Antifeedant activities of crude seed extracts of tropical African spices against Spodoptera littoralis (Lepidoptera: Noctuidae)

The antifeedant activities of Piper guineense Schum et Thonn (Piperaceae), Aframomum melegueta (Rosk) K. Schum (Zingiberaceae), Aframomum citratum (Pareira) K. Schum (Zingiberaceae) and Afrostyrax kamerunensis Perkins and Gilg (Huaceae) seed extracts were investigated in laboratory dual- and no-choice bioassays using third-instar Spodoptera littoralis (Boisduval) larvae. In the dual-choice test, the hexane and methanol extracts of A. melegueta showed potent dose-dependent antifeedant activity at concentrations of ≥300 ppm and the water extract at ≥500 ppm, as illustrated by significantly lower leaf consumptions. Aframomum citratum methanol and water extracts exhibited antifeedant activity at ≥300 and ≥1000 ppm, respectively, but the hexane and ethanol extracts did not affect feeding at any concentration. Piper guineense ethanol and water extracts showed dose-dependent antifeedant effects at ≥300 and ≥500 ppm, respectively, and the methanol extract was active only at 1000 ppm. None of the extracts of the highly aromatic A. kamerunensis exhibited antifeedant activity at any of the tested concentrations. In the no-choice bioassays, extracts with antifeedant activity in the dual-choice tests also showed dose-dependent feeding inhibition. The hexane and methanol extracts of A. melegueta were effective in the no-choice tests at ≥100 and ≥500 ppm, respectively, and the water extract at ≥300 ppm. Similarly, the A. citratum water and methanol extracts were active at ≥500 ppm and the P. guineense water and ethanol extracts at ≥100 ppm. GC/MS chromatography of A. melegueta hexane and methanol extracts revealed volatile constituents with known anti-insect activity. The hexane and methanol extracts of A. melegueta, the methanol extract of A. citratum and the water and ethanol extracts of P. guineense may have potential for use by subsistence farmers.

Selection for cuticular melanism reveals immune function and life-history trade-offs in Spodoptera littoralis

Several insect species show an increase in cuticular melanism in response to high densities. In some species, there is evidence that this melanism is correlated with an up-regulation of certain immune system components, particularly phenoloxidase (PO) activity, and with the down-regulation of lysozyme activity, suggesting a trade-off between the two traits. As melanism has a genetic component, we selected both melanic and nonmelanic lines of the phase-polyphenic lepidopteran, Spodoptera littoralis, in order to test for a causative genetic link between melanism, PO activity and lysozyme activity, and to establish if there are any life-history costs associated with the melanic response. We found that, in fact, melanic lines had lower PO activity and higher lysozyme activity than nonmelanic lines, confirming a genetic trade-off between the two immune responses, but also indicating a genetic trade-off between melanism and PO activity. In addition, we found that lines with high PO activity had slower development rates suggesting that investment in PO, rather than in melanism, is costly.

Heritability of immune function in the caterpillar Spodoptera littoralis

Phenoloxidase (PO) is believed to be a key mediator of immune function in insects and has been implicated both in non-self recognition and in resistance to a variety of parasites and pathogens, including baculoviruses and parasitoids. Using larvae of the Egyptian cotton leafworm, Spodoptera littoralis, we found that despite its apparent importance, haemolymph PO activity varied markedly between individuals, even amongst insects reared under apparently identical conditions. Sib-analysis methods were used to determine whether individuals varied genetically in their PO activity, and hence in one aspect of immune function. The heritability estimate of haemolymph PO activity was high (h 2 = 0.690 +/- 0.069), and PO activity in the haemolymph was strongly correlated with PO activity in both the cuticle and midgut; the sites of entry for most parasites and pathogens. Haemolymph PO activity was also strongly correlated with the degree to which a synthetic parasite (a small piece of nylon monofilament) was encapsulated and melanized (r = 0.622 +/- 0.142), suggesting that the encapsulation response is also heritable. The mechanism maintaining this genetic variation has yet to be elucidated.

Opposite Effects on Spodoptera littoralis Larvae of High Expression Level of a Trypsin Proteinase Inhibitor in Transgenic Plants1

This work illustrates potential adverse effects linked with the expression of proteinase inhibitor (PI) in plants used as a strategy to enhance pest resistance. Tobacco (Nicotiana tabacum L. cv Xanthi) and Arabidopsis [Heynh.] ecotype Wassilewskija) transgenic plants expressing the mustard trypsin PI 2 (MTI-2) at different levels were obtained. First-instar larvae of the Egyptian cotton worm (Spodoptera littoralis Boisd.) were fed on detached leaves of these plants. The high level of MTI-2 expression in leaves had deleterious effects on larvae, causing mortality and decreasing mean larval weight, and was correlated with a decrease in the leaf surface eaten. However, larvae fed leaves from plants expressing MTI-2 at the low expression level did not show increased mortality, but a net gain in weight and a faster development compared with control larvae. The low MTI-2 expression level also resulted in increased leaf damage. These observations are correlated with the differential expression of digestive proteinases in the larval gut; overexpression of existing proteinases on low-MTI-2-expression level plants and induction of new proteinases on high-MTI-2-expression level plants. These results emphasize the critical need for the development of a PI-based defense strategy for plants obtaining the appropriate PI-expression level relative to the pest's sensitivity threshold to that PI.

Combinación de diferentes estrategias de aplicación de hongos entomopatógenos para el control de Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) y Bemisia tabaci (Gennadius) (Hemiptera:Aleyrodidae)

Los ascomicetos mitospóricos entomopatógenos, que actúan por vía tegumentaria, han sido utilizados para el control microbiano de plagas mediante distintas estrategias que permiten el contacto de sus conidios con los insectos diana. Sin embargo, se conocen nuevos aspectos sobre su función ecológica tales como su capacidad secretora de compuestos con actividad insecticida, así como su sorprendente comportamiento como endófitos, que complementan su empleo clásico y que pueden pantag ermitir el desarrollo de nuevas estrategias de control de plagas. El empleo conjunto de estos hongos y sus extractos para el control de un insecto polífago de importancia mundial Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) es abordado en el capítulo II, donde se pone de manifiesto la existencia de variación intra- e interespecífica en la virulencia y estrategias patogénicas de las cepas de Beauveria bassiana y Metarhizium brunneum evaluadas frente a larvas del noctuido. La aplicación conjunta de cepas con distintas estrategias patogénicas junto con sus extractos a larvas del lepidóptero tuvo efecto aditivo, y alguno antagónico, de lo que depende su empleo conjunto. Los capítulos III y IV revelan la existencia de colonización endofítica transitoria tras la aplicación foliar de suspensiones de conidios de cepas de las especies mencionadas en alfalfa, tomate y melón, así como el impacto de esta colonización sobre dos fitófagos con diferentes hábitos de alimentación, uno masticador S. littoralis y otro picador-suctor Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). El capítulo III muestra la contribución aditiva de la mortalidad causada por la alimentación de larvas de S. littoralis a expensas de material vegetal colonizado endofíticamente por cepas de B. bassiana y M. anisopliae con la debida al tratamiento tópico de las mismas con las suspensiones fúngicas. El origen de la mortalidad iniciada a través de la vía digestiva en larvas de este lepidóptero permanece incierto para las cepas de B. bassiana, pero podría estar asociado a la presencia en la planta de metabolitos fúngicos en las cepas de M. brunneum como denota la presencia de destruxina (dtx) A en el 11,0% de los cadáveres. El capítulo IV revela que cepas de ambas especies fúngicas pueden iniciar ciclos de infección en insectos picadores-suctores cuando se alimentan a expensas de sustrato vegetal colonizado endofíticamente, si bien, ambas especies presentan estrategias diferentes. Así, B. bassiana muestra una gran capacidad para colonizar el melón, incluso con efecto traslaminar, que causa la infección de ninfas de B. tabaci por contacto con el tegumento. Sin embargo, la mortalidad de las mismas causada por M. brunneum, de crecimiento mucho más localizado en la hoja, está relacionada una vez mas con la presencia de dtx A en el 43,0% de los cadáveres. Estos resultados deben ser considerados para la evaluación del impacto real de los tratamientos con hongos entomopatógenos, y abren nuevas vías en el control de plagas.

Diurnal and nocturnal herbivore induction on maize elicit different innate response of the fall armyworm parasitoid, Campoletis flavicincta

Herbivore-attacked plants produce specific volatile substances that represent important cues for host finding by natural enemies. The fall armyworm Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is a voracious herbivore and usually feed on maize in all periods of the day. Given that plant needs light to synthesize de novo herbivore-induced volatiles, volatile blend may be changed depending on time of the day the plant is induced, what could interfere in natural enemy foraging. In this sense, the current study aimed to investigate differential attractiveness of maize elicited by fall armyworm regurgitant under light and dark conditions to its specialist larval parasitoid Campoletis flavicincta (Ashmead) (Hymenoptera: Ichneumonidae). All bioassays were conducted in Y-tube olfactometer to assess parasitoid response to odors from undamaged maize, mechanical damage, and regurgitant-treated plants at 0-1, 5-6, and 24-25 h after induction. The results showed that na < ve wasps were attracted to volatiles emitted by nocturnal regurgitant-treated maize at 5-6 h, but not to odors from diurnal regurgitant-treated plants. The differential attractiveness is likely due to blend composition as nocturnal regurgitant-treated plants emit aromatic compounds and the homoterpene (3E)-4,8-dimethyl-1,3,7-nonatriene in larger amounts than diurnal-treated plants.

Kymmenen teollisesti merkittävän puulajin kuoriuutteiden mahdollisuudet tuholaistorjunnassa

Metsäteollisuudesta kertyy vuosittain suuria määriä ylijäämämateriaalia, kuten puun kuorta ja oksia.Ylimääräinen aines käytetään pääasiassa energiantuotantoon, mutta uusia soveltamismahdollisuuksia kaivataan. Kuoren on havaittu olevan potentiaalinen lähde monille bioaktiivisille yhdisteille, joille olisi käyttöä esimerkiksi lääke- ja kemianteollisuudessa sekä maa-, metsä- ja puutarhatuotannon tuholaistorjunnassa. Tutkimus on osa Euroopan Unionin rahoittamaa ForestSpeCs-projektia, jonka tarkoituksena on selvittää metsäteollisuuden ylijäämämateriaalien vaihtoehtoisia käyttötapoja. Valittujen kymmenen teollisesti merkittävän pohjoisen puulajin (Abies nephrolepis, Betula pendula, Larix decidua, L. gmelinii, L. sibirica, Picea abies, P. ajanensis, P. pumila, Pinus sylvestris, Populus tremula) kuoresta uutettujen aineiden soveltuvuutta syönninestoaineeksi testattiin kaaliperhosen (Pieris brassicae L.) ja krysanteemiyökkösen (Spodoptera littoralis Boisduval) toukilla sekä osittain sinappikuoriaisella (Phaedon cochloreae Fabricius) ja idänlehtikuoriaisella (Agelastica alni L.). Uutteet valmistettiin yhteistyössä projektin ryhmien avulla tai itsenäisesti erilaisin menetelmin. Testaukset tehtiin laboratorio-oloissa käyttäen lehtikiekkojen valintabiotestiä sekä karkeilla uutteilla että niistä erotelluilla yksittäisillä yhdisteillä. Tehdyistä mittauksista laskettiin syönninestoindeksit (FDI). Tulosten perusteella lähes kaikki testatut uutteet vaikuttivat ainakin jossain määrin kohdehyönteisen syöntikäyttäytymiseen. Hieman yli puolet kaaliperhosella testatuista 46 uutteesta aiheuttivat yli 50 % syönnineston eli kaaliperhonen suosi kontrollilehtiä uutteella käsiteltyjä todennäköisemmin. Krysanteemiyökkösellä yli 50 %:n syönnineston aiheuttivat vain seitsemän testatuista 56 uutteesta. Lisäksi kolme uutetta lisäsi käsiteltyjen kiekkojen syöntiä merkittävästi. Idänlehtikuoriaistoukat ja -aikuiset karttoivat erityisesti abietiinihapolla käsiteltyjä lehtiä. Sinappikuoriaisella testatut uutteet toimivat myös lupaavasti. Testattujen puulajien kuoresta on mahdollista uuttaa biologisesti aktiivisia yhdisteitä, mutta tuholaistorjunnan kannalta oikeiden pitoisuuksien ja tehokkaiden uuttomenetelmien löytäminen vaatii jatkotutkimuksia. Kuoren sisältämien yhdisteiden laatu ja määrä vaihtelevat monien tekijöiden, kuten ympäristön ja genetiikan vaikutuksesta. Hyönteisten sietokyky vaihtelee myös paljon lajeittain ja yksilöidenkin välillä on eroja. Uutteista valmistettavia torjunta-aineita olisi kuitenkin mahdollista sisällyttää esimerkiksi integroituun torjuntaan muiden menetelmien rinnalle tulevaisuudessa.

Optimal foraging when regulating intake of multiple nutrients

There is growing evidence that, rather than maximizing energy intake subject to constraints, many animals attempt to regulate intake of multiple nutrients independently. In the complex diets of animals such as herbivores, the consumption of nutritionally imbalanced foods is sometimes inevitable, forcing trade-offs between eating too much of nutrients present in the foods in relative excess against too little of those in deficit. Such situations are not adequately represented in existing formulations of foraging theory. Here we provide the necessary theory to fit this case, using an approach that combines state-space models of nutrition with Tilman's models of resource exploitation (Tilman 1982, Resource Competition and Community Structure, Princeton: Princeton University Press). Our approach was to construct a smooth fitness landscape over nutrient space, centred on a 'target' intake at which no fitness cost is incurred, and this leads to a natural classification of the simple possible fitness landscapes based on Taylor series approximations of landscape shape. We next examined how needs for multiple nutrients can be assessed experimentally using direct measures of animal performance as the common currency, so that the nutritional strategies of animals can be mapped on to the performance surface, including the position of regulated points of intake and points of nutrient balance when fed suboptimal foods. We surveyed published data and conducted an experiment to map out the performance landscape of a generalist leaf-feeding caterpillar, Spodoptera littoralis. (C) 2004 Tire Association for the Study of Animal Behaviour. Poblished by Elsevier Ltd. All rights reserved.

The role of abscisic acid and water stress in root herbivore-induced leaf resistance

Herbivore-induced systemic resistance occurs in many plants and is commonly assumed to be adaptive. The mechanisms triggered by leaf-herbivores that lead to systemic resistance are largely understood, but it remains unknown how and why root herbivory also increases resistance in leaves. To resolve this, we investigated the mechanism by which the root herbivore Diabrotica virgifera induces resistance against lepidopteran herbivores in the leaves of Zea mays. Diabrotica virgifera infested plants suffered less aboveground herbivory in the field and showed reduced growth of Spodoptera littoralis caterpillars in the laboratory. Root herbivory did not lead to a jasmonate-dependent response in the leaves, but specifically triggered water loss and abscisic acid (ABA) accumulation. The induction of ABA by itself was partly responsible for the induction of leaf defenses, but not for the resistance against S. littoralis. Root-herbivore induced hydraulic changes in the leaves, however, were crucial for the increase in insect resistance. We conclude that the induced leaf resistance after root feeding is the result of hydraulic changes, which reduce the quality of the leaves for chewing herbivores. This finding calls into question whether root-herbivore induced leaf-resistance is an evolved response. © The Authors (2010). Journal compilation © New Phytologist Trust (2010).

A tritrophic signal that attracts parasitoids to host-damaged plants withstands disruption by non-host herbivores

Background: Volatiles emitted by herbivore-infested plants are highly attractive to parasitoids and therefore have been proposed to be part of an indirect plant defense strategy. However, this proposed function of the plant-provided signals remains controversial, and it is unclear how specific and reliable the signals are under natural conditions with simultaneous feeding by multiple herbivores. Phloem feeders in particular are assumed to interfere with plant defense responses. Therefore, we investigated how attack by the piercing-sucking cicadellid Euscelidius variegatus influences signaling by maize plants in response to the chewing herbivore Spodoptera littoralis.Results: The parasitoid Cotesia marginiventris strongly preferred volatiles of plants infested with its host S. littoralis. Overall, the volatile emissions induced by S. littoralis and E. variegatus were similar, but higher levels of certain wound-released compounds may have allowed the wasps to specifically recognize plants infested by hosts. Expression levels of defense marker genes and further behavioral bioassays with the parasitoid showed that neither the physiological defense responses nor the attractiveness of S. littoralis infested plants were altered by simultaneous E. variegatus attack.Conclusions: Our findings imply that plant defense responses to herbivory can be more robust than generally assumed and that ensuing volatiles convey specific information about the type of herbivore that is attacking a plant, even in complex situations with multiple herbivores. Hence, the results of this study support the notion that herbivore-induced plant volatiles may be part of a plant's indirect defense stratagem. © 2010 Erb et al; licensee BioMed Central Ltd.

Within-plant distribution of 1,4-benzoxazin-3-ones contributes to herbivore niche differentiation in maize

Plant defences vary in space and time, which may translate into specific herbivore-foraging patterns and feeding niche differentiation. To date, little is known about the effect of secondary metabolite patterning on within-plant herbivore foraging. We investigated how variation in the major maize secondary metabolites, 1,4-benzoxazin-3-one derivatives (BXDs), affects the foraging behaviour of two leaf-chewing herbivores. BXD levels varied substantially within plants. Older leaves had higher levels of constitutive BXDs while younger leaves were consistently more inducible. These differences were observed independently of plant age, even though the concentrations of most BXDs declined markedly in older plants. Larvae of the well-adapted maize pest Spodoptera frugiperda preferred and grew better on young inducible leaves irrespective of plant age, while larvae of the generalist Spodoptera littoralis preferred and tended to grow better on old leaves. In BXD-free mutants, the differences in herbivore weight gain between old and young leaves were absent for both species, and leaf preferences of S. frugiperda were attenuated. In contrast, S. littoralis foraging patterns were not affected. In summary, our study shows that plant secondary metabolites differentially affect performance and foraging of adapted and non-adapted herbivores and thereby likely contribute to feeding niche differentiation

Metabolomics reveals herbivore-induced metabolites of resistance and susceptibility in maize leaves and roots

Plants respond to herbivory by reprogramming their metabolism. Most research in this context has focused on locally induced compounds that function as toxins or feeding deterrents. We developed an ultra-high-pressure liquid chromatography time-of-flight mass spectrometry (UHPLC-TOF-MS)-based metabolomics approach to evaluate local and systemic herbivore-induced changes in maize leaves, sap, roots and root exudates without any prior assumptions about their function. Thirty-two differentially regulated compounds were identified from Spodoptera littoralis-infested maize seedlings and isolated for structure assignment by microflow nuclear magnetic resonance (CapNMR). Nine compounds were quantified by a high throughput direct nano-infusion tandem mass spectrometry/mass spectrometry (MS/MS) method. Leaf infestation led to a marked local increase of 1,3-benzoxazin-4-ones, phospholipids, N-hydroxycinnamoyltyramines, azealic acid and tryptophan. Only few changes were found in the root metabolome, but 1,3-benzoxazin-4-ones increased in the vascular sap and root exudates. The role of N-hydroxycinnamoyltyramines in plant–herbivore interactions is unknown, and we therefore tested the effect of the dominating p-coumaroyltyramine on S. littoralis. Unexpectedly, p-coumaroyltyramine was metabolized by the larvae and increased larval growth, possibly by providing additional nitrogen to the insect. Taken together, this study illustrates that herbivore attack leads to the induction of metabolites that can have contrasting effects on herbivore resistance in the leaves and roots.

A specialist root herbivore exploits defensive metabolites to locate nutritious tissues

he most valuable organs of plants are often particularly rich in essential elements, but also very well defended. This creates a dilemma for herbivores that need to maximise energy intake while minimising intoxication. We investigated how the specialist root herbivore Diabrotica virgifera solves this conundrum when feeding on wild and cultivated maize plants. We found that crown roots of maize seedlings were vital for plant development and, in accordance, were rich in nutritious primary metabolites and contained higher amounts of the insecticidal 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and the phenolic compound chlorogenic acid. The generalist herbivores Diabrotica balteata and Spodoptera littoralis were deterred from feeding on crown roots, whereas the specialist D. virgifera preferred and grew best on these tissues. Using a 1,4-benzoxazin-3-one-deficient maize mutant, we found that D. virgifera is resistant to DIMBOA and other 1,4-benzoxazin-3-ones and that it even hijacks these compounds to optimally forage for nutritious roots

Belowground ABA boosts aboveground production of DIMBOA and primes induction of chlorogenic acid in maize

Plants are important mediators between above- and belowground herbivores. Consequently, interactions between root and shoot defences can have far-reaching impacts on entire food webs. We recently reported that infestation of maize roots by the root feeding larvae of the beetle Diabrotica virgifera virgifera boosts shoot resistance against herbivores and pathogens. Root herbivory also induced DIMBOA levels and primed for enhanced induction of chlorogenic acid, two secondary metabolites that have been associated with biotic stress resistance. Interestingly, ABA emerged as a putative long-distance signal, possibly responsible for this effect. In this addendum, we investigate the role of root-derived ABA in the systemic regulation of aboveground DIMBOA, and the phenolic compounds chlorogenic acid, caffeic and ferulic acid. We discuss the relevance of the plant hormone in relation to defence against the leaf herbivore Spodoptera littoralis. Soil-drench treatment with ABA mimicked root herbivore-induced accumulation of DIMBOA in the leaves. Similarly, ABA mimicked aboveground priming of chlorogenic acid production, resulting in augmented accumulation of this compound upon subsequent shoot attack by S. littoralis. These findings confirm our notion that ABA acts as an important signal in the regulation of aboveground defence upon belowground herbivory. However, based on our previous finding that ABA alone is not sufficient to trigger aboveground resistance against S. littoralis caterpillars, the results suggest that the ABA-inducible effects on DIMBOA and chlorogenic acid are not solely responsible for root herbivore-induced resistance against S. littoralis. Full text HTML PDF

A physiological and behavioral mechanism for leaf-herbivore induced systemic root resistance

Indirect plant-mediated interactions between herbivores are important drivers of community composition in terrestrial ecosystems. Among the most striking examples are the strong indirect interactions between spatially separated leaf- and root-feeding insects sharing a host plant. Although leaf feeders generally reduce the performance of root herbivores, little is known about the underlying systemic changes in root physiology and the associated behavioral responses of the root feeders. We investigated the consequences of maize (Zea mays) leaf infestation by Spodoptera littoralis caterpillars for the root-feeding larvae of the beetle Diabrotica virgifera virgifera, a major pest of maize. D. virgifera strongly avoided leaf-infested plants by recognizing systemic changes in soluble root components. The avoidance response occurred within 12 h and was induced by real and mimicked herbivory, but not wounding alone. Roots of leaf-infested plants showed altered patterns in soluble free and soluble conjugated phenolic acids. Biochemical inhibition and genetic manipulation of phenolic acid biosynthesis led to a complete disappearance of the avoidance response of D. virgifera. Furthermore, bioactivity-guided fractionation revealed a direct link between the avoidance response of D. virgifera and changes in soluble conjugated phenolic acids in the roots of leaf-attacked plants. Our study provides a physiological mechanism for a behavioral pattern that explains the negative effect of leaf attack on a root-feeding insect. Furthermore, it opens up the possibility to control D. virgifera in the field by genetically mimicking leaf herbivore-induced changes in root phenylpropanoid patterns.