Effect of temperature on the transmission of Potato mop-top virus from seed tuber and by its vector, Spongospora subterranea

Potato mop-top virus (PMTV) causes disease in both the growing plant and tubers (spraing) of potato and is transmitted by the plasmodiophorid Spongospora subterranea , the cause of powdery scab. The effect of temperature during plant growth on the transmission of PMTV from infected seed tubers and from infested growing media was investigated in a series of glasshouse experiments. Symptoms developed on foliage of plants derived from infected seed tubers but none developed when PMTV was transmitted by S. subterranea in soil. The incidence of foliar symptoms was greatest on plants grown at 12°C, less at 16°C, few at 20°C and absent at 24°C. The transmission of PMTV from infected seed tubers was not significantly affected by temperatures between 12 and 24°C, but when the virus was transmitted by S. subterranea , minimal tuber infection occurred at 24°C and no differences were recorded at temperatures between 12 and 20°C. The incidence of powdery scab on tubers was greatest at 12 and 16°C and very low at 20 and 24°C. However, the incidence and severity of root galling caused by S. subterranea , was greatest at 20 and very low at 24°C. The incidence of powdery scab was greater on tubers of plants derived from infected seed tubers grown in a fluctuating temperature regime of 12 h at 20°C followed by 24 h at 12°C than on those grown at a constant 20°C, whereas the incidence of tuber infection by PMTV and spraing was similar for both regimes. This demonstrates that infection of roots can occur at a higher temperature than that for powdery scab on tubers and that this root infection can enable the transmission of PMTV into the potato plant.     

Germinate to exterminate: chemical stimulation of Spongospora subterranea resting spore germination and its potential to diminish soil inoculum

Hoagland's solution (HS), a defined nutrient supplement for plants, has been previously reported to stimulate zoospore release from resting spores of the potato pathogen Spongospora subterranea f. sp. subterranea. This study obtained direct empirical evidence for an increase in zoospore release with HS treatment, and identified Fe‐EDTA as the stimulant component of HS. Stimulation of resting spores by HS and Fe‐EDTA resulted in greater and earlier zoospore release compared to a distilled water control, and in the presence of a susceptible tomato host plant resulted in enhanced root infection. Given the labile nature of S. subterranea zoospores, it was postulated that stimulation of premature release of zoospores from the dormant resting spores in absence of susceptible hosts could reduce soil inoculum levels. In two glasshouse trials in the absence of host plants, both Fe‐EDTA and HS soil treatments reduced S. subterranea soil inoculum levels, providing proof of concept for the ‘germinate to exterminate’ approach to inoculum management.     

Root infection of potato by Spongospora subterranea: knowledge review and evidence for decreased plant productivity

Information is reviewed on root infection of potato by the plasmodiophorid Spongospora subterranea f. sp. subterranea. This pathogen has long been recognized as the cause of root galls (hyperplasia) and the economically important disease powdery scab on tubers (modified stolons). The significance for plant productivity of the zoosporangium stages of the pathogen in potato roots has only recently begun to be documented. Two experiments are described that assessed effects of S. subterranea root infection on potato plant root function and productivity. A greenhouse experiment measured root function and plant parameters for eight potato cultivars with markedly different susceptibilities to tuber powdery scab. Water uptake and plant growth were reduced by S. subterranea inoculation in all eight cultivars. The magnitudes of these negative effects, and intensities of root hyperplasia, differed among the cultivars, but were not related to respective susceptibilities to tuber powdery scab. A field trial assessed root function and plant productivity for a cultivar (Iwa) that is very susceptible to Spongospora tuber and root diseases. Soil water content beneath uninoculated plants was consistently less than for inoculated plants, indicating that inoculation reduced water uptake (root function). Inoculation reduced shoot and root dry weights, and reduced weight of tubers per plant by 42%. Spongospora subterranea causes three diseases of potato: root membrane dysfunction, root hyperplasia and tuber powdery scab. The root diseases caused by the pathogen are likely to be important both for powdery scab management and for deleterious effects on potato crop yields.     

Relationship between Spongospora subterranea f. sp. subterranea soil inoculum level,host resistance and powdery scab on potato tubers in the field

The relationship between initial soil inoculum level of Spongospora subterranea f. sp. subterranea (Sss) and the incidence and severity of powdery scab on potato tubers at harvest was investigated. In all experiments soil inoculum level of Sss (sporeballs/g soil) was measured using a quantitative real‐time PCR assay. Of 113 commercial potato fields across the UK, soil inoculum was detected in 75%, ranging from 0 to 148 Sss sporeballs/g soil. When arbitrary soil inoculum threshold values of 0, <10 and >10 sporeballs/g soil were set, it was observed that the number of progeny crops developing powdery scab increased with the level of inoculum quantified in the field soil preplanting. In four field trials carried out to investigate the link between the amount of inoculum added to the soil and disease development, disease incidence and severity on progeny tubers was found to be significantly (P < 0·01) greater in plots with increasing levels of inoculum incorporated. There was a cultivar effect in all years, with disease incidence and severity scores being significantly greater in cvs Agria and Estima than in Nicola (P < 0·01).     

Detection and Quantification of Spongospora subterranea f. sp. subterranea in Soils and on Tubers Using Specific PCR Primers

PCR-based methods were developed for the detection and quantification of the potato pathogen Spongospora subterranea f. sp. subterranea (S. subterranea) in peel, tuber washings and soil. A partial sequence was obtained for S. subterranea ribosomal DNA and specific PCR primers (Sps1 and Sps2) were chosen from the internal transcribed spacer regions. These primers amplified a 391bp product from S. subterranea DNA but did not amplify DNA from potato or a range of soil-borne microbes, including related species. Diluted S. subterranea DNA was detected at a concentration equivalent to 25×10^–5 cystosori or 1 zoospore per PCR. Amplification was detected from peel and washings of infected and apparently healthy tubers, but not from peel of Scottish classified seed potatoes or axenically micropropagated potatoes. A rapid method for extracting S. subterranea DNA from soils was developed. This yielded DNA pure enough for PCR within 3h and facilitated the detection of 1–5 cystosori per gram of soil. A PCR quantification technique was developed involving comparison of product ratios obtained after co-amplification of S. subterranea DNA along with an internal standard (competitor DNA fragment). This quantitative technique was also adapted for use in soil. PCR detection of S. subterranea in soil was considerably more sensitive than previously reported immunoassays and was quicker and easier than conventional bait plant bioassays. Such an assay could be useful for developing disease risk assessments for field soils and seed potato stocks and for future studies on the ecology and control of S. subterranea.     

Identification of differentially expressed genes in tolerant and susceptible potato cultivars in response to Spongospora subterranea f. sp. subterranea tuber infection

Powdery scab caused by Spongospora subterranea f. sp. subterranea (Sss) has recently become one of the most devastating potato diseases of economic importance in South Africa. The use of resistant cultivars has long been considered the most effective and sustainable strategy to manage the pathogen. However, little is known about the molecular mechanisms underlying resistance of potato tubers to Sss. Using RNA-sequencing (RNA-seq), 2058 differentially expressed genes (DEGs) were identified from two potato cultivars (tolerant and susceptible) in response to Sss infection. Analysis of the expression patterns of 10 selected defence-response genes was carried out at two different stages of tuber growth using RT-qPCR to validate the RNA-seq data. Several defence-related genes showed contrasting expression patterns between the tolerant and susceptible cultivars, including marker genes involved in the salicylic acid hormonal response pathway (StMRNA, StUDP and StWRKY6). Induction of six defence-related genes (StWRKY6, StTOSB, StSN2, StLOX, StUDP and StSN1) persisted until harvest of the tubers, while three other genes (StNBS, StMRNA and StPRF) were highly up-regulated during the initial stages of disease development. The results of this preliminary study suggest that the tolerant potato cultivar employs quantitative resistance and salicylic acid pathway hormonal responses against tuber infection by Sss. The identified genes have the potential to be used in the development of molecular markers for selection of powdery scab resistant potato lines in marker-assisted breeding programmes.     

云南省马铃薯粉痂病的发生与危害调查分析

粉痂病严重影响马铃薯的产量和品质,是我国马铃薯产业健康发展的重要制约因素之一.为进一步探明马铃薯粉痂病在云南省的发生、分布及危害情况,本研究于2018年-2019年,在云南省内马铃薯不同生态种植区,开展了马铃薯粉痂病的发生与危害情况调查.调查范围涉及19个县(区)、涵盖了19个当地马铃薯主栽品种(系).调查结果显示:粉...     

Effect of soil inoculum level and environmental factors on potato powdery scab caused by Spongospora subterranea

The effects of soil inoculum level and three environmental factors (soil type, soil moisture regime and temperature) on the incidence and severity of powdery scab caused by Spongospora subterranea were investigated in potato plants grown under controlled environmental conditions. Symptoms of powdery scab on tubers were assessed visually, after which DNA was extracted from tuber peelings and quantified in a real-time polymerase chain reaction assay using primers and a TaqMan® probe specific to S. subterranea to establish tuber infection levels. Soil inoculum concentration of S. subterranea did not significantly affect the incidence and severity of either tuber infection or powdery scab symptoms at maturity. No significant differences in disease incidence and severity were found between sandy, loamy and clay soils, although the two lighter soils yielded more powdery scab than clay soil. Constant dampness of the soil resulted in significantly more disease than a fluctuating moisture regime. Infection and disease levels were high at all three temperatures tested (9, 12 and 17°C), but symptoms were most severe at 12°C. The percentage of plants with infected tubers did not increase after tuber initiation, although the amount of S. subterranea DNA detected in tubers and the severity of powdery scab symptoms increased in mature plants. Latent tuber infections were found to be common, especially under conditions suboptimal for disease development. This new information may be important for the prevention of powdery scab in potato-growing areas around the world.     

云南马铃薯粉痂病发生情况初步研究

马铃薯粉痂病由Spongospora subterranea f.sp.subterranea引起,是典型的土传病害,主要危害块茎和根部。2004年调查了昭通市和会泽县7个乡镇,8个马铃薯品种的粉痂病发生情况。在调查的品种中,会-2发病最轻,平均发病率27%。会泽县马铃薯粉痂病平均发病率59%,昭通市马铃薯粉痂病平均发病率35%。分析土壤中氮、磷、钾、有机质含量及pH与发病率的关系。PB06发病率与土壤中氮、有机质呈正相关,与磷、钾、pH呈负相关,其中与钾含量相关系数为0.693。会-2发病率与土壤pH、钾含量呈负相关,相关系数分别为0.384、0.858,其中与钾含量相关性显著;与磷、有机质含量呈正相关,相关系数分别为0.727、0.323;与氮含量呈正相关,相关系数为0.993,相关性显著。     

Spongospora subterranea root infection assessed in two potato cultivars differing in susceptibility to tuber powdery scab

Infection by Spongospora subterranea of roots of two potato (Solanum tuberosum) cultivars, either very resistant or very susceptible to powdery scab on their tubers, was studied in a glasshouse experiment. Plants grown in sand/nutrient solution culture were inoculated with S. subterranea sporosori 2 weeks after planting. Plant parameters, the intensity of zoosporangium infection in roots, numbers of Spongospora root galls and amounts of Spongospora DNA in roots, measured using quantitative PCR (qPCR), were assessed at sequential harvests. Inoculation with S. subterranea reduced water use (56 days after planting) by 26% in the tuber resistant cultivar compared with uninoculated plants, and by 60% in the susceptible cultivar. Inoculation did not affect growth of the resistant cultivar, nor shoot mass of the susceptible cultivar, but caused a 38% reduction in root mass of the susceptible cultivar. The intensities of zoosporangium development in both cultivars were similar. The susceptible cultivar had approximately four times more Spongospora root galls g^?1 root mass than the resistant cultivar. Quantitative PCR detected S. subterranea DNA in roots 1 week after inoculation, and indicated a twofold greater amount of pathogen DNA in roots of the susceptible than the resistant cultivar. This study suggests that the S. subterranea zoosporangium stage in host roots is affected differently by host resistance factors than the sporosorus (root gall and tuber scab) stages. The study has also demonstrated the usefulness of qPCR for sensitive and consistent detection of S. subterranea across the duration of potato root infection.     

Detection and Quantification of Spongospora subterranea in Soil, Water and Plant Tissue Samples Using Real-Time PCR

A sensitive real-time polymerase chain reaction (PCR) assay was developed for the quantification of Spongospora subterranea, the cause of powdery scab and root galling in potato, and the vector of Potato mop top virus. A specific primer pair and a fluorogenic TaqMan® probe were designed to perform a quantitative assay for the detection of S. subterranea in soil, water and plant tissue samples. The assay was tested using DNA from cystosori, zoospores, plasmodia and zoosporangia of the pathogen. DNA was extracted directly from cystosori suspended in water and from clay soil with varying levels of added cystosori. DNA obtained from zoospores released into nutrient solution by cystosori in the presence of tomato bait plants was also tested, as was DNA from plasmodia and zoosporangia in infected tomato roots. In many cases, detection was successful even at low inoculum levels. This specific quantitative assay could therefore be a useful tool for studying the biology of S. subterranea, and for the optimisation of disease avoidance and control measures.     

甘肃省马铃薯粉痂病品种抗病性及防治药剂筛选

由马铃薯粉痂病菌Spongospora subterranea引起的粉痂病已成为甘肃省马铃薯生产上的重要病害之一,发生面积及危害程度均呈逐年增加的趋势,严重威胁马铃薯产业的健康可持续发展。研究马铃薯不同品种对粉痂菌的抗性并筛选高效防治药剂,是控制该病扩散蔓延最为直接和有效的手段,对马铃薯粉痂病的综合治理具有重要意义。本试验对甘肃省马铃薯主栽品种对粉痂病抗病性及田间防治药剂开展了系统研究,抗病性测定结果表明：22个品种块茎上均可感染粉痂病,发病率为25.00%～100.00%,病情指数为6.67～68.33,平均病情指数为29.45。从发病率和病情指数综合评价,‘冀张薯12号’对粉痂病有较好的抗性。田间药剂筛选试验结果表明：10亿cfu/g多黏类芽孢杆菌可湿性粉剂100倍液和0.3%四霉素水剂50倍液播前种薯浸种处理均对马铃薯粉痂病有较好的防治效果,防效分别为60.50%和60.22%,显著高于其他处理;0.3%四霉素水剂30倍液和10亿cfu/g多黏类芽孢杆菌可湿性粉剂45倍液于马铃薯花期时进行灌根处理,对粉痂病的防效分别为56.00%和52.87%,显著高于其他处理。因此,甘肃省马铃...     

Powdery scab resistance in Solanum tuberosum: an assessment of cultivar × environment effect

Powdery scab of potato caused by Spongospora subterranea is one of the main disease problems in many potato production regions of the world. However, no efficient and economically sound control method is currently available. Host resistance will be a key component of the integrated management of powdery scab, but there are discrepancies in published powdery scab resistance ratings of cultivars between countries. In order to identify the main factors causing such discrepancies, 10 reference cultivars thought to have a range of susceptibility to powdery scab and potato mop‐top virus were cropped over 4 years in four to six locations across Europe and disease levels on roots and tubers were assessed using standardized scoring scales. Soil contamination was tested using real‐time PCR and ELISA. The cultivars performed as expected according to previous characterization, with one exception. No relationship was found between tuber and root susceptibility. Assessment of powdery scab symptoms 1 month before harvest gave results comparable to those assessed 2 months after harvest. Neither real‐time PCR nor ELISA soil test results were closely related to disease index data. The field trial results indicate that different scoring methods are the main factor for the discrepancy in resistance ratings, and that environmental conditions and/or soil inoculum level play a minor role. Furthermore, there was either no difference between the pathogen populations in each location or the resistance of most of the cultivars is polygenic.     

生物有机肥GSJ-1对马铃薯土壤疮痂病病原菌分布影响及生防效果研究

为了确定生物有机肥 GSJ －1对连作马铃薯土壤疮痂病病原菌分布影响及对马铃薯疮痂病的防治效果,本试验进行了土壤疮痂病病原菌含量测定、马铃薯疮痂病病情田间调查。结果表明,连作土壤疮痂病病原菌在盛花期之前向根际富集,盛花期之后向匍匐茎和薯块富集。施用生物有机肥 GSJ －1显著降低了土壤疮痂病病原菌含量,比对照下降了19．6％～74．9％,且改变了土壤中疮痂病病原菌原有的分布规律：从连作处理的根际多于非根际改变为生物有机肥 GSJ －1处理的根际少于非根际,且对疮痂病的防效达到47．2％。     

不同覆盖种植模式对马铃薯土壤温度、水分及产量的影响

以陇薯6号为试验材料,研究了不同覆盖种植模式对甘肃定西马铃薯田土壤温度、水分和产量的影响,并对其经济效益进行分析比较。结果表明,全地面覆盖双垄垄作侧播和全地面覆盖垄上微沟穴播处理在马铃薯整个生育期0~30 cm平均土壤温度较CK增加2.2℃和2.5℃,秸秆带状覆盖处理平均土壤温度较CK降低了1.0℃。覆膜垄沟集雨和秸秆带状覆盖技术都能明显改善土壤水分状况,在马铃薯生长前期集雨保墒效果尤为明显,出苗期全地面覆盖双垄垄作侧播、全地面覆盖垄上微沟穴播、秸秆带状覆盖在0~100 cm平均土壤含水量分别较CK提高2.6%和2.2%和1.6%。各处理马铃薯土壤温度和含水量垂直变化随着土壤深度的增加而减弱。全地面覆盖双垄垄作侧播、全地面覆盖垄上微沟穴播和秸秆带状覆盖增产效果明显,产量分别较CK增加33.6%、28.6%和23.2%。全地面覆盖双垄垄作侧播纯收益最高,秸秆带状覆盖次之,纯收益较CK分别增加7 201.56元·hm~(-2)和5996.04元·hm~(-2)。因此,黑膜全覆盖双垄垄作侧播技术在改善土壤水温效应的同时能显著增产增收,是定西地区马铃薯种植的高效栽培模式。     

滴灌条件下加气灌溉对云南冬马铃薯根区生境因子和产量的影响

为了研究加气灌溉对云南冬马铃薯根区生境因子（土壤呼吸、土壤水分、土壤温度、土壤氧气摩尔分数）和产量的影响,采用水气分离方法进行加气灌溉,试验设置加气（Y）和不加气(N）2种处理以及高水（100%ET_c,W1）和低水（60%ET_c,W2）2个灌溉水平,共计4个处理。结果表明：加气处理和灌水量均对土壤呼吸速率影响显著（P<0.05）。加气灌溉对土壤含水率的影响较小,除块茎膨大期外,其他生育期加气灌溉对土壤含水率的影响均不显著,影响土壤含水率的主要因素是灌水量。加气处理和灌水量对根区土壤温度均有一定影响,W1Y处理下的土壤平均温度较W1N处理高1.68℃,W2Y处理较W2N处理高0.52℃,且在高水灌溉条件下,加气处理对土壤温度有显著影响（P<0.05）。在马铃薯生育期内土壤氧气摩尔分数的变化呈先增大后降低的特征,4种处理下土壤氧气摩尔分数的大小关系为W1Y>W2Y>W2N>W1N。同一灌溉水平下,加气处理的土壤氧气摩尔分数显著高于不加气处理（P<0.05）。同一加气条件下,高水处理的土壤氧气摩尔分数显著高于低水处理。加气处理和灌水量都对冬马铃薯产量和水分利用效率有显著影响,W1Y处理下冬马铃薯的产量和水分利用效率均最大。综合分析表明,W1Y（高水加气）处理有利于冬马铃薯根区形成良好的水气平衡状态。     

不同覆盖方式对旱地马铃薯产量和水分利用效率的影响

为研究秸秆和地膜覆盖栽培对马铃薯产量和水分利用效率的影响,旨在筛选出陕北黄土丘陵旱作区的最佳覆盖方式。通过2年(2012—2013)的田间定位试验,采用垄膜覆盖(LM)、双沟覆膜(GM)、全膜覆盖(QM)、秸秆覆盖(JG)和露地(CK)5种覆盖栽培方式,分析马铃薯生长、产量、土壤温度、土壤含水量变化。结果表明:(1)JG处理的马铃薯植株生长发育状况,其株高、SPAD值均优于其它处理,表现为株高JGCKGMLMQM,茎粗LMGMCKJGQM,SPAD值JGLMGMCKQM。(2)在整个生育期,地温均值表现为JGCKLMGMQM,JG、LM、GM及QM分别较CK提高-0.18℃、0.76℃、1.21℃及1.65℃。(3)全生育期,不同覆盖有增加土壤含水量的作用,均值表现为JGQMGMLMCK,JG、LM、GM及QM分别较CK提高17.18%、11.92%、5.65%和16.93%。(4)JG、QM、LM及GM分别比露地栽培平均增产19.75%、8.95%、7.00%及-0.25%,水分利用效率提高16.4%、7.4%、9.15%及1.65%。秸秆覆盖栽培方式是陕北旱作区马铃薯抗旱栽培推荐的栽培方式。     

Characterization of weeds and rotational crops as alternative hosts of Spongospora subterranea,the causal agent of powdery scab in Israel

Alternative hosts of Spongospora subterranea may allow multiplication and survival of the pathogen over time; thus, host range is important from an epidemiological aspect. Weeds and rotational crops, such as wheat and barley, were sampled from potato fields with a history of powdery scab (PS) and examined for the presence of S. subterranea by root staining followed by microscopic observations and by qPCR analysis after DNA extraction. The pathogen was detected in plants of 16 weed species from eight families and in volunteer plants of potato and wheat. The ability of the pathogen to infect weeds and rotational crops was further examined by artificial inoculations with sporosori in pot experiments. Successful inoculations occurred with 13 weed species from eight families and with 12 rotational crops from five families. The findings of this study indicate a wide host range in Israel; the families Malvaceae and Zygophyllaceae and the following species are reported for the first time as S. subterranea hosts: Solanum elaeagnifolium, Triticum aestivum, Cynodon dactylon, Phalaris paradoxa, Phalaris minor, Setaria verticillata, Rostaria cristata, Sinapis nigra, Arachis hypogaea, Medicago sativa, Astragalus hauraensis, Amaranthus albus, Chenopodium murale, Chenopodium opulifolium, Salsola soda, Malva nicaeensis, Chrysanthemum segetum, Verbesina encelioides, Ammi majus and Tribulus terrestris. Controlling weeds and avoiding the relevant rotational crops observed to be S. subterranea-positive and thus potential hosts, should be taken into consideration in the management of PS, to reduce pathogen inoculum build-up.     

干旱胁迫和磷肥用量对马铃薯根系形态及生理特征的影响

以马铃薯品种大西洋为材料,探究干旱胁迫条件下马铃薯根系生理及根系形态对磷肥不同增施量的响应,旨在确定干旱胁迫下提高马铃薯产量及抗旱性的最佳磷肥施用量。试验设2个干旱胁迫处理、1个充分灌溉处理,土壤含水量分别保持在9.5%、14.0%、32.0%左右;以及5个磷肥用量(以P₂O₅计),分别为:T1(27.3 kg·hm^-2)、T2(54.5 kg·hm^-2)、T3(81.8 kg·hm^-2)、T4(109.1 kg·hm^-2)、T5(136.3 kg·hm^-2),另设不施肥处理为对照(T0)。结果显示:与充分灌溉相比干旱对马铃薯造成了一系列不利影响,以轻度干旱为例,轻度干旱导致马铃薯根系含水量较充分灌溉处理组降低了5.54%,超氧阴离子■产生速率、丙二醛(MDA)含量分别升高了63.33%和180.47%,株高和匍匐茎数分别降低了11.74%、7.66%,马铃薯根重和块茎产量分别下降了9.85%和6.75%;比较5种磷肥增施量,以每公顷...     

The importance of the infected seed tuber and soil inoculum in transmitting Potato mop‐top virus to potato plants

Potato mop‐top virus (PMTV), the cause of spraing in potato tubers, is transmitted by Spongospora subterranea, the cause of powdery scab, and by planting infected seed tubers. This study was undertaken to determine the relative importance of these sources of infection in seed potato production in Scotland. The transmission of PMTV from tested seed tubers to daughter plants was examined over 2 years and six cultivars. The development of foliar symptoms varied with year and cultivar. Infection of daughter tubers derived from PMTV‐infected seed tubers was more prevalent on plants affected by foliar symptoms than those without symptoms. The rate of transmission of PMTV from infected seed tubers to daughter tubers ranged from 18 to 54%. Transmission was affected by cultivar and by origin of seed tubers used for a cultivar, but not by a cultivar's sensitivity to PMTV infection. The incidence of PMTV in daughter tubers of cv. Cara grown from seed potatoes from one source (common origin) by more than 25 seed producers was examined over two successive generations. The incidence of PMTV in daughter tubers was not correlated with that in the seed tubers but appeared to be strongly associated with soil inoculum. The incidence of PMTV was correlated with powdery scab in those crops in which both were present. There was some evidence from soil tests conducted in 2006 using a tomato bait plant and real‐time RT‐PCR that planting PMTV‐infected seed potatoes could increase the risk of introducing the virus into land not infested by PMTV.