The acquisition of a caulimovirus by different aphid species: comparison with a potyvirus

The acquisition and transmission of cauliflower mosaic virus (CaMV) by six aphid species and three clones of aphids was studied and compared with that of turnip mosaic virus (TuMV) with Myzus persicae. Two clones of Aphis fabae were unable to transmit CaMV, but the other species, Acyrthosiphon pisum, Brevicoryne brassicae, Megoura viciae, M. persicae and Rhopalosiphum padi transmitted in a bior multi-phasic manner. There was no statistical evidence of a bimodal transmission pattern. R. padi is recorded as a vector of CaMV for the first time. The transmission efficiency of CaMV varied with time of acquisition and suggested that accumulation of the virus occurred with two peaks of efficiency within the anterior region of the insect gut. The time at which these two peaks occurred varied between the species, but the basic pattern was common to all transmitting aphid species in this study. This pattern contrasted with that of TuMV. The transmission data are discussed in terms of bimodal transmission, the influence of feeding behaviour, the role of a helper protein associated with both TuMV and CaMV and the evidence for site specific attachment of CaMV.     

The prevalence and spatial distribution of viruses in natural populations of Brassica oleracea

We report a survey of four viruses (beet western yellows luteovirus (BWYV), cauliflower mosaic caulimovirus (CaMV), turnip mosaic potyvirus (TuMV), turnip yellow mosaic tymovirus (TYMV)) in five natural populations of Brassica oleracea in Dorset (UK). All four viruses were common; 43% of plants were infected with BWYV, 60% with CaMV, 43% with TuMV and 18% with TYMV. For each virus there were significant differences in the proportion of infected plants among populations, which were not completely explained by differences in the age of plants. Multiple virus infections were prevalent, with 54% of plants having two or more virus types. There were statistically significant associations between pairs of viruses. The CaMV was positively associated with the other three viruses, and BWYV was also positively associated with TuMV. There was no detectable association between BWYV and TYMV, whereas TuMV and TYMV were negatively associated. We suggest these associations result from BWYV, CaMV and TuMV having aphid vectors in common, as aphids are attracted to plants that already have a virus infection. Infected plants were distributed randomly or were very weakly aggregated within populations. The implications of widespread multiple virus infections in natural plant populations are discussed with respect to the release of transgenic plants expressing virus-derived genes.     

Quantitative studies on the uptake and retention of potato leafroll virus by aphids in laboratory and field conditions

Enzyme-linked immunosorbent assay (ELISA) was adapted for the efficient detection and assay of potato leafroll virus (PLRV) in aphids. Best results were obtained when aphids were extracted in 0.05 M phosphate buffer, pH 7.0, and the extracts incubated at 37 °C for 1 h before starting the assay. Using batches of 20 green peach aphids (Myzus persicae), about 0.01 ng PLRV/aphid could be detected. The virus could also be detected in single aphids allowed a 1-day acquisition access period on infected potato leaves. The PLRV content of aphids depended on the age of potato source-plants and the position of source leaves on them. It increased with increase in acquisition access period up to 7 days but differed considerably between individual aphids. A maximum of 7 ng PLRV/aphid was recorded but aphids more usually accumulated about 0.2 ng PLRV per day. When aphids were allowed acquisition access periods of 1–3 days, and then caged singly on Physalis floridana seedlings for 3 days, the PLRV content of each aphid, measured subsequently, was not strongly correlated with the infection of P. floridana. The concentration of PLRV in leaf extracts differed only slightly when potato plants were kept at 15, 20, 25 or 30 °C for 1 or 2 wk, but the virus content of aphids kept on leaves at the different temperatures decreased with increase of temperature. PLRV was transmitted readily to P. floridana at all temperatures, but by a slightly smaller proportion of aphids, and after a longer latent period, at 15 °C than at 30 °C. The PLRV content of M. persicae fed on infected potato leaves decreased with increasing time after transfer to turnip (immune to PLRV). The decrease occurred in two phases, the first rapid and the second very slow. In the first phase the decrease was faster, briefer and greater at 25 and 30 °C than at 15 and 20 °C. No evidence was obtained that PLRV multiplies in M. persicae. These results are compatible with a model in which much of the PLRV in aphids during the second phase is in the haemocoele, and transmission is mainly limited by the rate of passage of virus particles from haemolymph to saliva. The potato aphid, Macrosiphum euphorbiae, transmitted PLRV much less efficiently than M. persicae. Its inefficiency as a vector could not be ascribed to failure to acquire or retain PLRV, or to the degradation of virus particles in the aphid. Probably only few PLRV particles pass from the haemolymph to saliva in this species. The virus content of M. euphorbiae collected from PLRV-infected potato plants in the field increased from early June to early July, and then decreased. PLRV was detected both in spring migrants collected from the plants and in summer migrants caught in yellow water-traps. PLRV was also detected in M. persicae collected from infected plants in July and August, and in trapped summer migrants, but their PLRV content was less than that of M. euphorbiae, and in some instances was too small for unequivocal detection.     

The effects of temperature on the availability and acquisition of potato leafroll luteovirus by Myzus persicae

The concentration of potato leafroll luteovirus (PLRV) did not differ in potato plants with secondary infections grown at 15°C or 27°C. Detached leaves of plants grown at 15°C or 27°C were used as sources of PLRV for peach-potato aphids (Myzus persicae Sulz.) both at 15°C and 27°C. At comparable temperature during virus acquisition, aphids which fed on leaves of plants kept previously at 15°C contained more viral antigen detected by ELISA than aphids which fed on leaves of plants grown at 27°C. The aphids which acquired PLRV at 27°C contained evidently more viral antigen than those which acquired PLRV at 15°C. The greatest amount of PLRV was found in the aphids which acquired the virus at 27°C from the leaves of plants kept at 15°C. The ability of M. persicae to transmit PLRV to Physalis ftoridana Rydb. generally decreased with decrease in the amount of PLRV in vectors.     

Can plants use an entomopathogenic virus as a defense against herbivores?

It is by now well established that plants use various strategies to defend themselves against herbivores. Besides conventional weapons such as spines and stinging hairs and sophisticated chemical defenses, plants can also involve the enemies of the herbivores in their defense. It has been suggested that plants could even use entomopathogens as part of their defense strategies. In this paper, we show that Brassica oleraceae plants that are attacked by Myzus persicae aphids infected with an entomopathogenic parvovirus (M. persicae densovirus) transport the virus through the phloem locally and systematically. Moreover, healthy aphids that fed on the same leaf, but separated from infected aphids were infected via the plant. Hence, this is proof of the principle that plants can be vectors of an insect virus and can possibly use this virus as a defense against herbivores.     

Some effects of spraying with thiabendazole on the susceptibility of sugar beet to yellowing viruses and on their vector, Myzus persicae (Sulz.)

In a field experiment fewer sugar-beet plants became infected with aphid-transmitted yellowing viruses in plots that had been sprayed with solutions of thiabendazole lactate than in water-sprayed plots, after exposure to natural infestation with aphids. Subsequent glasshouse tests showed that foliar sprays of o·o1 % thiabendazole lactate in water significantly reduced the proportion of inoculated sugar-beet plants which became infected with beet yellows virus (BYV) or beet mild yellowing virus (BMYV) after inoculation with viruliferous Myzus persicae (Sulz.). This effect on virus transmission was not apparently due to a direct insecticidal action of thiabendazole, because adult aphids usually survived equally well on sprayed and unsprayed plants. Treatment of test plants with thiabendazole did not affect the transmission of beet mosaic virus to them by M. persicae. The fecundity of M. persicae was greatly reduced by transferring them to plants which had been sprayed with thiabendazole or by spraying them with thiabendazole before transfer to unsprayed plants. The fertility of adult Aphis fabae Scop, was also reduced by spraying with thiabendazole. The mechanisms whereby thiabendazole affected fecundity of aphids and transmission of viruses are not understood.     

The incidence of viruses in wild Brassica nigra in Dorset (UK)

Using enzyme‐linked immunosorbent assays, the frequency of occurrence of six viruses was determined in Brassica nigra collected from five coastal sites in Dorset, spanning approximately 24 km. During 1998–2000, the viruses detected were: Turnip mosaic virus (genus Potyvirus) (TuMV), Turnip yellow mosaic virus (genus Tymovirus) (TYMV), Turnip crinkle virus (genus Carmovirus) (TCV), Turnip rosette virus (genus Sobemovirus) (TRoV), Beet western yellows virus (genus Polerovirus) (BWYV) and Cauliflower mosaic virus (genus Caulimovirus) (CaMV). Multiple infections were detected in some individuals (48/447). TuMV was detected infrequently over the three‐year period (5/597). A representative isolate of each virus was tested for its effects on glasshouse‐grown individuals from different half‐sib families of B. nigra from four of the sites. Whether inoculated manually or via aphids (Myzus persicae), TuMV caused a rapid (within 10 days) lethal systemic necrosis in the B. nigra seedlings except when they were near flowering at the time of inoculation. Each of the other viruses invaded systemically but were not lethal. Indeed, BWYV systemically invaded 13/19 glasshouse‐grown B. nigra seedlings but did not produce any visible symptoms. Otherwise, the isolates tested differed in their pathogenicity and in the symptoms they produced in infected B. nigra. With TYMV or TCV viral antigen concentration was closely linked to pathogenicity; for TRoV or CaMV, there was little or no difference in virus concentration between plants with and without symptoms. Substantial and reproducible differences were observed in sensitivity/susceptibility among B. nigra genotypes from different sites in Dorset challenged with the same virus isolate.     

Aphid-injection experiments with carrot mottle virus and its helper virus, carrot red leaf

Carrot mottle virus (CMotV) and its helper virus, carrot red leaf (CRLV), were not transmitted by aphids (Cavariella aegopodii) that had fed through membranes on, or had been injected with, sap from mixedly infected chervil plants or partially purified preparations of CMotV. However, the viruses were transmitted by recipient aphids injected with haemolymph from donor aphids that had fed on mixedly infected plants but not by a second series of recipients injected with haemolymph from the first series. Some of the first series of recipients transmitted both viruses for up to 11 days but others transmitted erratically and many lost ability to transmit after a few days. The results confirm that both viruses are circulative but provide no evidence for multiplication in the vector. Non-viruliferous aphids, or aphids that had acquired CRLV by feeding, did not transmit CMotV when they were injected with haemolymph from aphids that had fed on a source of CMotV alone, confirming that they can only transmit CMotV when they acquire it from a mixedly infected plant. When extracts from donor aphids were treated with ether before injection, recipient aphids transmitted both CRLV and CMotV, although the infectivity of CMotV grown in Nicotiana clevelandii in the absence of CRLV is destroyed by ether treatment. CMotV particles acquired by aphids from mixedly infected plants therefore differed in some way from those in singly infected plants. A plausible explanation of these results, and of the dependence of CMotV on CRLV for aphid transmission, is that doubly infected plants contain some particles that consist of CMotV nucleic acid coated with CRLV protein.     

Cauliflower mosaic virus protein P6-TAV plays a major role in alteration of aphid vector feeding behaviour but not performance on infected Arabidopsis

Emerging evidence suggests that viral infection modifies host plant traits that in turn alter behaviour and performance of vectors colonizing the plants in a way conducive for transmission of both nonpersistent and persistent viruses. Similar evidence for semipersistent viruses like cauliflower mosaic virus (CaMV) is scarce. Here we compared the effects of Arabidopsis infection with mild (CM) and severe (JI) CaMV isolates on the feeding behaviour (recorded by the electrical penetration graph technique) and fecundity of the aphid vector Myzus persicae. Compared to mock-inoculated plants, feeding behaviour was altered similarly on CM- and JI-infected plants, but only aphids on JI-infected plants had reduced fecundity. To evaluate the role of the multifunctional CaMV protein P6-TAV, aphid feeding behaviour and fecundity were tested on transgenic Arabidopsis plants expressing wild-type (wt) and mutant versions of P6-TAV. In contrast to viral infection, aphid fecundity was unchanged on all transgenic lines, suggesting that other viral factors compromise fecundity. Aphid feeding behaviour was modified on wt P6-CM-, but not on wt P6-JI-expressing plants. Analysis of plants expressing P6 mutants identified N-terminal P6 domains contributing to modification of feeding behaviour. Taken together, we show that CaMV infection can modify both aphid fecundity and feeding behaviour and that P6 is only involved in the latter.     

The effect of mineral oil on stylet activities and potato virus Y transmission by aphids

Mineral oil sprayed onto potato virus Y (PVY) infected tobacco plants reduced acquisition of this potyvirus by Myzus persicae (Sulz.). Although the pre-penetration activities of aphids were longer on oil treated leaves, the inhibitory effect of the oil could not be attributed to differences in the duration of stylet penetration. Aphids were therefore made part of a DC circuit in order to investigate their stylet activities during penetration of PVY infected source plants and healthy test plants. Both acquisition and inoculation of the virus were reduced by the presence of oil on the plant surface, but these reductions could not be related to electrically recorded differences in plant penetration behaviour. In particular, stylet punctures of plant cell membranes were not reduced by mineral oil. Non-behavioural reasons are suggested to explain the mode of action of the oil.     

The transmission of potato virus Y by aphids of different vectoring abilities

Adult apterae of Myzus persicae and Macrosiphum euphorbiae that did not transmit potato virus Y^N (PVY^N) in a first test were as likely to transmit the virus in a subsequent test as those that did transmit on the first occasion. Only 16% of M. persicae that were allowed a single acquisition probe into a leaf infected with both PVY^O and PVY^N transmitted both strains, 37% transmitted either PVY^O or PVY^N and 47% did not transmit. There was no difference in the duration of probes that did or did not result in virus transmission. Statistical models were fitted to data on the frequency of transmission of PVY^O, PVY^N or both PVY^O and PVY^N by M. persicae and by aphids of poorer vector species, M. euphorbiae and Rhopalosiphum padi. Transmission of the two viruses ocurred independently of each other and consequently transmission of both was rare with M. euphorbiae and R. padi. Mineral oil applied to leaves infected with both strains diminished the frequency of transmission by M. persicae. Fitted models suggested that the aphids that probed through the oil droplets on leaves treated 30 min previously did not transmit virus, and that 24 h later, when the droplets had spread, aphids probing through them could transmit but with a decreased ability.     

球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响

在室内评价了球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响。分别在烟蚜茧蜂寄生桃蚜后不同时间进行高剂量（1900孢子/mm^{2）接菌,检测蚜虫感病率和寄生蜂形成的僵蚜率及僵蚜出蜂率。结果表明,球孢白僵菌对僵蚜率和僵蚜出蜂率的影响随接菌时间不同而变化。在烟蚜茧蜂寄生前1d、寄生当天和寄生后3d接菌,蚜虫感病率分别为59.6%、56.2%和34.8%;与对照相比,僵蚜率分别下降94%、59%和47%,僵蚜出蜂率分别减少83%、54%和49%。在寄生后5d或7d接菌,僵蚜率和僵蚜出蜂率不受明显影响,但蚜虫感病率降低到8.2%以下。对蚜尸内白僵菌菌体含量检测表明,随着烟蚜茧蜂寄生后接菌时间的推移,菌体数量迅速下降。寄生蜂寄生后5d或7d接菌,蚜尸内几乎检测不到菌体。直接喷雾接菌烟蚜茧蜂,成蜂寿命缩短4d左右,且81.8%的蜂尸受白僵菌感染。接菌后的寄生蜂对蚜虫寄生率几乎无影响,但寄生蜂在蚜虫体内的存活时间缩短了27.8%。}     

Infection of potato plants with potato leafroll virus changes attraction and feeding behaviour of Myzus persicae

Potato leafroll virus (PLRV; genus Polerovirus, family Luteoviridae) is a persistently transmitted circulative virus that depends on aphids for spreading. The primary vector of PLRV is the aphid Myzus persicae (Sulzer) (Homoptera: Aphididae). Solanum tuberosum L. potato cv. Kardal (Solanaceae) has a certain degree of resistance to M. persicae: young leaves seem to be resistant, whereas senescent leaves are susceptible. In this study, we investigated whether PLRV‐infection of potato plants affected aphid behaviour. We found that M. persicae's ability to differentiate headspace volatiles emitted from PLRV‐infected and non‐infected potato plants depends on the age of the leaf. In young apical leaves, no difference in aphid attraction was found between PLRV‐infected and non‐infected leaves. In fact, hardly any aphids were attracted. On the contrary, in mature leaves, headspace volatiles from virus infected leaves attracted the aphids. We also studied the effect of PLRV‐infection on probing and feeding behaviour (plant penetration) of M. persicae using the electrical penetration graph technique (DC system). Several differences were observed between plant penetration in PLRV‐infected and non‐infected plants, but only after infected plants showed visual symptoms of PLRV infection. The effects of PLRV‐infection in plants on the behaviour of M. persicae, the vector of the virus, and the implications of these effects on the transmission of the virus are thoroughly discussed.     

Effects of the antifeedant polygodial on plant penetration by aphids, assessed by video and electrical recording

Adult apterousMyzus persicae (Sulz.) discriminated within 2 min between mature Chinese cabbage (Brassica pekinensis L.) leaf halves treated with (±)-polygodial solution and solvent alone, and walked off leaf areas treated with polygodial faster than off the solvent-treated areas. However, when aphids were attached to a fine gold wire and stylet penetration of cabbage leaves was recorded electrically, polygodial treatment did not affect the number or duration of electrically-recorded penetrations, time taken to initiate a first penetration, or total penetration time. As the tethering of aphids causes behavioural restrictions which may negate the response to polygodial, indications of stylet penetration by freely-moving insects were sought. Simultaneous electrical recording and video monitoring showed that stylet penetration duration could be accurately inferred from antennal and body movements, enabling assessment of penetration times without attachment to the wire tether. When freely-movingM. persicae were video recorded during 15 min access to cabbage seedlings, polygodial treatment again had no apparent effect on stylet penetration. However, when aphids were presented with a choice of polygodial- and solvent-treated sides of floating mature cabbage leaf discs, video recordings revealed that the insects spent less time and made fewer penetrations on the polygodial-treated side. In addition to this rapid repellent effect, prolonged exposure to polygodial also produced behavioural changes. After being held for 24 h on polygodial-treated leaves or green paper prior to behavioural examination, aphids penetrated seedlings fewer times but for longer periods. The relevance of the results to virus transmission studies is discussed.     

Further studies on cucumber mosaic virus infection of narrow-leafed lupin (Lupinus angustifolius): seed-borne infection, aphid transmission, spread and effects on grain yield

Four field trials were done with narrow-leafed lupins (Lupinus angustifolius) in 1988 - 1989, to examine the effect of sowing seed with 5% and 0.5% cucumber mosaic virus (CMV) infection on subsequent virus spread, grain yield and percentage of infection in harvested seed. A proportion of the CM V-infected seed failed to produce established plants and thus, plots sown with 5% and 0.5% infected seed contained 1.5-2.9% and 0.2-0.3% of seed-infected plants respectively. The rate of virus spread by aphids was faster and resulted in more extensive infection at maturity in plots sown with 5% infected seed than with 0.5% infected seed. In three trials, sowing 5% infected seed resulted in yield losses of 34 - 53% and CMV infection in the seed harvested of 6 - 13%. The spread of CMV infection resulting from sowing 0.5% infected seed did not significantly decrease yield. However, late CMV spread in these plots caused > 1% seed infection. In the fourth trial, which was badly affected by drought, CMV spread only slowly, there was no significant effect of CMV on grain yield and the percentage of infected seed harvested was 3–5 times less than that in the seed sown. When CMV-infected seed was sown at different depths, target depths of 8 and 11 cm decreased the incidence of seed-infected plants by c. 15% and c. 50% respectively compared with sowing at 5 cm. However, in glasshouse tests, treatment with the pre-emergence herbicide simazine failed to selectively cull out seed-infected plants. The field trials were colonised by green peach (Myzus persicae), blue-green (Acyrthosiphon kondoi) and cowpea (Aphis craccivora) aphids. When the abilities of these aphid species and of the turnip aphid (Lipaphis erysimi) in transmitting CMV from lupins to lupins were examined in glasshouse tests, short acquisition access times favoured transmission. With 5–10 min acquisition access times, overall transmission efficiencies were 10.8%, 9.4%, 6.1% and 3.9% for the green peach, cowpea, blue-green and turnip aphids respectively.     

Cytopathology, Mode of Aphid Transmission and Search for the Casual Agent of Cotton Bunchy Top Disease

The cytopathological effects of cotton bunchy top (CBT) disease and its mode of transmission by Aphis gossypii Glover (cotton aphid), were studied. CBT infection affected the leaf epidermal layer producing a loose, ruptured and rough surface morphology with many stomata closed and misshapen. Roots of CBT‐infected plants showed reduced growth, small knots and a dark brown appearance. A single aphid per plant was capable of transmitting CBT at 5%, whereas three aphids per plant transmitted CBT to 50% of the cotton seedlings and 20 aphids per plant transmitted the disease agent to 80% of the cotton seedlings. Aphis gossypii acquired CBT after a minimum acquisition access period of 5 min and transmitted the agent after a minimum inoculation access period of 1 h. Both alate and apterous aphids and nymph instars 2, 3 and 4 of A. gossypii transmitted CBT. This preliminary data suggest that A. gossypii transmits CBT in a semi‐persistent manner. Myzus persicae Sulz (green peach aphid) was unable to transmit CBT. A comprehensive attempt to isolate the CBT agent, using a range of virological techniques including double‐stranded RNA extraction, two‐dimensional gel electrophoresis for viroid, circular DNA test, nanovirus polymerase chain reaction (PCR), luteovirus PCR and enzyme‐linked immunosorbent assay, phytoplasma test, nucleoprotein purification and electron microscopy, was unsuccessful, raising the possibility that CBT may be caused by a unique new pathogen.     

The effect of plant age and infection with virus yellows on the survival of Myzus persicae on sugar beet

Survival of Myzus persicae confined in clip-cages on mature leaves of sugar beet declined as the plants aged. Death of aphids was often preceded by the appearance of a black deposit in the aphids' stomachs, which may have been the cause of death. Both the rate of death and the proportion of aphids dying with black deposits was significantly less when plants were infected with beet yellows virus or beet mild yellowing virus, by comparison with healthy plants. The implication of these phenomena on the onset of mature plant resistance is discussed.     

Effects of sucrose sprays and darkness on aphid colonization of sugar beet and on aphid transmission of yellowing viruses

In the glasshouse, adult, apterous Myzus persicae (Sulz.) and Aphis fabae Scop, settled better and deposited more larvae on sucrose-sprayed sugar-beet plants than on water-sprayed plants. M. persicae settled badly and deposited few larvae on plants that were kept in the dark before or after infestation. The effects of darkness on aphids were reduced by spraying the host plants with 10% solutions of sucrose before infestation. Viruliferous M. persicae transmitted beet yellows virus (BYV) and beet mild yellowing virus (BMYV) less efficiently to dark-treated plants than to those grown in normal daylight. Spraying sugar beet with sucrose before inoculation with viruliferous M. persicae increased the proportion of successful BYV transmissions but only when the plants were dark-treated. The effects of sucrose and darkness on settling and larviposition of aphids and on virus transmission may be related to changes in the concentration of carbohydrates, particularly sugars, in the leaves.     

Seed transmission of turnip yellow mosaic virus in winter turnip and winter oilseed rapes

This is the first record of seed transmission of turnip yellow mosaic virus (TYMV) in oilseed and turnip rapes. The seed transmission of TYMV in a naturally infected winter turnip rape (Brassica napus var. silvestris) cultivar Perko PVH was investigated. By ELISA 1.6%, 3.2% and 8.3% seed transmission of the virus was found in seed of plants from three localities. The proportion of infected seeds produced by artificially infected plants of winter oilseed rape (Brassica napus ssp. oleifera) and winter turnip rape cultivars was determined. The virus transmission rate, expressed as the proportion of virus-infected plants which germinated from the seed was for the oilseed rape cvs Jet Neuf 0.1%, Solida 0.4%, Silesia 0.8%, Darmor 1.2%, SL-507 0.2%, SL-509 0.0% and for the winter turnip rape cv. Perko 1.5%. ELISA cannot be used in direct tests on bulk seed lots to estimate proportion of infected seed, but must be used on germinated seedlings.