Mycosphaerella Pinodes Research Articles

Spatiotemporal distribution of Ascochyta pinodes and Ascochyta pinodella during the winter growing season in France

Ascochyta blight of pea is caused by four related fungi, Ascochyta pisi, Phoma koolunga, Ascochyta pinodes and Ascochyta pinodella. The latter two taxa appear to be much more common and economically significant worldwide but the relative impact of each fungus on ascochyta blight epidemics is not well understood. To study the spatiotemporal distribution of A. pinodes and A. pinodella infecting pea in France, 368 isolates were sampled monthly, from February to May, at three locations (Rennes, Boigneville and Dijon) and molecular markers were used to genotype isolates. The aggressiveness of isolates from the fourth sampling date was estimated using a detached leaf assay on the winter cultivar Enduro. Disease was low during the sampling period as climatic conditions were generally not conducive to disease development (cold temperature, low rainfall). Population genetic analysis showed that 99% of the observed variation could be attributed to variation within populations compared to only 1% among populations. Both species were observed in each location, although A. pinodella was observed at a lower frequency (6–32%). Moreover, results showed that both species could develop on different nodes of the plant. Significant differences in aggressiveness were observed between species and among isolates within species with A. pinodes isolates being significantly more aggressive on average than A. pinodella isolates. These results emphasize the necessity to study the components of disease complexes in order to understand the impact of pathogen species interactions on disease and yield reduction as well as the dynamics of disease epidemics during the cropping season.

Identification and characterization of pathogens associated with root rot of winter peas grown under organic management in Germany

Root rots are limiting factor for pea production worldwide. This disease is caused by a pathogen complex and the role of single pathogens is unclear. This study aimed at identifying pathogens involved in a root rot of organically grown field pea in Germany, and establishing their importance in the disease complex. The potential of yard waste compost to suppress the diseased was also studied. Average disease severity index was similar in 2010 and 2011 (DI of 4.56 to 4.59, respectively) but it increased in 2012 to DI 5.8. Peyronellaea pinodella was most frequently isolated pathogen, with isolation frequency from 86%, 73% and 86% in 2010, 2011 and 2012, respectively. In addition, Didymella pinodes, Fusarium solani f. sp. pisi, F. oxysporum f. sp. pisi and F. avenaceum were the main fungi recovered from pea roots. In pathogenicity test all of the tested pathogens caused weak symptoms on the pigmented winter variety EFB33 and moderate to severe symptoms on the white flowering summer variety Santana. F. avenaceum was the most aggressive pathogen on Santana with DI of 7.4 followed by P. pinodella with DI of 5.7. The high aggressiveness combined with the wide host range highlights the possibility of F. avenaceum emerging as potential risk for organic crop rotation. High levels of resistance of EFB33 against all pathogens shows the potential of this variety to serve as a resource in further research for identification and development of new sources of resistance against root rot diseases of pea.

Rhizobium Impacts on Seed Productivity, Quality, and Protection of Pisum sativum upon Disease Stress Caused by Didymella pinodes: Phenotypic, Proteomic, and Metabolomic Traits.

In field peas, ascochyta blight is one of the most common fungal diseases caused by Didymella pinodes. Despite the high diversity of pea cultivars, only little resistance has been developed until to date, still leading to significant losses in grain yield. Rhizobia as plant growth promoting endosymbionts are the main partners for establishment of symbiosis with pea plants. The key role of Rhizobium as an effective nitrogen source for legumes seed quality and quantity improvement is in line with sustainable agriculture and food security programs. Besides these growth promoting effects, Rhizobium symbiosis has been shown to have a priming impact on the plants immune system that enhances resistance against environmental perturbations. This is the first integrative study that investigates the effect of Rhizobium leguminosarum bv. viceae (Rlv) on phenotypic seed quality, quantity and fungal disease in pot grown pea (Pisum sativum) cultivars with two different resistance levels against D. pinodes through metabolomics and proteomics analyses. In addition, the pathogen effects on seed quantity components and quality are assessed at morphological and molecular level. Rhizobium inoculation decreased disease severity by significant reduction of seed infection level. Rhizobium symbiont enhanced yield through increased seed fresh and dry weights based on better seed filling. Rhizobium inoculation also induced changes in seed proteome and metabolome involved in enhanced P. sativum resistance level against D. pinodes. Besides increased redox and cell wall adjustments light is shed on the role of late embryogenesis abundant proteins and metabolites such as the seed triterpenoid Soyasapogenol. The results of this study open new insights into the significance of symbiotic Rhizobium interactions for crop yield, health and seed quality enhancement and reveal new metabolite candidates involved in pathogen resistance.

Ultrastructural and Cytological Studies on Mycosphaerella pinodes Infection of the Model Legume Medicago truncatula.

Ascochyta (Mycosphaerella) blight on cultivated peas is primarily caused by infection through asexual spores (pycnospores) of Mycosphaerella pinodes (Berk. et Blox.) Vestergren [recently renamed Peyronellaea pinodes (Berk. & A. Bloxam) Aveskamp, Gruyter & Verkley]. Using a model pathosystem involving Medicago truncatula and Mycosphaerella pinodes strain OMP-1, we examined the histology and ultrastructure of early infection events and fungal development including penetration by appressoria, vegetative growth of infection hyphae, and host responses. On the susceptible ecotype R108-1, pycnospores germinated and grew over the surface of the epidermis, then formed an appressoria and penetrated the cuticle. Beneath the cuticle, the infection peg expanded into a hyphae that grew within the outer wall of the epidermis. Subsequently, the hyphae penetrated down within mesophyll cells and proliferated vigorously, eventually, forming asexual fruiting bodies (pycnidia). In contrast, successful penetration and subsequent growth of infection hyphae were considerably restricted in the ecotype Caliph. Detected by its reaction with cerium chloride (CeCl3) to generate electron-dense cerium perhydroxides in transmission electron micrographs, hydrogen peroxide (H2O2) accumulated in epidermal and mesophyll cells of Caliph challenged with pycnospores of M. pinodes. This intracellular localization was confirmed by energy-dispersive X-ray spectroscopy. Our observations thus indicate that the oxidative burst reaction leading to the generation of reactive oxygen species is associated with a local host defense response in Caliph, since no clear H2O2 accumulation was detectable in susceptible R108-1. Indeed, aberrant hyphae such as intrahyphal hyphae and dead hyphae, probably due to a local defense elicited by the fungus, were abundant in Caliph but not in R108-1. Our results on the cellular interactions between the fungus and host cells provide additional insights to understand foliar infection by M. pinodes on cultivated peas.

AAC Oriole red field pea

AAC Oriole is a semi-leafless red field pea (Pisum sativum L.) cultivar developed at the Lacombe Research and Development Centre, Agriculture and Agri-Food Canada, Lacombe, AB. It is shorter than most of the recently released yellow and green pea cultivars on the Canadian market with fair standability. It has early maturity, medium seed size, and round seed shape with a crude seed protein content of approximately 24%. AAC Oriole is resistant to powdery mildew caused by Erysiphe pisi DC. and moderately susceptible to mycosphaerella blight [caused by Mycosphaerella pinodes (Berk. & A. Blox.) Vestergr.] and fusarium wilt [caused by Fusarium oxysporum f. pisi (Linford) W.C. Snyder & H.N. Hansen]. It is adapted to all field pea growing regions in western Canada.

Expression of defence‐related genes in stems and leaves of resistant and susceptible field pea (Pisum sativum) during infection by Phoma koolunga

The differential expression of 13 defence‐related genes during Phoma koolunga infection of stems and leaves of susceptible versus resistant field pea (Pisum sativum) was determined using qRT‐PCR. Expression, in terms of relative mRNA level ratios, of genes encoding ferredoxin NADP oxidoreductase, 6a‐hydroxymaackiain methyltransferase (hmm6), chalcone synthase (PSCHS3) and ascorbate peroxidase in leaves and stems differed during 6–72 hours post‐inoculation (hpi) and reflected known host resistance levels in leaves versus stems. In comparison to the susceptible genotype, at 24, 48 and 72 hpi, two genes, hmm6 (122.43‐, 206.99‐ and 32.25‐fold, respectively) and PSCHS3 (175.00‐, 250.13‐ and 216.24‐fold, respectively), were strongly up‐regulated in leaves of the resistant genotype, highlighting that resistance against P. koolunga in field pea is governed by the early synthesis of pisatin. At 24 hpi, leaves infected by P. koolunga showed clear differences in expression of target genes. For example, the gene encoding a precursor of the defensin ‘disease resistance response protein 39’ was substantially down‐regulated in leaves of both the susceptible and the resistant genotypes inoculated with P. koolunga. This contrasts with other studies on another pea black spot pathogen, Didymella pinodes, where this same gene is strongly up‐regulated in leaves of resistant and susceptible genotypes. The current study provides the first understanding of defence‐related genes involved in the resistance against P. koolunga, opening novel avenues to engineer new field pea cultivars with improved leaf and stem black spot disease resistance as the basis for developing more effective and sustainable management strategies.

Key metabolic traits of Pisum sativum maintain cell vitality during Didymella pinodes infection: cultivar resistance and the microsymbionts' influence

Ascochyta blight causes severe losses in field pea production and the search for resistance traits towards the causal agent Didymella pinodes is of particular importance for farmers. Various microsymbionts have been reported to shape the plants' immune response. However, regardless their contribution to resistance, they are hardly included in experimental designs. We delineate the effect of symbionts (rhizobia, mycorrhiza) on the leaf proteome and metabolome of two field pea cultivars with varying resistance levels against D. pinodes and, furthermore, show cultivar specific symbiont colonisation efficiency. The pathogen infection showed a stronger influence on the interaction with the microsymbionts in the susceptible cultivar, which was reflected in decreased nodule weight and root mycorrhiza colonisation. Vice versa, symbionts induced variation of the host's infection response which, however, was overruled by genotypic resistance associated traits of the tolerant cultivar such as maintenance of photosynthesis and provision of sugars and carbon back bones to fuel secondary metabolism. Moreover, resistance appears to be linked to sulphur metabolism, a functional glutathione-ascorbate hub and fine adjustment of jasmonate and ethylene synthesis to suppress induced cell death. We conclude that these metabolic traits are essential for sustainment of cell vitality and thus, a more efficient infection response. SignificanceThe infection response of two Pisum sativum cultivars with varying resistance levels towards Didymella pinodes was analysed most comprehensively at proteomic and metabolomic levels. Enhanced tolerance was linked to newly discovered cultivar specific metabolic traits such as hormone synthesis and presumably suppression of cell death.

Seed health of organic peas and faba beans and its effects on the health of the harvested grains

A survey was conducted in Germany during 2009–2012 on seed health of organic pea and faba beans and their harvested progeny seed on up to 32 organic farms per year. Seeds were either basic seeds intended for production of certified seeds (Basic), certified organic seeds (Certified) and on-farm organically produced seeds but not certified (Uncertified). The frequency of Ascochytapisi or A. fabae, Didymella pinodes, Peyronellaea pinodella and Fusarium spp. was determined. For peas, 17 out of 71 and for faba beans 2 out of 48 seed lots had >10% infestation of Ascochyta blight complex pathogens, a level considered problematic. Ascochyta levels at harvest were >10% for 30 of the pea and four of the faba bean samples. As well for A. pisi as for D. pinodes, the mean percentage of infected seeds was significantly higher in uncertified pea seeds than in certified and basic seed lots. In most cases, the faba bean seeds were infected with one pathogen only, however, a combination of pathogens regularly occurred on peas. Overall, highly infested seed lots resulted in highly infested harvest lots. However, high infestations also occurred in lots originating from clean seed pointing to the importance of local soil- and air-borne infection sources. The relationships between the level of seed lot infestation, environmental factors and infestation levels of harvested progeny seeds were weak for both crops. For an overall improvement of pea and faba bean health in organic farming, there is a need to assure healthy seeds in addition to the reduction of soil- and potentially air-borne inoculum.

AAC Royce field pea

AAC Royce is a semi-leafless, green cotyledon field pea (Pisum sativum L.) cultivar developed at Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, Lacombe, in Alberta, Canada. It has a maturity of 105 d, 1000-seed weight of 254 g, and medium lodging resistance. AAC Royce is resistant to powdery mildew (caused by Erysiphe pisi Syd.) and moderately susceptible to mycosphaerella blight (caused by Mycosphaerella pinodes) and fusarium wilt (caused by Fusarium oxysporum). AAC Royce is adapted to all field pea growing regions in western Canada.

AAC Radius field pea

AAC Radius is a semi-leafless, green cotyledon field pea (Pisum sativum L.) cultivar developed at Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, Lacombe, Alberta, Canada. It is adapted to all field pea growing regions in western Canada. AAC Radius has medium maturity and seed size, and high seed coat integrity. AAC Radius has excellent bleaching resistance and high green seed color intensity. It is resistant to powdery mildew caused by Erysiphe pisi Syd., and moderately tolerant to mycosphaerella blight caused by Mycosphaerella pinodes [Berk. & Blox.] Vestergr, and fusarium wilt caused by Fusarium oxysporum Schlecht. emend. Snyd. & Hans. f. sp. pisi [van Hall] Snyd. & Hans.

Clarification on Host Range of Didymella pinodes the Causal Agent of Pea Ascochyta Blight.

Didymella pinodes is the principal causal agent of ascochyta blight, one of the most important fungal diseases of pea (Pisum sativum) worldwide. Understanding its host specificity has crucial implications in epidemiology and management; however, this has not been clearly delineated yet. In this study we attempt to clarify the host range of D. pinodes and to compare it with that of other close Didymella spp. D. pinodes was very virulent on pea accessions, although differences in virulence were identified among isolates. On the contrary, studied isolates of D. fabae, D. rabiei, and D. lentil showed a reduced ability to infect pea not causing macroscopically visible symptoms on any of the pea accessions tested. D. pinodes isolates were also infective to some extend on almost all species tested including species such as Hedysarum coronarium, Lathyrus sativus, Lupinus albus, Medicago spp., Trifolium spp., Trigonella foenum-graecum, and Vicia articulata which were not mentioned before as hosts of D. pinodes. On the contrary, D. lentil and D. rabiei were more specific, infecting only lentil and chickpea, respectively. D. fabae was intermediate, infecting mainly faba bean, but also slightly other species such as Glycine max, Phaseolus vulgaris, Trifolium spp., Vicia sativa, and V. articulata. DNA sequence analysis of the nuclear ribosomal internal transcribed spacer region (ITS) was performed to confirm identity of the isolates studies and to determine phylogenetic relationship among the Didymella species, revealing the presence of two clearly distinct clades. Clade one was represented by two supported subclusters including D. fabae isolates as well as D. rabiei with D. lentil isolates. Clade two was the largest and included all the D. pinodes isolates as well as Phoma medicaginis var. pinodella. Genetic distance between D. pinodes and the other Didymella spp. isolates was not correlated with overall differences in pathogenicity. Based on evidences presented here, D. pinodes is not specialized on pea and its host range is larger than that of D. fabae, D. lentil, and D. rabiei. This has relevant implications in epidemiology and control as these species might act as alternative hosts for D. pinodes.

Aggressiveness Changes in Populations of Didymella pinodes over Winter and Spring Pea Cropping Seasons.

Ascochyta blight, caused by the necrotrophic ascomycete Didymella pinodes, is responsible for severe losses in winter and spring pea crops. Despite different climatic conditions, epidemics on winter and spring crops are due to a single population of D. pinodes, suggesting gene flow either between the two crops or from reservoir sources during the cropping season. This should lead to similar pathogenicity characteristics in isolates sampled from the two crops. However, these hypotheses have never been formally tested. We therefore sampled a total of 520 D. pinodes strains throughout a growing season from winter and spring pea plots (WP and SP, respectively) and from winter and spring trap plants (TWP and TSP). Amplified fragment length polymorphism (AFLP) markers revealed high genetic diversity within subpopulations, whereas pathogenicity tests showed that mean aggressiveness increases over the course of an epidemic. These results support the idea that alloinoculum contributes to the carryover of epidemics between winter and spring crops and that the most aggressive isolates are selected as an epidemic progresses. Ascochyta blight, caused by Didymella pinodes, is responsible for severe losses in pea crops. While previous studies have shown that ascochyta blight epidemics on winter and spring crops are due to a single population of D. pinodes, suggesting that isolates from the two crops present similar pathogenicity characteristics, that hypothesis have never been tested. Genetic analysis of subpopulations sampled throughout a growing season from winter and spring pea plots revealed high genetic diversity within subpopulations, whereas pathogenicity tests showed that mean aggressiveness increases over the course of an epidemic.

Studies on the Control of Ascochyta Blight in Field Peas (Pisum sativum L.) Caused by Ascochyta pinodes in Zhejiang Province, China.

Ascochyta blight, an infection caused by a complex of Ascochyta pinodes, Ascochyta pinodella, Ascochyta pisi, and/or Phoma koolunga, is a destructive disease in many field peas (Pisum sativum L.)-growing regions, and it causes significant losses in grain yield. To understand the composition of fungi associated with this disease in Zhejiang Province, China, a total of 65 single-pycnidiospore fungal isolates were obtained from diseased pea samples collected from 5 locations in this region. These isolates were identified as Ascochyta pinodes by molecular techniques and their morphological and physiological characteristics. The mycelia of ZJ-1 could penetrate pea leaves across the stomas, and formed specific penetration structures and directly pierced leaves. The resistance level of 23 available pea cultivars was tested against their representative isolate A. pinodes ZJ-1 using the excised leaf-assay technique. The ZJ-1 mycelia could penetrate the leaves of all tested cultivars, and they developed typical symptoms, which suggested that all tested cultivars were susceptible to the fungus. Chemical fungicides and biological control agents were screened for management of this disease, and their efficacies were further determined. Most of the tested fungicides (11 out of 14) showed high activity toward ZJ-1 with EC50 < 5 μg/mL. Moreover, fungicides, including tebuconazole, boscalid, iprodione, carbendazim, and fludioxonil, displayed more than 80% disease control efficacy under the recorded conditions. Three biocontrol strains of Bacillus sp. and one of Pantoea agglomerans were isolated from pea-related niches and significantly reduced the severity of disease under greenhouse and field conditions. To our knowledge, this is the first study on ascochyta blight in field peas, and results presented here will be useful for controlling the disease in this area.

Microbial symbionts affect Pisum sativum proteome and metabolome under Didymella pinodes infection

The long cultivation of field pea led to an enormous diversity which, however, seems to hold just little resistance against the ascochyta blight disease complex. The potential of below ground microbial symbiosis to prime the immune system of Pisum for an upcoming pathogen attack has hitherto received little attention. This study investigates the effect of beneficial microbes on the leaf proteome and metabolome as well as phenotype characteristics of plants in various symbiont interactions (mycorrhiza, rhizobia, co-inoculation, non-symbiotic) after infestation by Didymella pinodes. In healthy plants, mycorrhiza and rhizobia induced changes in RNA metabolism and protein synthesis. Furthermore, metal handling and ROS dampening was affected in all mycorrhiza treatments. The co-inoculation caused the synthesis of stress related proteins with concomitant adjustment of proteins involved in lipid biosynthesis. The plant's disease infection response included hormonal adjustment, ROS scavenging as well as synthesis of proteins related to secondary metabolism. The regulation of the TCA, amino acid and secondary metabolism including the pisatin pathway, was most pronounced in rhizobia associated plants which had the lowest infection rate and the slowest disease progression. Biological significanceA most comprehensive study of the Pisum sativum proteome and metabolome infection response to Didymella pinodes is provided. Several distinct patterns of microbial symbioses on the plant metabolism are presented for the first time. Upon D. pinodes infection, rhizobial symbiosis revealed induced systemic resistance e.g. by an enhanced level of proteins involved in pisatin biosynthesis.

Ascochyta blight disease of pea (Pisum sativum L.): defence-related candidate genes associated with QTL regions and identification of epistatic QTL.

Advances have been made in our understanding of Ascochyta blight resistance genetics through mapping candidate genes associated with QTL regions and demonstrating the importance of epistatic interactions in determining resistance. Ascochyta blight disease of pea (Pisum sativum L.) is economically significant with worldwide distribution. The causal pathogens are Didymella pinodes, Phoma medicaginis var pinodella and, in South Australia, P. koolunga. This study aimed to identify candidate genes that map to quantitative trait loci (QTL) for Ascochyta blight field disease resistance and to explore the role of epistatic interactions. Candidate genes associated with QTL were identified beginning with 101 defence-related genes from the published literature. Synteny between pea and Medicago truncatula was used to narrow down the candidates for mapping. Fourteen pea candidate sequences were mapped in two QTL mapping populations, A26 × Rovar and A88 × Rovar. QTL peaks, or the intervals containing QTL peaks, for the Asc2.1, Asc4.2, Asc4.3 and Asc7.1 QTL were defined by four of these candidate genes, while another three candidate genes occurred within 1.0 LOD confidence intervals. Epistasis involving QTL × background marker and background marker × background marker interactions contributed to the disease response phenotypes observed in the two mapping populations. For each population, five pairwise interactions exceeded the 5% false discovery rate threshold. Two candidate genes were involved in significant pairwise interactions. Markers in three genomic regions were involved in two or more epistatic interactions. Therefore, this study has identified pea defence-related sequences that are candidates for resistance determination, and that may be useful for marker-assisted selection. The demonstration of epistasis informs breeders that the architecture of this complex quantitative resistance includes epistatic interactions with non-additive effects.

The complete mitogenome of Mycosphaerella pinodes (Ascomycota, Mycosphaerellaceae)

In this study, the complete mitochondrial genome of plant pathogenic fungus, Mycosphaerella pinodes, was sequenced. The nucleotide composition of the genome is: 36.0% of A, 15.0% of C, 14.6% of G and 34.5% of T. The mitochondrial genome is 55 973 bp in length and consists of 11 protein-coding genes, two ribosomal RNAs and 25 tRNA genes. The mitogenome analysis of M. pinodes provide a molecular basis for further studies on molecular systematics and evolutionary dynamics of Ascomycetes fungi especially belonging to Dothideomycetes.

Sensitivity of Mycosphaerella pinodes to Pyraclostrobin Fungicide.

Mycosphaerella blight, caused by Mycosphaerella pinodes, is a destructive disease of field pea that is managed using foliar fungicides. Strobilurin fungicides have been used in western Canada for disease management since 2003. To assess the baseline sensitivities of M. pinodes isolates to the strobilurin fungicide pyraclostrobin, the effective concentration to reduce mycelial growth by 50% (EC50) was determined for 70 isolates collected prior to 2003 from Alberta, Saskatchewan, North Dakota, and Washington State. Each of these isolates was sensitive to pyraclostrobin, with EC50 values ranging from 0.03 to 0.29 mg liter-1. The pyraclostrobin concentrations required to reduce conidia germination by 50% was lower, ranging from 0.008 to 0.041 mg liter-1. In all, 324 isolates collected in 2010 and 2011 were tested for high levels of insensitivity by examining mycelial growth using a discriminatory dose of 5 mg liter-1. Nineteen isolates were highly insensitive to pyraclostrobin, with EC50 values of 80 to 216 mg liter-1. Conidia of these isolates germinated when exposed to a discriminatory dose of 0.1 mg liter-1. Insensitive isolates infected field pea plants treated with pyraclostrobin but sensitive isolates did not. The identification of insensitive isolates indicates that insensitivity may be emerging in the pathogen population.

Pea black spot disease complex on field pea: dissecting the roles of the different pathogens in causing epicotyl and root disease

This study aimed to determine the relative importance of three well recognized devastating pathogens of field pea (Pisum sativum) on leaves and stems, Didymella pinodes, Phoma pinodella and P. koolunga, in also causing disease on epicotyls and roots. Two pea varieties, Kaspa and WAPEA2211, were inoculated with four isolates of each pathogen; subsequently epicotyl and root disease were scored accordingly. All pathogens caused epicotyl and root disease. D. pinodes isolates caused consistently severe disease on epicotyl, lateral root and tap root. Except for variable levels of disease on epicotyls, P. pinodella isolates also caused consistently high levels of disease on lateral and tap roots. While P. koolunga, generally caused less epicotyl and root disease than D. pinodes and P. pinodella, some isolates of P. koolunga caused severe tap root rot. For example, on WAPEA2211, P. koolunga (isolates#1 and #3) caused similar levels of tap root rot as with the two most virulent isolates of D. pinodes and as with the treatment with the three pathogens combined. The three pathogens combined treatment caused particularly severe disease on epicotyls, lateral and tap roots. For the first time, P. koolunga was shown to cause epicotyl and root disease on field pea, but less severe than the other two pathogens tested. Findings of this study highlight, for the first time, the relative roles of these three black spot pathogens, particularly P. koolunga; demonstrate a much wider and more significant adverse impact of the black spot complex than assumed historically; and underscore the additional challenges posed for pea breeders in developing black spot all-inclusive resistant varieties.

Sensitivity of Ascochyta Species Infecting Pea, Lentil, and Chickpea to Boscalid, Fluxapyroxad, and Prothioconazole.

Management of Ascochyta blight in pea, lentil, and chickpea relies on repeated fungicide applications, which has led to development of fungicide resistance and disease control failures in some systems. In vitro assays were conducted to determine baseline fungicide sensitivity in Mycosphaerella pinodes (Ascochyta pinodes), A. lentis, and A. rabiei populations to the demethylation-inhibiting fungicide prothioconazole and the succinate dehydrogenase-inhibiting fungicides boscalid and fluxapyroxad by determining the effective concentration at which 50% of germination or fungal growth was inhibited (EC50). Mean boscalid EC50 values from conidial germination assays were 0.669, 0.639, and 0.171 μg/ml and from mycelial growth assays were 0.258, 0.791, and 0.443 μg/ml for M. pinodes, A. lentis, and A. rabiei, respectively. Mean fluxapyroxad EC50 values were 0.050, 0.763, and 0.057 μg/ml for M. pinodes, A. lentis, and A. rabiei, respectively. Mean baseline EC50 values for prothioconazole with mycelial growth were 0.541, 0.604, and 0.283 μg/ml for M. pinodes, A. lentis, and A. rabiei, respectively. A single discriminatory fungicide concentration of 1 μg/ml was selected for all species. Established sensitivity profiles and discriminatory concentrations will be used to monitor sensitivity shifts in populations of Ascochyta spp. and to make effective disease management recommendations.

Using the Animal Model to Accelerate Response to Selection in a Self-Pollinating Crop.

We used the animal model in S0 (F1) recurrent selection in a self-pollinating crop including, for the first time, phenotypic and relationship records from self progeny, in addition to cross progeny, in the pedigree. We tested the model in Pisum sativum, the autogamous annual species used by Mendel to demonstrate the particulate nature of inheritance. Resistance to ascochyta blight (Didymella pinodes complex) in segregating S0 cross progeny was assessed by best linear unbiased prediction over two cycles of selection. Genotypic concurrence across cycles was provided by pure-line ancestors. From cycle 1, 102/959 S0 plants were selected, and their S1 self progeny were intercrossed and selfed to produce 430 S0 and 575 S2 individuals that were evaluated in cycle 2. The analysis was improved by including all genetic relationships (with crossing and selfing in the pedigree), additive and nonadditive genetic covariances between cycles, fixed effects (cycles and spatial linear trends), and other random effects. Narrow-sense heritability for ascochyta blight resistance was 0.305 and 0.352 in cycles 1 and 2, respectively, calculated from variance components in the full model. The fitted correlation of predicted breeding values across cycles was 0.82. Average accuracy of predicted breeding values was 0.851 for S2 progeny of S1 parent plants and 0.805 for S0 progeny tested in cycle 2, and 0.878 for S1 parent plants for which no records were available. The forecasted response to selection was 11.2% in the next cycle with 20% S0 selection proportion. This is the first application of the animal model to cyclic selection in heterozygous populations of selfing plants. The method can be used in genomic selection, and for traits measured on S0-derived bulks such as grain yield.