrates. In neuroblastoma cells, kirkiin inhibited protein synthesis and induced apoptosis at doses in the pM range. The biological characteristics of kirkiin make this protein a potential candidate for several experimental pharmacological applications both alone for local treatments and as component of immunoconjugates for systemic targeting in neurodegenerative studies and cancer therapy.

Evaluation of cytotoxic, immunomodulatory effects, antimicrobial activities and phytochemical constituents from various extracts of Passiflora edulis F. flavicarpa (Passifloraceae).

The leaves of P. edulis were subjected to physicochemical analysis, such as ion content, extractives, and structural molecules. The hexanic, ethanolic and ethyl acetate extracts were submitted to phytochemical analyzes by GC-MS, HPLC-MS, and spectrophotometry. In addition, antioxidant (DPPH, ABTS and TAA methods) potential, antimicrobial (MIC method) action, cytotoxicity and immunostimulant activity (flow cytometry analysis) were performed. The extracts showed a moderate antioxidant capacity and revealed the presence of several metabolites, mainly phenols, such as caffeic acid, p-coumaric acid and luteolin. The ethyl acetate and ethanolic extracts showed antifungal activity. In addition, the extracts did not affect splenocytes viability at 12.5 µg/mL and promoted the production of IL-6, IL-10, IL-17 and TNF-α cytokines. P. edulis extracts showed antifungal and antioxidant activity and were able to induce immunostimulatory action in splenocyte cultures in vitro.

Phenotypic plasticity in size of ant-domatia.

Ant-plants produce hollow structures called domatia to host protecting ants. Although size variation in domatia is well documented between related species, intraspecific variation is little explored. The central African ant-plant Barteria dewevrei exibits strong variation in domatium size, giving the opportunity to explore the mechanism underlying variation in a mutualistic trait. We showed that domatium size in Barteria dewevrei varies between sites. We transplanted individual plants between two sites in Gabon where plants have different domatium sizes. Domatium size of transplanted plants changed, revealing that variation in this mutualistic trait is driven by phenotypic plasticity. The two sites differed in their environmental conditions: highland open savanna on sandy soil vs lowland closed tropical rain forest on sandy-loam soil. However, as stomatal density and δ13C of leaves did not differ between sites or between branches produced before and after transplantation, we have no cue on the role of abiotic stress (such as light intensity and water availability) in domatium size variation. As the obligate Tetraponera ant symbionts are too large to fit in the small domatia, variation of the mutualistic trait in response to environmental change through phenotypic plasticity may impact this specialized mutualism.

Natural selection acting on integrated phenotypes: covariance among functional leaf traits increases plant fitness.

Plant functional strategies are usually accomplished through the simultaneous expression of different traits, and hence their correlations should be promoted by natural selection. The adaptive value of correlations among leaf functional traits, however, has not been assessed in natural populations. We estimated intraspecific variation in leaf functional traits related to the primary metabolism and anti-herbivore defence in a population of Turnera velutina. We analysed whether natural selection favoured the expression of individual traits, particular combinations of traits or leaf phenotypic integration. Patterns of covariation among traits were related to water and nitrogen economy, and were similar among genotypes, but the magnitude of their phenotypic integration differed by 10-fold. Although families did not differ in the mean values of leaf functional traits, directional selection favoured low nitrogen content and low chemical defence, high content of chlorophyll, sugar in extrafloral nectar and trichome density. Families with higher phenotypic integration among leaf traits grew faster and produced more flowers. We suggest that the coordinated expression of leaf traits has an adaptive value, probably related to optimisation in the expression of traits related to water conservation and nitrogen acquisition.

Economic Valuation of Bee Pollination Services for Passion Fruit (Malpighiales: Passifloraceae) Cultivation on Smallholding Farms in São Paulo, Brazil, Using the Avoided Cost Method.

This paper estimates the economic value of ecosystem services provided by Brazilian native bee, Xylocopa spp. Latreille (Hymenoptera: Apidae), pollination on a scale relevant to individual smallholder farmers that produce yellow passion fruit (Passiflora edulis Sims). The study areas are located in the vicinity of Pedro de Toledo and Itariri (Sao Paulo State-Brazil), in the Atlantic Forest region. The local economy is based on family farms, small stores, and ecotourism. The value was obtained using the ecological economics Avoided Cost Method, also known as replacement cost. Farms from this region informally hire temporary day laborers to supplement natural pollination with manual pollination of passion fruit flowers, so the cost of contracting temporary laborers was used to estimate the economic value of bee pollination. The value of pollination services was estimated at US$ 2,583.00 per hectare over 2 yr of P. edulis farming. Our estimates based on passion fruit farmer surveys and ecological valuation over 2 yr suggest that manual pollination accounts for 44-48% of production costs and results in a loss of ~58% of profits when wild bee pollination services are not available and manual pollination is required. We suggest that smallholder farmers follow the suggestions of previous studies and conserve adequate forest habitat for bee nesting and foraging, plan pesticide use around flowering and pollination, and supplement bee populations to maximize the benefit of the pollination ecosystem service and profits.

Etnobotánica de las flores de la pasión (Passiflora) en la provincia andina de Angaraes (Huancavelica, Perú) / Ethnobotany of passion flowers (Passiflora) in the Andean province of Angaraes (Huancavelica, Perú)

A study about the useful species of the genus Passiflora (Passifloraceae) found in a mountains province of Angaraes (Huancavelica, Perú) is presented. The methodology used was based on guided visits and semi-structured interviews to 50 informants who lived in the province of Angaraes. Seven species grouped in five categories of use were reported. Categories with a higher quantity of reported usage were food for humans (95 reports) and medicine (65 reports). The category with greater diversity of species was fodder (6 species). The part of the plant used most often was the fruit (80%). Two species were described as cultivated and five have been recorded as in wildlife state. The species with greater cultural importance were P. tripartita var. mollissima and P. pinnatistipula, since their fruits are much appreciated by the locals and commercialized in province fairs, demonstrating that the cultivation of these plant resources is the main livelihood in the region. Finally, a key is included in order to allow the identification of each of the species of Passiflora which inhabit in the study area.

Se presenta un estudio sobre las especies útiles del género Passiflora (Passifloraceae) de una provincia andina de Angaraes (Huancavelica, Perú). La metodología utilizada se basó en visitas guiadas y entrevistas semiestructuradas a 50 informantes de la provincia de Angaraes. Se reportaron siete especies agrupadas en cinco categorías de uso. Las categorías con mayores reportes de uso fueron alimento para humanos (95 reportes) y medicinal (65 reportes). La categoría con mayor riqueza de especies es alimento para animales (seis especies). La parte utilizada con mayor frecuencia es el fruto (80%). Dos especies fueron descritas como cultivadas y cinco han sido registradas en estado silvestre. Las especies con mayor importancia cultural fueron P. tripartita var. mollissima y P. pinnatistipula, siendo sus frutos muy apreciados y comercializados en las ferias de la provincia, lo cual demuestra que los pobladores dependen de sus recursos vegetales para su subsistencia. Finalmente, se incluye una clave que permite diferenciar las especies de Passiflora que habitan en la zona de estudio.

Identification of Soluble Epoxide Hydrolase Inhibitors from the Seeds of Passiflora edulis Cultivated in Vietnam

Soluble epoxide hydrolases (sEH) are enzymes present in all living organisms, metabolize epoxy fatty acids to 1,2-diols. sEH in the metabolism of polyunsaturated fatty acids plays a key role in inflammation. In addition, the endogenous lipid mediators in cardiovascular disease are also broken down to diols by the action of sEH that enhanced cardiovascular protection. In this study, sEH inhibitory guided fractionation led to the isolation of five phenolic compounds trans-resveratrol (1), trans-piceatannol (2), sulfuretin (3), (+)-balanophonin (4), and cassigarol E (5) from the ethanol extract of the seeds of Passiflora edulis Sims cultivated in Vietnam. The chemical structures of isolated compounds were determined by the interpretation of NMR spectral data, mass spectra, and comparison with data from the literature. The soluble epoxide hydrolase (sEH) inhibitory activity of isolated compounds was evaluated. Among them, trans-piceatannol (2) showed the most potent inhibitory activity on sEH with an IC₅₀ value of 3.4 µM. This study marks the first time that sulfuretin (3) was isolated from Passiflora edulis as well as (+)-balanophonin (4), and cassigarol E (5) were isolated from Passiflora genus.

Culture media to detect and criteria to evaluate and report the activity of extracellular enzymes produced by phytopathogenic fungi / Meios de cultura para detectar e critérios para avaliar e divulgar atividade de enzimas extracelulares produzidas por fungos fitopatogênicos

Extracellular enzymes are involved in the fungal pathogenesis in plants. Currently, culture media, data analyses, and data report related to extracellular enzymes produced in vitro conditions are different and therefore, lack standardization. This work aimed to compare the culture media cited on the literature (normal) with the potato-dextrose-agar (PDA) medium combined with a specific compound to produce extracellular enzymes through three soilborne phytopathogenic fungi (F. solani f. sp. passiflorae, S. rolfsii, and R. solani AG-4 HGI), as well as to analyze and report enzyme data based on five different criteria. The assay was randomized, with three factors (culture media, isolates, and enzymes) and six repetitions. The studied enzymes were amylase (AM), carboxymethylcellulase (CMCase), lipase (LP), laccase (LC), catalase (CT), and gelatinase (GT). The normal media detected more enzymes and was more precise compared to the PDA medium plus specific compound. The criteria that calculated the area of the circular crown of AM, CMCase, LP, and LC and measured the intensity (0 = absence, up to 4 = intense) of CT and GT adopting note scale were the best to evaluate and report the results of the enzymes. We suggest the normal media culture to study enzyme production, as well as the criteria mentioned to assess and report the data related to enzyme activities.(AU)

As enzimas extracelulares estão envolvidas na patogênese de fungos em plantas. Atualmente, não há uma padronização de meios de cultura, formas de analisar e divulgar os dados de enzimas extracelulares produzidas em condições in vitro. Assim, o presente trabalho objetivou comparar os meios de cultura específicos relatados na literatura (normal) com o meio de batata-dextrose-ágar mais adição do substrato específico para produção de enzimas extracelulares por três diferentes fungos fitopatogênicos habitantes de solo (Fusarium solani f. sp. passiflorae, Sclerotium rolfsii e Rhizoctonia solani AG-4 HGI), bem como avaliar os dados das enzimas por cinco critérios diferentes. O delineamento experimental adotado foi o inteiramente ao acaso, em esquema de três fatores (meios, isolados e enzimas), com seis repetições. As enzimas investigadas foram amilase, carboximetilcelulase, lipase, lacase, catalase e gelatinase. Os meios normais detectaram mais enzimas, e essa detecção foi mais precisa em comparação com os meios de batata-dextrose-ágar mais o substrato específico. Os critérios que calcularam a área da coroa circular para as enzimas amilase, carboximetilcelulase, lipase e lacase e adotaram a escala de notas para medir a intensidade (0=ausência até 4=intensa) de catalase e gelatinase foram os melhores para avaliar e divulgar os resultados das enzimas. Assim, sugere-se padronizar os meios normais para estudos de produção de enzimas, bem como os critérios citados para avaliar e divulgar os dados das atividades das referidas enzimas.(AU)

Purification, characterization and fine sugar specificity of a N-Acetylgalactosamine specific lectin from Adenia hondala.

Plant lectins are gaining interest because of their interesting biological properties. Several Adenia species, that are being used in traditional medicine to treat many health ailments have shown presence of lectins or carbohydrate binding proteins. Here, we report the purification, characterization and biological significance of N-Acetyl galactosamine specific lectin from Adenia hondala (AHL) from Passifloraceae family. AHL was purified in a single step by affinity chromatography on asialofetuin Sepharose 4B column, characterized and its fine sugar specificity determined by glycan array analysis. AHL is human blood group non specific and also agglutinates rabbit erythrocytes. AHL is a glycoprotein with 12.5% of the carbohydrate, SDS-PAGE, MALDI-TOF-MS and ESI-MS analysis showed that AHL is a monomer of 31.6 kDa. AHL is devoid of DNase activity unlike other Ribosome inactivating proteins (RIPs). Glycan array analysis of AHL revealed its highest affinity for terminal lactosamine or polylactosamine of N- glycans, known to be over expressed in hepatocellular carcinoma and colon cancer. AHL showed strong binding to human hepatocellular carcinoma HepG2 cells with MFI of 59.1 expressing these glycans which was effectively blocked by 93.1% by asialofetuin. AHL showed dose and time dependent growth inhibitory effects on HepG2 cells with IC50 of 4.8 µg/ml. AHL can be explored for its clinical potential.

Self-discrimination in vine tendrils of different plant families.

Previous study reported a novel type of self-discrimination in the tendrils of the vine Cayratia japonica (Vitaceae). However, whether self-discrimination in tendrils is common in vine plant species has not been elucidated. Here, we investigated whether tendrils of Momordica charantia var. pavel (Cucurbitaceae), Cucumis sativus (Cucurbitaceae) and Passiflora caerulea (Passifloraceae) can discriminate self and non-self plants. We also investigated whether the tendrils of M. charantia and C. sativus can discriminate differences in cultivars to determine the discrimination ability for genetic similarity. We found that tendrils of the M. charantia and P. caerulea were more likely to coil around non-self plant than self plants, but not in C. sativus. Our findings support the common occurrence of self-discrimination in tendrils in different plant taxa, although some species lacked it. Furthermore, tendrils of M. charantia more rapidly coil around different cultivars than around same cultivars. The tendrils of M. charantia may can discriminate differences in cultivars.

Passifloraceae seeds from the late Eocene of Colombia.

PREMISE OF THE STUDY: The plant fossil record for the neotropics is still sparse and temporally discontinuous. The location and description of new fossil material are fundamental for understanding evolutionary and biogeographic patterns of lineages. A new fossil record of Passifloraceae from the late Eocene of Colombia is described in this study. METHODS: Plant fossils were collected from a new locality from the Eocene Esmeraldas Formation. Eighteen fossil seeds were selected, described, and compared with fossil and extant angiosperm seeds based on the literature and herbarium collections. Taxonomic affinities of the fossil seeds within Passifloraceae s.l. were evaluated by comparing morphological characters of the seeds in a phylogenetic context. Stratigraphic information associated with the fossil locality was used to interpret the environment and taphonomic processes associated with fossil deposition. KEY RESULTS: A new seed fossil genus and species, Passifloroidesperma sogamosense gen. and sp. nov., is described and associated with the subfamily Passifloroideae based on the presence of a foveolate seed surface, ruminate endosperm, and a seed coat with prismatic palisade cells. The depositional environment of the locality is described as a floodplain associated with river channels. CONCLUSIONS: A detailed review of the Passifloraceae fossil record indicates that P. sogamosense is the oldest confirmed record of Passifloraceae. Its late Eocene age provides a minimum age that can be used as a calibration point for the crown Passifloroideae node in future dating analyses that together with its neotropical geographic location can shed light on the origin and diversification of the subfamily.