Characterization and distribution of digestive proteases of the stalk corn borer,Sesamia nonagrioides Lef. (Lepidoptera: Noctuidae)

Ortego, Félix; Novillo, Concepción; Castañera, Pedro

doi:10.1002/(sici)1520-6327(1996)33:2<163::aid-arch6>3.0.co;2-z

Cited by 73 publications

(24 citation statements)

References 43 publications

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“…APNs play a prominent role in proteolysis in Lepidoptera, as shown by the significant reduction of azocasein proteolysis by adding a specific APN inhibitor (Ortega et al, 1996). Thus it is not surprising that APNs are so strongly expressed in larval gut.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization and RNA interference of three midgut aminopeptidase N isozymes from Bacillus thuringiensis-susceptible and -resistant strains of sugarcane borer, Diatraea saccharalis

Yang

Zhu

Ottea

et al. 2010

Insect Biochemistry and Molecular Biology

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Section: Discussionmentioning

confidence: 99%

Molecular characterization and RNA interference of three midgut aminopeptidase N isozymes from Bacillus thuringiensis-susceptible and -resistant strains of sugarcane borer, Diatraea saccharalis

Yang

Zhu

Ottea

et al. 2010

Insect Biochemistry and Molecular Biology

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“…Carboxypeptidases A and B-like activities were assayed in 0.5 ml of reaction buffer containing 30 lg of protein extracts, using 1 mM HPA (hippuryl-phenylalanine) for carboxypeptidase A-like and 1 mM HA (hippuryl-L-arginine) for carboxypeptidase B-like as described by Ortego et al (1996).…”

Section: Enzyme Activity Assaysmentioning

confidence: 99%

A barley cysteine-proteinase inhibitor reduces the performance of two aphid species in artificial diets and transgenic Arabidopsis plants

et al. 2010

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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 (LC(50) = 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 species.

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“…Non-specific protease activity was assayed with 0.1% sulfanilamide-azocasein as substrate. Other protease assays were as follows: trypsin-like activity using 1 mM BApNa (Na-benzoyl-DL-arginine p-nitroanilide), chymotrypsin-like activity with 0.25 mM Sa 2 PPpNa (N-succinyl-(alanine) 2 -prolinephenylalanine-p-nitroanilide), and elastase-like activity with 0.25 mM SA 3 pNa (N-succinyl-(alanine) 3 -pnitroanilide), as describe by Ortego et al (1996). Cathepsin D-like activity was measured with 0.2% haemoglobin as substrate, and cathepsin B-like activity was assayed with 50 lM ZAA 2 MNA (N-carbobenzoxy-alanine-arginine-arginine 4-methoxy-b-naphthyl amide), as described by Novillo et al (1997).…”

Section: Plant Materialsmentioning

confidence: 99%

Prey mediated effects of Bt maize on fitness and digestive physiology of the red spider mite predator Stethorus punctillum Weise (Coleoptera: Coccinellidae)

et al. 2008

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The present study investigated prey-mediated effects of two maize varieties expressing a truncated Cry1Ab, Compa CB (event Bt176) and DKC7565 (event MON810), on the biology of the ladybird Stethorus punctillum. Although immuno-assays demonstrated the presence of Cry1Ab in both prey and predator collected from commercial maize-growing fields, neither transgenic variety had any negative effects on survival of the predator, nor on the developmental time through to adulthood. Furthermore, no subsequent effects on ladybird fecundity were observed. As a prerequisite to studying the interaction of ladybird proteases with Cry1Ab, proteases were characterised using a range of natural and synthetic substrates with diagnostic inhibitors. These results demonstrated that this predator utilises both serine and cysteine proteases for digestion. In vitro studies demonstrated that T. urticae were not able to process or hydrolyze Cry1Ab, suggesting that the toxin passes through the prey to the third trophic level undegraded, thus presumably retaining its insecticidal properties. In contrast, S. punctillum was able to activate the 130 kDa protoxin into the 65 kDa fragment; a fragment of similar size was also obtained with bovine trypsin, which is known to cleave the protoxin to the active form. Thus, despite a potential hazard to the ladybird of Bt-expressing maize (since the predator was both exposed to, and able to proteolytically cleave the toxin, at least in vitro), no deleterious effects were observed.

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Characterization and distribution of digestive proteases of the stalk corn borer,Sesamia nonagrioides Lef. (Lepidoptera: Noctuidae)

Cited by 73 publications

References 43 publications

Molecular characterization and RNA interference of three midgut aminopeptidase N isozymes from Bacillus thuringiensis-susceptible and -resistant strains of sugarcane borer, Diatraea saccharalis

Molecular characterization and RNA interference of three midgut aminopeptidase N isozymes from Bacillus thuringiensis-susceptible and -resistant strains of sugarcane borer, Diatraea saccharalis

A barley cysteine-proteinase inhibitor reduces the performance of two aphid species in artificial diets and transgenic Arabidopsis plants

Prey mediated effects of Bt maize on fitness and digestive physiology of the red spider mite predator Stethorus punctillum Weise (Coleoptera: Coccinellidae)

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