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GUT PROTEINASE ACTIVITY IN INSECT PESTS OF CANOLA1

Published online by Cambridge University Press:  31 May 2012

Robert T. Rymerson  and
Robert P. Bodnaryk
Robert T. Rymerson
Affiliation:
Agriculture and Agri-Food Canada, Research Station, 195 Dafoe Road, Winnipeg, Manitoba, Canada R3T 2M9
Robert P. Bodnaryk
Affiliation:
Agriculture and Agri-Food Canada, Research Station, 195 Dafoe Road, Winnipeg, Manitoba, Canada R3T 2M9
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Abstract

The digestive proteinases of three important pests of canola, Brassica napus L. and B. rapa L., in western Canada were characterized by assessing the proteolytic activity of homogenates of their midguts against azocasein or azoalbumin at various pH levels and in the presence of diagnostic proteinase inhibitors. The midgut of larvae of the bertha armyworm, Mamestra configurata Wlk., had maximum proteolytic activity at pH 10.5 which was inhibited 45–60% by serine proteinase inhibitors such as the soybean trypsin inhibitor. The midgut of larvae of the diamondback moth, Plutella xylostella L., had maximum proteolytic activity at pH 10 which was inhibited 56–75% by serine proteinase inhibitors. The two lepidopterans thus use a serine-like proteinase in digestion. The midgut of adults of the flea beetle, Phyllotreta cruciferae Goeze, exhibited maximum proteolytic activity at pH 5 which was inhibited 33–61% by specific cysteine proteinase inhibitors such as cystatin and trans-epoxysuccinyl-L-leucylamido (4-guanidino)-butane (E-64) and was activated strongly by L-cysteine. Aspartic proteinase inhibitors such as pepstatin A also decreased proteolytic activity by 21–50%. Serine proteinase inhibitors were without effect. Therefore, P. cruciferae appears to use both cysteine- and aspartic-like proteinases in digestion. Cotyledons and first true leaves of canola, B. napus cv. Westar, contained inhibitory activity against serine, cysteine, and aspartic proteinases when tested against bovine trypsin, papain, or porcine pepsin, but the level of antiproteinase activity is insufficient to provide significant resistance against any of these pests.

Résumé

Les protéinases digestives de trois parasites importants du colza, Brassica napus L. et B. rapa L., dans l’ouest du Canada ont été analysées par évaluation de l’activité protéolytique d’homogénats de leur intestin moyen contre l’azocaséine ou l’azoalbumine à différents pH et en présence d’inhibiteurs spécifiques des protéinases. Dans l’intestin moyen des larves de la Légionnaire bertha, Mamestra configurata Wlk., l’activité protéolytique est maximale à pH 10,5 et elle est inhibée de 45–60% par les inhibiteurs de la serine protéinases, tels l’inhibiteur de la trypsine de la fève soja. Chez les larves de la Fausse Teigne des crucifères, Plutella xylostella L., l’activité protéolytique est maximale à pH 10 et elle est inhibée à 56–75% par les inhibiteurs de la sérine protéinase. Les deux lépidoptères possèdent donc une protéinase digestive de type sérine. L’activité protéolytique maximale dans l’intestin moyen des adultes de Phyllotreta cruciferae Goeze, l’Altise des crucifères, se produit à pH 5 et est inhibée à 33–61% par des inhibiteurs spécifiques de la cystéine protéinase, tels la cystatine et le trans-époxysuccinyl-L-leucylamido (4-guanidino)-butane (E-64); elle est fortement activée par la L-cystéine. Les inhibiteurs de la protéinase aspartique, tels la pepstatine A, diminuent également l’activité protéolytique de 21–50%. Les inhibiteurs de la sérine protéinase sont restés sans effet. Les adultes de P. cruciferae utilisent donc des protéinases de types cystéine et de type aspartique dans leur digestion. Les cotylédons et les premières vraies feuilles du colza B. napus cv. Westar ont une activité inhibitrice contre la sérine protéinases, la cystéine protéinase et la protéinase aspartique en présence de trypsine de bovin, de papaïne ou de pepsine de porc, mais l’intensité de l’activité antiprotéinase est insuffisante pour procurer à la plante une résistance de quelque importance contre ces parasites.

[Traduit par la Rédaction]

Type
Articles
Information
The Canadian Entomologist , Volume 127 , Issue 1 , February 1995 , pp. 41 - 48
Copyright
Copyright © Entomological Society of Canada 1995

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