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1.
Insect Mol Biol ; 29(3): 337-351, 2020 06.
Article in English | MEDLINE | ID: mdl-32065441

ABSTRACT

In moths, sex pheromones play a key role in mate finding. These chemicals are transported in the antennae by odorant-binding proteins (OBPs). Commonly, males encounter conspecific females; therefore, several OBPs are male-biased. Less is known, however, about how the olfactory system of moths has evolved toward inverse sexual communication, ie where females seek males. Therefore, the objective of this study was to identify the profile of OBPs and their expression patterns in the bee hive pest, Galleria mellonella, a moth that uses inverse sexual communication. Here, OBP-related transcripts were identified by an RNA Sequencing (RNA-Seq) approach and analysed through both Reverse Transcription Polymerase Chain Reaction (RT-PCR) in different tissues and quantitative real-time PCR for two states, virgin and postmating. Our results indicate that G. mellonella has 20 OBPs distributed amongst different tissues. Interestingly, 17 of the 20 OBPs were significantly down-regulated after mating in females, whereas only OBP7 was up-regulated. By contrast, 18 OBP transcripts were up-regulated in males after mating. Additionally, binding assays and structural simulations showed general odorant-binding protein 2 (GOBP2) was able to bind sex pheromone components and analogues. These findings suggest a possible role of OBPs, especially GOBPs, in the inverse sexual communication of G. mellonella, with gene expression regulated as a response to mating.


Subject(s)
Animal Communication , Gene Expression Regulation , Insect Proteins/genetics , Moths/physiology , Receptors, Odorant/genetics , Sexual Behavior, Animal , Animals , Female , Insect Proteins/metabolism , Ligands , Male , Receptors, Odorant/metabolism
2.
Neotrop Entomol ; 48(3): 422-432, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30414018

ABSTRACT

In the sensory system of insects, olfactory sensilla constitute important functional elements for discriminating odors. Therefore, we used light microscopy and scanning electron microscopy to investigate the morphology and distribution of sensilla in the antennae of Lobesia botrana (Denis & Schiffermüller). In addition, we studied the expression of the gene encoding for pheromone-binding protein 1 (LbotPBP1) by in situ hybridization. Lobesia botrana antennae are filiform and are subdivided into three segments: scape, pedicel, and flagellum. The number of flagellum and their overall length were significantly higher and longer in males than in females. Six morphological types of sensilla (trichodea, chaetica, coeloconica, auricillica, basiconica, and styloconica) were identified on the antennae of both sexes. Trichodea sensilla were the most abundant on the antennae of L. botrana, and three subtypes, discerned by their lengths, were observed. However, sensilla trichodea subtype III was only present in male antennae. Moreover, LbotPBP1 expression was restricted to this type of sensilla, thus confirming its olfactory role, specifically under the context of sexual pheromone perception.


Subject(s)
Arthropod Antennae/anatomy & histology , Carrier Proteins/metabolism , Insect Proteins/metabolism , Moths , Pheromones/metabolism , Sensilla/ultrastructure , Animals , Arthropod Antennae/ultrastructure , Female , Male , Microscopy, Electron, Scanning , Smell
3.
J Appl Microbiol ; 123(4): 886-895, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28718996

ABSTRACT

AIMS: The aim of this study was to evaluate the synthesis of ligninolytic enzymes and some diffusible antifungal compounds by white-rot fungi (WRF) using peels or discarded potato as the sole nutrient source. METHODS AND RESULTS: The strain Trametes hirsuta Ru-513 highlighted for its laccase activity (595 ± 33 U l-1 ), which is able to decolourize 87% of an anthraquinone dye using potato peels as the sole nutritional support. A native polyacrylamide gel of laccase proteins showed the presence of two isoenzymes, corresponding to proteins of 56 and 67 kDa, which were detected by SDS-PAGE. The antifungal activity of ethyl acetate extracts was evaluated by the agar diffusion method, where Anthracophyllum discolor Sp4 and Inonotus sp. Sp2 showed the highest inhibition zones of Mucor miehei. The fungal extracts also inhibited Fusarium oxysporum and Botrytis cinerea growth, with inhibition zones of up to 18 mm. The extract with the highest antifungal activity, from A. discolor Sp4 grown in discarded potato medium, was analysed using a gas chromatograph coupled to a mass spectrometer. Among the identified compounds, chlorinated aromatic compounds and veratryl alcohol were the most abundant compounds. CONCLUSIONS: The results revealed the relevance of potato waste valorization for the sustainable production of ligninolytic enzymes and antifungal compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the synthesis of ligninolytic enzymes and diffusible antifungal compounds by WRF using potato wastes as the sole nutrient source and suggests a relationship between the enzymatic activity and the synthesis of antifungal compounds. These compounds and the synthesis of halogen compounds by WRF using agro-industrial wastes have been poorly studied before.


Subject(s)
Agaricales/metabolism , Antifungal Agents/metabolism , Fungal Proteins/metabolism , Laccase/metabolism , Solanum tuberosum/microbiology , Solid Waste/analysis , Trametes/enzymology , Agaricales/chemistry , Agaricales/growth & development , Coloring Agents/metabolism , Culture Media/metabolism , Electrophoresis, Polyacrylamide Gel , Fungal Proteins/chemistry , Industrial Waste/analysis , Trametes/chemistry , Trametes/growth & development , Trametes/metabolism
4.
Neotrop Entomol ; 44(1): 77-83, 2015 Feb.
Article in English | MEDLINE | ID: mdl-26013015

ABSTRACT

The raspberry weevil, Aegorhinus superciliosus (Guérin) (Coleoptera: Curculionidae), is an economically important pest of blueberry in southern Chile. The digestive protease activity of adult insects was investigated using general and specific substrates and inhibitors. Enzymatic assays demonstrated the presence of trypsin- and chymotrypsin-like serine proteinases. Furthermore, in vitro assays using phenylmethylsulfonyl fluoride (PMSF) and soybean trypsin inhibitor (SBTI) at 0.01 and 0.1 mM showed percentages of enzymatic inhibition between 0 and 16% for PMSF and 67 to 76% for SBTI, whereas in vivo assays indicated that SBTI caused between 50 and 90% mortality in males and between 80 and 100% in females. Our data indicate the presence of serine proteases and suggest that digestive proteases could be a target for the design and development of strategies to control the raspberry weevil.


Subject(s)
Protease Inhibitors/pharmacology , Proteolysis/drug effects , Weevils/drug effects , Weevils/metabolism , Animals , Digestive System/drug effects , Digestive System/enzymology , Female , Male
5.
Neotrop Entomol ; 44(3): 279-85, 2015 Jun.
Article in English | MEDLINE | ID: mdl-26013273

ABSTRACT

Aegorhinus nodipennis (Hope) (Coleoptera: Curculionidae) is an important native pest in fruit crops that is mainly found in European hazelnut fields in the south of Chile. We investigated the behavioral response of A. nodipennis to volatile compounds released from the essential oil of Achillea millefolium and its main constituent using olfactometric bioassays. Gas chromatographic and mass spectral analysis of the A. millefolium essential oil revealed the presence of 11 compounds. Monoterpene ß-thujone (96.2%) was the main component of the oil. Other compounds identified were α-thujone, 1,8-cineole, p-cymene, and 4-terpineol, all with percentages below 1%. Both A. millefolium essential oil and thujone exhibited a repellent activity against this insect at the higher doses tested (285.7 ng/cm(2)), demonstrating their potential as repellents for this species.


Subject(s)
Achillea , Insect Repellents/pharmacology , Oils, Volatile/pharmacology , Weevils/drug effects , Animals , Chile , Oils, Volatile/metabolism , Weevils/metabolism
6.
Neotrop Entomol ; 43(3): 260-5, 2014 Jun.
Article in English | MEDLINE | ID: mdl-27193622

ABSTRACT

Considering that sensilla constitute important functional elements of sensory systems in insects, the aim of this study was to determine the type and distribution of sensilla in the antennae of Hylamorpha elegans Burmeister examined by scanning electron microscopy. Hylamorpha elegans antennae are lamellate and consist of the scape, pedicel, and flagellum. The antennal club of this beetle consists of three terminal plates: proximal, middle, and distal lamellae. Four types of sensilla were observed in the lamellae from both sexes: sensilla trichoidea, chaetica, coeloconica, and placodea. Antennal length was larger in males than in females, and significant sexual variation in the number of sensilla placodea and sensilla coeloconica was observed.


Subject(s)
Arthropod Antennae , Coleoptera/anatomy & histology , Animals , Female , Male , Microscopy, Electron, Scanning , Sensilla
7.
Neotrop Entomol ; 43(3): 266-75, 2014 Jun.
Article in English | MEDLINE | ID: mdl-27193623

ABSTRACT

The European grapevine moth Lobesia botrana (Denis & Schiffermüller) is an economically important insect in Europe. The species invaded vineyards in Chile, Argentina, and California during 2008-2010 causing severe problems. A major component of the sex pheromone, (E,Z)-7,9-dodecadienyl acetate (E7,Z9-12:Ac), is used in a mating disruption technique when grapevine moth populations are low or to monitor pest numbers. It is thought that these sexual pheromones are blends of volatiles that typically are specific to a species and are transported in the insect antenna by pheromone-binding proteins (PBPs) across the sensillar lymph to the olfactory receptors. Currently, an increasing number of Lepidopteran PBPs are being identified and cloned. However, there are no studies of the olfactory system and of proteins involved in the olfactory perception of L. botrana at the molecular level. In the present study, we report, for the first time, the sequence of a PBP from L. botrana (LbotPBP), which was determined using reverse transcription technology. Homology modeling was used to generate the three-dimensional protein structure. The model suggests that PBP consists of six α-helices as follows: Lys2-Met23 (α1), Thr28-Phe36 (α2), Arg46-Leu59 (α3), His70-Asn80 (α4), Glu84-Asn100 (α5), and Cys108-Lys125 (α6), held together by three disulfide bridges, Cys19-Cys54, Cys50-Cys108, and Cys97-Cys117. Docking simulations based on this model suggested that Trp114 is a key residue in the recognition of acetate pheromones, such as E7,Z9-12:Ac. In silico results in this study are consistent with previous findings in which E7,Z9-12:Ac acts as the most active compound in behavioral and electroantennographic assays.


Subject(s)
Carrier Proteins/genetics , Moths/genetics , Pheromones/genetics , Animals , Argentina , Chile , Computer Simulation , Insect Proteins , Moths/physiology
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