Frédéric Francis

Identification of aphid salivary proteins: a proteomic investigation of Myzus persicae

The role of insect saliva in the first contact between an insect and a plant is crucial during feeding. Some elicitors, particularly in insect regurgitants, have been identified as inducing plant defence reactions. Here, we focused on the salivary proteome of the green peach aphid, Myzus persicae. Proteins were either directly in‐solution digested or were separated by 2D SDS‐PAGE before trypsin digestion. Resulting peptides were then identified by mass spectrometry coupled with database investigations. A homemade database was constituted of expressed sequence tags from the pea aphid Acyrtosiphon pisum and M. persicae. The databases were used to identify proteins related to M. persicae with a nonsequenced genome. This procedure enabled us to discover glucose oxidase, glucose dehydrogenase, NADH dehydrogenase, α‐glucosidase and α‐amylase in M. persicae saliva. The presence of these enzymes is discussed in terms of plant–aphid interactions.

Olfactory responses to aphid and host plant volatile releases: (E)-beta-farnesene an effective kairomone for the predator Adalia bipunctata.

The volatiles released from several aphid and host plant species, alone or associated, were studied for their infochemical role in prey location. Using a four-arm olfactometer, the attraction of several combinations of three aphid (Myzus persicae, Acyrthosiphon pisum, and Brevicoryne brassicae) and three plant (Vicia faba, Brassica napus, and Sinapis alba) species toward Adalia bipunctata larvae and adults was observed. Both predatory larvae and adults were attracted only by A. pisum and M. persicae when they were crushed, whatever the host plant. (E)-β-Farnesene, the aphid alarm pheromone, was the effective kairomone for the ladybird. Plant leaves alone (V. faba, B. napus, and S. alba) or in association with nonstressed whole aphids (the three species) did not have any attraction for the predator. The B. brassicae specialist aphid is the only prey that was not attracted to A. bipunctata larvae and adults, even if they were crushed. Release of B. brassicae molecules similar to the host plant allelochemicals was demonstrated by GC–MS analysis. The lack of behavioral response of the ladybird at short distance toward the cruciferous specialist aphid was related only to the absence of (E)-β-farnesene in the aphid prey volatile pattern.

Effects of Allelochemicals from First (Brassicaceae) and Second (Myzus persicae and Brevicoryne brassicae) Trophic Levels on Adalia bipunctata

Three Brassicaceae species, Brassica napus (low glucosinolate content), Brassica nigra (including sinigrin), and Sinapis alba (including sinalbin) were used as host plants for two aphid species: the generalist Myzus persicae and the specialist Brevicoryne brassicae. Each combination of aphid species and prey host plant was used to feed the polyphagous ladybird beetle, Adalia bipunctata. Experiments with Brassicaceae species including different amounts and kinds of glucosinolates (GLS) showed increased ladybird larval mortality at higher GLS concentrations. When reared on plants with higher GLS concentrations, the specialist aphid, B. brassicae, was found to be more toxic than M. persicae. Identification of GLS and related degradation products, mainly isothiocyanates (ITC), was investigated in the first two trophic levels, plant and aphid species, by high-performance liquid chromatography and gas chromatography–mass spectrometry, respectively. While only GLS were detected in M. persicae on each Brassicaceae species, high amounts of ITC were identified in B. brassicae samples (allyl-ITC and benzyl-ITC from B. nigra and S. alba, respectively) from all host plants. Biological effects of allelochemicals from plants on predators through aphid prey are discussed in relation to aphid species to emphasize the role of the crop plant in integrated pest management in terms of biological control efficacy.

Is the (E)-β-farnesene only volatile terpenoid in aphids?: (E)-β-farnesene: the only volatile terpenoid in aphids

Abstract:  Herbivore insects use a broad range of chemical cues to locate their host to feed or to oviposit. Whether several plant volatiles are effective allelochemicals for insects, the latter also emit molecules which have infochemical role. The (E)‐β‐farnesene (EBF) is a well‐known aphid alarm pheromone commonly found in all previously tested species. Analysis of the released molecules from 23 aphid species, mainly collected on their natural host plant from May to July, was performed by gas chromatography–mass spectrometry. While EBF was identified as the main volatile substance in 16 species, alone or associated with other molecules, the alarm pheromone was only a minor component of the volatile molecule pattern of five other species. Moreover, two species, Euceraphis punctipennis and Drepanosiphum platanoides, did not release EBF at all but other terpenes were identified. This original observation raised the question on the utility and the source of the non‐EBF volatiles. Are these potential infochemical substances produced by the aphid or only absorbed from the host plant? Here we determined that terpenes released by insects were not only provided by the host plants. Indeed, Megoura viciae emitted additional molecules than the ones from several aphid species reared on the same host plant. Moreover, no systematic relation between the feeding behaviour of the aphid species and the volatile releases was observed. Aphid terpene composition and proportion would provide reliable cues to identify the emitting organism, plant or insect. The next step of this work will be to determine the infochemical role of terpenes found in the range of tested aphid samples to better understand the relations between the different tritrophic levels.

Electrophysiological and Behavioral Responses of the Multicolored Asian Lady Beetle, Harmonia axyridis Pallas, to Sesquiterpene Semiochemicals

The role of two volatile sesquiterpenes, (E)-β-farnesene and (−)-β-caryophyllene, in the chemical ecology of the multicolored Asian lady beetle, Harmonia axyridis Pallas, was investigated by using both electrophysiological and behavioral techniques. (E)-β-Farnesene is the major component of the alarm pheromone of most aphid species, which are preyed on by H. axyridis. (−)-β-Caryophyllene was previously isolated from the headspace volatiles above overwintering and aggregated H. axyridis females. These sesquiterpenes elicited significant electroantennogram (EAG) activity from both H. axyridis male and female antennae. In a four-arm olfactometer, male and female H. axyridis were highly attracted toward (E)-β-farnesene, whereas only males were attracted to (−)-β-caryophyllene. In a bioassay technique that used a passively ventilated plastic box, both male and female H. axyridis aggregated in the (−)-β-caryophyllene-treated side of the box. These results support the potential usefulness of (E)-β-farnesene and (−)-β-caryophyllene in push–pull strategies that use H. axyridis as a biological control agent in aphid-infested sites or to control this new urban pest in residential structures.

Microorganisms from aphid honeydew attract and enhance the efficacy of natural enemies

Aphids are one of the most serious pests of crops worldwide, causing major yield and economic losses. To control aphids, natural enemies could be an option but their efficacy is sometimes limited by their dispersal in natural environment. Here we report the first isolation of a bacterium from the pea aphid Acyrthosiphon pisum honeydew, Staphylococcus sciuri, which acts as a kairomone enhancing the efficiency of aphid natural enemies. Our findings represent the first case of a host-associated bacterium driving prey location and ovipositional preference for the natural enemy. We show that this bacterium has a key role in tritrophic interactions because it is the direct source of volatiles used to locate prey. Some specific semiochemicals produced by S. sciuri were also identified as significant attractants and ovipositional stimulants. The use of this host-associated bacterium could certainly provide a novel approach to control aphids in field and greenhouse systems.

Attacks by a piercing-sucking insect (Myzus persicae Sultzer) or a chewing insect (Leptinotarsa decemlineata Say) on potato plants (Solanum tuberosum L.) induce differential changes in volatile compound release and oxylipin synthesis

Plant defensive strategies bring into play blends of compounds dependent on the type of attacker and coming from different synthesis pathways. Interest in the field is mainly focused on volatile organic compounds (VOCs) and jasmonic acid (JA). By contrast, little is known about the oxidized polyunsaturated fatty acids (PUFAs), such as PUFA-hydroperoxides, PUFA-hydroxides, or PUFA-ketones. PUFA-hydroperoxides and their derivatives might be involved in stress response and show antimicrobial activities. Hydroperoxides are also precursors of JA and some volatile compounds. In this paper, the differential biochemical response of a plant against insects with distinct feeding behaviours is characterized not only in terms of VOC signature and JA profile but also in terms of their precursors synthesized through the lipoxygenase (LOX)-pathway at the early stage of the plant response. For this purpose, two leading pests of potato with distinct feeding behaviours were used: the Colorado Potato Beetle (Leptinotarsa decemlineata Say), a chewing herbivore, and the Green Peach Aphid (Myzus persicae Sulzer), a piercing-sucking insect. The volatile signatures identified clearly differ in function with the feeding behaviour of the attacker and the aphid, which causes the smaller damages, triggers the emission of a higher number of volatiles. In addition, 9-LOX products, which are usually associated with defence against pathogens, were exclusively activated by aphid attack. Furthermore, a correlation between volatiles and JA accumulation and the evolution of their precursors was determined. Finally, the role of the insect itself on the plant response after insect infestation was highlighted.

Salivary glucose oxidase from caterpillars mediates the induction of rapid and delayed-induced defenses in the tomato plant.

Caterpillars produce oral secretions that may serve as cues to elicit plant defenses, but in other cases these secretions have been shown to suppress plant defenses. Ongoing work in our laboratory has focused on the salivary secretions of the tomato fruitworm, Helicoverpa zea. In previous studies we have shown that saliva and its principal component glucose oxidase acts as an effector by suppressing defenses in tobacco. In this current study, we report that saliva elicits a burst of jasmonic acid (JA) and the induction of late responding defense genes such as proteinase inhibitor 2 (Pin2). Transcripts encoding early response genes associated with the JA pathway were not affected by saliva. We also observed a delayed response to saliva with increased densities of Type VI glandular trichomes in newly emerged leaves. Proteomic analysis of saliva revealed glucose oxidase (GOX) was the most abundant protein identified and we confirmed that it plays a primary role in the induction of defenses in tomato. These results suggest that the recognition of GOX in tomato may represent a case for effector-triggered immunity. Examination of saliva from other caterpillar species indicates that saliva from the noctuids Spodoptera exigua and Heliothis virescens also induced Pin2 transcripts.

Aphid alarm pheromone: An overview of current knowledge on biosynthesis and functions

Aphids are important agricultural and forest pests that exhibit complex behaviors elicited by pheromonal signals. The aphid alarm pheromone--of which (E)-β-farnesene is the key (or only) component in most species--plays important roles in mediating interactions among individuals as well as multitrophic interactions among plants, aphids, and aphid natural enemies. Though many important questions remain to be answered, a large body of research has addressed various aspects of the biology, physiology, and ecology of aphid alarm pheromones. Here we review recent advances in our understanding of (a) the identity and composition of aphid alarm signals; (b) their biosynthesis and production; (c) their effects on conspecifics; (d) their role as cues for other insect species; and (e) their potential application for the management of pest organisms.

Role of terpenes from aphid‐infested potato on searching and oviposition behavior of Episyrphus balteatus

To cope with pathogen and insect attacks, plants develop different mechanisms of defence, in both direct (physical and chemical) and indirect ways (attractive volatiles to entomophagous beneficials). Plants are then able to express traits that facilitate “top‐down” control of pests by attracting herbivore predators. Here we investigate the indirect defence mechanism of potato plants by analyzing the volatile patterns of both healthy and aphid‐infested plants. Important changes in the emitted terpene pattern by the Myzus persicae infested host plant were observed. Using Solid Phase Micro Extraction (SPME) and GC‐MS, the (E)‐β‐farnesene (EBF) appeared to be emitted by aphid‐infested potato and not by healthy plants. To assess the infochemical role of these volatile releases after aphid damage on the aphidophagous predators Episyrphus balteatus, the hoverfly foraging behavior was assessed using the Observer 5.0 software (Noldus, Wageningen, The Netherlands). Aphid‐free potato plants were also used as a control volatile source in the predator behavioral study. While aphid‐infested plants induced efficient searching and acceptation behaviors leading to egg‐laying, no kairomonal effect of healthy potato plants was observed, leading to longer immobility durations and shorter searching periods in the net cage. High oviposition rate of E. balteatus was observed when aphid‐infested potato was used (mean of 48.9 eggs per laying and per female). On the other hand, no egg was produced by the hoverfly on healthy aphid‐free plants. The E. balteatus foraging and reproductive behaviors according to the volatile emission from aphid‐infested plants are discussed in relation to the potential use of active infochemical molecules in integrated aphid pest management.