Toxicity,behavior impairment,and repellence of essential oils from pepper‐rosmarin and patchouli to termites

Plant essential oils are potential sources of insecticidal compounds, but have rarely been explored for their effect on termites. In the present study, we assessed the chemical composition of essential oils of Lippia sidoides Cham. (pepper‐rosmarin; Verbenaceae) and Pogostemon cablin (Blanco) Benth. (patchouli; Lamiacaeae) and evaluated their toxicity, behavioral impairment, and repellence to termite species of the genera Amitermes and Microcerotermes (Isoptera: Termitidae: Termitinae). The main components of essential oils of L. sidoides and P. cablin were thymol (44.6%) and patchouli alcohol (36.6%), respectively. The essential oil of P. cablin was most potent against Amitermes cf. amifer Silvestri and had the lowest LD₅₀ (0.63 μg mg^?1). There was no difference in toxicity for Microcerotermes indistinctus Mathews between the essential oils of L. sidoides (LD₅₀ = 1.49 μg mg^?1) and P. cablin (LD₅₀ = 1.67 μg mg^?1). Pogostemon cablin essential oil was the most toxic to M. indistinctus (LC₅₀ = 0.32 μl ml^?1) and A. cf. amifer (LC₅₀ = 0.29 μl ml^?1). The essential oils analyzed exhibited high toxicity and repellence to the termites, in addition to reducing behavioral interactions among individuals, thus constituting potential termiticides.     

RUN and FYVE domain–containing protein 4 enhances autophagy and lysosome tethering in response to Interleukin-4

Autophagy is a key degradative pathway coordinated by external cues, including starvation, oxidative stress, or pathogen detection. Rare are the molecules known to contribute mechanistically to the regulation of autophagy and expressed specifically in particular environmental contexts or in distinct cell types. Here, we unravel the role of RUN and FYVE domain–containing protein 4 (RUFY4) as a positive molecular regulator of macroautophagy in primary dendritic cells (DCs). We show that exposure to interleukin-4 (IL-4) during DC differentiation enhances autophagy flux through mTORC1 regulation and RUFY4 induction, which in turn actively promote LC3 degradation, Syntaxin 17–positive autophagosome formation, and lysosome tethering. Enhanced autophagy boosts endogenous antigen presentation by MHC II and allows host control of Brucella abortus replication in IL-4–treated DCs and in RUFY4-expressing cells. RUFY4 is therefore the first molecule characterized to date that promotes autophagy and influences endosome dynamics in a subset of immune cells.     

Demonstration of Lignin-to-Peroxidase Direct Electron Transfer: A TRANSIENT-STATE KINETICS,DIRECTED MUTAGENESIS,EPR, AND NMR STUDY*

Versatile peroxidase (VP) is a high redox-potential peroxidase of biotechnological interest that is able to oxidize phenolic and non-phenolic aromatics, Mn²⁺, and different dyes. The ability of VP from Pleurotus eryngii to oxidize water-soluble lignins (softwood and hardwood lignosulfonates) is demonstrated here by a combination of directed mutagenesis and spectroscopic techniques, among others. In addition, direct electron transfer between the peroxidase and the lignin macromolecule was kinetically characterized using stopped-flow spectrophotometry. VP variants were used to show that this reaction strongly depends on the presence of a solvent-exposed tryptophan residue (Trp-164). Moreover, the tryptophanyl radical detected by EPR spectroscopy of H₂O₂-activated VP (being absent from the W164S variant) was identified as catalytically active because it was reduced during lignosulfonate oxidation, resulting in the appearance of a lignin radical. The decrease of lignin fluorescence (excitation at 355 nm/emission at 400 nm) during VP treatment under steady-state conditions was accompanied by a decrease of the lignin (aromatic nuclei and side chains) signals in one-dimensional and two-dimensional NMR spectra, confirming the ligninolytic capabilities of the enzyme. Simultaneously, size-exclusion chromatography showed an increase of the molecular mass of the modified residual lignin, especially for the (low molecular mass) hardwood lignosulfonate, revealing that the oxidation products tend to recondense during the VP treatment. Finally, mutagenesis of selected residues neighboring Trp-164 resulted in improved apparent second-order rate constants for lignosulfonate reactions, revealing that changes in its protein environment (modifying the net negative charge and/or substrate accessibility/binding) can modulate the reactivity of the catalytic tryptophan.     

Evidence of incipient speciation in the Neotropical mangrove Pelliciera rhizophorae (Tetrameristaceae) as revealed by molecular,morphological, physiological and climatic characteristics

Two variants of the Neotropical mangrove species Pelliciera rhizophorae distributed along both sides of the Isthmus of Panama were detected by different colouration of the floral bracts and the size of the floral and vegetative structures. These findings raised questions concerning a possible speciation event in P. rhizophorae, for which a series of macro‐ and microscopic morphological traits (reproductive and vegetative structures), molecular markers from plastid DNA and climatic profiles were analyzed. Samples of P. rhizophorae were collected in three localities from the Panamanian Caribbean and Pacific coasts. The data obtained from molecular markers and morphological traits showed significant differences between the variants. The climatic profiles showed contrasting characteristics of rainfall and temperature in their habitats: variant A is found in wetter zones and variant B occupies drier zones. Evidence suggesting that a process of incipient speciation has occurred in P. rhizophorae in response to ecogeographical isolation due to climatic factors is presented. The presence of two geographically separate genetic‐morphological groups, adapted to contrasting climatic conditions, will be the basis for suggesting the existence of incipient lineages in Pelliciera. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 499–510.     

Fruits of Heterocoma (Vernonieae‐Lychnophorinae): taxonomic significance and a new pattern of phytomelanin deposition in Asteraceae

Heterocoma is a Brazilian endemic genus resulting from the dismemberment of Sipolisiinae, in which only representatives with fruit containing phytomelanin were included in the genus. As the fruits of Asteraceae are known to be systematically important at various taxonomic levels and Heterocoma fruit has not been described previously, we studied the morphology and anatomy of the cypselas of all species of the genus, comparing them with other fruits in the family containing phytomelanin and evaluating the systematic potential at the specific and tribal levels. The fruits were analysed by scanning electron microscopy (SEM) and light microscopy. The morphological features of the fruit, including the carpopodium, ribs and pappi, varied in the genus and demonstrated potential for species discrimination. The anatomy showed a pattern for the genus with a uniseriate exocarp, the outer mesocarp composed of fibres arranged in several layers, the inner mesocarp composed of several layers of parenchyma, the endocarp, and phytomelanin deposited between the inner and outer mesocarp. This anatomical pattern of phytomelanin deposition differs from that of other Asteraceae with phytomelanin in their fruit. Heterocoma is also the only genus in Vernonieae that has phytomelanin deposition in the cypselas. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 255–265.     

Xylanase production by endophytic Aspergillus niger using pentose-rich hydrothermal liquor from sugarcane bagasse

Fungal xylanases have been widely studied and various production methods have been proposed using submerged and solid-state fermentation. This class of enzyme is used to supplement cellulolytic enzyme cocktails in order to enhance the enzymatic hydrolysis of plant cell walls. The present work investigates the production of xylanase and other accessory enzymes by a recently isolated endophytic Aspergillus niger DR02 strain, using the pentose-rich liquor from hydrothermal pretreatment of sugarcane bagasse as carbon source. Batch and fed-batch submerged cultivation approaches were developed in order to minimize the toxicity of the liquor and increase enzyme production. Maximum xylanase activities obtained were 458.1 U/mL for constant fed-batch, 428.1 U/mL for exponential fed-batch, and 264.37 U/mL for pulsed fed-batch modes. The results indicated that carbon-limited fed-batch cultivation can reduce fungal catabolite repression, as well as overcome possible negative effects of toxic compounds present in the pentose-rich liquor. Enzymatic panel and mass spectrometric analyses of the fed-batch A. niger secretome showed high levels of xylanolytic enzymes (GH10, GH11, and GH62 Cazy families), together with cellobiohydrolase (G6 and GH7), β-glucosidase, β-xylosidase (GH3), and feruloyl esterase (CE1) accessory enzyme activities. The yields of glucose and xylose from enzymatic hydrolysis of hydrothermally pretreated sugarcane bagasse increased by 43.7 and 65.3%, respectively, when a commercial cellulase preparation was supplemented with the A. niger DR02 constant fed-batch enzyme complex.