Characterization of a Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae) - derived chitinase and its potential for pest control

The larval endoparasitoid Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae) has a toolbox of biological weapons to secure for host colonization and the successful parasitization of its host Heliothis virescens (F.) (Lepidoptera: Noctuidae). The cDNA of a putative chitinase has been previously isolated and initially characterized from teratocytes of this parasitoid among the plethora of molecules available in the venom and calyx fluids injected by females, oral and/or anal secretions released by the parasitoid larvae and/or produced by the expression of genes of the symbiotic associated polydnavirus. This putative chitinase has been initially associated with the host cuticle digestion to allow for parasitoid egression and with the asepsis of the host environment, acting as an antimicrobial. As chitinases are commonly expressed in plants against plant pathogens, the chitinase derived from the teratocytes of T. nigriceps is a potential tool for the development of insect pest control methods based on the disruption of the perithrophic membrane of herbivores. Therefore, we aimed to characterize the activity of the putative chitinase from teratocytes of T. nigriceps (Tnchi) produced using the Escherichia coli expression system and its potential to control H. virescens larvae when expressed into transgenic tobacco plants. The purified E. coli-produced Tnchi protein showed no chitinolitic activity, but was active in binding with colloidal and crystalline chitins in water and with colloidal chitin in buffered solution (pH = 6.74). Transgenic tobacco plants showed no enhanced chitinolitic activity relative to control plants, but survival of three-day old larvae of H. virescens was severely affected when directly fed on transgenic tobacco leaves expressing the recombinant Tnchi protein. Some properties of the Tnchi protein and the potential use of Tnchi-transgenic plants to control plant pests are discussed. (c) 2012 Elsevier Inc. All rights reserved.

Gradients in N-cycling attributes along forestry and agricultural land-use systems are indicative of soil capacity for N supply

Indicators of soil quality associated with N-cycling were assessed under different land-use systems (native forest NAT, reforestation with Araucaria angustifolia or Pinus taeda and agricultural use AGR) to appraise the effects on the soil potential for N supply. The soil total N ranged from 2 to 4 g/kg (AGR and NAT, respectively), and the microbial biomass N ranged from 80 to 250 mg/kg, being higher in NAT and A. angustifolia, and lower in P. taeda and AGR sites. Activities of asparaginase (ca. 50200 mg NH4+-N/kg per h), glutaminase (ca. 200800 mg NH4+-N/kg per h) and urease (ca. 80200 mg NH4+-N/kg/h) were also more intense in the NAT and A. angustifolia-reforested soils, indicating greater capacity for N mineralization. The NAT and AGR soils showed the highest and the lowest ammonification rate, respectively (ca. 1 and 0.4 mg NH4+-N/kg per day), but the inverse for nitrification rate (ca. 12 and 26%), indicating a low capacity for N supply, in addition to higher risks of N losses in the AGR soil. A multivariate analysis indicated more similarity between NAT and A. angustifolia-reforested sites, whilst the AGR soil was different and associated with a higher nitrification rate. In general, reforestation with the native species A. angustifolia had less impact than reforestation with the exogenous species P. taeda, considering the soil capacity for N supply. However, AGR use caused more changes, generally decrease in indicators of N-cycling, showing a negative soil management effect on the sustainability of this agroecosystem.

Status of Aceria guerreronis Keifer (Acari: Eriophyidae) as a Pest of Coconut in the State of Sao Paulo, Southeastern Brazil

The coconut mite, Aceria guerreronis Keifer, is one of the main pests of coconut palms (Cocos nucifera) in northeastern Brazil. The objective of this study was to evaluate the levels of the coconut mite and other mites on coconut palms in the state of So Paulo and to estimate the possible role of predatory mites in the control of this pest. The effect of cultivated genotypes and sampling dates on the mite populations was also estimated. We sampled attached fruits, leaflets, inflorescences, and fallen fruits. The coconut mite was the main phytophagous mite found on attached and fallen fruits, with average densities of 110.0 and 20.5 mites per fruit, respectively. The prevalent predatory mites on attached and fallen fruits were Proctolaelaps bulbosus Moraes, Reis & Gondim Jr. and Proctolaelaps bickleyi (Bram), both Melicharidae. On leaflets, the tenuipalpids Brevipalpus phoenicis (Geijsks) and Tenuipalpus coyacus De Leon and the tetranychid Oligonychus modestus (Banks) were the predominant phytophagous mites. On both leaflets and inflorescences, the predominant predatory mites belonged to the Phytoseiidae. Neoseiulus baraki (Athias-Henriot) and Neoseiulus paspalivorus (De Leon), predators widely associated with the coconut mite in northeastern Brazil and several other countries, were not found. The low densities of the coconut mite in So Paulo could be related to prevailing climatic conditions, scarcity of coconut plantations (hampering the dispersion of the coconut mite between fields), and to the fact that some of the genotypes cultivated in the region are unfavorable for its development.

EVALUATION OF IN VITRO AND IN VIVO EFFECTS OF SEMIPURIFIED PROTEINASE INHIBITORS FROM THEOBROMA SEEDS ON MIDGUT PROTEASE ACTIVITY OF LEPIDOPTERAN PEST INSECTS

We have characterized in vitro and in vivo effects of trypsin inhibitors from Theobroma seeds on the activity of trypsin- and chymotrypsin-like proteins from Lepidopteran pest insects. The action of semipurified trypsin inhibitors from Theobroma was evaluated by the inhibition of bovine trypsin and chymotrypsin activities determined by the hydrolysis of N-Benzoyl-DL-Arginine-p-Nitroanilide (BAPA) and N-Succinyl-Ala-Ala-Pho-Phe p-Nitroanilide (S-(Ala)2ProPhe-pNA). Proteinase inhibitor activities from Theobroma cacao and T. obovatum seeds were the most effective in inhibiting trypsin-like proteins, whereas those from T. obovatum and T. sylvestre were the most efficient against chymotrypsin-like proteins. All larvae midgut extracts showed trypsin-like proteolytic activities, and the putative trypsin inhibitors from Theobroma seeds significantly inhibited purified bovine trypsin. With respect to the influence of Theobroma trypsin inhibitors on intact insects, the inclusion of T. cacao extracts in artificial diets of velvet bean caterpillars (Anticarsia gemmatalis) and sugarcane borer (Diatraea saccharalis) produced a significant increase in the percentage of adult deformation, which is directly related to both the survival rate of the insects and oviposition.

Dispersal strategies of Aceria guerreronis (Acari: Eriophyidae), a coconut pest

The dispersal of plant-feeding mites can occur involuntarily, through transportation of infested plant parts, or voluntarily, by walking to new plant parts or to suitable spots where biotic (phoresis) or abiotic (wind, agricultural tools, etc.) factors carry them over long distances. Elucidating the dispersal mechanisms of the coconut mite, Aceria guerreronis Keifer, is important for understanding the process of colonization of new fruits of a same or different plants, essential for the improvement of control strategies of this serious coconut pest. Thus, the objective of this work was to investigate the voluntary dispersal mechanisms of this mite. The hypothesis that the coconut mite disperses by walking, phoresis or wind were tested. The coconut mite was shown to be able to walk short distances between fruits of the same bunch or between bunches of the same plant. Phoresis on insects of the orders Hymenoptera (Apidae), Coleoptera (Curculionidae) and Lepidoptera (Phycitidae) was evaluated in the laboratory and in the field. Although in the laboratory mites were shown to be able to climb onto honeybees, field investigations failed to show these insects as important carriers of the pest, corroborating findings of previous works; however, both laboratory and field investigations suggested the curculionid Parisoschoenus obesulus Casey to be able to transport the coconut mite between plants. Similarly, laboratory and field investigations suggested wind to be important in the dispersal of the coconut mite between plants.

Natural occurrence of Zoophthora radicans (Entomophthorales: Entomophthoraceae) on Thaumastocoris peregrinus (Heteroptera: Thaumastocoridae), an invasive pest recently found in Brazil

Occurrence of Zoophthora radicans infecting nymphs and adults of Thaumastocoris peregrinus Carpintero and Dellape, 2006 is reported in Brazil. This is a new record of host for this fungal species and the first fungal pathogen associated with this pest worldwide. Infection of Z. radicans on T. peregrinus populations on commercial Eucalyptus plantation (Eucalyptus spp.) reached up to 100%, and low insect densities were associated with high levels of fungal infection in three out of seven plots. This pathogen seems to be virulent against T. peregrinus and may play an important role in population regulations of this invasive pest through naturally induced epizootics. (c) 2012 Elsevier Inc. All rights reserved.