Structural insights regarding an insecticidal Talisia esculenta protein and its biotechnological potential for Diatraea saccharalis larval control

Talisin is a seed-storage protein from Talisia esculenta that presents lectin-like activities, as well as proteinase-inhibitor properties. The present study aims to provide new in vitro and in silico biochemical information about this protein, shedding some light on its mechanistic inhibitory strategies. A theoretical three-dimensional structure of Talisin bound to trypsin was constructed in order to determine the relative interaction mode. Since the structure of non-competitive inhibition has not been elucidated, Talisin-trypsin docking was carried out using Hex v5.1, since the structure of non-competitive inhibition has not been elucidated. The predicted non-coincidence of the trypsin binding site is completely different from that previously proposed for Kunitz-type inhibitors, which demonstrate a substitution of an Arg(64) for the Glu(64) residue. Data, therefore, provide more information regarding the mechanisms of non-competitive plant proteinase inhibitors. Bioassays with Talisin also presented a strong insecticide effect on the larval development of Diatraea saccharalis, demonstrating LD50 and ED50 of ca. 2.0% and 1.5%, respectively. (C) 2011 Elsevier Inc. All rights reserved.

Intraspecific competition in Zabrotes subfasciatus: Physiological and behavioral adaptations to different amounts of host

The effects of competition on populations of the bean weevil Zabrotes subfasciatus were analyzed during 41 generations under different competition levels. Three competition environments were established by maintaining the number of couples (6) and varying the amount of available host seeds: HC, high (limited availability of host: 1.35 g); IC, intermediate (intermediate availability of host: 6 g); and LC, low competition (abundance of host: 36 g). It was found that the distribution of the eggs laid on grains was different among treatments: in LC, for example, although females showed high fecundity (35.4 +/- 5.6 eggs/female) the number of eggs laid on each grain was small (1.2 +/- 0.4 eggs on each seed), thus avoiding larval competition of their offspring; whereas in HC treatment, females showed low fecundity (27.04 +/- 4.5 eggs/female) but laid many eggs on each grain (15.03 +/- 4.3 eggs). There were no changes in the ability to respond to different amounts of host via oviposition behavior (egg distribution) during 41 generations. However, HC females had more offspring than LC females under HC conditions. This suggests that HC insects evolved toward higher fitness in crowded conditions. In addition, after inverting the competition level, insects behaved independently of the treatment conditions they experienced through generations, thus showing that oviposition behavior is flexible. Taken together, our results show that Z. subfasciatus presents a broad range of behavioral and physiological responses which allows for quick and reversible adjustments to sudden changes in the amount of resources.

Recombinant expression, localization and in vitro inhibition of midgut cysteine peptidase (Sl-CathL) from sugarcane weevil, Sphenophorus levis

A cDNA coding for a digestive cathepsin L, denominated Sl-CathL, was isolated from a cDNA library of Sphenophorus levis larvae, representing the most abundant EST (10.49%) responsible for proteolysis in the midgut. The open reading frame of 972 bp encodes a preproenzyme similar to midgut cathepsin L-like enzymes in other coleopterans. Recombinant Sl-CathL was expressed in Pichia pastoris, with molecular mass of about 42 kDa. The recombinant protein was catalytically activated at low pH and the mature enzyme of 39 kDa displayed thermal instability and maximal activity at 37 degrees C and pH 6.0. Immunocytochemical analysis revealed Sl-CathL production in the midgut epithelium and secretion from vesicles containing the enzyme into the gut lumen, confirming an important role for this enzyme in the digestion of the insect larvae. The expression profile identified by RT-PCR through the biological cycle indicates that Sl-CathL is mainly produced in larval stages, with peak expression in 30-day-old larvae. At this stage, the enzyme is 1250-fold more expressed than in the pupal fase, in which the lowest expression level is detected. This enzyme is also produced in the adult stage, albeit in lesser abundance, assuming the presence of a different array of enzymes in the digestive system of adults. Tissue-specific analysis revealed that Sl-CathL mRNA synthesis occurs fundamentally in the larval midgut, thereby confirming its function as a digestive enzyme, as detected in immunolocalization assays. The catalytic efficiency of the purified recombinant enzyme was calculated using different substrates (Z-Leu-Arg-AMC, Z-Arg-Arg-AMC and Z-Phe-Arg-AMC) and rSl-CathL exhibited hydrolysis preference for Z-Leu-Arg-AMC (k(cat)/K-m = 37.53 mM S-1), which is similar to other insect cathepsin L-like enzymes. rSl-CathL activity inhibition assays were performed using four recombinant sugarcane cystatins. rSl-CathL was strongly inhibited by recombinant cystatin CaneCPI-4 (K-i = 0.196 nM), indicating that this protease is a potential target for pest control. (C) 2011 Elsevier Ltd. All rights reserved.

Molecular cloning and insecticidal effect of Inga laurina trypsin inhibitor on Diatraea saccharalis and Heliothis virescens

Native Inga laurina (Fabaceae) trypsin inhibitor (ILTI) was tested for anti-insect activity against Diatraea saccharalis and Heliothis virescens larvae. The addition of 0.1% ILTI to the diet of D. saccharalis did not alter larval survival but decreased larval weight by 51%. The H. virescens larvae that were fed a diet containing 0.5% ILTI showed an 84% decrease in weight. ILTI was not digested by the midgut proteinases of either species of larvae. The trypsin levels were reduced by 55.3% in the feces of D. saccharalis and increased by 24.1% in the feces of H. virescens. The trypsin activity in both species fed with ILTI was sensitive to the inhibitor, suggesting that no novel proteinase resistant to ILTI was induced. Additionally, ILTI exhibited inhibitory activity against the proteinases present in the larval midgut of different species of Lepidoptera. The organization of the ilti gene was elucidated by analyzing its corresponding genomic sequence. The recombinant ILTI protein (reILTI) was expressed and purified, and its efficacy was evaluated. Both native ILTI and reILTI exhibited a similar strong inhibitory effect on bovine trypsin activity. These results suggest that ILTI presents insecticidal properties against both insects and may thus be a useful tool in the genetic engineering of plants. (c) 2012 Elsevier Inc. All rights reserved.

Description of the larva of Potamophilops cinereus (Blanchard) from Southeastern Brazil (Coleoptera, Elmidae, Larainae)

Larvae of Potamophilops cinereus (Blanchard, 1841) from Brazil, Sao Paulo, Ribeirao Grande (Fazenda Intervales, Rio Carmo) are described for the first time and illustrated. The larvae were found associated with adults. The larva of P. cinereus is most similar to larvae of species of the Mexican and Central American genera Disersus Sharp and Hispaniolara Brown, but can be easily distinguished chiefly by the head being not visible when seen from above, due to being concealed by the anterior projection of the pronotum. Comparisons of the three genera are presented. Photographs of the adult habitus of P. cinereus are also furnished. A key to the larvae of the genera of Larainae of Brazil is included.

A taxonomic revision and revalidation of Nycterilampus Montrouzier (Coleoptera: Elateridae, Agrypninae)

Nycterilampus Montrouzier, 1860, from Oceania, is removed from junior synonymy with Tetrigus Candeze, 1857, and is redescribed and revalidated. The genus includes two species, N. lifuanus Montrouzier, 1860, and N. velutinus Fleutiaux, 1891 both from New Caledonia. A comparative study of the morphological characters of males and females, including the reproductive organs of the Nycterilampus species and Tetrigus parallelus Candeze, 1857 (type-species) is presented. A key to Nycterilampus species and their separation from Tetrigus parallelus is given.

A new Brazilian species of Loxandrus LeConte, 1852, with description of immatures and notes on natural history (Coleoptera: Carabidae: Loxandrini)

Loxandrus oophagus sp. nov. is described (type-locality: Uberlandia, State of Minas Gerais, Brazil). Larvae, pupae and adults of the new species of the carabid beetle were collected on foam nests of the anuran Leptodactylus fuscus (Schneider, 1799) in the surroundings of Uberlandia, 18 degrees 55S, 48 degrees 17W (Brazil, Minas Gerais), at 750 m altitude. The new species is compared with the similar Loxandrus quinarius Will & Liebherr, 1997, only known from Santa Cruz, Bolivia, differing by the morphology of tarsomeres. Larvae prey on anuran eggs. Description of the immatures and the natural history of the species are provided. The larva differs from the known larvae of Loxandrus species mainly by being eruciform, glabrous and depigmented, its small head and legs, and the lack of stemmata and urogomphi, representing an unusual last instar type among the Carabidae.

New species of Triplonychus Candeze and Globothorax Fleutiaux from Brazil (Coleoptera, Elateridae, Cardiophorinae)

Three new species of Triplonychus Candeze, T. cruspinosus sp. nov., T. crassifemoris sp. nov. and T. tibialatus sp. nov., and a new species of Globothorax Fleutiaux, G. latidens sp. nov., from Brazil are described. Illustrations, photographic and SEM images are presented. A review of the diagnostic generic characters and a key to the species of Triplonychus and Globothorax from Brazil are also given.

New species of Rhinotragini and redescription of two species of Stultutragus Clarke (Coleoptera, Cerambycidae, Cerambycinae)

Agaone amazonica sp. nov. and Stultutragus endoluteus sp. nov. are described from Brazil, and variation in A. peruviensis Fisher, is discussed. Stultutragus nigricornis is elevated to species rank and re-described. Updated keys to Agaone and Stultutragus are provided.

Notes and new taxa in Acanthoderini (Coleoptera: Cerambycidae: Lamiinae). V. Three new genera and notes on Irundisaua.

Generic characters of Irundisaua Martins & Galileo, 2005 are expanded (antennal formula, width of prosternal process inferior or equal to the diameter of procoxa, protibia enlarged and flattened) and two species are transferred from Acanthoderes Audinet-Serville, 1835: I. forsteri (Tippmann, 1960) comb. nov from Peru and Brazil (Amazonas) and I. ucayalensis (Tippmann,1960) comb. nov from Ecuador, Peru and Brazil (Amazonas).Three new genera are described: (1) Catuana gen. nov., type species, C. spinicornis (Tippmann, 1960) comb. nov, characterized by the mesosternal tubercle; (2) Mundeu gen. nov, type species, M. maculicollis (Bates, 1861) comb. nov, with rounded sides of protorax and expanded protibiae; (3) Urangaua gen. nov, with eyes divided and finelly granulated, length of antenomeres III-IV subequal to V-XI; the genus is proposed of two species: U. analis Melzer, 1935 comb. nov, type species, and U. subanalis (Zajciw,1964) comb. nov. A key to the species of Urangaua is added.

New taxa and new record of Hemilophini (Coleoptera: Cerambycidae: Lamiinae).

New species described and illustrated: Adesmus nigrolineatus sp. nov from Mexico (Oaxaca). From Costa Rica: A, moruna sp. nov. (Heredia); Corcovado bezarki sp. nov. (Guanacaste); Alampyris fuscus sp. nov. (Guanacaste), Cariua gen. nov type species C. sulphurea sp. nov. (Guanacaste). From Bolivia: Phoebemima albomaculata sp. nov. (Cochabamba); Ipepo gen. nov type species I. dilatatus sp. nov. (Santa Cruz). From Brazil: Adesmus facetus sp. nov and Canarana arguta sp. nov. (Rondonia). A new record from Costa Rica of Piruanycha pitilla Galileo & Martins, 2005 is added. The three new species of Adesmus are recognized: A. nigrolineatus by the longitudinal black stripes on elytra; A. moruna by the elytra entirely black; A. facetus by the white belts behind the middle of the elytra and white maculae on apical quarter. Phoebemima albomaculata is characterized by the white macula on the elytral suture. Corcovado bezarki sp. nov is distinguished by the black antennal scape and whitish flagelomeres. Canarana arguta sp. nov has prothorax and urosternites I-IV covered by dense yellowish pubescence. Alampyris fusca sp. nov differs from A. cretaria by the antenomere III longer than scape. Cariua sulphurea sp. nov is distinguished by the urosternites covered by compact white pubescence and Ipepo dilatatus is characterized by the elytra with three carinae.

New taxa of Hemilophini (Coleoptera, Cerambycidae, Lamiinae) of the Americas

New taxa of Hemilophini (Coleoptera, Cerambycidae, Lamiinae) of the Americas. The new genus Mexicoscylus is described to include: M. rosae sp. nov., type species of the genus, and M. bivittatus (Gahan, 1892), comb. nov. both from Mexico. A key to the species of Mexicoscylus is added. More three species are described: Cotycuara villosa sp. nov., from Costa Rica; Phoebe parvimacula sp. nov., from Bolivia and Adesmus beruri sp. nov., from Brazil (Amazonas). All new species are illustrated.

Novos táxons de Cerambycidae (Coleoptera) das Américas Central e do Sul

Novos táxons descritos: em Cerambycinae, Bomarion caudatum sp. nov., da Bolívia (Ectenessini); Nephalius levigatus sp. nov., do Brasil, Paraíba (Elaphidiini); Bothriospila pulcherrima sp. nov., do Brasil, Bahia (Bothriospilini); Cycnidolon rufescens sp. nov., do Brasil, Paraíba (Neoibidionini). Em Lamiinae: Cicatricallia gen. nov., espécie-tipo: C. cicatricosa sp. nov. de Trindade e Tobago (Calliini); Mimasyngenes clarkei sp. nov., de Trinidad e Tobago e Mimasyngenes fonticulus sp. nov., do Brasil, Piauí; Ibypeba gen. nov., espécie-tipo: I. camiri sp. nov., da Bolívia; Micratelodesmis gen. nov., espécie-tipo: M. minor sp. nov. (Desmiphorini); Lycidola affinis sp. nov. de Trinidad e Tobago (Hemilophini). Novos registros: Lycidola beltii Bates, 1872 para a Costa Rica e Alampyris fuscus Martins & Galileo, 2008 para o Panamá (Hemilophini).

Descriptions of Macrolygistopterus subparallelus Pic, 1930 immatures from southeastern Brazil (Coleoptera, Lycidae, Lycinae, Calochromini)

Mature larva and pupa of Macrolygistopterus subparallelus Pic are described and compared to known Calochromini immatures. Larvae were collected alive inside dead trunk in the Atlantic Forest at Estacao Biologica de Boraceia, Salesopolis, Sao Paulo, Brazil. They were maintained in laboratory conditions, and the pupal period was 12 days (one observation). The pupa of this genus is described for the first time. This larva differs from known Calochromini larvae mainly because of its almost cylindrical campodeiform body and longer urogomphi.

Aspisoma lineatum (Gyllenhal) (Coleoptera: Lampyridae) Firefly: Description of the Immatures, Biological, and Ecological Aspects

(Gyllenhal) is a common firefly in the Southeastern region of Brazil. Adults and larvae were collected in the municipality of Campinas, state of So Paulo, Brazil, and the immature stages were described and reared in the laboratory. Four generations were reproduced in the laboratory, and a method for its rearing was established. The life cycle usually lasts 6 months, but under optimal laboratory conditions, it lasted from 2 to 4 months. Larvae were fed with and snails since the beginning of the larval stage. This species was found to be easily adapted to environments under anthropic influence, such as urban areas and farms.