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- 저자 : F. BERAN (검색결과 4 건)
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F. BERAN,T. Sporer,S.-J. Ahn,F. Betzin,C. Paetz,G. Kunert,M. Reichelt 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10
Many insects are able to feed on crucifers despite the presence of a potent activated defense system known as the mustard oil bomb. In damaged tissue, mustard oil glucosides (glucosinolates) are hydrolyzed by the enzyme myrosinase to form toxic mustard oils (isothiocyanates). Here, we analyzed how the the cabbage stem flea beetle Psylliodes chrysocephala, a key pest of oilseed rape, copes with this chemical defense. First, we found that P. chrysocephala prevents the activation of ingested glucosinolates by two different strategies, a) by sequestering glucosinolates and b) by converting glucosinolates to desulfo-glucosinolates. Our next aim was to identify the sulfatase enzyme(s) responsible for the detoxification of glucosinolates in P. chrysocephala. Nine arylsulfatase-like genes were identified in the transcriptome of P. chrysocephala, and five of them showed glucosinolate sulfatase activity upon heterologous expression in Sf9 cells. By using RNAi, we confirmed that PcGSS1 and PcGSS2 are active towards benzenic and indolic glucosinolates in P. chrysocephala adults in vivo. However, in feeding experiments, the proportion of sequestered and desulfated glucosinolates ranged from 26 to 35% which suggests that these strategies alone are likely not sufficient to overcome the chemical plant defense. Indeed, P. chrysocephala additionally conjugates isothiocyanates to glutathione and metabolizes them via the conserved mercapturic acid pathway. In summary, the cabbage stem flea beetle avoids isothiocyanate formation by specialized strategies (sequestration and desulfation), but also relies on a conserved detoxification pathway to prevent toxicity of isothiocyanates.
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How insect herbivores hijack plant chemical defenses for their own defense against natural enemies
F. Beran,Z.-L. Yang,T. Sporer 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10
Many herbivorous insects sequester plant defense compounds from their host plants to protect themselves from natural enemies. In plants, these defense compounds are often stored as protoxins separated from their activating enzymes. A well-known example is the glucosinolate-myrosinase defense system in plants of the order Brassicales. When plant tissue is ingested by herbivores, glucosinolates are hydrolyzed by the enzyme myrosinase to form highly reactive isothiocyanates. We previously reported that flea beetles of the genus Phyllotreta selectively sequester high amounts of glucosinolates from their crucifer host plants, and convergently evolved their own myrosinase which enables them to utilize sequestered glucosinolates for their own purposes (Beran et al., 2014). The presence of intact glucosinolates in these beetles suggests that despite tissue damage, ingested glucosinolates are not activated by the plant myrosinase. Rapid and efficient glucosinolate uptake from the gut lumen into gut epithelial cells can prevent hydrolysis and thus might be crucial to overcome this activated plant defense. We use the horseradish flea beetle Phyllotreta armoraciae as a model to study the molecular basis of sequestration in insects. In short-term feeding experiments, we showed that ingested glucosinolates are rapidly sequestered in the foregut. To identify the transporters that mediate glucosinolate import from the foregut lumen into gut epithelial cells, we focused on the MFS transporter family, which is known to transport a wide range of substrates. A phylogenetic analysis of putative MFS transporter sequences identified in P. armoraciae and other beetles revealed several specifically expanded clades in P. armoraciae. Out of 21 candidate genes that were heterologously expressed in Sf9 cells, nine showed glucosinolate transport activity in vitro. Interestingly, most candidate genes were exclusively expressed in the malpighian tubules, and two genes were additionally expressed in the foregut. We currently elucidate the function of these transporters in glucosinolate sequestration in vivo using RNAi. To better understand the function of sequestered glucosinolates, we performed bioassays with P. armoraciae larvae and the generalist predatory ladybird Harmonia axyridis. Upon predator attack, P. armoraciae larvae emitted high amounts of isothiocyanates and ladybird larvae stopped feeding within a few seconds and were highly irritated. However, silencing myrosinase gene expression in P. armoraciae larvae led to increased mortality compared to control larvae in survival assays with ladybird larvae. Our results demonstrate how Phyllotreta use plant defense metabolites to defend themselves against predators.
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