Detection and characterization of QoI resistance in Pyrenophora tritici-repentis populations causing tan spot of wheat in Argentina

Tan spot, caused by the fungus Pyrenophora tritici-repentis (Ptr), is a disease that has become more prevalent and intense in wheat crops in Argentina in recent years. Failure to control the disease with strobilurin fungicides, which were once effective, has been observed in different zones where wheat is grown. However, whether or not true resistance is present in the pathogen population in the region is not scientifically confirmed. This study evaluated the sensitivity of numerous Ptr isolates to representative QoI fungicides used in Argentina through in vitro and in planta assays, as well as through molecular analysis. Eighty-two monosporic isolates obtained in different locations in the north and south of Buenos Aires province in 2014, 2016, and 2018 were tested to determine sensitivity to selected QoI fungicides in conidial germination and mycelial inhibition assays, as well as in molecular analysis. Conidial germination was not inhibited at 1 µg/ml of azoxystrobin, trifloxystrobin, and pyraclostrobin. On the other hand, mycelial growth was inhibited by 59%, 56%, and 86% at 100 µg/ml of azoxystrobin, trifloxystrobin, and pyraclostrobin, respectively. The molecular analysis detected the G143A mutation in the cytb gene of all the 82 Ptr isolates, but the F129L and G137R substitutions were not present. This study documents the G143A mutation conferring QoI resistance in Ptr in South America. The findings of this study are key for future decisions regarding use of fungicide and rotation in the region.     

Pathogenic and molecular variability among isolates of Pyrenophora tritici-repentis, causal agent of tan spot of wheat in Argentina

Tan spot, caused by Pyrenophora tritici-repentis, is a common disease of wheat (Triticum aestivum) responsible for economic losses in some wheat growing areas worldwide. In this study the pathogenic and genetic diversity of 51 P. tritici-repentis isolates collected from different ecological regions of Argentina were analyzed. Virulence tests were conducted on 10 selected wheat cultivars: Buck Halcón, Chris, Gabo, Glenlea, Klein Dragón, Klein Sendero, Max, ND 495, ProInta Guazú and ProInta Imperial. Data revealed significant differences between all main factors evaluated and the interactions for 19 of the isolates analyzed. Based on the reaction type of each isolate/cultivar combination, 48 different pathogenic patterns were detected. The molecular analysis using Inter-Simple Sequence Repeats (ISSR) revealed the existence of 36 different haplotypes among 37 isolates of P. tritici-repentis originally selected for this study. These results indicate that P. tritici-repentis on wheat in Argentina is a heterogeneous fungus, implying that screening wheat germoplasm for resistance for tan spot disease requires a wide range of pathogen isolates.     

Emergence of tan spot disease caused by toxigenic Pyrenophora tritici-repentis in Australia is not associated with increased deployment of toxin-sensitive cultivars

The wheat disease tan (or yellow leaf) spot, caused by Pyrenophora tritici-repentis, was first described in the period 1934 to 1941 in Canada, India, and the United States. It was first noted in Australia in 1953 and only became a serious disease in the 1970s. The emergence of this disease has recently been linked to the acquisition by P. tritici-repentis of the ToxA gene from the wheat leaf and glume blotch pathogen, Stagonospora nodorum. ToxA encodes a host-specific toxin that interacts with the product of the wheat gene Tsn1. Interaction of ToxA with the dominant allele of Tsn1 causes host necrosis. P. tritici-repentis races lacking ToxA give minor indistinct lesions on wheat lines, whereas wheat lines expressing the recessive tsn1 are significantly less susceptible to the disease. Although the emergence and spread of tan spot had been attributed to the adoption of minimum tillage practices, we wished to test the alternative idea that the planting of Tsn1 wheat lines may have contributed to the establishment of the pathogen in Australia. To do this, wheat cultivars released in Australia from 1911 to 1986 were tested for their sensitivity to ToxA. Prior to 1941, 16% of wheat cultivars were ToxA-insensitive and hence, all other factors being equal, would be more resistant to the disease. Surprisingly, only one of the cultivars released since 1940 was ToxA insensitive, and the area planted to ToxA-insensitive cultivars varied from 0 to a maximum of only 14% in New South Wales. Thus, the majority of the cultivars were ToxA-sensitive both before and during the period of emergence and spread of the disease. We therefore conclude that the spread of P. tritici-repentis in Australia cannot be causally linked to the deployment of ToxA-sensitive cultivars.     

Seed transmission ofPyrenophora tritici-repentis,causal fungus of tan spot of wheat

Seed transmission ofPyrenophora tritici-repentis, a common foliar pathogen of wheat, was investigated in soft white winter wheat cv. Frankenmuth and found to be non-systemic; the emerging coleoptile was infected externally by hyphal growth from the infected pericarp. Hyphae from the infected coleoptile then infected the first and second seedling leaves as they emerged. Coleoptile symptoms ranged from tiny brown streaks or spots to large, brown necrotic areas accompanied by cracking and distortion of the coleoptile. Small brown spots sometimes occurred on the first and rarely on the second seedling leaves, often accompanied by leaf distortion. Pseudothecial initials of the fungus were present within or on the seed remnants. Seed transmission efficiency was as high as 92%in vitro and 60% in potting soil outdoors. Seed infection did not affect germinationin vitro, but slightly reduced emergence in potting soil. Seedling weight and height were reduced significantly. In potting soil, seed infection also resulted in delayed plant growth and increased tan spot severity at later stages of plant development. Under controlled conditions, seed transmission efficiency and incidence of pseudothecia on seed were negatively correlated with seed germination temperature in the range of 9 to 21 °C, whereas frequency of recovery of the fungus from symptomatic coleoptiles and leaves was positively correlated with seed germination temperature.These results suggest that infected seed may serve as a source of inoculum for tan spot epidemics and for dispersal of strains of the fungus to new areas.     

Infection of resistant and susceptible cultivars of wheat by Pyrenophora tritici-repentis

Conidial germination, appressorial formation. penetration of epidermal walls, formation of intracellular vesicles and growth of intracellular hyphae in epidermal cells occurred within 12 h of inoculation. Hyphae then grew slowly between mesophyll cells for the next 12 h. Some papillae formed beneath appressoria and most infected epidermal cells retained stain by 24 h after inoculation, indicating major changes in cellular physiology. Slight differences between cultivars in some of these events were not related to resistance.
On the second day. intercellular hyphae emerged more extensively from the infection sites into the mesophyll of the susceptible cultivar Banks, and formed significantly larger mycelia than in the resistant cultivar BH1146 by 3-5 days from inoculation. Rapid intercellular growth then continued in the susceptible cultivar but not in the resistant cultivar. Necrotic lesions expanded faster in the susceptible cultivar from day 3. By day 10. most lesions in this cultivar were large and light brown with a conspicuous chlorotic margin but those in the resistant cultivar were small and dark brown with inconspicuous chlorosis.     

Silicon suppresses tan spot development on wheat infected by <Emphasis Type="BoldItalic">Pyrenophora tritici-repentis</Emphasis>

Tan spot caused by Pyrenophora tritici-repentis is the main foliar diseases of wheat in Brazil. The effect of silicon (Si) on the components of resistance of a susceptible (Fundacep Horizonte) and a moderately resistant (Quartzo) wheat cultivar was studied in a controlled environment. Silicon was supplied as calcium silicate in the soil 30 days before sowing. At the booting stage, a conidial suspension of the fungus was sprayed onto the flag leaves of potted plants, which were incubated under moist conditions for 48 h. Afterwards, inoculated leaves were assessed for: incubation period (IP), infection efficiency (IE), area under lesion size curve (AULSC), lesion size (LS), severity (SEV) and area under severity curve (AUSC). Foliar Si concentrations were quantified at the end of the evaluations. Si supply to plants increased leaf Si concentration in 233% for Fundacep Horizonte (from 4.8 to 16.0 g kg^?1 of dry matter) and 211% for Quartzo (from 5.3 to 16.5 g kg^?1 of dry matter). In the Si + treatments, IP was longer by 24 and 17 h, IE declined by 53.5 and 65.5%, LS (at 264 h after inoculation) by 4.6 mm (from 9.5 to 4.9 mm) and 5.9 mm (from 8.2 to 2.3 mm), and SEV by 53% (from 54.4 to 18.8%) and 88% (from 47.7 to 5.5%) respectively, for the Fundacep Horizonte and Quartzo cultivars. The Si x cultivar interaction was not significant for AULSC and AUSC, and these variables were reduced by 55.8 and 80.8%, respectively, in plants supplied with Si. In conclusion, Si enhanced the resistance of wheat plants to tan spot development by affecting several resistance components, regardless of the resistance level of the cultivar. However, the greatest reduction in tan spot development by Si supply was observed when using a moderately resistant cultivar.     

Genetic analysis of resistance to Pyrenophora tritici-repentis races 1 and 5 in tetraploid and hexaploid wheat

Tan spot of wheat, caused by the fungus Pyrenophora tritici-repentis, is a destructive disease worldwide that can lead to serious losses in quality and quantity of wheat grain production. Resistance to multiple races of P. tritici-repentis was identified in a wide range of genetically diverse genotypes, including three different species Triticum aestivum (AABBDD), T. spelta (AABBDD), and T. turgidum (AABB). The major objectives of this study were to determine the genetic control of resistance to P. tritici-repentis races 1 and 5 in 12 newly identified sources of resistance. The parents, F(1), F(2), and F(2:3) or F(2:5) families of each cross were analyzed for the allelism tests and/or inheritance studies. Plants were inoculated at the two-leaf stage under controlled environmental conditions and disease reaction was assessed based on lesion-type rating scale. A single recessive gene controlled resistance to necrosis caused by P. tritici-repentis race 1 in both tetraploid and hexaploid resistant genotypes. The lack of segregation in the inter- and intra-specific crosses between the resistant tetraploid and hexaploid genotypes indicated that they possess the same genes for resistance to tan necrosis and chlorosis induced by P. tritici-repentis race 1. A single dominant gene for chlorosis in hexaploid wheat and a single recessive gene for necrosis in tetraploid wheat, controlled resistance to P. tritici-repentis race 5.     

Vavilov wheat accessions provide useful sources of resistance to tan spot (syn. yellow spot) of wheat

Host genetic resistance is the most effective and sustainable means of managing tan spot or yellow spot of wheat. The disease is becoming increasingly problematic due to the adoption of minimum tillage practices, evolution of effector‐mediated pathogenicity, and widespread cultivation of susceptible cultivars from a narrow genetic base. This highlights the importance of broadening the diversity of resistance factors in modern breeding germplasm. This study explored 300 genetically diverse wheat accessions, originally sourced from the N. I. Vavilov Institute of Plant Genetic Resources (VIR), St Petersburg, Russia. The collection was screened for resistance to tan spot at seedling and adult stage under controlled conditions, and in the field across 2 years. The phenotypic datasets, coupled with ToxA bioassay screening, identified a number of accessions with useful sources of resistance. Seedling disease response corresponded well with ToxA sensitivity (r = 0.49, P < 0.000), but not adult responses (r = ?0.02 to ?0.19, P < 0.002), and overall reactions to ToxA appeared to show poor correspondence with disease response at the adult stage. ToxA‐insensitive accessions were generally found resistant across different growth stages (all‐stage resistance, ASR) in all experiments (seedling and adult stage under controlled conditions and field). ToxA‐sensitive accessions that were susceptible at seedling stage, but resistant at both adult‐plant stages, were deemed to carry adult‐plant resistance (APR). This study provides detailed information on the degree of tan spot resistance in the Vavilov wheat collection and discusses strategies to harness these sources to boost the diversity of resistance factors in modern wheat breeding germplasm.     

Comparative analyses of spot blotch and tan spot epidemics on wheat under optimum and late sowing period in South Asia

In South Asia, foliar blight of wheat (Triticum aestivum L.) is a disease complex caused by Cochliobolus sativus (the spot blotch pathogen) and Pyrenophora tritici-repentis (the tan spot pathogen) which can reduce yields by >30?%. Little is known about the effects of wheat genotypes and their planting time on foliar blight epidemics. Field experiments were conducted to determine the infection potential and epidemic development of C. sativus and P. tritici-repentis on two susceptible (Sonalika and BL1473) and two tolerant (NL750 and Milan/Shanghai-7) wheat genotypes under optimum (November 26) and late (December 11 and December 26) planting conditions. The dynamics of airborne conidia were studied using air samplers. The highest aerial concentrations of conidia and disease incidence of both pathogens on all four wheat genotypes were detected during the first 3?weeks of March under both optimum and late seeding conditions in both years. Compared to optimum sowing time (i.e. November 26), wheat genotypes had higher disease severity when planted late on December 11 and 26. The disease complex reduced grain yield by 20.5, 27.2 and 37.3?% for November 26, December 11 and December 26 plantings, respectively in 2004. The corresponding differences were 17.7, 23.6 and 30.2?% in 2005. The findings of this study provide valuable information on the epidemiology of spot blotch and tan spot, which could help in developing strategies for managing these diseases in South Asian region through the selection of suitable genotypes and planting dates.     

Distribution and Pathogenic Characterization of Pyrenophora tritici-repentis and Stagonospora nodorum in Ohio

ABSTRACT To determine the distribution of Stagonospora nodorum and Pyrenophora tritici-repentis on wheat in Ohio, flag leaves with lesions were collected from wheat-producing counties in 2002 and 2003. Counties were arbitrarily grouped into seven regions. Log-linear analysis of pathogen presence within regions indicated that the presence of S. nodorum was independent of the presence of P. tritici-repentis. A logistic analysis revealed that the occurrence of both pathogens varied by region in one or both years. The aggressiveness of S. nodorum isolates was determined by inoculating two susceptible genotypes with a subsample of isolates from each region from both years. S. nodorum isolates obtained from northeast Ohio, with fewer wheat fields, were less aggressive than those from other regions. Isolates obtained from west-central Ohio, surrounded by regions with high wheat production annually, were significantly more aggressive than those obtained in the remaining five regions. Isolates from the five other regions did not differ significantly (P > 0.05) in aggressiveness. Races 1 and 2, and a few race 3 isolates, of P. tritici-repentis were detected in Ohio. The distribution of P. tritici-repentis races 1 and 2 was not associated with any region, although the prevalence of race 1 was three times greater than race 2. The rarer race 3 was associated with three dispersed regions. Results indicate that S. nodorum was the major wheat leaf-blotching pathogen. There were no positive or negative associations of S. nodorum and P. tritici-repentis or individual races of P. tritici-repentis in any of the tested regions, which indicates that neither pathogen can be used to predict the presence of the other. The isolated northeastern corner of Ohio appeared to contain isolates of S. nodorum with unique characteristics and potentially only one race of P. tritici-repentis, indicating that this area may be genetically isolated from the remaining tested areas of the state.     

Forensic pathology of canadian bread wheat: the case of tan spot

ABSTRACT Pyrenophora tritici-repentis causes necrosis and chlorosis in its wheat host. Susceptibility to races 2 (necrosis) and 5 (chlorosis) of the pathogen is known to be mediated by Ptr ToxA and Ptr ToxB, respectively. Sensitivity to each toxin is controlled by a single dominant and independently inherited gene. We used sensitivity to Ptr ToxA and Ptr ToxB as two genetic markers to investigate the origin and the state of tan spot susceptibility in Canadian Western Red Spring (CWRS) wheat over a period of more than a century. Sensitivity to Ptr ToxA, the toxin produced by nearly all isolates of the pathogen collected in the past 20 years in western Canada, appears to have been present in the first major cultivar, Red Fife, grown massively in the late 1800s. Sensitivity then was transmitted unknowingly into Canadian wheat lines through extensive use of backcrossing to maintain the Marquis-Thatcher breadmaking quality. Sensitivity to Ptr ToxA, which nearly disappeared from cultivars grown in western Canada in the 1950s, was reintroduced in the 1960s and unintentionally bred into many of the present-day cultivars. Sensitivity to Ptr ToxB, a toxin rarely found in isolates from western Canada, appeared with the release of Thatcher in 1934 and was transferred to many cultivars through backcross programs. In spite of large areas planted to Ptr ToxAand Ptr ToxB-sensitive cultivars over decades, tan spot epidemics remained sporadic until the 1970s. The results of this study raise the problem of the narrowing genetic base of CWRS wheat lines and the potential for unanticipated threats from plant pathogens. The intercrossing of genetically diverse material in one Canadian wheat breeding program resulted in the release of several modern cultivars with resistance to tan spot. The absence of wild-type Ptr ToxB-producing isolates in western Canada remains unexplained, given that sensitivity to Ptr ToxB was present continuously in western Canadian cultivars grown on vast areas for more than 70 years.     

Structural and Physical Properties of a Necrosis-Inducing Toxin from Pyrenophora tritici-repentis

ABSTRACT Cultivar-specific toxic metabolites of Pyrenophora tritici-repentis are involved in the appearance of necrotic and chlorotic foliar lesions characteristic of tan spot. A P. tritici-repentis necrosis-inducing toxin, Ptr necrosis toxin, was purified from isolate 86-124, sequenced by gas-phase amino acid microsequencing, and characterized by circular dichroism (CD) spectroscopy and isoelectric focusing. The purified protein had a similar amino acid composition and molecular weight as previously reported. Analysis of the CD spectrum from 178 to 250 nm indicated a protein consisting of 13% alpha-helix, 36% antiparallel beta-sheet, 25% turns, and 25% other structures. The Ptr necrosis toxin from isolate 86-124 has an isoelectric point near pH 10. Using overlapping proteolytic fragments obtained from the toxin, a sequence of 101 continuous amino acids was obtained, but the amino terminus was blocked and 9 to 16 amino acids could not be sequenced. Secondary structure prediction based on the amino acid sequence indicated a beta-sheet protein with little alpha-helix, which is in agreement with the structure determined by CD spectroscopy. Sequence analysis indicated the presence of a possible membrane adhesion site and several possible phosphorylation sites that may be involved in phytotoxicity.     

Epidemiology of foliar blights (spot blotch and tan spot) of wheat in the plains bordering the himalayas

ABSTRACT Helminthosporium leaf blight (HLB), a complex of spot blotch caused by Cochliobolus sativus and of tan spot caused by Pyrenophora tritici-repentis, is a major wheat disease in South Asia. This 2-year study elucidated HLB development and its impact on yield. Symptoms caused by C. sativus and P. tritici-repentis were first observed at the seedling and tillering stages, respectively. The number of airborne conidia and leaves infected by the two pathogens remained low for several weeks under lower temperatures, followed by a sharp rise as temperatures increased. The number of airborne conidia of C. sativus and incidence of infection by C. sativus were higher compared with P. tritici-repentis. The disease complex caused an average 30% reduction in yield, with greater losses under delayed seeding. Delayed seeding increased disease severity even in resistant genotypes and caused higher yield losses. 'Milan/Shanghai-7' was the most resistant among six genotypes evaluated. Despite higher disease severity, 'BL 1473' showed relatively lower yield losses, indicating its tolerance to foliar blight. The findings of this study bear implications for integrated foliar blight management in the warmer areas of South Asia by combining optimum seeding date, seed treatment and foliar spray of fungicides, and resistant wheat genotypes.     

Neural networks that distinguish infection periods of wheat tan spot in an outdoor environment

ABSTRACT Tan spot of wheat, caused by Pyrenophora tritici-repentis, provided a model system for testing disease forecasts based on an artificial neural network. Infection periods for P. tritici-repentis on susceptible wheat cultivars were identified from a bioassay system that correlated tan spot incidence with crop growth stage and 24-h summaries of environmental data, including temperature, relative humidity, wind speed, wind direction, solar radiation, precipitation, and flat-plate resistance-type wetness sensors. The resulting data set consisted of 97 discrete periods, of which 32 were reserved for validation analysis. Neural networks with zero to nine processing elements were evaluated 20 times each to identify the model that most accurately predicted an infection event. The 200 models averaged 74 to 77% accuracy, depending on the number of processing elements and random initialization of coefficients. The most accurate model had five processing elements and correctly predicted 87% of the infection periods in the validation set. In comparison, stepwise logistic regression correctly predicted 69% of the validation cases, and multivariate discriminant analysis distinguished 50% of the validation cases. When wetness-sensor inputs were withheld from the models, both the neural network and logistic regression models declined 6% in prediction accuracy. Thus, neural networks were more accurate than statistical procedures, both with and without wetness-sensor inputs. These results demonstrate the applicability of neural networks to plant disease forecasting.     

Occurrence of halo spot in barley caused by Pseudoseptoria donacis in Argentina

Barley is an important malting crop extensively cultivated in the province of Buenos Aires (AR). In 1992, an unusual disease was detected with a prevalence of 8% on 36 commercial fields examined in this province. The pattern of the disease, the characteristics of the pathogen and the test for pathogenicity indicated that the causal agent was Pseudoseptoria donacis.     

Influence of site factors on yield response of winter wheat to fungicide programmes in England and Wales, 1979–1987

Results of 460 field experiments between 1979 and 1987 incorporating one, two and three fungicide sprays, with a range of active ingredients, were analysed according to site factors including previous crop, sowing date and geographical area. The average untreated yield was 7–48 t/ha, although this fluctuated widely from 6 15 t/ha in 1985 to 9 411 ha in 1984. A single spray at flag-leaf emergence gave an average yield increment of 0 59 t/ha over plots receiving no fungicide. The addition of a second spray at the first-node growth stage gave an extra benefit of 0.22 t/ha, while an additional third spray at ear emergence gave a further 0.28t/ha. The effects on yield of different spray timings were modified by other factors, of which sowing date and previous crop were most important to the first-node spray, and cultivar and geographical area to the ear emergence spray. The data suggest that all crops would benefit economically from a broad-spectrum fungicide applied at flag-leaf emergence. Additionally, crops sown in September, those following winter wheat and those in regions bordering the North Sea respond well at the first-node stage to a fungicide active against eyespot.