Characterisation of digestive protease in the rose sawfly,Arge rosaeLinnaeus (Hymenoptera: Argidae)

Abstract

Insect midgut proteases are excellent targets for insecticidal agents such as protease inhibitors. These inhibitors are used for producing transgenic plants, resistant to pests. For achieving this goal, it is necessary to find the nature of specific proteases and their properties for adopting possible pest management procedure. Therefore, characterisation of the enzymes in the gut of the rose sawfly, Arge rosae (Hymenoptera: Argidae), responsible for proteolysis, was performed using a range of synthetic substrates and specific inhibitors. The optimum conditions for general proteases and trypsin were achieved at pH 10. The highest activity for general proteases was obtained at a temperature of 458C. The use of specific inhibitors and SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) provided evidence to suggest that most of the proteases belonged to the serine group because of high inhibitory effect of phenyl methane sulfonyl fluoride on total proteolytic activity. Also, inhibition assays and zymogram analysis showed that metalloproteases are present in A. rosae digestive system. These results indicated that A. rosae larvae mainly used serine proteases for protein digestion, with chymotrypsin as the dominant form. The kinetic parameters of trypsin-like proteases using N-benzoyl-DL-arg-p-nitroanilide as substrate indicated that the K m and V max values of trypsin in the gut of the fifth instar larvae were 730 + 17.3 mM and 456 + 13.85 nmol min 71 mg 71 protein, respectively.

New parasitoid records of Arge rosae (Linnaeus, 1758) (Hymenoptera: Argidae) from Diyarbakır province: Tetrastichus hylotomarum (Bouché, 1834) (Hymenoptera: Eulophidae) and Boethus thoracicus (Giraud, 1872) (Hymenoptera: Ichneumonidae)

“…Arge rosae (Linnaeus, 1758) (Hymenoptera: Argidae) (Figure 2) Distribution in world: Bulgaria (Margina et al, 1999), Persia (Sahragard and Heydari, 2001;Sharifi et al, 2012;Khosravi et al, 2015) Distribution in Türkiye: Antalya, Diyarbakır, Isparta, İstanbul, İzmir, Manisa (Zümreoğlu, 1972;Oğurlu et al, 1996;Özbek and Çalmaşur, 2005;Demirözer and Karaca, 2011;Bolu et al, 2021;Baki et al, 2021). Host plants: Rosa canina Linnaeus, 1753, R. damascena Miller (Margina et al, 1999;Özbek and Çalmaşur, 2005;Bolu et al, 2021).…”

“…Material examined: 10♂♂, 10♀♀ (multiple samples), Locality: Diyarbakır (Sur district), 37°53'31.8"N, 40°16'22.2"E, 666 m. As a result; it was determined that the NC of turfgrass can be estimated with similar prediction performance using both hand-held NDVI meter and hand-held chromameter in a more objective and economic way. Arge rosae is one of the pests (Figure 1) which causes economic damage to roses (Sharifi et al, 2012;Bolu et al, 2021;Baki et al, 2021). This damage occurs in two ways.…”

“…Many insect species have been reported to cause damage to different parts of the rose plant. Amongs the rose pests, Arge rosae (Hymenoptera: Argidae) causes significant damage to the rose plant in worldwide including Diyarbakır province of Türkiye (Sharifi et al, 2012;Khosravi et al, 2015;Bolu et al, 2021). The Ichneumonidae is a widespread parasitoid family attacking a large host spectrum, including species of Lepidoptera, Diptera, Coleoptera and Hymenoptera.…”

Identification and characterisation of gut proteases in the fig tree skeletoniser moth, Choreutis nemorana Hübner (Lepidoptera: Choreutidae)

“…BApNA was used as substrate at a final concentration range of 0.0156 to 1mM. The experiments were performed in triplicate (Sharifi et al 2012b) Effect of protease inhibitors. To analyse the effect of protease inhibitors, 10 µl of each freshly prepared protease inhibitor including 5mM PMSF, 1mM TLCK, 1mM TPCK, 2mM EDTA, and 5mM iodoacetate were added to 15 µl enzyme.…”

Biochemical characterization and structural analysis of trypsin from Plodia interpunctella midgut: implication of determinants in extremely alkaline pH activity profile

“…Proteases provide essential amino acids for insect pests by hydrolyzing ingested proteins and, consequently, they can influence both the insect growth and development. Therefore, targeting digestive enzymes with appropriate inhibitors could be a novel alternative for chemical pesticides (Sharifi et al, 2012). The inhibition of insect proteases may also lead the hyperproduction of enzymes and the depletion of the sulphur amino acid pool, thus weakening the pest (Shukle & Murdock, 1983).…”