Glen Powell

The cabbage aphid: a walking mustard oil bomb

The cabbage aphid, Brevicoryne brassicae, has developed a chemical defence system that exploits and mimics that of its host plants, involving sequestration of the major plant secondary metabolites (glucosinolates). Like its host plants, the aphid produces a myrosinase (β-thioglucoside glucohydrolase) to catalyse the hydrolysis of glucosinolates, yielding biologically active products. Here, we demonstrate that aphid myrosinase expression in head/thoracic muscle starts during embryonic development and protein levels continue to accumulate after the nymphs are born. However, aphids are entirely dependent on the host plant for the glucosinolate substrate, which they store in the haemolymph. Uptake of a glucosinolate (sinigrin) was investigated when aphids fed on plants or an in vitro system and followed a different developmental pattern in winged and wingless aphid morphs. In nymphs of the wingless aphid morph, glucosinolate level continued to increase throughout the development to the adult stage, but the quantity in nymphs of the winged form peaked before eclosion (at day 7) and subsequently declined. Winged aphids excreted significantly higher amounts of glucosinolate in the honeydew when compared with wingless aphids, suggesting regulated transport across the gut. The higher level of sinigrin in wingless aphids had a significant negative impact on survival of a ladybird predator. Larvae of Adalia bipunctata were unable to survive when fed adult wingless aphids from a 1% sinigrin diet, but survived successfully when fed aphids from a glucosinolate-free diet (wingless or winged), or winged aphids from 1% sinigrin. The apparent lack of an effective chemical defence system in adult winged aphids possibly reflects their energetic investment in flight as an alternative predator avoidance mechanism.

Phloem specific aphid resistance in Cucumis melo line AR 5 : effects on feeding behaviour and performance of Aphis gossypii

The feeding behaviour, excretion rate, and life history traits of the cotton‐melon aphid, Aphis gossypii (Glover) (Homoptera, Aphididae), were measured on a resistant melon, Cucumis melo L., breeding line, AR 5. The site of resistance detection by the aphids was determined using the electrical penetration graph (EPG) technique. EPG recordings showed that resistance is expressed within the host plant, rather than on its surface, because the time to first stylet penetration was not significantly different between AR 5 and the closely related susceptible breeding line, PMR 5. EPG patterns associated with stylet pathway activities of the aphids were not significantly different between the resistant and susceptible lines. Significant behavioural differences were observed only after stylets contacted phloem sieve elements. On AR 5, the duration of salivation after sieve element puncture (waveform E1) was significantly longer, and the number of aphids showing phloem sap ingestion (waveform E2) was significantly reduced. We conclude that the resistance mechanism producing the effects seen in this study acts within the phloem sieve elements. Monitoring of excretion rates on the two genotypes showed that aphid feeding was delayed and greatly reduced on the resistant genotype. Comparisons of aphid life history traits and population development between host plant genotypes showed that the effects of resistance act throughout aphid development and are highly effective at slowing down population increase.

Aphid stylet activities during potyvirus acquisition from plants and anin vitro system that correlate with subsequent transmission

The behavioural events associated with acquisition of tobacco etch potyvirus by starvedMyzus persicae during single, electrically-recorded penetrations of plants or a Parafilm membrane were compared. Twenty nine percent of aphids acquired virus from plants and subsequently transmitted to test plants. Stylet puncture of the plasmalemma, indicated by a potential drop (pd) to the intracellular signal voltage level, occurred during 84% of penetrations, and virus transmission was always associated with this behavioural event during acquisition. Periods of intracellular stylet tip location, known as pd phase II, ranged from 3.6–12.2s, and always comprised three consecutive sub-phases, designated II1, II2 and II3. Ninety six percent of pds included distinct pulses during phase II3. A waveform which closely resembled these pulses was produced by 59% of aphids that probed a virus suspension through a Parafilm membrane; nineteen percent of the aphids subsequently transmitted membrane-acquired virus and transmission was significantly associated with the occurrence of the phase II3-like pulses during acquisition. The duration of occurrence of recorded phase II3 pulses, either on plants or thein vitro system, did not influence the virus transmission efficiency of aphids. The association of virus uptake from aqueous suspension with a particular behavioural activity is discussed as evidence for the ‘ingestion-egestion’ hypothesis for nonpersistent transmission. Starved aphids acquiring virus from infected leaf tissue or thein vitro system had significantly higher transmission efficiencies than non-starved aphids. Starved and non-starved insects were electrically-recorded penetrating the artificial membrane, and again there was a clear difference in transmission efficiency (starved aphids, 26%; non-starved aphids, 2%). The higher transmission efficiency of starved insects could not be explained by behavioural differences, and the results lend support to the hypothesis that non-behavioural factors determine the enhancement of potyvirus transmission by preacquisition starvation.

Cucumber mosaic virus and its 2b RNA silencing suppressor modify plant-aphid interactions in tobacco.

The cucumber mosaic virus (CMV) 2b protein not only inhibits anti-viral RNA silencing but also quenches transcriptional responses of plant genes to jasmonic acid, a key signalling molecule in defence against insects. This suggested that it might affect interactions between infected plants and aphids, insects that transmit CMV. We found that infection of tobacco with a 2b gene deletion mutant (CMVΔ2b) induced strong resistance to aphids (Myzus persicae) while CMV infection fostered aphid survival. Using electrical penetration graph methodology we found that higher proportions of aphids showed sustained phloem ingestion on CMV-infected plants than on CMVΔ2b-infected or mock-inoculated plants although this did not increase the rate of growth of individual aphids. This indicates that while CMV infection or certain viral gene products might elicit aphid resistance, the 2b protein normally counteracts this during a wild-type CMV infection. Our findings suggest that the 2b protein could indirectly affect aphid-mediated virus transmission.

Aphid responses to non-host epicuticular lipids.

Initiation of stylet penetration by aphids depends on the sensory assessment of a number of plant surface features including colour, texture and phytochemicals (volatile and non-volatile). Video recording behaviour of the black bean aphid, Aphis fabae, showed that these insects rapidly inserted their stylets following contact with host plants (beans), but were reluctant to penetrate non-hosts (oats). However, when epicuticular waxes were stripped from oats, using cellulose acetate, aphids penetrated the plant surface significantly earlier than on oats with the wax layer intact. Chloroform extraction of epicuticular lipids, followed by coupled gas chromatography-mass spectrometry, revealed a complex blend of wax components on beans, whereas one compound (1-hexacosanol) predominated on oats. Epicuticular lipids were applied to artificial (glass) substrates in order to investigate their behavioural activity. Initiation of a stylet penetration attempt by A. fabae was delayed when the oat extract or pure 1-hexacosanol were applied, but the bean extract had no behavioural effect. The results suggest that epicuticular lipids play an important role in the early stages of host-plant selection by A. fabae.

Accumulation of Glucosinolates by the Cabbage Aphid Brevicoryne brassicae as a Defense Against Two Coccinellid Species

Brassica nigra plants, characterized by high levels of sinigrin, and artificial aphid diets to which sinigrin was selectively added were used to rear the crucifer specialist, Brevicoryne brassicae. Aphids were provided as a food source to two species of polyphagous ladybird, Adalia bipunctata and Coccinella septempunctata. First instar A. bipunctata were unable to survive when fed with B. brassicae reared on B. nigra or diets containing 0.2% sinigrin, but when fed with aphids reared on diets containing 0% sinigrin, survival rates were high. By contrast, first instar C. septempunctata were able to survive when fed with aphids reared on B. nigra or artificial diets containing up to 1% sinigrin. However, the presence of sinigrin in the aphid diet decreased larval growth and increased the time necessary for larvae to reach second instar for this species of ladybird. These results indicate that the presence of sinigrin in the diet of B. brassicae makes this aphid unsuitable as a food source for A. bipunctata but not for C. septempunctata, although for this ladybird species, there appear to be costs associated with feeding on aphids that contain this secondary metabolite.

ROLE OF THE HELPER COMPONENT IN VECTOR-SPECIFIC TRANSMISSION OF POTYVIRUSES

Four aphid species were tested for their ability to transmit tobacco etch (TEV) and turnip mosaic (TuMV) potyviruses. Myzus persicae and Aphis gossypii transmitted both viruses efficiently from infected plants, whereas Lipaphis erysimi transmitted only TuMV and Myzus ascalonicus was a poor or non-transmitter of either virus. Similar electrically monitored probing patterns were produced by M. persicae, L. erysimi and M. ascalonicus, ruling out behavioural differences as the cause of differential transmission. Transmission results similar to those from infected plants were obtained when these aphids acquired homologous virus/helper component (HC) mixtures through membranes. With heterologous virus/HC mixtures, M. persicae remained a highly efficient vector and M. ascalonicus a non-vector, but L. erysimi became an efficient vector of TEV if acquired in the presence of TuMV HC and A. gossypii transmitted both viruses less efficiently when acquired with TuMV HC. Transmission was highly correlated with the retention of virus in the stylets, as determined by autoradiography of 125I-labelled virions. The results show that constituent(s) of or in the food canal of different aphid species differ in their ability to interact with specific HCs, leading to qualitative or quantitative differences in ability to retain and subsequently transmit specific potyviruses.

Host-selection behaviour by genetically identical aphids with different plant preferences

The behaviour of summer and autumn winged forms of the black bean aphid, Aphis fabae Scopoli (Homoptera: Aphididae), was compared on two plants utilized at different stages of the insect’s life cycle. Adult autumn migrants (gynoparae) are monophagous, colonizing spindle (Euonymus europaeus), whereas polyphagous summer winged aphids (alate virginoparae) are associated with a variety of herbaceous plants, including broad bean (Vicia faba). When aphids from a single clone were given access to a spindle leaf and a bean seedling in choice tests, many virginoparae settled and larviposited on both plant species over 24 h. By contrast, gynoparae showed a clear preference for spindle, with 93.5% of settled adults and 98.3% of larvae on this plant species. Close‐up video monitoring showed that gynoparae discriminated beans from spindle within a 5‐min period, whereas virginoparae behaved similarly on both plant species. For gynoparae, the major behavioural difference on the two plants appeared after a brief (epidermal) stylet penetration, with many insects taking flight within a few seconds of stylet withdrawal from bean. Factors detected during stylet insertion by gynoparae must therefore inhibit take‐off on spindle. Electrical recording experiments showed that aphids often punctured a cell membrane during brief probes on both plant species, and intracellular stylet activities always included a waveform associated with ingestion. When gynoparae puncture spindle cells their behaviour is probably modified by intracellular metabolites detected via gustation of ingested epidermal cell sap. These cues may inhibit the take‐off reflex which otherwise follows probing.

Do plant viruses facilitate their aphid vectors by inducing symptoms that alter behavior and performance

Abstract Aphids can respond both positively and negatively to virus-induced modifications of the shared host plant. It can be speculated that viruses dependent on aphids for their transmission might evolve to induce changes in the host plant that attract aphids and improve their performance, subsequently enhancing the success of the pathogen itself. We studied how pea aphids [Acyrthosiphon pisum (Harris)] responded to infection of tic beans (Vicia faba L.) by three viruses with varying degrees of dependence on this aphid for their transmission: pea enation mosaic virus (PEMV), bean yellow mosaic virus (BYMV), and broad bean mottle virus (BBMV). BYMV has a nonpersistent mode of transmission by aphids, whereas PEMV is transmitted in a circulative-persistent manner. BBMV is not aphid transmitted. When reared on plants infected by PEMV, no changes in aphid survival, growth, or reproductive performance were observed, whereas infection of beans by the other aphid-dependent virus, BYMV, actually caused a reduction in aphid survival in some assays. None of the viruses induced A. pisum to increase production of winged progeny, and aphids settled preferentially on leaf tissue from plants infected by all three viruses, the likely mechanism being visual responses to yellowing of foliage. Thus, in this system, the attractiveness of an infected host plant and its quality in terms of aphid growth and reproduction were not related to the pathogen’s dependence on the aphid for transmission to new hosts.

Application of DL-β-aminobutyric acid (BABA) as a root drench to legumes inhibits the growth and reproduction of the pea aphid Acyrthosiphon pisum (Hemiptera: Aphididae)

DL-beta-aminobutyric acid (BABA) is a non-protein amino acid that is an effective inducer of resistance against a variety of plant pathogens. However, examples of BABA-induced resistance against insect herbivores have not been reported. We applied BABA as a soil drench to legumes and monitored its effects on the pea aphid Acyrthosiphon pisum (Harris). On tic bean (Vicia faba var. minor), BABA increased aphid mortality, caused a reduction in the mean relative growth rate of individual insects and lessened the intrinsic rate of population increase (rm). BABA also caused significant reductions in the growth rate of A. pisum on pea (Pisum sativa), broad bean (Vicia faba var. major), runner bean (Phaseolus coccineus), red clover (Trifolium pratense) and alfalfa (Medicago sativa). No direct toxic effects of BABA against A. pisum were found, and no phytotoxic effects that may have caused a reduction in aphid performance were detected. Possible mechanisms behind this BABA-induced inhibition of aphid performance are discussed.