Analogues of virus resistance genes map to QTLs for resistance to sharka disease in <Emphasis Type="Italic">Prunus davidiana</Emphasis>

Plum pox virus (PPV), the causative agent of sharka disease in Prunoideae, is one of the most serious problems affecting stone fruit production in Europe and America. Resistance to PPV was previously described in a Prunus davidiana clone, P1908, and introduced into peach (Prunus persica) genotypes. Genetic resistance to PPV displays a complex pattern of quantitative inheritance. An analysis of quantitative trait loci (QTLs) for resistance was performed on an F1 interspecific peach population obtained from a cross between the susceptible nectarine cultivar Summergrand and P. davidiana. The hybrids were graft-inoculated with PPV in duplicate following a classical procedure. The incidence of infection was evaluated four times, over two vegetative cycles, by symptom observation and enzyme-linked immunoadsorbent assays (ELISA). Restriction of systemic downward movement of the PPV virus was also evaluated by testing the susceptible rootstocks. Using both analysis of variance and non-parametric tests, six genomic regions involved in PPV resistance were detected. Depending on the scoring data considered, between 22 and 51% of the phenotypic variance could be explained by the quantitative model. One QTL, located in the distal region of linkage group 1, maps in a genomic region that is syntenic to the location of a resistance gene previously identified in the apricot cv. Goldrich. Some QTLs appeared to be temporally specific, reflecting the environmental dependence of PPV-resistance scoring. Candidate gene fragments were amplified by PCR, isolated and mapped on the peach interspecific linkage map. We report here the co-localization of three analogues of virus resistance genes with two distinct genomic regions linked to PPV resistance in P. davidiana.Electronic Supplementary Material Supplementary material is available for this article at     

The distribution of poplar mosaic virus in hybrid poplars and virus detection by ELISA

Purified poplar mosaic virus (PMV) at a concentration of 8 ng/ml was readily detected by enzyme-linked immunosorbent assay (ELISA). Bioassay in Nicotiana megalosiphon was more sensitive (detecting 1–4 ng/ml) and latex flocculation less sensitive (c. 25 ng/ml) than ELISA assays. While the foliar sap of fresh, naturally-infected poplars (e.g. Populus. euramericana cv. Robusta) was not infective at dilutions greater than 2 . 10–², ELISA easily detected PMV antigen when sap was diluted 4 . 10–³ in buffer or when one part of infected tissue was triturated with 99 parts healthy leaf. Furthermore, although sap from poplar leaves stored at -20 °C for 6 months was not infective, PMV was still detectable in ELISA tests. PMV antigen in poplar leaves was not all pelleted after centrifugation for 2.5 h at 130 000 g yet parallel tests using unbuffered sap from systemically infected Nicotiana megalosiphon foliage showed that infectivity was restricted to the pellet. In poplar foliage, the concentration of PMV antigen was generally greatest where symptoms were most obvious; least antigen was detected in the overwintering leaves located at the bases of long shoots. In winter, when root and inner bark tissue in the trunk was an erratic source of PMV, the virus was readily detected in buds, the concentration being greatest in the bases, including the meristem, of terminal buds. Propagation from single node cuttings of P. euramericana cv. Regenerata allowed the selection of clones that consistently showed either ‘severe’ or ‘mild’ foliar symptoms. The associated virus isolates also infected another poplar clone causing symptoms characteristic of their source. ELISA consistently detected less PMV antigen in field-grown cv. Regenerata than in cv. Robusta foliage, but this was reversed when the associated virus isolates were propagated in Nicotiana glutinosa at 24 °C. During 6 yr, 21 out of 127 poplars at a site in Western England, became infected with PMV. By contrast, in Eastern England, none of 46 were infected. The aphids Pterocomma populea and Myzus persicae did not transmit PMV.     

Effects of fungicides and insecticides applied to spring barley sown on different dates in 1976-79

Factorial experiments in 1976–1979 investigated the effects of sowing date, fungicides (ethirimol seed treatments and tridemorph sprays) and insecticides (phorate applied to the soil, and menazon or dimethoate sprays) on powdery mildew, aphids, barley yellow dwarf virus (BYDV) and grain yield of spring barley (cv. Julia in 1976 and 1977; cv. Wing in 1978 and 1979). Late sowing usually increased the severity of powdery mildew, numbers of aphids and incidence of BYDV and generally decreased yield. Responses to pesticides were commonly greater on the late-sown than on the early-sown barley. Response to fungicides are principally attributed to the control of powdery mildew (Erysiphe graminis f. sp. hordei; the target species) but responses to insecticides cannot be attributed to virus control and seem unlikely to be due solely to control of aphids, whose numbers were relatively small. There were some effects of fungicides on aphids and insecticides on mildew, but they were inconsistent and too small to affect crop protection strategies.     

Restricted distribution of potato leafroll virus antigen in resistant potato genotypes and its effect on transmission of the virus by aphids

Enzyme-linked immunosorbent assay was used to measure the concentration of potato leafroll virus (PLRV) antigen in different parts of field-grown secondarily infected plants of three potato genotypes known to differ in resistance to infection. The antigen concentration in leaves of cv. Maris Piper (susceptible) was 10–30 times greater than that in cv. Pentland Crown or G 7445(1), a breeder's line (both resistant). Differences between genotypes in antigen concentration were smaller in petioles and tubers (5–10-fold) and in above-ground stems (about 4-fold), and were least in below-ground stems, stolons and roots (about 2-fold). PLRV antigen, detected by fluorescent antibody staining of tissue sections, was confined to phloem companion cells. In Pentland Crown, the decrease in PLRV antigen concentration in leaf mid-veins and petioles, relative to that in Maris Piper, was proportional to the decrease in number of PLRV-containing companion cells; this decrease was greater in the external phloem than in the internal phloem. The spread of PLRV infection within the phloem system seems to be impaired in the resistant genotypes. Green peach aphids (Myzuspersicae) acquired < 2800 pg PLRV/aphid when fed for 4 days on infected field-grown Maris Piper plants and < 58% of such aphids transmitted the virus to Physalis floridana test plants. In contrast, aphids fed on infected Pentland Crown plants acquired <120 pg PLRV/aphid and <3% transmitted the virus to P. floridana. The ease with which M. persicae acquired and transmitted PLRV from field-grown Maris Piper plants decreased greatly after the end of June without a proportionate drop in PLRV concentration. Spread of PLRV in potato crops should be substantially decreased by growing cultivars in which the virus multiplies to only a limited extent.     

Quantitative trait loci analysis of <Emphasis Type="Italic">Plum pox virus</Emphasis> resistance in <Emphasis Type="Italic">Prunus davidiana</Emphasis> P1908: new insights on the organization of genomic resistance regions

No valuable source of resistance to Plum pox virus (PPV), the causative agent of sharka disease, has been found in peach (Prunus persica), but polygenic resistance to PPV was described in Prunus davidiana clone P1908. Two previous studies using F₁ and F₂ populations derived from the nectarine cv. Summergrand and P. davidiana P1908 identified a total of six P. davidiana quantitative trait loci (QTLs) involved in PPV resistance (Marcus strain). In an effort to verify the QTL stability in a large progeny and to search for possible interactions of the genetic backgrounds, the current study evaluated the incidence of PPV infection in an F₁ population derived from the susceptible peach cv. Rubira and P. davidiana P1908 over three growth periods using an improved method of PPV phenotyping referred to as “heavy test.” The phenotypic dataset was analyzed using similar methods as the previous studies and a newly developed simple-sequence-repeat-based P. davidiana map. Nine regions involved in differential symptom expression were identified among which six were common between studies. However, the level of resistance observed in the population was very low compared to the other studies, and the main QTL previously identified in linkage group 6 was not conserved, suggesting strong interaction of the genetic background of the susceptible parent with that of P. davidiana 1908. Consequently, this could be a limiting factor for developing resistant cultivars derived from P. davidiana P1908.     

Interspecific transfer of resistance to Plum pox virus from almond to peach by grafting

Grafting almond variety ‘Garrigues’ onto ‘GF305’ peach seedlings heavily infected by Plum pox virus (PPV) progressively produces the disappearance of viral symptoms and drastically reduces virus accumulation in ‘GF305’ rootstock, in most cases to undetectable levels. This response appears to be specific between almond and peach, as it was not consistently observed by grafting ‘Garrigues’ onto other Prunus species such as plum (‘Adesoto’) or apricot (‘Real Fino’). The ability to induce resistance to PPV in ‘GF305’ was transmitted to the sexual descendants of Garrigues. Furthermore, grafting ‘Garrigues’ onto ‘GF305’ before PPV inoculation completely prevented virus infection, showing that the resistance is constitutive and not induced by the virus. This fact suggests that resistance may be due to the transfer of a defence factor from ‘Garrigues’ almond through the graft union and its interaction with specific factors of ‘GF305’ peach to produce the antiviral response. These results open new avenues to potential protection against PPV in peach, the most economically important species among stone fruits.     

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.     

Susceptibility of Prunus rootstocks to natural infection of Plum pox virus and effect of mineral oil treatments

The use of rootstocks that are less susceptible or resistant to natural Plum pox virus (PPV) infection and/or the application of mineral oil treatments are two possible strategies to reduce viral incidence in nursery plots. We evaluated the susceptibility of Prunus rootstocks used in the Spanish stone fruit industry and the effect of mineral oil treatment (Sunspray Ultrafine at 1%) on the spread of the virus at two different localities in Valencia, Spain, under different natural PPV inoculum pressures (high inoculum pressure, 2006–08; low inoculum pressure, 2006–07). Samples from both plots were analysed by double-antibody sandwich indirect enzyme-linked immunosorbent assay (DASI-ELISA) and spot real-time RT-PCR. Under high inoculum pressure, the assayed rootstocks exhibited significant differences in their susceptibility to natural infection. The most susceptible rootstocks at the end of the experiment were Adesoto 101 and Mariana GF8-1. Cadaman and Garnem rootstocks presented the fewest PPV-infected plants; these infections could be detected only by spot real-time RT-PCR. No differences among the assayed rootstocks were found under low PPV inoculum pressure. Aphid species were monitored using Moericke yellow water traps and sticky-plant methods at both localities in May 2006 and 2007. Aphis spiraecola was the most abundant aphid species monitored by both methods at both localities. The average percentage of A. spiraecola carrying PPV PCR-amplifiable targets was 30.37% in the plot with high PPV inoculum pressure and only 7.98% in the plot with low inoculum pressure. We found significant differences in PPV incidence between Mariana GF8-1 plots that were treated with mineral oil and those that were not treated after one year under natural conditions and high PPV inoculum pressure.     

Effects of infection with the cabbage black ringspot strain of turnip mosaic virus on turnip as a host to Myzus persicae and Brevicoryne brassicae

When turnip plants with 3–7 leaves were inoculated with cabbage black ringspot virus (CBRSV) on the 3rd rough-leaf, symptoms only appeared on leaves that had been less than 15 mm long at the time of inoculation, although infection decreased the area and both fresh and dry weight of all leaves. Leaves were ‘aged’ by their appearance and placed in Leaf Age Categories (LACs). Leaves with symptoms senesced (‘aged’) prematurely. CBRSV-infection of cv. Green Top White did not change the distribution of populations of Myzus persicae between LACs, but increased the proportion of the plant suitable for colonisation. All suitable LACs were quickly colonised by adult apterae and nymphs. On CBRSV-infected plants the nymphal period was shorter, F₁ adults deposited larvae more frequently and the live body weight and tibial length of the F₂ generation was greater, than on healthy plants. The distribution of Brevicoryne brassicae populations on cv. Green Top White differed from that of M. persicae but was also unchanged by CBRSV-infection. On healthy plants the largest colonies were on mature leaves, so that on virus-infected plants premature senescence shortened the life of the colony. On CBRSV-infected plants the nymphal period was prolonged and the live weight of F₁ and F₂ adult apterae was less than on healthy plants. The differences between the biology of M. persicae and B. brassicae on CBRSV-infected cv. Green Top White were associated with the accelerated senescence of CBRSV-infected leaves. The possibility that CBRSV-infection might reduce the resistance of turnips to aphid infestation was tested. M. persicae and B. brassicae were cultured on two favourable and two less favourable cultivars. No improvement in population growth rate was found when the less favourable host cultivars were infected with CBRSV, but both aphid species weighed less and/or had smaller nymphal populations on cultivars showing the severest symptoms. These results are discussed in relation to the evolution of non-persistent virus transmission by aphids.     

Differential Responses of Guayule (Parthenium argentatum Gray) Genotypes to Culture Filtrate and Toxin from Macrophomina phaseolina (Tassi) Goidanich

The potential for using cell-free culture filtrate (CFCF) and toxin (phaseolinone) from Macrophomina phaseolina for rapid and effective screening procedures for charcoalrot resistance in guayule (Parthenium argentatum) germplasm was assessed. The CFCF and partially purified phaseolinone were incorporated into modified Murashige and Skoog solid medium aat the rates of 0-100% (v/v) and 0-1000 μg ml^-1 respectively. The medium pH was adjusted to 5.8 before solidifying with 0.8% agar. Fourweek-old seedlings of 10 guayule genotypes were plantanted in the medium, incubated and rated for phytotoxic symptoms and tissue damage over a 15-day period. In a green-house study, seedling growth, phytotoxicity and damage severity were compared in 12-week-old guayule seedlings root-inoculated with M. phaseolina microsclerotia. There were significant differences (P = 0.05) in genotypic responses to the fungus, the filtrate and the toxin inoculations. Time until phytotoxic symptoms developed was inversely related to the concentrations of CFCF and the toxin. Phytotoxic symptoms were produced 6 days after exposure to 50% CFCF and 48 h after exposure to 1000μg ml^-1 of partially purified phaseolinone. A comparison of photomicrographs of the control and toxintreated root tissues revealed no damage to the control roots and extensive damage to epidermal layers of the treated roots, which was evident 48 h after exposure to 100μg ml^-1 level of phaseolinone. Significant correlations were found between tolerance to the fungus and insensitivity to the culture filtrate (r = 0.89, P = 0.05) and the toxin (r = 0.95, P = 0.001) suggesting the possibility of screening for resistance to M. phaseolina using CFCF or phaseolinone. The genotypic reactions to the CFCF were also correlated with reactions to the toxin (r = 0.90, P = 0.05). Guayule breeding lines‘UC101 and‘P3-1 exhibited the greatest tolerance to the pathogen and insensitivity to the CFCF or the toxin whereas‘Ca16′,‘Cal7′,‘N576′,‘N9-5′, 11605’and‘N6-5’were very susceptible to the pathogen and sensitive to the CFCF or the toxin.     

Transmission and differentiation of six viruses infecting hogweed (Heracleum sphondylium) in Scotland

Six viruses, code-named HV1-HV6, were transmitted manually and/or by aphids (Cavariella spp. from symptomless wild plants of hogweed (Heracleum sphondylium) in Scotland. HV1 was identified as parsnip yellow fleck virus (PYFV); anthriscus yellows virus, on which it depends for transmission by aphids, was presumably also present in the hogweed plants. HV2 was transmitted manually and by aphids and had very flexuous filamentous particles c. 700–750 nm long; it has affinities with the closteroviruses, and the name heracleum latent virus is proposed. HV3, HV4 and HV5 were transmitted manually, HV3 and HV5 also by aphids, but their particle morphology is unknown. HV6 was transmitted only by aphids and has very flexuous particles up to 1400 nm long; it is presumably a closterovirus distinct from HV2. All the viruses infected cultivated umbelliferous species experimentally but only PYFV is known to infect umbelliferous crops.     

Detection and Identification of ‘Candidatus Phytoplasma prunorum’, ‘Candidatus Phytoplasma mali’ and ‘Candidatus Phytoplasma pyri’ in Stone Fruit Trees in Poland

A survey was made to determine the incidence of phytoplasmas in 39 sweet and sour cherry, peach, nectarine, apricot and plum commercial and experimental orchards in seven growing regions of Poland. Nested polymerase chain reaction (PCR) using the phytoplasma‐universal primer pairs P1/P7 followed by R16F2n/R16R2 showed the presence of phytoplasmas in 29 of 435 tested stone fruit trees. The random fragment length polymorphism (RFLP) patterns obtained after digestion of the nested PCR products separately with RsaI, AluI and SspI endonucleases indicated that selected Prunus spp. trees were infected by phytoplasmas belonging to three different subgroups of the apple proliferation group (16SrX‐A, ‐B, ‐C). Nucleotide sequence analysis of 16S rDNA fragment amplified with primers R16F2n/R16R2 confirmed the PCR/Restriction Fragment Length Polymorphism (RFLP) results and revealed that phytoplasma infecting sweet cherry cv. Regina (Reg), sour cherry cv. Sokowka (Sok), apricots cv. Early Orange (EO) and AI/5, Japanese plum cv. Ozark Premier (OzPr) and peach cv. Redhaven (RedH) was closely related to isolate European stone fruit yellows‐G1 of the ‘Candidatus Phytoplasma prunorum’ (16SrX‐B). Sequence and phylogenetic analyses resulted in the highest similarity of the 16S rDNA fragment of phytoplasma from nectarine cv. Super Queen (SQ) with the parallel sequence of the strain AP15 of the ‘Candidatus Phytoplasma mali’ (16SrX‐A). The phytoplasma infecting sweet cherry cv. Kordia (Kord) was most similar to the PD1 strain of the ‘Candidatus Phytoplasma pyri’ (16SrX‐C). This is the first report of the occurrence of ‘Ca. P. prunorum’, ‘Ca. P. mali’ and ‘Ca. P. pyri’ in naturally infected stone fruit trees in Poland.     

Nucleosides VIII: 1 Synthesis of 2′, 3′-Dideoxy- and 2′, 3′-Didehydro-2′, 3′-Di Deoxyisoguanosine as Potential Antiretroviral Agents

Abstract

2′, 3′-Didehydro-2′, 3′-dideoxyisoguanosine (2) and 2′, 3′- dideoxyisoguanosine (3) have been synthesized by utilizing the Corey-Winter approach starting from isoguanosine. The 6-amino and 5′-hydroxy biprotected isoguanosine derivative was converted to the corresponding 2′, 3′- thionocarbonate, which was heated with triethyl phosphite to afford the 2′,3′- olefinic product. Either a tert-butyldimethylsilyl or a 4, 4′-dimethoxytrityl group was used in the protection of 5′-hydroxy function. Compounds 2 and 3 were found inactive against human immunodeficiency virus (HIV), human cytomegalovirus (HCMV), and herpes simplex virus type 1 (HSV-1).

     

Horticultural mineral oil influences Plum pox virus transmission by Myzus persicae

The residual activity of horticultural mineral oil (HMO) on the ability of green peach aphids, Myzus persicae (Sulzer), (GPA) to transmit Plum pox virus (PPV) to peach was measured by infection rates of detached leaves from plants sprayed with either HMO or water as a control that were inoculated using transfer of 25 viruliferous aphids per leaf at 0, 2, 4, 7, 9, 11 and 14 days after treatment (DAT). Persistent effects of HMO residue on the probing and feeding behaviours of GPA were also monitored with the electrical penetration graph (EPG) system. For glasshouse‐grown peach seedlings, the residual activity of HMO reduced PPV infection rates by more than 58% for up to 4 DAT following an initial reduction of approximately 81%. EPG recordings of GPA feeding behaviour showed that HMO significantly delayed first feeding probes and first intracellular punctures by more than 50 min without changing the ensuing stylet penetration behaviour. Applying HMO reduced virus infection rates for up to a week depending on the environmental conditions. EPG monitoring of aphid probing showed that HMO reduced the mean duration and mean number of potential drop (PD) phase feeding occurrences, compared with the water control. A reduction in the PD that has been shown to be related to the transmission of non‐persistently transmitted viruses may partly explain the reduction in PPV infection rates.     

The role of the helper virus, anthriscus yellows, in the transmission of parsnip yellow fleck virus by the aphid Cavariella aegopodii

Transmission of parsnip yellow fleck virus (PYFV) by the aphid Cavariella aegopodii occurs only when the aphids are also carrying the helper virus, anthriscus yellows (AYV). None of five other viruses tested was able to act as helper. In experiments in which aphids were allowed to feed through membranes on crude or treated extracts from infected plants, aphids already carrying AYV acquired PYFV, but virus-free aphids failed to acquire either AYV or PYFV. PYFV was not transmitted by insects injected with haemolymph from aphids carrying both viruses, or with purified preparations of PYFV. PYFV was transmitted when AYV-carrying aphids, except those whose stylets had been removed, were contaminated externally with PYFV preparations. Ultraviolet irradiation of infected leaves did not prevent aphids from acquiring AYV, presumably because it is confined to deeply-lying tissues. AYV-carrying aphids could acquire PYFV from u.v.-irradiated leaves after acquisition access times of 2 h but not after feeds of only 2 or 15 min (which are adequate on unirradiated leaves), suggesting that PYFV is present in all parts of the leaf. No ‘helper agent’ distinct from AYV itself was detected in these experiments or in experiments on minimum acquisition feeding time or maximum period of persistence in the aphid. U.v.-inactivated PYFV competed with infective PYFV for retention sites in AYV-carrying aphids, whereas AYV apparently did not. It is suggested that there is no helper agent for PYFV, other than AYV particles. The possibility that there is one for AYV is not excluded.     

Aphid behavioral responses to virus‐infected plants are similar despite divergent fitness effects

We compared the settling preferences and reproductive potential of an oligophagous herbivore, the pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae), in response to pea plants, Pisum sativum L. cv. ‘Aragorn’ (Fabaceae), infected with two persistently transmitted viruses, Pea enation mosaic virus (PEMV) and Bean leaf roll virus (BLRV), that differ in their distribution within an infected plant. Aphids preferentially oriented toward and settled on plants infected with PEMV or BLRV in comparison with sham‐inoculated plants (plants exposed to herbivory by uninfected aphids), but aphids did not discriminate between plants infected with the two viruses. Analysis of plant volatiles indicated that plants inoculated with either virus had significantly higher green leaf volatile‐to‐monoterpene ratios. Time until reproductive maturity was marginally influenced by plant infection status, with a trend toward earlier nymph production on infected plants. There were consistent age‐specific effects of plant infection status on aphid fecundity: reproduction was significantly enhanced for aphids on BLRV‐infected plants across most time intervals, though mean aphid fecundity did not differ between sham and PEMV‐infected plants. There was no clear pattern of age‐specific survivorship; however, mean aphid lifespan was reduced on plants infected with PEMV. Our results are consistent with predictions of the host manipulation hypothesis, extended to include plant viruses: non‐viruliferous A. pisum preferentially orient to virus‐infected host plants, potentially facilitating pathogen transmission. These studies extend the scope of the host manipulation hypothesis by demonstrating that divergent fitness effects on vectors arise relative to the mode of virus transmission.     

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.     

Silencing of Plum pox virus 5′UTR/P1 sequence confers resistance to a wide range of PPV strains

An effective disease-control strategy should protect the host from the major economically important and geographically widespread variants of a pathogen. Plum pox virus (PPV) is the causal agent of sharka, the most devastating viral disease of Prunus species. We have shown previously that the hairpin RNA expression driven by h-UTR/P1, h-P1/HCPro, h-HCPro and h-HCPro/P3 constructs, derived from the PPV-M ISPaVe44 isolate, confers resistance to the homologous virus in Nicotiana benthamiana plants. Since the production of transgenic stone fruits and their evaluation for PPV resistance would take several years, the ISPaVe44-resistant plant lines were used to evaluate which construct would be the best candidate to be transferred to Prunus elite cultivars. To do that, nine PPV isolates of the D, M, Rec, EA and C strains originally collected from five Prunus species in different geographical areas, were typed by sequencing and used to challenge the transgenic N. benthamiana lines; 464 out of 464 virus-inoculated plants of lines h-UTR/P1, h-HCPro and h-HCPro/P3 showed complete and long-lasting resistance to the seven PPV isolates of D, M and Rec strains. Moreover, the h-UTR/P1 plants were also fully resistant to PPV-C and -EA isolates. Our data suggest that the h-UTR/P1 construct is of particular practical interest to obtain stone fruit plants resistant to the sharka disease.