Ueli Merz

Variation for neutral markers is correlated with variation for quantitative traits in the plant pathogenic fungus Mycosphaerella graminicola

We compared genetic variation and population differentiation at RFLP marker loci with seven quantitative characters including fungicide resistance, temperature sensitivity, pycnidial size, pycnidial density, colony size, percentage of leaves covered by pycnidia (PLACP) and percentage of leaves covered by lesions (PLACL) in Mycosphaerella graminicola populations sampled from four regions. Wide variation in population differentiation was found across the quantitative traits assayed. Fungicide resistance, temperature sensitivity, and PLACP displayed a significantly higher QST than GST, consistent with selection for local adaptation, while pycnidial size, pycnidial density and colony size displayed a lower or significantly lower QST than GST, consistent with constraining selection. There was not a statistical difference between QST and GST in PLACL. We also found a positive and significant correlation between genetic variation in molecular marker loci and quantitative traits at the multitrait scale, suggesting that estimates of overall genetic variation for quantitative traits in M. graminicola could be derived from analysis of the molecular genetic markers.

Review: Powdery Scab of Potato—Increased Knowledge of Pathogen Biology and Disease Epidemiology for Effective Disease Management

The importance of the potato tuber disease powdery scab, caused by the zoosporic pathogen Spongospora subterranea f. sp. subterranea, has increased worldwide, and the disease is one of the most important problems facing potato production in some regions. This soilborne pathogen produces many resting spores which can remain dormant for long periods, are highly resistant to environmental stresses and can spread the disease on seed potatoes and in contaminated soil. The enigmatic nature of this organism exacerbates the development of effective powdery scab control methods. Substantial knowledge has been gained in the last decade on the biology of the pathogen and the epidemiology of the disease, but no single effective control measure is, or is likely to be, available. An integrated approach to powdery scab management is the aim, with host resistance as a substantial and sustainable component. Further research on the epidemiology of powdery scab and population genetics of the pathogen is urgently required. All stakeholders involved in the potato industry must become aware that solution of the powdery scab problem is likely to be a long-term goal. When resistant cultivars with all the other characteristics demanded for marketing high-quality potatoes are available, and when disease risk from the pathogen on seed and/or in soil can be accurately determined, then the mission to develop effective powdery scab control will become achievable.

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.

Improved immunological detection of Spongospora subterranea

The genus Spongospora has two members which are important pathogens of vegetables, S. subterranea f.sp. subterranea (Sss) and S. subterranea f.sp. nasturtii (Ssn). The close taxonomic relationship of these formae speciales is based on similar cystosori morphology. The potato disease powdery scab, caused by Sss, is difficult to control. The key control measure is avoidance, aimed at planting clean seed in clean soil. For the development of routine tests for the presence of the pathogen on tubers and in soil, a monoclonal antibody (MAb) was developed using Sss cystosori as immunogen. It detected less than one Sss cystosorus and recognised Sss material from many parts of the world. No cross-reactions with other Plasmodiophoromycetes including Plasmodiophora brassicae, Polymyxa betae, Polymyxa graminis and different Streptomyces species causing common and netted scab of potatoes were observed. A novel tuber sample test method was developed using a kitchen peeling machine. This detected two tubers with one powdery scab lesion each in a sample including eighteen uninfected tubers. When soil samples spiked with cystosori were tested with the MAb, different Sss infestation levels could be discriminated. Ssn cystosori gave absorbance values in ELISA as high as Sss cystosori, whereas fresh crook roots of watercress containing Ssnzoosporangia and plasmodia or mud from an Ssninfected watercress bed gave low absorbance values or no reaction. The potential of these findings for the development of a disease control management are discussed.

Incidence, Distribution, and Association of Spongospora subterranea and Potato mop-top virus in Costa Rica

A survey was conducted in 39 potato (Solanum tuberosum) fields in Costa Rica to determine incidence and association of Spongospora subterranea f. sp. subterranea and Potato mop-top pomovirus (PMTV). The fields were located in Costa Ricas two major potato-production regions and were further characterized by their altitude. In all, 633 paired samples of leaf tissue and corresponding tubers were collected, assessed visually for disease, and subsequently assayed by enzyme-linked immunosorbent assay (ELISA). S. subterranea presence in tuber tissue was tested by double-antibody sandwich (DAS)-ELISA and PMTV presence in leaf and tuber tissues was tested by triple-antibody sandwich (TAS)-ELISA. Moreover, soil samples were collected from 10 fields surveyed and were evaluated for both pathogens via ELISA and bioassay. The incidence of both diseases ranged from 0 to 100% within individual fields, with incidences lower than 40% occurring in more than 70% of the fields. Higher incidences were found in fields located at higher altitudes. Of the 633 paired samples, 179 and 146 were positive for PMTV and S. subterranea, respectively, according to ELISA in either the foliage or tubers. A low correlation was found for PMTV visual symptoms and ELISA test results. Only 14 of the 81 foliar samples testing positive for PMTV had visual symptoms; the remaining 67 samples were asymptomatic. Conversely, comparison of visual evaluation with detection of S. subterranea by ELISA on tubers showed that 70% of the results were coincident. S. subterranea was detected in 4 of 10 soil samples tested by ELISA. Soilborne PMTV was detected by ELISA in roots of bait plants sown in these soil samples. Co-occurrence of both pathogens was detected in 64 samples. A significant but low degree of association for vector and virus was determined, and data suggests that S. subterranea is participating in the transmission of PMTV in Costa Rica in low frequency.

Global Genetics and Invasion History of the Potato Powdery Scab Pathogen, Spongospora subterranea f.sp. subterranea

Spongospora subterranea f. sp. subterranea (Sss) causes two diseases on potato (Solanum tuberosum), lesions on tubers and galls on roots, which are economically important worldwide. Knowledge of global genetic diversity and population structure of pathogens is essential for disease management including resistance breeding. A combination of microsatellite and DNA sequence data was used to investigate the structure and invasion history of Sss. South American populations (four countries, 132 samples) were consistently more diverse than those from all other regions (15 countries, 566 samples), in agreement with the hypothesis that Sss originated in South America where potato was domesticated. A substantial genetic differenciation was found between root and tuber-derived samples from South America. Estimates of past and recent gene flow suggested that Sss was probably introduced from South America into Europe. Subsequently, Europe is likely to have been the recent source of migrants of the pathogen, acting as a “bridgehead” for further global dissemination. Quarantine measures must continue to be focussed on maintaining low global genetic diversity and avoiding exchange of genetic material between the native and introduced regions. Nevertheless, the current low global genetic diversity of Sss allows potato breeders to select for resistance, which is likely to be durable.

Powdery scab effect on the potato Solanum tuberosum ssp. andigena growth and yield

Powdery scab affects most potato production zones in the world. The causal organism Spongospora subterranea f. sp. subterranea forms galls in roots and lesions on tubers during its replication. Recent researches suggest that S. subterranea could cause harmful effects on the host plant and potentially on yield.In order to quantify the disease impact on plant growth and yield, potato Solanum tuberosum ssp. andigena, cv. Diacol Capiro was grown in a soil infested with S. subterranea and compared to a healthy control soil. Data was taken at the flowering stage and at harvest. S. subterranea caused a plant length reduction of 23%, a foliar dry weight reduction of 32% and a tuber weight reduction of 30%, whereas S. subterranea did not reduce the number of tubers or stems.

Morphological Enumeration of Resting Spores in Sporosori of the Plant Pathogen Spongospora subterranean

Plasmodiophorid sporosori (aggregations of resting spores) reach their most complex form in Spongospora subterranea f. sp. subterranea, the biotrophic plant pathogen which causes the economically important disease powdery scab of potato ( Solanum tuberosum ). Resting spores are the perennation life cycle stage of plasmodiophorids, allowing them to survive for long periods and infect subsequent host generations. Light microscopy was used to measure resting spores and sporosori of Sp. subterrane a, and enumerate resting spores in individual sporosori. Mean resting spore diameters differed for two sporosorus collections, being 4.0 μm (from New Zealand) and 4.3 μm (from Switzerland). Counts of resting spores in 4 μm thick serial sections of sporosori from one collection gave a mean of 667 (range 155 to 1,526) resting spores per sporosorus. Number of resting spores per sporosorus was closely related to sporosorus volume, and could be accurately estimated using the formula; number of resting spores = 0.0081 × sporosorus volume (assuming sporosori to be spheroids). Using this formula, mean numbers of resting spores in sporosori from 37 Sp. subterranea collections from field-grown potato tubers from 13 countries were determined to range from 199 to 713. Differences in numbers of resting spores between the collections were statistically significant (P < 0.05), and independent of country or host cultivar of origin, indicating that enumeration should be carried out for individual sporosorus collections to accurately quantify inoculum. Morphology, using scanning electron microscopy, also showed that between 2 and 51% (average 20%) of resting spores released zoospores after exposure to roots of host plants. The formula for resting spore enumeration validated in this study can be used to standardise Sp. subterranea resting spore inoculum for plant pathology studies, and possibly to assist determination of soil inoculum potential for disease risk evaluations.

Evaluation of the Sss AgriStrip rapid diagnostic test for the detection of Spongospora subterranea on potato tubers

Spongospora subterranea, f.sp. subterranea (Sss), which causes powdery scab, is mainly spread through infected seed tubers and survives in contaminated soil for many years. The visual assessment of tuber lots by inspectors carries the risk of misidentification due to the difficulty of distinguishing lesions caused by either Sss or Streptomyces spp.. To avoid this, the “Sss AgriStrip”, a rapid and lab-independent test tool based on a lateral flow immunoassay has been developed, and we assessed its accuracy and sensitivity for detecting Sss. The Sss AgriStrip performed as well as other lab-based identification methods. The Sss AgriStrip, microscopy, ELISA, PCR, and real-time PCR techniques identified infection with S. subterranea in all tubers with typical powdery scab lesions. When lots with tubers showing a mixture of typical and atypical (suspicious) symptoms were tested, the presence of S. subterranea was confirmed in all lesions by all methods. The DNA content was generally lower in atypical than in typical lesions. Diverse and suspicious symptoms, which were difficult to assign to either powdery or common scab, tested negative with Sss AgriStrip and the other methods. This was despite microscopic observation of sporosori-like structures in some samples. Isolation and molecular identification confirmed that these lesions were mostly caused by Streptomyces spp. The Sss AgriStrip is as sensitive as DAS-ELISA with a detection limit between 1 and 10 sporosori per ml buffer. It is ideal for rapid and selective detection of Sss on farms and border inspection points to prevent spread of the pathogen.

Not in your usual Top 10 – protists that infect plants and algae

Summary Fungi, nematodes and oomycetes belong to the most prominent eukaryotic plant pathogenic organisms. Unicellular organisms from other eukaryotic lineages, commonly addressed as protists, also infect plants. This review provides an introduction to plant pathogenic protists, including algae infecting oomycetes, and their current state of research.