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In-field management of Potato virus Y (PVY) faces challenges caused by the changing availability and environmental acceptability of chemical agents to control aphid vectors of the virus and by proliferation of PVY strains with different symptoms and rates of spread. From 2018 to 2020, foliar spray treatments were compared in field experiments in New Brunswick, Canada, to measure effectiveness at reducing spread of PVYO, PVYN:O, and PVYNTN strains. Mineral oil, insecticide, combined oil and insecticide spray, and a biopesticide (i.e., LifeGard WG) were compared. Insecticide-only and mineral oil–only treatments were not effective, but several combined oil and insecticide treatments and biopesticide treatments significantly reduced PVY spread. The biopesticide was proportionately more effective with recombinant PVYN:O and PVYNTN strains, possibly by exciting the plant’s hypersensitive resistance response, caused naturally only in cultivar ‘Goldrush’ by PVYO. Pesticide residue analysis showed that mineral oil increased the retention of pyrethroid insecticide in the potato foliage longer than with insecticide applied alone, which may explain the beneficial synergistic effect of combined sprays for reducing PVY spread. Tuber yields were generally unchanged in chemical insecticide treatments but were slightly lower in biopesticide treatment. The cost per PVY treatment was competitive across all effective treatments, including biopesticide; however, there was some revenue loss from lower yield with the biopesticide. This biopesticide is certified organic, however, and thus a small premium on the price for organic production could offset this yield deficit.

Potato virus A (PVA) and potato virus Y (PVY) are two members of genus Potyvirus infecting potato crops worldwide. Host resistance offers an economical and effective means for the control or management of these viruses. In this study, 20 potato clones were screened for their resistance against PVA and PVY by mechanical or graft inoculation assay, and were explored for the relationship between extreme resistance genes Ra and Ry by the detection of molecular markers linked to Ryadg, Rysto, and Rychc. Six clones, including Barbara, Jizhangshu 8, Longshu 7, Longshu 8, M6, and Solara, were found to be extremely resistant to both PVA and PVY; three clones (AC142, Eshu 3, and Shepody) were deemed to be extremely resistant to PVA but susceptible to PVY. To further reveal the inheritance of the extreme resistance (ER) against PVA, a tetraploid F1 population of Barbara × F58050 (susceptible to both PVY and PVA) and a tetraploid BC1 population of BF145 (a PVA-resistant but PVY-susceptible progeny of Barbara × F58050) × F58050 were obtained. Phenotyping of the F1 and BC1 populations by graft inoculation with PVA showed segregation ratios of 3:1 and 1:1 (resistant:susceptible), respectively. These results suggest that two independent loci control ER against PVA in Barbara: one confers ER to both PVA and PVY and the other confers ER to PVA only. The deduced genotype of Barbara is RyryryryRararara.

Potato virus Y (PVY) exists as several strains with distinct symptomology and tuber yield effects in different potato varieties. Recently, new recombinant strains have proliferated and dominated local populations around the world. In this study, PVYO, PVYN:O, PVYN-Wi, and PVYNTN strains were tracked across Canada from 2014 to 2017, showing rapid evolution of populations away from the traditionally dominant PVYO to recombinants PVYN-Wi (western Canada) and PVYNTN (eastern Canada). Simultaneously, 30 potato varieties were inoculated with PVYO, PVYN:O, and PVYNTN in controlled greenhouse experiments. Foliar symptoms of primary (mechanical inoculation mimicking aphid infection) and secondary (tuber seedborne) infection were cataloged, and tuber yield measured. On average, and generally similar in primary and secondary infection, symptom expression and yield reduction were most severe with PVYO, followed by PVYN:O and PVYNTN. Strong mosaic symptoms were most commonly expressed with PVYO infection, and only seen with PVYN:O or PVYNTN in 15 and 3 varieties, respectively. Across variety–strain combinations, yield reduction was correlated with symptom severity, most strongly in PVYO-infected plants (e.g., AC Chaleur, Beljade, Envol, Norland, and Pacific Russet), and four varieties exhibited tuber necrotic ringspot disease with PVYNTN (AC Chaleur, Envol, Pacific Russet, and Yukon Gold).

Potato virus Y (PVY) is an economically important disease agent in potatoes with worldwide distribution. It is mainly transmitted vegetatively by tubers and between plants by an aphid vector. PVY management is primarily through reducing viral inoculum in seed tubers and other sources and controlling transmission by aphids. Seed certification programs are designed to comprehensively test and restrict levels of PVY within a region, enforcing low levels of PVY in marketed seed lots and allowing growers informed choice to plant low-PVY crops. Sanitation techniques on the farm can also reduce volunteer, weed, and neighboring field sources of PVY. Major techniques to reduce aphid-mediated PVY spread include mineral oil foliar sprays, especially combined with insecticides. Spray timing is important, starting early after first plants emerge, continuing weekly through season to vine-kill, with additional sprays during rapid growth and periods of aphid abundance. Other practices for PVY reduction include avoiding mechanical transmission by field equipment, planting crop borders or intercropping as a barrier or sink for PVY, breeding PVY-resistant potato varieties, and roguing symptomatic plants from fields. Major remaining challenges in combatting PVY are management complacency, proliferation of PVY strains circumventing resistance and roguing, and informal trade of untested potatoes.

Potato virus Y (PVY) transmission was studied in six commercial potato fields in 2015 and 2016 in New Brunswick, Canada. Plants emerged from PVY-positive tubers, or PVY-free plants that were artificially inoculated with PVY shortly after emergence, were used as PVY inoculum plants in the study. In all trials, equal numbers of PVY inoculum plants from each of three strains common in the region, PVYO, PVYN:O and PVYNTN, were used. PVY inoculated into tractor-traffic rows showed 2 to 7 times as much PVY spread to previously virus-free plants by the end of the season (up to 48.5% in one tractor row, compared to a maximum of 16.3% in a distant control row unaffected by tractor traffic). Evidence supporting a hypothesis that tractor traffic enhances PVY transmission through aphid disturbance was observed by PVY spread in both directions along the rows, not biased in the direction of tractor travel, and that the ratio of spread of the three strains was nearly indentical in control and tractor rows. However, the lack of spread to immediately adjacent rows, and statistically significant spatial pattern matching the circumference of tractor wheels specific to each field support the hypothesis that direct mechanical transmission of wounded plant sap could also be a factor in the enhanced PVY transmission.

Potato virus Y (PVY) is a major cause of yield and quality loss in potato crops worldwide. Recently, populations of PVY strains have shifted dramatically toward recombinant strains such as PVYNTN and PVYN:O. A 2010 to 2016 survey of PVY strains in commercial fields of New Brunswick (NB), Canada, and five field trials tracking PVY spread in NB and Manitoba, were conducted to study the current status of PVY strains and their relative rates of spread. In NB, PVYO dropped from 82% of infections in 2010 to 14% in 2016, replaced mostly by PVYNTN (64%) and PVYN:O (22%).In field trials with Russet Burbank and Gold rush varieties, PVYNTN spread most effectively compared to PVYN:O and PVYO. Strain-specific PVY spread varied with the potato variety, possibly due to selective PVYO resistance in Goldrush, mostly expressed at the plant-to-plant transmission level with little difference in transduction to tubers in infected plants. Relevance of in-field differences in spread of strains to changes in regional PVY populations, and potential mechanisms responsible, are discussed.

In the 2014 and 2015 crop seasons, the efficacies of different types, rates and combinations of mineral oil and insecticide foliar sprays for reducing Potato virus Y (PVY) spread were tested in controlled field trials in New Brunswick (NB), Canada. Experimental plots were planted with certified PVY-free Goldrush, supplemented with known virus-infected seed to raise PVY inoculum to 2.3% and 3% at the beginning of the 2014 and 2015 seasons, respectively. Treatments consisted of mineral oil-only sprays at different application rates, insecticide-only sprays of differing numbers, and several combined mineral oil and insecticide spray regimes, all compared to a no-spray control treatment. PVY spread to 18% (2014) and 22% (2015) of initially virus-free plants in no-spray control plots, with significant reductions observed in PVY spread in several treatments. Greatest PVY reductions, as low as 4% (2014) and 12% (2015), were in combined mineral oil and insecticide spray treatments, followed by oil-only sprays; while insecticide-only sprays did not significantly reduce PVY spread. As well as measuring PVY spread to marked test plants and randomly collected post-harvest tuber sample from the plots, exhibited similar treatment pattern for PVY incidence. Multiple logistic regression modeling confirmed the relative efficacy of combined oil and insecticide sprays for reducing PVY spread, while accounting for variable inoculum and aphid factors. Modeling also highlighted the importance of planting low-PVY seed initially, and of early application of foliar sprays. Local best management practice recommendations for reduction of in-field PVY spread were discussed.

Molecular markers reported to be linked to extreme resistance (ER) against Potato virus Y (PVY) were evaluated in potato germplasm. YES3-3A and YES3-3B, markers linked to Rysto, were detected in ‘Barbara’ and its three descendants that exhibit ER to PVY; RYSC3, a marker linked to Ryadg, was detected in breeding clones NY121 and NY123. Assessment of RYSC3 as a marker for selection for Ryadg-mediated ER validated its efficacy in identification of selections with ER to PVY.

In this study, on-farm within-season spread of Potato virus Y (PVY) was measured in 56 seed, processing and tablestock potato fields in New Brunswick, Canada between 2010 and 2014. These represented 13 potato varieties, managed by 16 growers employing a wide range of PVY control techniques. Many aspects of management techniques were quantified, as well local aphid abundances and climatological data. PVY spread, measured in leaves and tubers (by ELISA and RT-PCR), was tracked through the season in individually marked plants. Over the five seasons under study, on-farm PVY spread overall has declined substantially. Across all 56 fields, however, it has varied widely, from 10 fields not showing any spread during the crop season, up to as high as 76 % spread in one 2012 field. Factors correlated with increased PVY spread included seed-borne PVY inoculum planted in the field, aphid abundance early in the season, and to a lesser degree temperatures in July and over the preceding winter. Factors correlated with decreased spread included numbers of foliar mineral oil and insecticide sprays (especially of the lambda-cyhalothrin and flonicamid types), later crop planting and earlier first spraying dates. A mid-season leaf test for PVY was shown to be strongly indicative of ultimate PVY spread at harvest, and may prove a useful test in advising growers. A set of locally-relevant best management practices based on these results is discussed.

Surveys of commercial and seed potato fields for virus diseases (1998 to 2002) in Manitoba established that Potato virus Y (PVY) is of concern in seed potato production. To determine the prevalence of PVY strains, PVY-infected tubers identified by reverse transcription-polymerase chain reaction (RT-PCR) from surveys (2000 to 2001) were grown for symptom expression and strain characterization by strain-specific RT-PCR, bioassays, and serological assays. Of the samples collected (2000 to 2001) and tested by RT-PCR, 4.0% contained PVY. Further analysis of the PVY-positive samples by a duplex RT-PCR facilitating the simultaneous detection of common (PVYO) and tobacco veinal necrosis strains (PVYN/NTN) indicated that 37.5% contained PVYO and 63.5% contained PVYN-type isolates. Analysis of the PVYN-type samples using three monoclonal antibodies (MAbs) showed that all reacted with only the PVYO MAbs and not with the PVYN-specific MAb. Partial nucleotide sequences of both ends of PVY-RNA showed that the PVYN-type isolates resembled those reported in 1996 from Manitoba. These isolates are designated as PVYN:O. In view of the increased incidence of PVYN:O in one production area, seed tubers imported from other provinces of Canada and the neighboring United States were analyzed for PVYN:O. The PVYN:O was detected in imported seeds from Minnesota, Montana, and North Dakota.

The responses of 14 potato cultivars to five Potato virus Y (PVY) isolates belonging to four strains (ordinary [PVYO], tobacco veinal necrosis [PVYN], N:O group [PVYN:O], and nonrecombinant potato tuber necrotic [PVYNTN]) were studied in primary and secondary infections. For the primary infection experiments, foliage symptoms were monitored daily after mechanical inoculation with a PVY isolate until harvest; and, for the secondary infection experiments, foliage symptoms were monitored regularly from plant emergence until harvest. Tuber symptoms (namely, tuber necrotic ringspots) were checked at harvest and monthly postharvest for up to 4 months. In both infections, symptoms varied significantly depending on potato cultivar and virus strain or isolate. In primary infections, local lesions occurred on inoculated leaves of ‘AC Chaleur’, ‘Eramosa’, ‘Goldrush’, ‘Jemseg’, ‘Katahdin’, ‘Ranger Russet’, and ‘Yukon Gold’ after inoculation with PVYO isolates, followed by systemic necrosis on latterly emerged uninoculated leaves. In contrast, plants of ‘CalWhite’, ‘La Rouge’, ‘Red LaSoda’, ‘Russet Burbank’, ‘Russet Norkotah’, and ‘Superior’ did not exhibit any visible symptoms on inoculated leaves but developed mild to severe mosaic on latterly emerged leaves after infection with PVYO isolates. In all cultivars, near-symptomless to mild mosaic was induced by PVYN and mild to severe mosaic by PVYN:O. PVYNTN induced mild to severe mosaic in plants of all cultivars except AC Chaleur, ‘Cherokee’, and Yukon Gold, which developed visible systemic necrosis. Necrotic ringspots were observed in tubers of PVYNTN-infected plants of AC Chaleur, Cherokee, and Yukon Gold. The tuber symptoms were also incited by PVYN-Jg on Cherokee. In secondary infections, the symptoms were generally more severe than primary infections even though the symptom types did not alter. As in the greenhouse, a clear symptom severity pattern (PVYO-FL > PVYO-RB > PVYNTN-Sl > PVYN:O-Mb58 > PVYN-Jg) was observed in AC Chaleur, Cherokee, Eramosa, Goldrush, Jemseg, Katahdin, Ranger Russet, and Yukon Gold in the field.

Potato virus Y (PVY) undergoes continuous nucleotide mutation and genome recombination, leading to novel strains with varying pathogenicity on host plants. Little is known about how tomato plants respond to infections with different PVY strains, especially the emerging strains such as PVYNTN and PVYN:O, and in combination with other viral species such as Potato virus X (PVX). In this study, the response of tomato (cultivar Rutgers) plants to single and mixed infections with PVY and PVX was investigated. Plants infected singly with PVYO, PVYN:O, PVYN or PVYNTN developed mosaic symptoms and leaf deformation; whereas plants infected with PVX developed mild local lesion and varying degrees of mosaic and leaf deformation symptoms. Mixed PVX+PVY infections induced more severe symptoms, which include severe local and systemic leaf necrosis, leaf drop and severe leaf deformation, compared to single infections with PVX or PVY, indicating PVX-PVY synergism. The level of PVX-PVY synergism was affected significantly by PVY strains, with PVYNTN being the greatest effector. Quantitative analysis of viral titre indicated that mixed PVX+PVY infections elevated PVX level and reduced PVY level. A comparative transcriptional analysis of 46 key genes involved in signal molecule synthesis and signal transduction using NanoString technology revealed diverse gene expression profiles in both single and mixed infections. Two hierarchical expression clusters, one up-regulated and the other down-regulated, were observed during the infections. In general, mixed infections led to more genes being differentially regulated than single infections. The expression alteration of genes involved in oxidative stress and salicylic acid (SA)-pathways is particularly noteworthy, as the mixed infections, especially PVX+PVYNTN infections, led to a further elevation of reactive oxygen species (ROS) and SA producing abilities and a reduction of ROS scavenging ability over single infections.

Aphids are viral vectors in potatoes, most importantly of Potato virus Y (PVY), and insecticides are frequently used to reduce viral spread during the crop season. Aphids collected from the potato belt of New Brunswick, Canada, in 2015 and 2016 were surveyed for known and novel mutations in the Na-channel (para) gene, coding for the target of synthetic pyrethroid insecticides. Specific genetic mutations known to confer resistance (kdr and skdr) were found in great abundance in Myzus persicae (Sulzer) (Hemiptera: Aphididae), which rose from 76% in 2015 to 96% in 2016. Aphids other than M. persicae showed lower frequency of resistance. In 2015, 3% of individuals contained the resistance mutation skdr, rising to 13% in 2016 (of 45 species). Several novel resistance mutations or mutations not before reported in aphids were identified in this gene target. One of these mutations, I936V, is known to confer pyrethroid resistance in another unrelated insect, and three others occur immediately adjacent and prompt similar chemical shifts in the primary protein structure, to previously characterized mutations associated with pyrethroid resistance. Most novel mutations were found in species other than M. persicae or others currently tracked individually by the provincial aphid monitoring program, which were determined by cytochrome C oxidase I (cox1) sequencing. Through our cox1 DNA barcoding survey, at least 45 species of aphids were discovered in NB potato fields in 2015 and 2016, many of which are known carriers of PVY.

Potato cultivar Exploits and breeding clone F87084 have been considered resistant to Potato virus Y (PVY). To further explore the degree of resistance and whether the resistance is strain specific, these materials, together with cultivar Rochdale Gold-Doree and breeding line F02010, were investigated for their response to different PVY strains including PVYO, PVYN:O, PVYNTN and PVYN. Both F02010 and Rochdale Gold-Doree were readily infected with all tested PVY strains after either mechanical or graft inoculation, indicating susceptibility of the materials to PVY. F87084 was unable to be infected by any of the tested PVY strains as no ELISA-detectable level of PVY were found in plants after either mechanical or graft inoculation, demonstrating extreme resistance in F87084 to all strains of PVY tested. Exploits was infected with PVYN:O/PVYNTN/PVYN after either mechanical or graft inoculation, indicating susceptibility of the cultivar to these PVY strains. The cultivar was also infected readily with PVYO after graft inoculation. However, despite induction of local lesions on the inoculated leaves, mechanical inoculation with PVYO may or may not lead to systemic symptoms and ELISA-detectable level of PVY, depending on temperature. At low temperature (e.g., 22 °C), no visible systemic symptoms or ELISA-detectable level of PVY was found in the plants; whereas at high temperature (e.g., 30 °C), systemic symptoms and high level of PVY were detected in the plants. These results demonstrate that Exploits possesses temperature-dependent hypersensitive resistance to PVYO. Analysis of a segregating population of F87084 × F02010 revealed that the ER in F87084 is controlled by a dominant resistance gene.

The current-season spread of Potato virus Y (PVY) was monitored in 19 fields under various management practices in New Brunswick, Canada, through the 2011 and 2012 growing seasons. The focus of this study was to evaluate the role of seedborne PVY inoculum, aphid vector abundance, and the numbers, timing, and types of insecticide and mineral oil sprays, and to confirm the reliability and forecasting capacity of midseason PVY testing. In each field, 100 to 110 virus-free plants were identified shortly after emergence and were assessed four times from early July to early September (after top-kill) with enzyme-linked immunosorbent assay (ELISA) and reverse-transcription polymerase chain reaction (RT-PCR) to track PVY spread. In addition, tubers harvested during development in August and after top-kill were grown-out in the greenhouse for ELISA testing. PVY spread to selected virus-free plants varied widely, ranging from 0 to 76.2% across all studied fields. Of the 19 fields over two seasons, 10 fields were planted with no detectable seedborne PVY, and they showed 0 to 8.7% (mean 2.9%) PVY spread by harvest. The remaining nine study fields with 0.9 to 5.8% seedborne PVY showed 1 to 76.2% (mean 15.2%) PVY spread by harvest. PVY spread was detected in most fields during midseason testing with ELISA and RT-PCR; all tests correlated well with final PVY rates after top-kill, though RT-PCR detection in developing tubers was most sensitive and correlated. Logistic regression modeling was used to identify major factors in PVY spread, including seedborne PVY, early-season aphid abundance, and the numbers of insecticide and mineral oil sprays. The best-fitting model, constructed using these factors as well as a measurement of July PVY incidence (ELISAJuly), strongly explained PVY spread by harvest, with the most significant management factor being the number of mineral oil sprays supplemented with insecticide used during the growing season. A similar model fitted without the ELISAJuly did not adequately predict ultimate PVY spread. The analysis suggests that mineral oil alone was effective at lowering PVY spread, and more effective when combined with insecticide, particularly when used early in the season. No evidence was found for differences in PVY spread across the eight cultivars used or across the range of mineral oil application rates, whereas some evidence was found for differences in the effectiveness of different insecticide types.

Post-harvest screening of potato ‘Kennebec’ revealed a Potato virus Y (PVY) incidence of 15.8%, a rate that is unusually high for a cultivar possessing a high level of field resistance to the virus. Randomly selected tubers were planted in a field plot and the resulting plants were monitored. Approximately 16% of plants developed symptoms ranging from mild mosaic symptoms to severe necrosis/rugosity/stunting. ELISA and RT–PCR analysis revealed that infections with Potato virus S (PVS), Potato virus X (PVX) and PVY, mostly in mixed-infections, occurred commonly in 14 sampled plants. Two strains, namely the common strain (PVY°) and the recombinant tuber necrotic strain (PVYNTN) were identified in the PVY-positive plants. In general, mild mosaic was associated with infections with PVX and PVS; intermediated mosaic was associated with PVS and PVYNTN infections; whereas severe leaf deformation/necrosis/drop symptoms were associated with PVYNTN and PVX co-infections, or with PVY° and either PVS or PVX ...

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.

International audience; The management of Potato virus Y (PVY) in potato crops poses a con-tinual challenge due to the non-persistent mode of transmission of the virus and the propagation of seed potato tubers over several generations in the field. While PVY- resistant cultivars remain the most efficient way to protect potato crops against PVY, a vast majority of cultivars grown do not display significant resistance to PVY. Due to the short time period for PVY transmission by non-colonising aphid vectors, efficient control of PVY relies on preventing aphids landing on a crop and on adopting precautionary measures by ensuring that crops are grown in areas of low aphid and low virus pressure and limiting field generation. Prophylactic mea-sures such as roguing and early haulm destruction limit PVY spread but are not efficient alone. Among all existing control methods, spraying potato crops with mineral oils can offer significant protection against PVY spread, but their efficacy do vary in field conditions. The combination of several control methods such as mineral oil treatments, crop borders, intercropping, straw mulching or insecticide treatments can increase protection. These emphasise the importance of controlling virus through appropriate monitoring methods and crop management enforced by seed certification schemes through the use of ‘clean’ input seed and, when possible, the segregation of seed and ware crops to minimise the risk of virus transmission. This chapter presents and discusses the most widely used techniques of control and management of PVY, their effectiveness and their mode of action. This chapter also presents the history, objectives and principles of seed potato certification schemes and their role in minimising the spread of viruses within potato crops worldwide.

Significant progress has been made in recent years in understanding pathological, serological and molecular properties of various strains of PVY and the aphid-mediated transmission. PVYO and PVYN appear to be the basic strain groups. Through genome recombination between these two groups, progeny groups whose genome possess one (e.g., PVYN:O or PVYN-Wi) to three (e.g., recombinant PVYNTN or European-PVYNTN) recombinant joints of PVYO and PVYN emerged. PVYN:O causes PVYN-like veinal necrosis in tobacco, but reacts to PVYO-specific antibody. PVYNTN causes potato tuber necrotic ringspot diseases in sensitive potato cultivars, and PVYN-like necrosis in tobacco plants, and reacts to PVYN-specific antibody. Through single nucleotide mutation(s), non-recombinant PVYNTN (or North American PVYNTN) also emerged from PVYN. It is also noteworthy that PVYN isolates originated from North America and Europe may have evolved separately; and to date most recombinant strains appear to be progenies of Eu-PVYN and PVYO. Several RT-PCR-based methodologies have been developed to characterize and detect various strains of PVY. A field survey revealed that PVYN:O has become a predominant strain in Manitoba and neighbouring states in USA. Moreover, three distinct variant groups inciting severe, intermediate and mild veinal/petiole/stem necrosis, respectively, on tobacco plants were observed within the PVYN:O isolates collected in Manitoba. Pathological and molecular diversity within PVYO strain group were revealed in New Brunswick, represented by PVYO-FL as a severe variant and by PVYO-RB as a mild variant. Studies on the transmission of PVY by various species of aphids revealed that aphid behavior plays an important role in the vector-mediated transmission. Application of mineral oil on the growing crop, especially in combination with use of crop borders, reduces aphid-mediated PVY transmission. Based on recent PVY research studies, Bartlett Superior 70 Oil was approved in 2011 for application to potato crops in Canada.

A TaqMan real-time reverse transcription-PCR (real-time RT-PCR) procedure was developed, optimized, and compared with other routine methods to detect Potato virus Y (PVY) in dormant potato tubers. Three pairs of primers and probes were designed and evaluated for their suitability to facilitate the real-time RT-PCR detection of PVY for all strain groups including PVYO, PVYN, PVYN:O (= PVYN-Wi) and PVYNTN. Among the primer and probe combinations tested, the combination PVY-1 produced the lowest threshold cycle (Ct) value of 25.75. The procedure was further optimized by adjusting various parameters including primer/probe concentration, reaction volume, amplification cycles, and master mixes from different sources. The real-time RT-PCR was then employed to detect PVY from dormant tubers of different cultivars and potato fields, and the results were compared with those obtained from conventional RT-PCR, enzyme-linked immunosorbent assay (ELISA) on sprouts and grow-out testing. Out of 1,069 single-virus infected (PVY only) tubers tested, both formats of RT-PCR detected PVY in 52 samples, ELISA on sprouts in 45, ELISA on leaves in 54 and visual observations in 53. However, in 61 multiple-virus infected tubers tested, both formats of RT-PCR, and ELISA on both sprouts and leaves detected a similar number of positives, thus, making all the methods equally sensitive. Considering that ELISA requires sprouting of dormant potato tubers for PVY testing, grow-out testing takes approximately 6–8 weeks to obtain results, and conventional RT-PCR needs post-PCR processing, real-time RT-PCR offers a speedy alternative for large scale detection of PVY from dormant tubers. The method is therefore recommended for testing of PVY in potato tubers on a commercial scale in a diagnostic laboratory.

The strain diversity and variant population of Potato virus Y (PVY) was investigated in potato lots with high PVY incidence in 2009 in New Brunswick, Canada, using reverse transcription (RT)-PCR, enzyme-linked immunosorbent assay (ELISA) and biological assays. Twenty lots with PVY infection rates >3 % were chosen following post-harvest testing for further investigation. Assays using multiplex RT-PCR revealed the existence of the common (PVYO), the recombinant N:O (PVYN:O) and the recombinant (European) potato tuber necrosis (Eu-PVYNTN) strains. PVYO was the predominant strain while PVYN:O and Eu-PVYNTN were widespread in these potato lots. PVYN:O and Eu-PVYNTN, either present alone or mixed with PVYO, were found in 19 and 13 of the 20 lots, respectively. Neither the tobacco veinal necrosis strain (PVYN) nor the non-recombinant (North American) PVYNTN was detected in the potatoes. Among the PVYO strain group, PVYO-Oz/-FL variant type was the predominant followed by the uncharacterized PVYO and the PVYO-139/-RB types. Biological assays of representative PVY positive-samples using tobacco and Yukon Gold were consistent with the findings of RT-PCR and serological assays.

A field isolate of Potato virus Y (PVY) was collected in New Brunswick, Canada in 2007 due to unusual symptoms observed on different potato cultivars. To unveil the PVY strain identity, tobacco and potato bioassays, PVYO and PVYN-specific antibody-based enzyme-linked immunosorbent assays, and reverse-transcription polymerase chain reaction (PCR)-based genotyping were carried out. All the assays demonstrated that the isolate, designated as PVYO-FL in this study, belonged to the PVYO strain group. Greenhouse tests with the potato cvs. FL 1533 and Jemseg confirmed the severe nature of infection by PVYO-FL. The complete genome sequences of PVYO-FL and PVYO-RB, the latter a mild PVYO isolate, were determined. BLAST analysis revealed that the two isolates shared 97 and 98% sequence identities at the nucleotide and polyprotein levels, respectively. Further BLAST analysis unveiled that PVYO-FL shared 99.7% nucleotide sequence identity with PVYO-Oz, an isolate reported in New York, United States, whereas the PVYO-RB isolate shared 99.2% sequence identity with PVYO-139, a PVYO isolate reported in New Brunswick, Canada. A phylogenetic tree of available, full-length sequences of PVY isolates demonstrated two subgroups within the PVYO branch, one clustered with PVYO-RB and the other with PVYO-FL. Group-specific sense primers for differentiation of the two subgroups were developed and evaluated. A limited survey of potato tubers collected from a field plot at the Potato Research Centre, Agriculture and Agri-Food Canada, using the newly developed PCR primers, indicated that 65.3 and 2.4% of the PVYO-positive tubers were infected with PVYO isolates belonging to the PVYO-FL and PVYO-RB subgroups, respectively. Assessment of the pathogenicity of three representative isolates from each subgroup on the potato cv. Jemseg demonstrated that severe and mild symptoms were induced by the PVYO-FL-like and PVYO-RB-like isolates, respectively.

BACKGROUND: The objectives of this work were to determine if the control of potato virus Y (PVY, genus Potyvirus, family Potyviridae) in seed potato could be improved by combining border crops and mineral oil sprays, and if the border crop acts as a barrier or a virus sink. RESULTS: Field tests over 3 years confirmed that mineral oils alone are an effective barrier to PVY, and showed that borders alone act as a PVY sink. Combining the familiar mineral oil and the more recent crop border methods was almost twice as effective in reducing PVY incidence as either one used alone. The combination provided consistently high PVY control compared with the variable and often lower level of control by either method alone. The contribution of the oil to PVY reduction was similar whether it was applied to the border, the center seed plot, or both. Oil application to the border alone should not affect efficacy and would help keep control costs down. CONCLUSION: Combining border and oil provided the best reduction in PVY incidence 3 years out of 3, providing producers with a tool to reduce year-to-year variation in the effectiveness of crop borders or oil sprays used separately.

Previous studies have identified a recombinant isolate group of the Potato virus Y (PVY), PVYN:O, which exhibits a PVYO serotype and a PVYN pathotype. Molecular fingerprinting indicated that the isolates possess a European (Eu)–PVYN/NTN-like P1 gene and one PVYN-PVYO recombinant joint at the HC/Pro-P3 site. In this study, the complete genomic RNA sequence of two isolates of PVYN:O was determined. The overall identities at the polyprotein level between PVYN:O and other isolates ranged from 94.1% to 97.2%. However, the identities varied from 70.9% to 100% when individual mature proteins were compared. Scanning the nucleotide sequences revealed that PVYN:O comprises a recombinant genome in which the segment of nucleotides 1 to ~2400 is from Eu-PVYN, and the remainder from PVYO. Analysis of a population of PVYN:O isolates indicated the existence of pathological variants that induced different symptoms level in tobacco plants: severe, 16.7%; intermediate, 66.7%; and mild, 16.7%. Three representative isolates w...

A rapid and simple protocol for preparing viral RNA from aphids ( Myzus persicae ) and potato ( Solanum tuberosum L.) tissue (leaves, sprouts, and tubers) for reverse transcription–polymerase chain reaction (RT–PCR) was developed. The four-step method involves: (1) preparing plant crude sap or aphid macerates in a buffered detergent (Triton XL-80N) solution; (2) immobilizing clarified sap on a nitrocellulose membrane; (3) performing reverse transcription using eluted water extract from cut-out spot from membrane; and (4) amplifying cDNA through PCR. The entire procedure from tissue grinding to RT–PCR can be completed within 6 h. The protocol was applied successfully for the detection of individual potato viruses: carlavirus Potato virus S (PVS), potexvirus Potato virus X (PVX), potyvirus Potato virus Y (PVY), and polerovirus Potato leafroll virus (PLRV). PLRV was also detected from individual viruliferous aphids or composites of viruliferous and healthy aphids. PVY and PLRV were detected from extracts immobilized on nitrocellulose membranes, stored for more than 65–273 days at room temperature (25 °C). The protocol was companed with the ‘long protocols’ involving enzyme and phenol extraction for aphids or sodium sulfite and phenol extraction for tubers. The simplified protocol was found comparable in sensitivity to these long procedures, and is especially suitable for those regions where specialized PCR laboratories are only available in centralized locations. Viral RNA immobilized membranes can be mailed out for detection by RT–PCR to these centralized laboratories from remote areas of the country.

Potato virus Y (PVY) is a major threat to potato crops around the world. It is an RNA virus of the family Potyviridae, exhibiting many different strains that cause a range of symptoms in potato. ELISA detection of viral proteins has traditionally been used to quantify virus incidence in a crop or seed lot. ELISA, however, cannot reliably detect the virus directly in dormant tubers, requiring several weeks of sprouting tubers to produce detectable levels of virus. Nor can ELISA fully discriminate between the wide range of strains of the virus. Several techniques for directly detecting the viral RNA have been developed which allow rapid detection of PVY in leaf or tuber tissue, and that can be used to easily distinguish between different strains of the virus. Described in this chapter are several protocols for the extraction of RNA from leaf and tuber tissues, and three detection methods based upon reverse-transcription-PCR (RT-PCR). First described is a traditional two-step protocol with separate reverse transcription of viral RNA into cDNA, then PCR to amplify the viral cDNA fragment. Second described is a one-step RT-PCR protocol combining the cDNA production and PCR in one tube and one step, which greatly reduces material and labor costs for PVY detection. The third protocol is a real-time RT-PCR procedure which not only saves on labor but also allows for more precise quantification of PVY titre. The three protocols are described in detail, and accompanied with a discussion of their relative advantages, costs, and possibilities for cost-saving modifications. While these techniques have primarily been developed for large-scale screening of many samples for determining viral incidence in commercial fields or seed lots, they are also amenable to use in smaller-scale research applications.

Detection of the common strain of PVY (PVYO) in field-grown dormant potato tubers, by nucleic acid spot hybridization (NASH) and reverse-transcription polymerase chain reaction (RT-PCR) was compared with diagnosis by visual symptoms and enzyme linked immunosorbent assay (ELISA) on leaves of eight potato cultivars, differing in their symptom expression and virus content. NASH and RT-PCR readily detected PVYO in dormant tubers. The percent agreement between NASH and RT-PCR ranged from 77.6 to 100 for cvs. included in this study (AC Novachip, Atlantic, Norchip, Red Pontiac, Russet Burbank, Russet Norkotah, Shepody and Superior). ELISA on leaves was almost as good as NASH and RT-PCR on dormant tubers, but required dormancy break. Only two cultivars, namely Red Pontiac and Russet Burbank, could accurately be assessed, based on the visual symptoms. A very high degree of discrepancy was observed for PVYO diagnosis between visual symptoms and other detection procedures for varieties like Russet Norkotah and Shepody, which are known to be latent carriers of PVYO. The use of NASH and RT-PCR in large-scale post-harvest testing is feasible.

Polyphenolics are a widely distributed class of plant constituents implicated in the inhibition of the polymerase chain reaction (PCR). Since chlorogenic acid forms the bulk of phenolics in the potato tuber, a range of chlorogenic acid concentrations were tested for inhibition of reverse transcription (RT) and PCR steps. The RT step was inhibited at chlorogenic acid concentrations of 6.0 to 7.5 microg, while the PCR step was inhibited at much lower concentrations of 1.2 to 1.8 microg. Incorporation of 1.2% citric acid at the nucleic acid extraction step prevented darkening of the extract and neutralized the inhibitory effect of chlorogenic acid.

(1996). Nucleotide sequence and genome organization of a Canadian isolate of the common strain of potato virus Y (PVY°) Canadian Journal of Plant Pathology: Vol. 18, No. 3, pp. 209-224.

A reverse transcription polymerase chain reaction (RT-PCR) protocol was developed using two 20-mer primers located in nuclear inclusion genes NIa and NIb of potato virus Y (PVY). A 1017 bp PCR-product was detected in dormant potato tubers, infected with PVY O , but not in tubers from healthy plants. The PCR product was specific to PVY, as determined by Southern blot detection by hybridization with a PVY O -specific probe. As little as 1 pg of purified PVY O -RNA can be detected after RT-PCR amplification. The presence of phenolics or polysaccharides in tuber nucleic acids inhibited PVY O amplification, which was eliminated by diluting nucleic acid preparations prior to cDNA synthesis, modifying the nucleic acid extraction procedure by isopropanol precipitation and using phosphate-buffered saline-Tween in the cDNA mix. Potato cultivars differed in PVY O concentration in tubers as much as 128-fold. Tuber parts used for nucleic acid extractions were important in potato cultivars with low virus titres and combining of saps from bud and stem ends improved the PVY O detection. Storage of tubers up to 90 days at 4 °C did not result in reduced detection of PVY O by both nucleic acid spot hybridization and RT-PCR, but RT-PCR band intensity was lower at longer storage periods. The primer pair developed in this study exhibited broad specificities with field isolates from Peru, Scotland and North America.

An isolate (Y139) of the potato virus Y strain PVYO, common in the potato-growing regions of eastern Canada, was cloned and segments of about 1800 bases from the 3'-end of the PVY-genome were sequenced. These clones were used to prepare molecular detection probes by labelling with digoxigenin. Five methods reported to yield nucleic acid preparations from tuber tissue were compared. Two of the methods failed to yield nucleic acids from tubers suitable for use in dot-blot hybridization. The other three methods yielded satisfactory nucleic acids which could be successfully detected with the PVYO probes. A method of nucleic acid extraction from tuber tissues was selected which is simple, reproducible and does not use phenol. Using this standardized procedure, PVY was detected in freshly harvested, greenhouse-grown tubers of 8 potato cultivars; the probe was specific to PVY; and the sensitivity of detection using purified viral RNA was down to 10 pg. The sensitivity of detection in tuber extracts was down to a dilution of 1:64, while that from potato leaves was to a dilution of 1:1024. Nucleic acids prepared by the standardized procedure and denatured prior to storage at 4 degrees C, -20 degrees C or at -70 degrees C remained suitable for hybridization for over 2 weeks.

Test conditions for the simultaneous detection of potato leafroll virus (PLRV) and potato virus Y (PVY) in dormant tubers and leaves by reverse transcription-polymerase chain reaction (RT-PCR) were optimized. Various factors optimized at the reverse transcription (RT) stage rather than at the amplification (PCR) stage affected the outcome. In the simplex RT-PCR a onefold dNTPs concentration (0.5 mM) was sufficient in yielding a PLRV or PVY band. In contrast, the duplex RT-PCR required a minimum twofold dNTPs concentration (1.0 mM) during RT to produce distinct bands in PCR. Similarly, various proportions of antisense primers of PLRV and PVY used during RT affected subsequent duplex RT-PCR. Optimal amplification of both viruses were obtained at a ratio of 0.90:0.49 microM of PLRV:PVY antisense primers. An interaction of dNTPs and RNA template concentration was observed. A higher concentration of RNA was required at onefold dNTPs concentration than at twofold dNTPs. Dilutions down to 1:300 of RNA template yielded distinct bands of both viruses at twofold dNTPs concentration. At optimized conditions of the duplex RT-PCR both viruses were reliably detected in composite samples at a ratio of one part infected sap mixed with 399 parts of sap from healthy tubers. Application of optimized conditions to singly- and doubly-infected tubers detected both viruses from naturally infected field-grown tubers. A nearly perfect correlation (r(2)=0.99) was observed between visible plant symptoms and the virus detection from leaves and tubers by the duplex RT-PCR.