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.     

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).     

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.     

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.     

Enhanced resistance to common scab of potato through somatic cell selection in cv. Iwa with the phytotoxin thaxtomin A

A somatic cell selection approach, developed using thaxtomin A as a positive selection agent, recovered 113 potato cell colonies of cv. Iwa from which 39 regenerated plants within a single subculture period. In glasshouse pathogenicity trials, 13 of these regenerated variants exhibited significantly greater resistance to common scab disease than the unselected parent cultivar, both in mean tuber surface coverage with lesions and the proportion of disease free tubers. The best variant had on average an 85–86% lower disease score (or 91–92% less estimated tuber surface cover) than the unselected parent cultivar. The disease-resistant variants varied in their response to thaxtomin A in detached leaflet and tuber slice bioassays. This suggested that the cell selection procedure and thaxtomin A screening had limited success in selecting for toxin tolerance, and that thaxtomin A tolerance was not necessarily the driving factor for induction of disease resistance observed. Variation in relative necrosis scores between leaflet and tuber assays for individual variants possibly suggests differential expression of thaxtomin A tolerance in the different tissues. Yield estimates from the glasshouse grown potatoes showed the majority of disease-resistant variants had tuber yields equivalent to the unselected parent cultivar. This suggests that selection for disease resistance was not associated with negative tuber yield attributes and that these variants may have commercial merit and are worthy of further agronomic testing.     

<Emphasis Type="Italic">Spongospora subterranea</Emphasis> soil contamination and its relationship to severity of powdery scab on potatoes

We investigated soil contamination by Spongospora subterranea f. sp. subterranea (Sss) and disease severity of powdery scab in 29 potato fields in Hokkaido, Japan, using a hydroponic culture method with tomato seedlings as bait plants. The quantity of Sss infection on the roots of bait plants was evaluated using the polymerase chain reaction (PCR) and expressed in terms of the infection potential in the soil. The infection potential was positively correlated with the disease severity of harvested tubers, whereas the spore ball density determined using PCR had an indistinct relationship with disease severity. The infection potential can be useful in evaluating soil contamination and in applying countermeasures against powdery scab.     

The potential for the Rapid Screening of Potato Cultivars (Solanum tuberosum) for Resistance to Powdery Scab (Spongospora subterranea) using a Laboratory Bioassay

Powdery scab of potato, once established in a field, is difficult to control because of the longevity of the resting spores (cystosori) of the causal organism, Spongospora subterranea f.sp. subterranea. Host resistance is likely to be the most efficient in a long-term control strategy for preventing build-up of field inoculum and spread of the disease. Resistance screening of potato cultivars is mostly done in laborious field trials where disease development is likely to be unpredictable. A bioassay with potato tissue cultured plantlets and cystosori as inoculum is described and was tested for its potential to screen potato cultivars at an early stage for their relative susceptibility to powdery scab by comparing the lab results with field data. With cystosori inoculum of Swiss origin, the laboratory test showed clear differences between the potato cultivars in the severity of zoosporangial root infection which correlated better with ranked tuber infection data, compared to root galling. There are apparent differences in the relative trends in susceptibility between roots and tubers of five selected cultivars when using naturally infested soil instead of prepared cystosori as inoculum in the lab bioassay. Furthermore, differences in the severity of zoosporangial root infection of two selected cultivars were found when cystosori from different countries where used as inoculum. A possible host genotype × pathogen interaction is discussed. The bioassay has the potential to screen and select for resistant material at an early breeding stage thus making field trials not unnecessary but more economical. It will allow the use of a standard set of pathogen collections and facilitate testing for inoculum virulence in infested soils.     

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.     

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:粉痂病在云南发生普遍,发病率在0%~51.72%,平均发病率和病情指数分别为15.01%和5.32。其中春作区马铃薯粉痂病的发生较为普遍,平均发病率为25.08%,病情指数为8.24。冬作区马铃薯粉痂病在部分产区有发生,平均发病率为4.93%,病情指数为2.39。与2004年-2008年云南省粉痂病调查数据相比,粉痂病发病率下降,病害有所减轻。     

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.     

Biocontrol of powdery scab of potato by seed tuber application of an antagonistic fungus, <Emphasis Type="Italic">Aspergillus versicolor</Emphasis>, isolated from potato roots

An environmentally friendly measure to control potato powdery scab caused by a protozoan pathogen Spongospora subterranea f.sp. subterranea (Sss) was developed by focusing on antagonistic microorganisms that were considered compatible with potato root. Five hundred and eight soil fungi, isolated from potato root cultivated in soil suspensions from four potato fields in Hokkaido, were screened for suppressiveness of root infection by Sss in a hydroponic culture system and for powdery scab severity in greenhouse and field experiments. Antagonistic isolate Im6-50, identified as Aspergillus versicolor, was selected as a potent biological control agent. In a 3-year field test, A. versicolor Im6-50 suppressed powdery scab with a protection value of 54–70 (100?=?complete protection) when applied directly on seed tubers compared with a protection value of 77–93 by the synthetic fungicide fluazinam. A. versicolor Im6-50 was detected from the surface of daughter tubers and from the soil in which the inoculated seed tubers were cultivated by PCR using species-specific primers. The establishment of A. versicolor Im6-50 on the stolon of inoculated potato plants and in the rhizosphere is considered to contribute to the mechanism for disease suppression.     

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。     

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.     

Increased abundance of patatins,lipoxygenase and miraculins in a thaxtomin A-habituated potato Russet Burbank somaclone with enhanced resistance to common scab

Potato common scab caused by the actinobacterium Streptomyces scabiei is characterized by the formation of corky lesions on tubers that reduce their marketability. Management of common scab is very complex and often ineffective under various environmental conditions. Using potato varieties that are more resistant to common scab remains one of the most efficient strategies to control this disease. However, very little is known about the factors associated with resistance to common scab. Somaclone RB9 regenerated from thaxtomin A-habituated potato Russet Burbank calli produced tubers more resistant to common scab than the original variety. Comparison of the RB9 tuber proteome with that of Russet Burbank using label-free quantitative proteomic analysis revealed changes in the accumulation of defence-related proteins from the patatin and lipoxygenase (LOX) families, which are involved in the metabolism of lipids, and of two miraculins of the Kunitz-type protease inhibitors family. The implication of LOX during common scab infection was studied using synchronized minitubers developed from leaf-bud cuttings. S. scabiei infection stimulated the accumulation of LOX in both Russet Burbank and RB9 minitubers, but this accumulation was intensified in RB9 minitubers. Infection also increased LOX activity in Russet Burbank and RB9 minitubers. However, LOX activity measured in noninfected RB9 minitubers was similar to that of infected Russet Burbank minitubers, indicating endogenous activation of LOX activity in RB9 minitubers. We discuss how increased LOX abundance and activity in the somaclone RB9 may contribute to improving tuber defence against common scab.     

Prevalence and distribution of Potato mop‐top virus in Scotland

This study was undertaken to determine the current occurrence in Scottish seed potato crops of Potato mop‐top virus (PMTV), which is transmitted by Spongospora subterranea and causes spraing (brown arcs and lines) in the flesh of potato tubers, rendering them unsaleable. In 2004, a stratified survey of four commonly grown cultivars was conducted, while in 2007 and 2008, only samples from powdery scab‐affected crops were collected. The incidence of crops in which infection by PMTV was present was 37·5% in the stratified survey in 2004, but was greater in surveys in which tubers with powdery scab were tested (47·2% in 2007 and 44·6% in 2008). Similarly, the frequency of crops with incidences of more than 10% tuber infection was lower (9·4%) in 2004 than in 2007 (25·4%) and in 2008 (26·2%). Significant differences in crop infection were found amongst the four major seed‐producing regions and the counties within these regions. The incidence of crop and tuber infection was least for class Pre‐basic seed potatoes and greatest for class Super Elite 3 and Elite seed potatoes. The results indicate that the prevalence of PMTV has not increased since surveys in the early 1970s.     

Prevalence of fungal and fungus‐like diseases on potato seed tubers imported from Europe to Jordan

A total of 109 samples of potato seed tubers imported to Jordan from France, Netherlands and Denmark during the 2007/2008 growing seasons were surveyed for 10 different diseases caused by fungi and fungus‐like organisms to determine the prevalence of different tuberborne diseases and to evaluate the efficacy of the current visual examination procedures in detecting different potato diseases. The results demonstrated that most potato seed tuber lots imported to Jordan were infected with one or more of the following potato pests: Colletotrichum coccodes (black dot) (66.0%), Rhizoctonia solani (black scurf) (42.4%), Helminthosporium solani (silver scurf) (92.6%), and Spongospora subterranea (powdery scab) (13.8%). Just over a quarter (25.7%) of all lots examined in this study were found to exceed the acceptance limits for one or more potato diseases according to the Jordanian standards. This study suggests that the current visual examination procedure is not adequate to detect all tuberborne diseases of potato and to estimate their incidence and severity precisely.     

Relationship of resistance to common scab disease and tolerance to thaxtomin A toxicity within potato cultivars

Thaxtomin A has a central and implicit role in common scab disease expression in potato. Thaxtomin A tolerance has been suggested as a possible rapid means for screening potato germplasm for disease resistance, during breeding selections. We have tested a range of genetically diverse cultivars with varying resistances to common scab disease in both pot and field based studies and measured their mean necrosis response to thaxtomin A. We found no association between resistance to common scab disease and tolerance to thaxtomin A toxicity. For example, disease resistant cultivars ‘Russet Burbank’ and ‘Atlantic’ were sensitive and tolerant to thaxtomin A toxicity respectively. Similarly; disease susceptible cultivars ‘Bismark’ and ‘Tasman’ showed susceptibility and tolerance to thaxtomin A. This demonstrates that whilst thaxtomin A is critical to disease expression, reaction to this toxin is only one component influencing resistance to common scab disease and many other anatomical, physiological or biochemical factors are critical to defence against this disease.     

Differential interaction of Phytophthora infestans on tubers of potato cultivars with different levels of blight resistance

Differential interactions in tuber blight attack between potato cultivars and Phytophthora infestans isolates were studied using whole tuber and tuber slice assays. Tuber blight incidence and severity were studied in a whole tuber assay, whilst necrosis and mycelium coverage were evaluated in a tuber slice assay. The overall defence reaction of the potato cultivars tested varied considerably. Cultivars like Kartel and Producent showed resistant reactions, whilst Bintje and, to a lesser extent, Astarte reacted more susceptibly after inoculation with aggressive strains of P. infestans . A highly significant cultivar by year interaction was observed when tuber blight incidence was evaluated in two successive years. Differential responses were revealed by changing ranked order of cultivars after exposure to aggressive isolates of P. infestans . The results show that cultivar by isolate interactions existed for all components of tuber blight resistance studied. The quantitative nature of the observed resistance responses suggests the presence of quantitative trait loci governing resistance to tuber blight. The consequences of differential interactions in relation to the stability of tuber resistance are discussed.     

Temporal association of potato tuber development with susceptibility to common scab and Streptomyces scabiei‐induced responses in the potato periderm

Using hydroponics and novel non‐destructive pot culture systems which enable inoculation at specific tuber development stages, the dynamics of common scab infection patterns in potato were studied in order to provide more precise identification of tuber physiological factors associated with susceptibility. At the whole‐tuber level, infection percentages were greatest when Streptomyces scabiei inoculation occurred early; at 2 weeks after tuberization (WAT) 68% of tubers became infected, contrasting with late inoculation (8 WAT), when only 4% infection occurred. The first‐formed internodes were most susceptible to infection, whilst later‐forming and slower‐expanding internodes were less susceptible. Detailed tuber physiological examination of internode 2 showed that pathogen‐induced changes, including increased phellem (periderm) thickness, cell layers and phellem suberization (key physiological features believed critical to S. scabiei infection) were promoted through S. scabiei inoculation. Sequential harvesting showed enhanced phellem suberization (28% greater than the control) within 7 days of pathogen exposure, while phellem thickness and layer responses were also initiated early in the infection process (10–14 days after pathogen exposure) and these responses were independent of symptom expression. Differences in cultivar response were observed, with greater phellem suberization observed 10 days after tuberization (DAT) in the common‐scab‐tolerant cv. Russet Burbank than in the susceptible cv. Desiree. Likewise, Russet Burbank had thicker and more numerous cell layers in the phellem (up to eight cell layers) during early tuber growth (20–30 DAT) than Desiree (up to six cell layers).