The flavonoid pattern of larvae of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) reared on the leaves of tronchuda
cabbage was analysed by HPLC-DAD-MS/MS-ESI. Twenty flavonoids were identified or characterised, namely 16 kaempferol
and four quercetin derivatives. Kaempferol 3-O-sophoroside, a minor component of tronchuda cabbage, was found to be the main component
in P. brassicae (15.8%). Apart from this, only two other flavonoids present in significant amounts in tronchuda cabbage (kaempferol
3-O-sophoroside-7-O-glucoside and kaempferol 3-O-sophoroside-7-O-sophoroside) were found in the larvae. The larvae have high
amounts of quercetin derivatives (18.5%), which were present only in trace amounts in tronchuda cabbage extracts, suggesting that
P. brassicae is able to selectively sequester these flavonoids. The occurrence of a high content of flavonoids not detectable in tronchuda
cabbage extracts indicates that P. brassicae larvae are able to metabolize dietary flavonoids.
The phenolic profiles of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) at different development stages (larvae, exuviae, and butterfly), its excrements, and its host plant Brassica rapa
var. rapa L. were determined by high performance liquid chromatography- diode-array detector-mass
spectrometry/mass spectrometry-electrospray ionization (HPLC-DAD-MS/MS-ESI). Twenty-five
acylated and nonacylated flavonoid glycosides and ferulic and sinapic acids were identified in host
plant, from which only 12 compounds were found in the excrements. In addition, the excrements
showed the presence of sulfate flavonoids and other flavonoid glycosides that were not detected in
the leaves. In the larvae kept without food for 12 h, only 3 compounds common to the plant material
and 2 others, also present in the excrements, were characterized. The results indicate that deacylation,
deglycosylation, and sulfating steps are involved in the metabolic process of P. brassicae and that
its excrements may constitute a promising source of bioactive compounds, which could be used to
take profit of this common pest of Brassica cultures.
The antioxidant potential of an aqueous extract obtained from Pieris brassicae larvae reared on
Brassica oleracea L. var. costata DC was evaluated against 2,2-diphenyl-1-picrylhydrazyl radical and
several reactive oxygen species. The results revealed an effective concentration-dependent protective
activity against superoxide and hydroxyl radicals, being superior to that of the host plant. In addition,
the larvae extract also exhibited a strong inhibitory effect on xanthine oxidase that was not observed
for B. oleracea var. costata. A weak scavenging ability was noticed for hypochlorous acid. Several
phenolic compounds with complex chemical structures that are hard to synthesize in the laboratory
were found in P. brassicae extract. This is the first time that an insect has been tested for its xanthine
oxidase inhibitory capacity, which proved to be very high. These findings are interesting considering
that they can be used by food or pharmaceutical industries to prevent the oxidation of their products,
to increase the dietary supply of antioxidants, or for prevention of free radical-mediated diseases,
Phenolic and organic acid profiles of aqueous extracts from Pieris brassicae material and the host kale
(Brassica oleracea L. var. acephala) leaves were determined by HPLC/UV–DAD/MSn-ESI and HPLC–UV,
respectively. The identified phenolics included acylated and nonacylated flavonoid glycosides, hydroxycinnamic
acyl gentiobiosides, and sulphate phenolics. Kale exhibited the highest content (11 g/kg lyophilized
extract), while no phenolics were identified in the butterflies or exuviae. Nine different organic
acids were characterized in the materials, with kale showing the highest amount (112 g/kg lyophilized
extract). With the exception of the exuviae extract, the rest were screened for bioactivity. Using spectrophotometric
microassays, all exhibited antiradical capacity against DPPH and NO in a concentrationdependent
way, whereas only kale and excrement extracts were active against superoxide. All displayed
activity on intestinal smooth muscle, albeit with distinct relaxation–contraction profiles. Larvae and butterfly
extracts were more efficacious for intestinal relaxation than was kale extract, whereas excrement
extract evoked only contractions, thus evidencing their different compositions. Collectively, these results
show that P. brassicae sequesters and metabolizes kale’s phenolic compounds. Moreover...
Changes in the phenolics composition of Pieris brassicae larvae fasted for distinct periods (1, 2, 4,
6, and 8 h) and their excrements and of Brassica oleracea L. var. costata DC leaves were
determined by high-pressure liquid chromatography/UV-photo diode array detector/mass spectrometry-
electrospray ionization. This is the first report following phenolics’ metabolism by P.
brassicae through time. The results evidence that P. brassicae sequesters and metabolizes the
phenolic compounds from the host plant. In a general way, deacylation was the main metabolic
reaction that took place, but deglycosylation and sulfate conjugation reactions also occur. Additionally,
several kaempferol derivatives containing rhamnose, which is not common in Brassica, were
found in the host plant. Attending to the bioactivities recognized for the type of identified compounds,
the different materials may constitute an interesting source of bioactive compounds, namely, of
highly glycosylated and acylated kaempferol and quercetin derivatives, constituting an economic
advantage for producers who have great losses caused by this pest. In addition, a deeper
understanding of phenolics metabolism in insects was pursued.
Seeds of Brassica oleracea var. acephala (kale) were analyzed by HPLC/UV-PAD/MSn-ESI.
Several phenolic acids and flavonol derivatives were identified. The seeds of this B. oleracea
variety exhibited more flavonol derivatives than those of tronchuda cabbage (Brassica oleracea var.
costata), also characterized in this paper. Quercetin and isorhamnetin derivatives were found only in
kale seeds. Oxalic, aconitic, citric, pyruvic, malic, quinic, shikimic, and fumaric acids were the
organic acids present in these matrices, malic acid being predominant in kale and citric acid in
tronchuda cabbage seeds. Acetylcholinesterase (AChE) inhibitory activity was determined in
aqueous extracts from both seeds. Kale leaves and butterflies, larvae, and excrements of Pieris
brassicae reared on kale were also evaluated. Kale seeds were the most effective AChE inhibitor,
followed by tronchuda cabbage seeds and kale leaves. With regard to P. brassicae material,
excrements exhibited stronger inhibitory capacity. These results may be explained by the presence
of sinapine, an analogue of acetylcholine, only in seed materials. A strong concentration-dependent
antioxidant capacity against DPPH, nitric oxide, and superoxide radicals was observed for kale
The influence of Pieris brassicae feeding on kale was monitored, by evaluating its effect on the volatiles
released by the plant through time. This is the first study applying headspace solid-phase microextraction
(HS-SPME) and gas chromatography/ion trap-mass spectrometry to an isolated insect, as most studies
analyse the insect–plant system as a whole, being unable to evaluate the contribution of the insect itself.
Substantial differences were noticed between the volatiles composition of kale before and after the
insect’s attack. More than 60 compounds were found, including terpenes, lipoxygenase pathway by-products,
ketones, norisoprenoids, etc. After insect attack, l-camphor, sabinene and a-thujene were found and
limonene and eucalyptol suffered a noticeable increase. A considerable rise in (Z)-3-hexenyl acetate was
also observed. In vivo accumulation of limonene and camphor by the insect was detected. The findings
contribute to the knowledge of the ecological interactions between the two species.
Fonte: American Society of PharmacognosyPublicador: American Society of Pharmacognosy
Tipo: Conferência ou Objeto de Conferência
Relevância na Pesquisa
Pieris brassicae is a specialist on crucifers and its interactions with some of its host plants have
been increasingly studied given their importance in pest management, chemical ecology and
entomology. Plants are known to respond to insect attack by releasing volatiles that can either
warn neighbour plants to the presence of a predator or attract insect parasitoids, thus reducing
the attack efficiency. The influence of P. brassicae feeding on Brassica oleracea L. var. acephala
(kale) was investigated, namely, its effect in the volatiles released by the plant through time.
Substantial differences, mainly quantitative, were found between the volatiles composition of B.
oleracea var. acephala before the attack and after insect feeding. More than 40 compounds were
found, between terpenes (monoterpenes and sesquiterpenes) and lipoxygenase pathway byproducts
(alcohols and aldehydes), ketones, norisoprenoids, among others. After insect attack,
monoterpenes like limonene, eucalyptol, camphor and caryophyllene increased noticeably.
These compounds have been regarded as markers in local tissue responses and in this study an
increase in their concentration was detected at 1 and 4 hours after feeding, with subsequent
reduction after 8 hours. Menthol...
A lagarta da couve, Pieris brassicae (L.), é uma praga que ataca com frequência diferentes
espécies de Brassica. Nos estádios de desenvolvimento mais avançados, esta lagarta apresenta
uma forte voracidade, consumindo grande quantidade de alimento e produzindo quantidade
apreciável de excrementos. Nos últimos anos a procura de novas moléculas com interesse
farmacêutico tem tido grande impulso, constituindo os produtos naturais fontes importantes
desses compostos. Assim, o presente trabalho reúne informação sobre a possibilidade da P.
brassica ser fornecedora de compostos bioactivos.
IV Encontro Nacional de Protecção Integrada, 3–4 Outubro, 1997, Angra do Heroísmo, Açores.; Apanteles militaris (Walsh) é o mais importante parasitóide larvar de Mythimna
unipuncta (Haworth) em todas as ilhas dos Açores. Pretendendo-se saber do modo de
actuação deste parasitóide larvícola, procedeu-se ao estudo semanal da sua abundância e à do seu hospedeiro natural. Numa tentativa de compreender onde se encontra este parasitóide durante o Invermo, realizámos ensaios de parasitismo, em condições controladas, utilizando como hospedeiro larvas de Noctua pronuba L., Peridroma saucia (Hb), Xestia c-nigrum (L.), Spodoptera litoralis B., Agrotis ipsilon (Hüf.), Heliothis armigera (Hb.),
Sesamia nonagrioides (Lef.), Autographa gamma (L.) e Pieris brassicae azorensis Rebel. Estes Lepidópteros foram seleccionados por existirem nos biótopos que são ocupados por M. unipuncta, apresentarem uma densidade importante nas condições ambientais dos Açores ou por serem hospedeiros de parasitóides pertencentes ao mesmo género. Para além de M. unipuncta, apenas S. nonagrioides e A. gamma apresentaram uma percentagem muito reduzida de larvas efectivamente parasitadas, 10 e 12,5%, respectivamente. Assim, a população açoreana de A. militaris apresenta uma grande especificidade em relação ao seu hospedeiro natural.
III Simpósio Fauna e Flora das Ilhas Atlânticas / Symposium Fauna and Flora of the Atlantic Islands, Universidade dos Açores, Ponta Delgada (Açores), 21 - 25 September, 1998.; A influência dos factores abióticos sobre o desenvolvimento de espécies de Lepidópteros, nomeadamente no estado larvar, foi e será objecto de numerosos estudos, contribuindo assim para o conhecimento dos seus
limites teóricos, duração/velocidade de desenvolvimento e percentagem de
mortalidade no estado larvar. Analisou-se, em condições controladas, a duração do desenvolvimento de cada fase do ciclo de vida das diferentes espécies em estudo. Estes Lepidópteros foram seleccionados de acordo com os níveis de abundância registados nas armadilhas luminosas. Para cada espécie, observou-se diferentes tipos larvares (5, 6, 7 e 8). Nas espécies
Mythimna unipuncta (HAW.), Agrotis ipsilon(HUF.), Xestia c-nigrum (L.) e Heliothis armigera (HBN.) a duração do desenvolvimento dos diferenles tipos larvares foi significativarnente diferente. 0 desenvolvimento total foi significativamente diferente nas especies M. unipuncta, X. c-nigrum e A. ipsilon. A proporção verificada para as tipos larvares dependeu da alimentação e, provavelmente da sua informação genética.
Cabbage plants respond to caterpillar (Pieris brassicae) herbivory by releasing a mixture of volatiles that makes them highly attractive to parasitic wasps (Cotesia glomerata) that attack the herbivores. Cabbage leaves that are artificially damaged and subsequently treated with gut regurgitant of P. brassicae caterpillars release a volatile blend similar to that of herbivore-damaged plants. We demonstrate the presence of beta-glucosidase in P. brassicae regurgitant. Leaves treated with commercial beta-glucosidase (from almonds) release a volatile blend similar to that of leaves treated with P. brassicae regurgitant. In a flight bioassay, leaves treated with almond beta-glucosidase are highly attractive to the parasitic wasp C. glomerata. Furthermore, the wasps do not discriminate between cabbage leaves treated with almond beta-glucosidase and leaves treated with larval regurgitant. beta-Glucosidase was also recorded in cabbage leaf extract, but this is not as effective as caterpillar beta-glucosidase in eliciting the volatile production. Caterpillars that feed on a beta-glucosidase-free diet secrete the enzyme, and their regurgitant is an effective elicitor of the plant response. These findings show that beta-glucosidase is a P. brassicae-secreted elicitor of the defense response of cabbage plants to herbivore injury...
Parasitoid wasps lay female eggs or a high proportion of female eggs in favourable host insects because female wasps require many more resources during their development. Many studies have tested the effects of host physiological status on the sex allocation of parasitoids, but few have attempted to test the effects of host behavioural traits. Cotesia glomerata is a gregarious parasitoid wasp that lays eggs in caterpillars of pierid butterflies. The brood sex ratio in C. glomerata females that attacked aggressive host caterpillars was compared with that in females that attacked less aggressive hosts. The male ratio was higher when C. glomerata attacked aggressive Pieris brassicae caterpillars than when it attacked less aggressive Pieris rapae crucivora caterpillars. However, when C. glomerata females were induced to oviposit in anaesthetized P. brassicae caterpillars, the male ratio in their offspring was significantly lower than when they attacked unanaesthetized caterpillars. C. glomerata was attacked by aggressive host caterpillars during oviposition bouts. It is likely that this aggressive host behaviour disturbed the fertilization process in ovipositing C. glomerata females. These results suggest that a behavioural defence by host caterpillars affects sex allocation in the parasitoid wasp C. glomerata.
Porphobilinogen represents a key building block of tetrapyrroles serving as functional ligands of many vitally important proteins. Here we report the first purification of porphobilinogen synthase (PBGS) from whole insects by sequentially employing two modes of native electrophoresis on polyacrylamide gels subsequent to more conventional procedures. Using adults of Pieris brassicae L. (Lepidoptera: Pieridae) we achieved ∼10,000-fold purification with final yields of up to 25% of electrophoretically pure PBGS with a specific activity of ∼160 µmol PBG h-1 mg-1 at 37°C and an affinity of 0.36 mM to its substrate 5-aminolevulinic acid. Enzyme activity was inhibited by the substrate mimics, levulinic acid and succinylacetone, and by chelating agents. PBGS behaved as a relatively heat-stable octameric complex of 292.3 kDa composed of 36.5 kDa subunits. Most general features of this insect PBGS were comparable to those published for other animal PBGS enzymes, while remarkable differences were found to the reported recombinant Drosophila enzyme. Moreover, rabbit antiserum directed against purified Pieris PBGS revealed significant immunological differences among insect PBGS enzymes from a wide range of orders contrasting to the overall evolutionary conserved features of this enzyme.
Superparasitism occurs in Cotesia glomerata L. (Hymenoptera: Braconidae), a gregarious endoparasitoid of Pieris spp. (Lepidoptera: Pieridae). The responses of Pieris brassicae L. larvae to superparasitism were examined in order to elucidate the ecological significance of this behaviour. Models of tritrophic interactions often imply that attraction of herbivore natural enemies by the plant constitutes a defence. Parasitoid attack on herbivores is assumed to result in a reduction in herbivory and or an increase in plant fitness. Coupled with the active involvement of the plant in producing signals, this can be seen as an indirect mediation of wound induced defence. The results show that superparasitism of P. brassicae by the parasitoid C. glomerata reduced survivorship but increased food consumption and weight growth in P. brassicae larvae. The duration of host larval development was found prolonged as the number of oviposition increased and superparasitized larvae (three to five time parasitized) grew slower than unparasitized larvae or larvae parasitized one or two times.
Pieris brassicae L. is a serious pest of cultivated crucifers in several parts of the world. Larvae of P. brassicae also feed prolifically on garden nasturtium (Tropaeolum majus L., of the family Tropaeolaceae). Proteolytic digestion was studied in larvae feeding on multiple hosts. Fourth instars were collected from cauliflower fields before transfer onto detached, aerial tissues of selected host plants in the lab. Variable levels of midgut proteases were detected in larvae fed on different hosts using protein substrates (casein and recombinant RBCL cloned from cauliflower) and diagnostic, synthetic substrates. Qualitative changes in midgut trypsin activities and quantitative changes in midgut chymotrypsin activities were implicated in physiological adaptation of larvae transferred to T. majus. Midgut proteolytic activities were inhibited to different extents by serine protease inhibitors, including putative trypsin inhibitors isolated from herbivore-attacked and herbivore-free leaves of cauliflower (CfTI) and T. majus (TpTI). Transfer of larvae to T. majus significantly influenced feeding parameters but not necessarily when transferred to different tissues of the same host. Results obtained are relevant for devising sustainable pest management strategies...
The use of diagnostic polymerase chain reaction (PCR) is a valuable approach to study host–parasitoid interactions. It circumvents problems inherent to rearing parasitoids from field-collected hosts or host dissection. Here, we present a PCR-based detection method for Diadegma semiclausum, Cotesia glomerata, and Cotesia rubecula, which are key-parasitoids of the three lepidopteran cabbage pests Plutella xylostella, Pieris brassicae, and Pieris rapae, respectively. Primer pairs for the three parasitoid species and Pl. xylostella were developed: they were used either separately in singleplex PCR or combined in multiplex PCR to (1) screen simultaneously for the two Cotesia species or to (2) detect the parasitoid D. semiclausum and identify its host Pl. xylostella in one PCR. The new idea to simultaneously identify parasitoid and host by molecular markers is useful when the host, in our case early-instar larvae of Pl. xylostella, is morphologically difficult to distinguish from other host species also occurring in the same habitat. Concentration–response trials revealed comparable detection sensitivity of singleplex and multiplex PCR, with detection limits ranging from 0.03 to 2.2 pg of parasitoid DNA/μl PCR. Furthermore, the different developmental time of immature D. semiclausum and C. glomerata did not influence parasitoid detection success in either assay type. Based on multiplex PCR screening of field-collected caterpillars...
10 pages, 6 figures, 3 tables.; The phenolic profiles of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) at different development stages (larvae, exuviae, and butterfly), its excrements, and its host plant Brassica rapa var. rapa L. were determined by high performance liquid chromatography− diode-array detector−mass spectrometry/mass spectrometry−electrospray ionization (HPLC-DAD-MS/MS-ESI). Twenty-five acylated and nonacylated flavonoid glycosides and ferulic and sinapic acids were identified in host plant, from which only 12 compounds were found in the excrements. In addition, the excrements showed the presence of sulfate flavonoids and other flavonoid glycosides that were not detected in the leaves. In the larvae kept without food for 12 h, only 3 compounds common to the plant material and 2 others, also present in the excrements, were characterized. The results indicate that deacylation, deglycosylation, and sulfating steps are involved in the metabolic process of P. brassicae and that its excrements may constitute a promising source of bioactive compounds, which could be used to take profit of this common pest of Brassica cultures.; The authors are grateful to Fundação para a Ciência e a Tecnologia (PTDC/AGR-AAM/64150/2006) for financial support
of this work.; Peer reviewed
7 pages, 4 tables, 4 figures.-- El PDF es la copia de autor.; The flavonoid pattern of larvae of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) reared on the leaves of tronchuda cabbage was analysed by HPLC-DAD-MS/MS-ESI. Twenty flavonoids were identified or characterised, namely 16 kaempferol and four quercetin derivatives. Kaempferol 3-O-sophoroside, a minor component of tronchuda cabbage, was found to be the main component in P. brassicae (15.8%). Apart from this, only two other flavonoids present in significant amounts in tronchuda cabbage (kaempferol 3-O-sophoroside-7-O-glucoside and kaempferol 3-O-sophoroside-7-O-sophoroside) were found in the larvae. The larvae have high amounts of quercetin derivatives (18.5%), which were present only in trace amounts in tronchuda cabbage extracts, suggesting that P. brassicae is able to selectively sequester these flavonoids. The occurrence of a high content of flavonoids not detectable in tronchuda cabbage extracts indicates that P. brassicae larvae are able to metabolize dietary flavonoids.; [Graphical abstract]: The flavonoid pattern of larvae of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) reared on the leaves of tronchuda cabbage was analysed by HPLC-DAD-MS/MS-ESI. Twenty kaempferol and quercetin derivatives were identified or characterised. The results suggest that P. brassicae is able to selectively sequester quercetin derivatives and to metabolize dietary flavonoids.; The authors are grateful to Fundaçäo para a Ciência e Tecnologia (PTDC/AGR-AAM/64150/2006) for financial support of this work.; Peer reviewed
The main function of floral emissions of volatile organic compounds (VOCs) in entomophilous plants is to attract pollinators. Floral blends, however, can also contain volatile compounds with defensive functions. These defensive volatiles are specifically emitted when plants are attacked by pathogens or herbivores. We characterized the changes in the floral emissions of Diplotaxis erucoides induced by folivory and florivory by Pieris brassicae. Plants were continually subjected to folivory, florivory and folivory + florivory treatments for two days. We measured floral emissions with proton transfer reaction/mass spectroscopy (PTR-MS) at different times during the application of the treatments. The emissions of methanol, ethyl acetate and another compound, likely 3-butenenitrile, increased significantly in response to florivory. Methanol and 3-butenenitrile increased 2.4- and 26-fold, respectively, in response to the florivory treatment. Methanol, 3-butenenitrile and ethyl acetate increased 3-, 100- and 9-fold, respectively, in response to the folivory + florivory treatment. Folivory alone had no detectable effect on floral emissions. All VOC emissions began immediately after attack, with no evidence of delayed induction in any of the treatments. Folivory and florivory had a synergistic effect when applied together...