Potato Virus Y from Petunia can cause Symptoms of Potato Tuber Necrotic Ringspot Disease (PTNRD)

A potyvirus known to be an important agent involved in causing a disease of trailing petunias, was identified as being a member of the necrotic strain of potato virus Y (PVY) using a number of monoclonal antibodies. The sequence of the coat protein gene for the PVY isolate was determined and when compared with sequences for other PVY strains it was shown to cluster closely with isolates of PVY^NTN and to have a recombination point present within the coat protein common with other isolates of PVY^NTN. When inoculated onto potato tuber necrotic ringspot disease (PTNRD) susceptible potato cultivars the petunia isolate was found to be capable of causing necrotic tuber symptoms, consistent with those caused by other isolates of PVY^NTN. Due to the number of similarities it is thought the petunia isolate belongs to the PVY^NTN group of isolates. Out of 24 species of bedding and pot plant crops tested, 19 were shown by mechanical inoculation to be susceptible to PVY, highlighting not only a clear risk to a number of commercially important plant species from PVY^NTN infected trailing petunias, but also other susceptible crops grown in these areas.     

A novel type of Potato virus Y recombinant genome,determined for the genetic strain PVYE

Two Potato virus Y (PVY) isolates collected in Brazil, PVY‐AGA and PVY‐MON, were identified as recombinants between two parent genomes, PVY^NTN and PVY‐NE‐11, with a novel type of genomic pattern. The new recombinants had an ordinary PVY^NTN genome structure for approximately 6·7‐kb from the 5′‐end of the genome whereas the 3′‐terminal 3·0‐kb segment had two fragments of NE‐11‐like sequence separated by another small PVY^NTN‐like fragment. PVY strains are defined based on the hypersensitive resistance (HR) response in potato indicators. Both PVY‐AGA and PVY‐MON isolates did not induce the HR in potato cultivars carrying Ny, Nc, or (putative) Nz genes and thus were able to overcome all known resistance genes to PVY. Only one of the two isolates, PVY‐AGA, induced a vein necrosis reaction in tobacco. The biological responses of the potato indicators and tobacco defined PVY‐MON as an isolate of the PVY^E strain. To distinguish PVY‐AGA and PVY‐MON from other PVY^NTN isolates, an RT‐PCR test was developed utilizing new specific primers from the capsid protein gene area and producing a characteristic 955‐bp band. Serological profiling of these PVY isolates with three monoclonal antibodies revealed an unusual reactivity, where one of the two commercial PVY^N‐specific monoclonal antibodies did not recognize PVY‐AGA. The ability of these new PVY recombinants to overcome resistance genes in potato producing mild or no symptoms, combined with the lack of serological reactivity towards at least one PVY^N‐specific antibody may present a significant threat posed by these isolates to seed potato production areas.     

Occurrence of Potato virus Y strain PVYNTN in foundation seed potatoes in Japan,and screening for symptoms in Japanese potato cultivars

In 2008 and 2009 seasons, a sudden increase in Potato virus Y (PVY) incidence was recorded in foundation seed potatoes in Hokkaido, northern Japan. This increase was obvious during the field inspection and the postharvest indexing. Molecular typing revealed that besides the previously reported strains of PVY^O and PVY^NA‐N, the most common strain identified was the recombinant PVY^NTN, with three characteristic recombinant junctions at the HC‐Pro, VPg and CP regions. No potato tuber necrotic ringspot disease (PTNRD) was observed in foundation seed potatoes in correlation with the presence of PVY^NTN. Moreover, an isolate with a typical PVY^NTN recombinant genome, namely Eu‐12Jp, did not induce PTNRD in 62 Japanese potato cultivars tested in both primarily and secondarily infected plants. Two cultivars carrying the extreme resistance gene Ry_chc were resistant to the infection with Eu‐12Jp, which presents potential sources of resistance to PVY^NTN. Eu‐12Jp induced systemic mottle in potato cultivars Desiree and King Edward carrying resistance genes Ny and Nc, respectively, but induced a hypersensitive reaction in potato cultivar Maris Bard, with the Nz hypothetical resistance gene typical of the PVY^Z strain group. Therefore, based on the genome structure and the reaction of the potato N resistance genes, Eu‐12Jp should be classified as PVY^Z‐NTN, as described for isolates from Idaho, USA recently. This is the first report of PVY^Z‐NTN in Japan and the sudden and increased occurrence of PVY^NTN/PVY^Z‐NTN represents a potential risk of PTNRD developing and increases the significance of PVY in Japan.     

Incidence of potato viruses and characterisation of <Emphasis Type="Italic">Potato virus Y</Emphasis> variability in late season planted potato crops in Northern Tunisia

Surveys were conducted of symptomatic potato plants in late season crops, from the major potato production regions in Northern Tunisia, for infection with six common potato viruses. The presence of Potato leafroll virus (PLRV), Potato virus Y (PVY), Potato virus X (PVX), Potato virus A (PVA), Potato virus S (PVS) and Potato virus M (PVM) was confirmed serologically with virus infection levels up to 5.4, 90.2, 4.3, 3.8, 7.1 and 4.8%, respectively. As PVY was prevalent in all seven surveyed regions, further biological, serological and molecular typing of 32 PVY isolates was undertaken. Only one isolate was shown to induce PVY^O-type symptoms following transmission to tobacco and to react only against anti-PVY^O-C antibodies. Typical vein necrosis symptoms were obtained from 31 samples, six of which reacted against both anti-PVY^N and anti-PVY^O-C antibodies showing they contained mixed isolates, while 25 of them reacted only with anti-PVY^N antibodies. An immunocapture RT-PCR molecular test using a PVY^NTN specific primer pair set in the 5’NTR/P1 genomic region and examination of recombinant points in three genomic regions (HC-Pro/P3, CI/NIa and CP/3’NTR) showed that all 25 serotype-N PVY isolates were PVY^NTN variants with similar recombinations to the standard PVYNTN-H isolate. This is the first report of the occurrence of the PVY^NTN variant and its high incidence in late season potatoes in Tunisia.     

Molecular characterization of potato virus YN isolates by PCR-RFLP

Based on the sequence polymorphism in the 5 terminal part of the viral genome, a range of PVY^N isolates were characterized by polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP). Three pairs of primers selected in the 5 non-translated and P1 protein region were tested. Two of them yielded PCR products of about 1Kb from all isolates tested. Restriction analysis of the PCR products gave two distinct electrophoretic patterns, whichever of the three enzymes was used. In this way, the 18 isolates were separated into two easily identifiable subgroups. All tuber necrosing isolates (PVY^NTN) were clustered in the same subgroup.     

The movement of potato virus Y (PVY) in the vascular system of potato plants

Potato virus Y (PVY) is responsible for major viral diseases in most potato seed areas. It is transmitted by aphids in a non-persistent manner, and it is spread in potato fields by the winged aphids flying from an infected source plant to a healthy one. Six different PVY strains groups affect potato crops: PVY^C, PVY^N, PVY^O, PVY^N:O, PVY^NTN, and PVY^N-Wi. Nowadays, PVY^NTN and PVY^N-Wi are the predominant strains in Europe and the USA. After the infection of the leaf and accumulation of the virus, the virus is translocated to the progeny tubers. It is known that PVY^N is better translocated than PVY^O, but little is known about the translocation of the other PVY strains. The translocation of PVY occurs faster in young plants than in old plants; this mature plant resistance is generally explained by a restriction of the cell-to-cell movement of the virus in the leaves. The mother tuber may play an important role in explaining mature plant resistance. PVY is able to pass from one stem to the other stems of the same plant through the vascular system of the mother tuber, but it is unknown whether this vascular link between stems is permanent during the whole life of the plant. Two greenhouse trials were set up to study the spread of PVY in the vascular system of the potato plant. The PVY-susceptible cultivar Charlotte was used for both trials. It was demonstrated that all stems growing from a PVY-infected tuber will become infected sooner or later, and that PVY^N-Wi translocates more efficiently to progeny tubers than PVY^NTN. It was also demonstrated that the progressive decay of the mother tuber in the soil reduces the possibility for virus particles to infect healthy stems through the vascular system of the mother tuber. This new element contributes to a better understanding of the mechanism of mature plant resistance.     

Characteristics of a resistance-breaking isolate of potato virus Y causing potato tuber necrotic ringspot disease

An Austrian isolate of potato virus Y^NTN, the causal agent of potato tuber necrotic ringspot disease (PTNRD), was serologically compared with seven Dutch PVY^N isolates. Using polyclonal and monoclonal antibodies, it was found indistinguishable from PVY^N. Determination of the nucleotide sequence of the coat protein cistron and comparison of the deduced amino acid sequence with coat protein sequences of other potyviruses revealed a high level of homology with PVY^N coat protein sequences. This confirmed the close taxonomic relationship of PVY^NTN with the PVY^N subgroup of potato virus Y. PVY^NTN is able to overcome all resistance genes known so far in commercial potato cultivars. Remarkably, transgenic PVY-protected tobacco plants are also resistant to PVY^NTN infection upon mechanical and aphid-mediated inoculation. These experiments indicate that genetically engineered resistance offers great potential in protection of potato to new aggressive strains of PVY^N.     

The effects of co‐infection by different Potato virus Y (PVY) isolates on virus concentration in solanaceous hosts and efficiency of transmission

The influence of co‐infection on concentration and accumulation of genetically different isolates of Potato virus Y (PVY) in potato and tobacco plants and the efficiency of transmission by Myzus persicae of PVY isolates from doubly versus singly infected plants were evaluated. The vector ability to simultaneously transmit two virus isolates was examined. Eight PVY isolates represented three strain groups: PVY^O (pathotype and serotype O), PVY^NW (pathotype N and serotype O), and PVY^NTN (pathotype and serotype N). Different diagnostic methods, including DAS‐ELISA, multiplex RT‐PCR, aphid transmission tests and bioassays, were applied to detect the presence of PVY isolates in source and assay plants. Significant reductions in concentrations of certain PVY isolates during co‐infection with other isolates were found both in potato and tobacco plants. The observed effects were both isolate‐ and host‐dependent in form. The highest rates of virus transmission by single aphids were recorded with PVY^NTN isolates, and the lowest ones with PVY^O isolates. Individual aphids of M. persicae were able to simultaneously transmit two PVY isolates. The frequency of transmission was generally low, but it reached as high as 20% for one of the isolate combinations. The findings presented in the work provide proof for antagonistic within‐plant interactions between isolates of PVY, with some implications of these interactions for virus transmission by aphid vectors. Consequently, this research contributes to a better understanding of the epidemiology of the disease caused by PVY.     

Characterization of potato Y potyvirus isolates from tomato crops in Islas Canarias (Spain)*

A disease caused by potato Y potyvirus (PVY) affects tomato plantations with variable severity in Tenerife Island. Affected plants show diverse symptoms such as necrotic lesions or mild to severe mosaic in leaves and whitish spots in green fruits that remain after ripening. Tomato PVY isolates and few potato and capsicum PVY isolates have been characterized on the basis of biological, serological and molecular criteria. All PVY isolates reacted positively to monoclonal antibodies specific for PVY^O/C or PVY^N strains, and nearly 50% of tomato PVY isolates were recognized by both. Differentiation of PVY strains according to the response of inoculated experimental plants was confusing due to the variability of viral aggressiveness and symptomatology induced. RFLP analysis of the CP gene and 3’untranslated region (UTR) revealed high variability. In addition to mixed infection by different PVY strains, the biological and molecular properties of those tomato PVY isolates that react to both monoclonal antibodies could be explained as the result of RNA recombination between distinct PVY strains which infect the same host plant.     

Aggressive and mild Potato virus Y isolates trigger different specific responses in susceptible potato plants

Differences in the early responses of two potato cultivars, Igor and Nadine, to two isolates of Potato virus Y (PVY), the aggressive PVY^NTN and the mild PVY^N, were monitored. Microarray and quantitative real‐time PCR analyses were carried out to identify differentially expressed genes after inoculation with each virus isolate. Additionally, symptom severity and development was observed and the amount of virus isolate accumulated in systemically infected leaves was evaluated, where a significantly higher amount of PVY^NTN was detected. Microarray analysis revealed 572, 1288 and 1706 differentially expressed genes at 0·5, 12 and 48 h post‐inoculation, respectively in cv. Igor, with a similar pattern observed in cv. Nadine. Microarray and quantitative real‐time PCR results implied an earlier accumulation of sugars and lower photosynthesis in leaves inoculated with the aggressive isolate than in leaves inoculated with the mild isolate. The PVY^NTN isolate did not activate early differential expression of the Fe‐superoxide dismutase and pectin methylesterase inhibitor (PMEI) genes, indicating a delay in plant response relative to that following PVY^N inoculation. Differences in the expression of the β‐glucanase‐I gene were also observed in early plant responses to inoculation with each virus isolate.     

Improving Potato virus Y strain nomenclature: lessons from comparing isolates obtained over a 73‐year period

Biological and whole genome properties were compared between eight historical European (1943–1984) and five Australian (2003–2012) Potato virus Y (PVY) isolates. Based on eliciting hypersensitivity genes Nc, Ny or Nz, the former belonged to biological strain groups PVY^C (CT, CRM1), PVY^O (CRN, KE, RS) or PVY^Z (CM2, CRM2, DS). The latter were inoculated to differential and other potato cultivars, tobacco and tomato. Two belonged to PVY^O (BL, DEL3), one to PVY^Z (ATL1), and one (KIP1) to suggested strain group (PVY^D) which elicited putative hypersensitivity gene Nd. Tomato isolate CN1 (and unsequenced CN2), which were poorly adapted to infect potato, were not grouped. Next‐generation sequencing (NGS) of samples containing all isolates except CN2, yielded 13 complete sequences of 9592–9700 nucleotides (nt), and one partial sequence of 9002 nt, none being recombinants. Comparing the former with 60 other PVY complete genomes, found one (CRM2) in phylogenetic subgroup Y^O, eight in Y^O5 (CM2, CRN, DS, KE, RS, ATL1, BL, DEL3), three in Y^C2 (CRM1, CT, KIP1) and one in Y^C1 (CN1). Thus, biologically defined PVY^O (5) and PVY^Z (4) isolates were within phylogenetic subgroups Y^O or Y^O5, biological PVY^C isolates (2) within Y^C2, biological PVY^D isolate KIP1 in Y^C2 and tomato isolate CN1 in Y^C1. NGS identified KIP1 and partial sequence KIP from mixed infection. KIP was in Y^O5. Grouping of four PVY^Z isolates within phylogenetic PVY^O, and the PVY^D isolate within phylogenetic PVY^C, reveals disagreement between current biological and phylogenetic PVY nomenclature systems. Using Latinized numerals for phylogenetic group names resolved this.     

两个马铃薯Y病毒黑龙江马铃薯分离物株系鉴定

为明确分离自黑龙江省克山县马铃薯上的2个病毒分离物KS4和KS7的分类地位,通过RTPCR扩增、克隆获得其基因组序列,利用重组分析程序包和最大似然法分别进行重组分析和系统发育分析。结果显示,分离物KS4和KS7的开放阅读框均有9 186个核苷酸,编码3 061个氨基酸,分离物KS4的核苷酸和氨基酸序列均与马铃薯Y病毒(potato virus Y,PVY)分离物Mb112一致率最高,分别为96.9%和98.4%;分离物KS7的核苷酸序列与PVY分离物12-94一致率最高,为97.4%,其氨基酸序列与PVY分离物SYR-Ⅱ-Be1一致率最高,为97.8%。重组分析表明,分离物KS4和KS7均为分离物N-605和Oz的重组体,其中KS4基因组5′-端的2 392个核苷酸来自分离物N-605,其余核苷酸来自分离物Oz;KS7基因组的第800～2 227个核苷酸和第5 637～8 950个核苷酸来自分离物N-605,其余核苷酸来自分离物Oz。系统发育分析发现,分离物KS4被聚类到N:O株系(PVY~(N:O)),分离物KS7被聚类到NTN株系(PVY~(NTN))b型。     

New aphid vectors of potato virus YN

During three successive years, 1983, 1984 and 1985, winged aphids were caught alive in a potato field with a conical net and with transportable suction traps.One hundred and one aphid species or species groups were checked for their ability and efficiency in transmitting potato virus Y^N (PVY^N) from potato to potato. Seventy-eight species or species groups were found unable to transmit PVY^N, whereas twenty-three species did transmit, among them beingAphis nasturtii, Brachycaudus helichrysi, Cryptomyzus galeopsidis, Cryptomyzus ribis, Hyadaphis foeniculi, Hyalopterus pruni, Hyperomyzus lactucae, Sitobion avenae andSitobion fragariae. All species, with the exception ofA. nasturtii, are recorded the first time as vectors for PVY^N.In transmission experiments alatae caught with a conical net yielded better results than did those caught with a suction trap.Samenvatting Gedurende drie opeenvolgende jaren (1983, 1984, 1985) werden gevleugelde exemplaren van 101 bladluissoorten levend gevangen met een fuik en met verplaatsbare zuigvallen, en in een kas getoetst op hun vermogen om aardappelvirus Y^N (PVY^N) van aardappel naar aardappel over te brengen.Achtenzeventig soorten brachten het PVY^N niet over. Naast de reeds algemeen bekende vectorsoorten werden nog enkele soorten gevangen die in staat bleken PVY^N over te brengen in het laboratorium, te weten:Aphis nasturtii, Brachycaudus helichrysi, Cryptomyzus galeopsidis, Cryptomyzus ribis, Hyadaphis foeniculi, Hyalopterus pruni, Hyperomyzus lactucae, Sitobion avenae enSitobion fragariae.Met bladluizen gevangen in de fuik werden betere overdrachtsresultaten behaald dan met de bladluizen gevangen in de zuigvallen.     

Enzyme-linked immunosorbent assay (ELISA) for the detection of potato viruses A and Y in potato leaves and sprouts

With the enzyme-linked immunosorbent assay (ELISA) potato virus A (PVA) could be detected reliably in potato sprouts, especially when these were young and sappy. The detection of this virus in leaves of glasshouse-grown potato plants was less reliable. The tobacco veinal necrosis strain of potato virus Y (PVY^N) was readily demonstrated in foliage of glass-house-grown potato plants using an antiserum to this strain. Plants infected with the common strain (PVY^O) did not react in ELISA with this antiserum. In young sappy sprouts, using the PVY^N antiserum, PVY^N could be detected reliably when samples with PVY^O were excluded, as the reaction of samples infected with the latter virus was intermediate between PVY^N-diseased and PVY-free samples. PVY was also detected in plants inadvertently infected during the experiments.     

Problems associated with potyviruses in potato certification-field inspection and serological testing1

In UK, the tobacco veinal necrosis strain of potato virus Y (PVY^N), potato virus A (PVA) and potato virus V (PVV) each occur in the field only in limited ranges of potato cultivars in which they mostly cause mild symptoms or even symptomless infection; little is known about incidence of strain C of PVY (PVY^C). The ordinary strain of PVY (PVY°), however, is widespread causing symptoms ranging in severity from very severe through to very mild, depending on cultivar sensitivity/tolerance. During field inspections, very mild potyvirus symptoms may be missed, so inspectors are trained to be particularly vigilant when examining problem cultivars which react in this way. PVA is almost invariably treated, along with PVX, as a mild kind of virus infection, but infections with PVY°, PVY^N and PVV are treated as severe with stricter tolerances being applied for them (especially for PVY^N) regardless of symptom severity. Wide variation within the same cultivar in the behaviour of variants within the PVY° strain group also sometimes causes difficulties in interpretation at inspection. To detect PVY, PVA and PVV in routine serological testing on potato certification samples, it is necessary to employ specific antisera to each of them. PVY^N-specific monoclonal antibodies can be used in ELISA to distinguish PVY^N from PVY°.     

Spread of potato virus Y in potato cultivars (Solanum tuberosum L.) with different levels of sensitivity

The aim of this work was to correlate the appearance of the symptoms, multiplication and spread of virus after mechanical inoculation of potato (Solanum tuberosum L.) cultivars showing different levels of susceptibility and sensitivity to Potato virus Y^NTN (PVY^NTN). The potato cultivars used were the resistant cultivar Sante and susceptible cultivars Igor, Pentland squire and Désirée. The spread of the virus PVY^NTN in infected plants was monitored using different methods: DAS-ELISA, tissue printing, immuno-serological electron microscopy and real-time PCR. In all three susceptible cultivars, the virus was detected in the inoculated leaves 4–5 days after inoculation. From there virus spread rapidly, first into the stem, then more or less simultaneously to the upper leaves and roots. Real-time PCR was shown to be very sensitive and enabled viral RNA to be detected in non-inoculated leaves of susceptible cultivar Igor earlier than other methods. Therefore, for exact studies of plant–virus interaction, a combination of methods which detect viruses on the basis of their different properties (coat protein, morphology or RNA) should be used to monitor the spread of viruses.     

Effect of temperature on resistance to Potato virus Y in potato cultivars carrying the resistance gene Rychc

The effect of cultivation temperatures on the resistance reaction to three Potato virus Y strains (PVY^O, PVY^N and PVY^NTN) in potato cultivars carrying Ry_chc was examined. When potato plants carrying Ry_chc were cultivated at 22 °C, a few small necrotic spots developed on inoculated leaves by 5 days after mechanical inoculation (dpi), and systemic infection of a few symptomless plants was confirmed at 28 dpi by IC‐RT‐PCR. At 28 °C, distinct necrotic spots developed on inoculated leaves by 5 dpi, and systemic symptoms occasionally appeared at 28 dpi. Thus, high temperature weakens Ry_chc‐conferred resistance. However, the incidence of systemic infection and the titre of virus in resistant cultivars at 28 °C were lower than in a susceptible cultivar. In graft inoculation under high summer temperatures, some plants developed necrosis on the leaves and stem, but PVY was barely detected by RT‐PCR in leaves on potato carrying Ry_chc. When seedlings from progeny tubers of plants that were inoculated with PVY and grown in a greenhouse at >30 °C in the daytime were examined by ELISA and IC‐RT‐PCR, PVY was not detected in cultivars carrying Ry_chc. These results show that Ry_chc confers an extreme resistance to PVY strains occurring in Japan.     

Aphid vectors of potato virus YN

BesideMyzus persicae a dozen other species were found to be vectors of potato virus Y^N. Eleven other species did not transmit the virus.White Burley tobacco and A6 potato are equally suitable as test plant to monitor the efficency ofRhopalosiphum padi as vector of PVY^N, but as PVY^N source tobacco is not suitable for this aphid species.Between some aphid species rather large differences exist in retention periods of PVY^N. WithR. insertum andAphis fabae transmission after a 1 h starvation period was still 50% of that without starvation. WithPhorodon humuli, M. certus andM. persicae this value was only 15, 30 and 30%, respectively.Samenvatting Van 12 bladluissoorten werd vastgesteld dat zij, evenalsMyzus persicae, vectoren van het aardappelvirus Y^N (PVY^N) zijn. Van 11 andere soorten kon dit niet worden vastgesteld. Nicotiana tabacum cv. White Burley enSolanum tuberosum cv. A6 bleken beide goed bruikbaar als toetsplant voor het vaststellen van de efficiëntie vanRhopalosiphum padi als vector van het PVY^N; voor deze bladluissoort is tabak ongeschikt als bron van PVY^N.De retentieperiode van het PVY^N lijkt bij verschillende bladluissoorten aanzienlijk te variëren. BijRhopalosiphum insertum enAphis fabae bracht één uur vasten na de acquisitie de overbrenging terug tot 50% van die welke zonder vasten werd verkregen. BijPhorodon humuli was de reductie in overbrenging na één uur vasten 85%, bijMyzus certus enM. persicae was deze 70%.