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- 저자 : Yu Okamura (검색결과 1 건)
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Chemo-analytical case study in an insect: Salicinoid metabolism in Cerura vinula
Florian Schnurrer,Yu Okamura,Juil Kim,Heiko Vogel,Nathalie Wielsch,Christian Paetz 한국응용곤충학회 2019 한국응용곤충학회 학술대회논문집 Vol.2019 No.04
Plants of the Salicacea genus have in common that they utilize phenolic glycosides, so called salicinoids, as protection against leaf chewing herbivores. The caterpillar Cerura vinula only feeds on poplar and willow trees that belong to the family of Salicacea. For this reason, Cerura vinula is viewed as a specialist herbivore that is adapted to the salicinoid defense. Taking advantage of chemical and biochemical techniques we want to trace the salicinoid metabolism pathway, the location of transformation and the involved enzymes. To gain insights into the salicinoid metabolism the caterpillars were put on a specially designed diet. It consisted either of one salicinoid in large excess or one 13C labelled compound, applied together with fresh poplar leaves. As a next step, we are going to identify the new compounds formed in the caterpillar by MS and NMR techniques. To determine the location and mechanism of the transformation we started with a dissection of the caterpillar and a check of the tissue pH value. Afterwards, we incubated the midgut, hindgut and salivary gland tissues with salicin as model substrate and analyzed the transformation products by LC-MS. The transformation products resulted from deglycosilation of the substrate and consecutive oxidation and conjugation of the aglycon. With the gained knowledge we then aimed to identify the enzymes, which are involved in the metabolism. We successfully proofed the existence of glucosidases in the midgut by isoelectric focusing and incubation of the gel with the model substrate 4-methylumbelliferyl β-D-glucopyranosid. Additionally, we performed RNA sequencing on the caterpillar tissue. The transcripts and enzymes involved in the salicinoid metabolism are currently analyzed. During our studies we could determine the mid gut as the place for the deglycosilaton, oxidation and conjugation of salicinoids. Further we identified a new for Cerura vinula undescribed metabolite and proofed the presence of β-glucosidases in the midgut.
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