Expression of a barley cystatin gene in maize enhances resistance against phytophagous mites by altering their cysteine-proteases

Phytocystatins are inhibitors of cysteine-proteases from plants putatively involved in plant defence based on their capability of inhibit heterologous enzymes. We have previously characterised the whole cystatin gene family members from barley (HvCPI-1 to HvCPI-13). The aim of this study was to assess the effects of barley cystatins on two phytophagous spider mites, Tetranychus urticae and Brevipalpus chilensis. The determination of proteolytic activity profile in both mite species showed the presence of the cysteine-proteases, putative targets of cystatins, among other enzymatic activities. All barley cystatins, except HvCPI-1 and HvCPI-7, inhibited in vitro mite cathepsin L- and/or cathepsin B-like activities, HvCPI-6 being the strongest inhibitor for both mite species. Transgenic maize plants expressing HvCPI-6 protein were generated and the functional integrity of the cystatin transgene was confirmed by in vitro inhibitory effect observed against T. urticae and B. chilensis protein extracts. Feeding experiments impaired on transgenic lines performed with T. urticae impaired mite development and reproductive performance. Besides, a significant reduction of cathepsin L-like and/or cathepsin B-like activities was observed when the spider mite fed on maize plants expressing HvCPI-6 cystatin. These findings reveal the potential of barley cystatins as acaricide proteins to protect plants against two important mite pests

Control of Pyrethroid-Resistant Chagas Disease Vectors with Entomopathogenic Fungi

Chagas disease, also known as American Trypanosomiasis, is the most relevant parasitic disease in Latin America, being a major burden that affects mostly poor human populations living in rural areas. The kissing-bugs of the Triatominae family transmit the parasite Trypanosoma cruzi by infectious blood-sucking; Triatoma infestans is the vector of major relevance in the southern Cone of South America. Current control strategies, heavily based on residual insecticide spraying, are threatened by the emergence of pyrethroid-resistant bug populations. Furthermore, ensuring the long-term and sustainable control of this overwhelming disease remains a major challenge. Here we show the utility of a simple, low-cost, biological control methodology against T. infestans bugs, regardless of their susceptibility to pyrethroid insecticides. It is based on the understanding of the initial contact interactions between a mycoinsecticide agent—the fungus Beauveria bassiana—and the host defense barrier, the bug cuticle. The proposed methodology is also supported by present data showing a relationship between the triatomine cuticle width and its hydrocarbon surface components, with insecticide resistance. These results will help to provide a safe and efficient alternative to overcome pyrethroid-resilience of these noxious bugs. A high transfer potential to immediate application in rural communities located in remote areas inaccessible to sanitary control teams, and to the control of other Chagas disease vectors as well, is also envisaged

A barley cysteine-protease inhibitor reduces teh performance of two aphid species in artificial diets and transgenic arabidopsis plants

Cystatins from plants have been implicated in plant defense towards insects, based on their role as inhibitors of heterologous cysteine-proteinases. We have previously characterized thirteen genes encoding cystatins (HvCPI-1 to HvCPI-13) from barley (Hordeum vulgare), but only HvCPI-1 C68 → G, a variant generated by direct-mutagenesis, has been tested against insects. The aim of this study was to analyze the effects of the whole gene family members of barley cystatins against two aphids, Myzus persicae and Acyrthosiphon pisum. All the cystatins, except HvCPI-7, HvCPI-10 and HvCPI-12, inhibited in vitro the activity of cathepsin L- and/or B-like proteinases, with HvCPI-6 being the most effective inhibitor for both aphid species. When administered in artificial diets, HvCPI-6 was toxic to A. pisum nymphs (LC50 = 150 μg/ml), whereas no significant mortality was observed on M. persicae nymphs up to 1000 μg/ml. The effects of HvCPI-6 ingestion on A. pisum were correlated with a decrease of cathepsin B- and L-like proteinase activities. In the case of M. persicae, there was an increase of these proteolytic activities, but also of the aminopeptidase-like activity, suggesting that this species is regulating both target and insensitive enzymes to overcome the effects of the cystatin. To further analyze the potential of barley cystatins as insecticidal proteins against aphids, Arabidopsis plants expressing HvCPI-6 were tested against M. persicae. For A. pisum, which does not feed on Arabidopsis, a combined diet-Vicia faba plant bioassay was performed. A significant delay in the development time to reach the adult stage was observed in both species. The present study demonstrates the potential of barley cystatins to interfere with the performance of two aphid specie

The whole genome sequence of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), reveals insights into the biology and adaptive evolution of a highly invasive pest species

The Mediterranean fruit fly (medfly), Ceratitis capitata, is a major destructive insect pest due to its broad host range, which includes hundreds of fruits and vegetables. It exhibits a unique ability to invade and adapt to ecological niches throughout tropical and subtropical regions of the world, though medfly infestations have been prevented and controlled by the sterile insect technique (SIT) as part of integrated pest management programs (IPMs). The genetic analysis and manipulation of medfly has been subject to intensive study in an effort to improve SIT efficacy and other aspects of IPM control

Risk factors associated with Trypanosoma cruziexposure in domestic dogs from a rural community in Panama

Chagas disease, caused by Trypanosoma cruzi infection, is a zoonosis of humans, wild and domestic mammals,including dogs. In Panama, the main T. cruzi vector is Rhodnius pallescens, a triatomine bug whose main naturalhabitat is the royal palm, Attalea butyracea. In this paper, we present results from three T. cruzi serological tests(immunochromatographic dipstick, indirect immunofluorescence and ELISA) performed in 51 dogs from 24 housesin Trinidad de Las Minas, western Panama. We found that nine dogs were seropositive (17.6% prevalence). Dogswere 1.6 times more likely to become T. cruzi seropositive with each year of age and 11.6 times if royal palms wherepresent in the peridomiciliary area of the dog’s household or its two nearest neighbours. Mouse-baited-adhesivetraps were employed to evaluate 12 peridomestic royal palms. All palms were found infested with R. pallescens withan average of 25.50 triatomines captured per palm. Of 35 adult bugs analysed, 88.6% showed protozoa flagellates intheir intestinal contents. In addition, dogs were five times more likely to be infected by the presence of an additionaldomestic animal species in the dog’s peridomiciliary environment. Our results suggest that interventions focused onroyal palms might reduce the exposure to T. cruzi infection

Insecticidal effects of Moroccan plant extracts on development, energy reserves and enzymatic activities of Plodia interpunctella

10 p.-3 tab.The aim of this work was to study the effects of methanol extracts of ten plant species used in traditional medicine in Morocco (Peganum harmala, Ajuga iva, Rosmarinus officinalis, Lavandula stoechas, Lavandula dentata, Cistus ladanifer, Cistus salviaefolius, Cistus monspeliensis, Centaurium erythraea and Launaea arborescens) on Plodia interpunctella Hübner (Lepidoptera: Pyralidae) larvae. Firstly, we studied the effects of the ingestion of these extracts at 500 ppm on post-embryonic development parameters. Most plant extracts provoked a notable decrease of larval weight 8 days after treatment (up to 33% weight loss with C. erythraea) and caused significant alterations on pupation (ranging from 5% to 85%) and adult emergence (below 2.5% with R. officinalis, C. erythraea and A. iva). The plant extracts that showed strongest effects on post-embryonic development were selected to test their effects on the following physiological parameters: larval reserve substances (at 500 ppm); and midgut activities of hydrolytic and detoxification enzymes (at 500, 750 and 1,000 ppm). All treatments provoked a significant reduction of protein and carbon hydrate larval contents, the inhibition of proteases and α-amylase activities in a dose depended manner, and the induction of glutathione Stransferase and esterase (using MtB as substrate) activities, whereas the activity of cytochrome P450 monooxygenases and esterases (using 1-NA as substrate) increase or decrease depending on the extract concentration and the plant analyzed.This work was supported by the cooperation program: CNRST (580/08)/CSIC (2007MA0011) and MAEC-AECI fellowship (Nº 0000536790 II.E Program).Peer reviewe