Pathogenicity of fungi associated with wheat and barley seedling emergence and fungicide efficacy of seed treatment

Presented study focused on the influence of Cochliobolus sativus isolates origin on pathogenicity towards wheat and barley seedlings in comparison with pathogenicity of certain Fusarium species and Microdochium nivale. The efficacy of fungicide seed treatment against C. sativus was estimated. The C. sativus isolates were collected from different locations and were isolated from wheat, barley and sunflower seeds. The pathogenicity of C. sativus, Fusarium species and M. nivale towards germinating seedlings were expressed as germination (GA) retardation and coleoptile growth rate retardation (CGR). Of wheat only, the CGR was significantly influenced by the isolate origin. The C. sativus isolates obtained from sunflower seeds were the most aggressive. Of the barley seeds, the barley isolates were the most aggressive. Barley was significantly more susceptible to damage by C. sativus isolates than wheat. The pathogenicity of tested fungal species declined in the order: F. culmorum, F. graminearum, C. sativus, F. avenaceum, M. nivale, F. poae for both barley and wheat. The results highlighted high pathogenicity potential of C. sativus equal to that of F. avenaceum and M. nivale. The symptoms of C. sativus on coleoptile and roots were very similar or the same as the symptoms caused by Fusarium species and M. nivale, except of white, pink or red colours. Of wheat sprouts, the fungicide efficacy (FE) against C. sativus declined in the order: tebuconazole + thiram, carboxin + thiram, quazatine, difenoconazole, iprodione + triticonazole (in term of GA) and carboxin + thiram, iprodione + triticonazole, tebuconazole + thiram, difenoconazole, quazatine (in term of CGR). In barley, the FE declined in the order: carboxin + thiram, iprodione + triticonazole, tebuconazole + thiram, difenoconazole, quazatine (in term of GA) and carboxin + thiram, tebuconazole + thiram, difenoconazole, iprodione + triticonazole, quazatine (in term of CGR).     

Morphological and Pathogenic Characterization of Genetically Diverse Sclerotinia Isolates from European Red Clover Crops (Trifolium Pratense L.)

Clover rot, an important disease in European red clover crops, is caused by Sclerotinia trifoliorum or Sclerotinia sclerotiorum. Until today, little is known about the variation in aggressiveness among Sclerotinia isolates from red clover. Aggressiveness has never been correlated with morphological characteristics. Rapidly growing isolates may be more aggressive, but this was never investigated in S. trifoliorum before. Also nothing is known about the link between sclerotia production and aggressiveness. Oxalic acid is an important pathogenicity factor in Sclerotinia species, but its effect on aggressiveness is unknown in S. trifoliorum isolates. For this study, we selected 30 Sclerotinia isolates from 25 locations Europe: 26 S. trifoliorum isolates and 4 S. sclerotiorum isolates from two locations in France (Fr.A and Fr.B). For each isolate, the in vitro growth speed, sclerotia production, oxalate production and aggressiveness were analysed and correlations were estimated between aggressiveness and the other characteristics. Aggressiveness was assessed in vitro on detached leaves and in a greenhouse on young plants. Our isolates differed significantly in growth speed, sclerotia production, oxalate production and aggressiveness. The infections on detached leaves and young plants revealed interaction between isolates and plant genotypes and between isolates and cultivars, but there was no indication that pathotypes exist. In vitro growth speed and in vitro aggressiveness on detached leaves were positively correlated with aggressiveness on young plants, while sclerotia production was negatively correlated with aggressiveness on young plants. These factors can be used as predictors of aggressiveness of Sclerotinia isolates from red clover crops.     

Genetic Structure of Cochliobolus sativus Populations Sampled from Roots and Leaves of Barley and Wheat in North Dakota

Common root rot (CRR) and spot blotch, caused by Cochliobolus sativus (Ito and Kurib.) Drechsl. ex Dast., are important diseases of barley (Hordeum vulgare L.) and wheat (Triticum aestivum L.) worldwide. However, the population biology of C. sativus is still poorly understood. In this study, the genetic structure of three C. sativus populations, consisting of isolates sampled respectively from barley leaves (BL), barley roots (BR) and wheat roots (WR) in North Dakota, was analysed with amplified fragment length polymorphism (AFLP) markers. A total of 127 AFLP loci were generated among 208 C. sativus isolates analysed with three primer combinations. Gene diversity (H = 0.277–0.335) were high in all three populations. Genetic variation among C. sativus individuals within population accounted for 74%, whereas 26% of the genetic variation was explained among populations. Genetic differentiation was high (ØPT = 0.261, corrected = 0.39), whereas gene flow (Nm) ranged from 1.27 to 1.56 among the three populations analysed. The multilocus linkage disequilibrium (LD) (= 0.076–0.117) was moderate in C  sativus populations. Cluster analyses indicate that C. sativus populations differentiated according to the hosts (barley and wheat) and tissues (root and leaf) although generalists also exist in North Dakota. Crop breeding may benefit from combining genes for resistance against both specialists and generalists of C. sativus.     

Effect of additional carbon source and moisture level on xylanase production by <Emphasis Type="Italic">Cochliobolus sativus</Emphasis> in solid fermentation

The fungus Cochliobolus sativus has been shown to be an efficient producer of xylanase from an industrial point of view. The addition of extra carbon sources and the initial moisture content of the solid-state fermentation were found to have a marked influence on the xylanase production by C. sativus Cs6 strain. Xylan and starch resulted in an increased xylanase production (1469.4 and 1396.56 U/g, respectively) after 8 days of incubation. Optimal initial moisture content for xylanase production was 80%. The cultivation systems can easily be modified to enhance the productivity of the enzyme formation by C. sativus Cs6, which will facilitate the scale up processes for mass production.     

In vitro assessment of Fusarium head blight spp. on wheat cultivars

Aggressiveness in four isolates of Fusarium head blight (FHB) species (F. culmorum, F. solani, F. verticillioides and F. equiesti) was studied in vitro on six wheat cultivars using a modified Petri-dish test. Results showed differences between cultivars inoculated with FHB isolates and control for three aggressiveness criteria: germination rate reduction, standardised area under disease progress curve (AUDPC_standard), and coleoptile length reduction. Regarding AUDPC_standard and Petri-dish aggressiveness index, significant differences were detected among fungal isolates. The other two aggressiveness criteria: germination rate reduction and coleoptile length reduction did not distinguish between FHB isolates. The Petri-dish test was repeatable and stable method to assess aggressiveness of four FHB species for all tested wheat cultivars. The current study confirmed the suitability of in vitro modified Petri-dish method to be used as fast and reliable test to analyse aggressiveness in FHB species.     

Untersuchungen zur Bildung extrazellulärer Hemicellulasen durch Pseudocercosporella herpotrichoides in vitro

Investigations on the production of extracellular hemicellulases by Pseudocercosporella herpotrichoides in vitro For all 15 isolates of Pseudocercosporella herpotrichoides investigated, xylanase as well as arabanase activity could be demonstrated. After cultivation of 3 weeks, the activity of the enzymes reached a peak. The activity of xylanase was considerably increased by addition of xylan in comparison to Maltzin as the sole source of carbohydrate. Also the arabanase activity could be increased significantly by addition of araban or xylan as compared to the Maltzin variant. The optimum temperature with regard to activity and stability of xylanase ranged at 50°C. The pH-optimum for xylanase activity was found to be at pH 5.0, and the enzyme was stable in ° range between pH4.0 and 8.0 (9.0). In case of arabanase, the temperature optimum varied between 40 and 50°C; up to this temperature, the enzyme was also stable. At pH 5.0, the arabanase activity reached its optimum; stability was observed in - pH range between 4.0 and 9.0. In extracts prepared from autoclaved wheat coleoptiles which were inoculated with Pseudocercosporella herpotrichoides, the presence of the enzymes xylanase, arabanase, cellulase and polymethylgalacturonase could be demonstrated. The enzyme activities of the inoculated samples were considerably higher than those of non-inoculated controls. The differences, in most cases, were statistically significant. Der Deutschen Forchungsgemeinschaft danken wir für finanzielle Unterstützung.     

Molecular karyotyping and chromosome length polymorphism in Cochliobolus sativus

Fungi are known to have variable genomes that can generate new virulence types capable of attacking important crop plants. To assess chromosome length polymorphisms in the barley spot blotch pathogen (Cochliobolus sativus), we analyzed the karyotypes of 16 isolates using contour-clamped homogeneous electric field (CHEF) electrophoresis. The collection of isolates studied were from diverse regions of the world (USA, Canada, Japan, Brazil, Uruguay, and Poland) and included representatives comprising the three known C. sativus pathotypes of 0, 1, and 2. Under two different running conditions, the number of CHEF bands observed ranged from 8 to 13 with a size range of 0.85 to 3.80 mega-bases (Mb). Each of the 16 isolates showed a unique banding pattern, except for two North Dakota isolates ND90Pr and ND91-Bowman, which were very similar. Single-copy DNA probes, previously assigned to each of the 15 chromosomes identified in reference isolate ND93-1, were hybridized to Southern blots of CHEF-separated chromosomes and revealed highly polymorphic chromosomes among isolates. Chromosomal rearrangements (translocations, deletions, duplications) were found in several isolates. DNA markers previously found linked to VHv1, a gene in pathotype 2 isolates conferring virulence on barley cultivar Bowman, also were used as probes in hybridizations with the CHEF blots. The results showed that the chromosome carrying the virulence gene in pathotype 2 isolates is larger than its counterpart without the gene in other isolates. This suggests that the genomic region carrying the virulence locus VHv1 is unique to pathotype 2 isolates. This study provides useful information on genome structure and divergence, which is essential for advancing our understanding of the genetics and biology of C. sativus.     

Variation in Aggressiveness Components in the Hemileia vastatrix Population in Brazil

The Pucciniomycete fungus Hemileia vastatrix causes leaf rust on coffee trees. The pathogen is responsible for considerable yield losses in susceptible coffee cultivars if appropriate management strategies are not implemented. Rapid spread and epidemics of rust fungi are usually associated with the emergence of new races of the pathogen that overcome resistance or with the emergence of more aggressive populations of the pathogen. In Brazil, coffee production is dominated by susceptible cultivars of Coffea arabica and Coffea canephora. We assessed aggressiveness in 46 populations of H. vastatrix from Minas Gerais and Espírito Santo, two of the most important coffee‐producing states in Brazil. We observed a significant difference in the incubation period between the populations from Minas Gerais and Espírito Santo when 183 single‐pustule isolates were inoculated onto Catuaí Vermelho IAC 44, a susceptible C. arabica cultivar. Variation in aggressiveness components was observed between and within localities. Isolates with longer incubation periods also tended to have longer latent periods, although there was only a low correlation between these two aggressiveness components (r² = 0.34, P = 2.2 × 10^?16). Low‐sporulating isolates also had significantly longer incubation and latent periods. The H. vastatrix population from Minas Gerais and Espírito Santo is structured by the formation of groups of individuals with differential level of aggressiveness. Our results indicate that the variation in aggressiveness of the Brazilian H. vastatrix population may be associated with the geographic coffee‐producing areas.     

Transgenic barley expressing a fungal xylanase gene in the endosperm of the developing grains

The feasibility of producing plant cell wall polysaccharide-hydrolysing feed enzymes in the endosperm of barley grain was investigated. The coding region of a modified xylanase gene (xynA) from the rumen fungus, Neocallimastix patriciarum, linked with an endosperm-specific promoter from cereal storage protein genes was introduced into barley by Agrobacterium-mediated transformation. Twenty-four independently transformed barley lines with the xylanase gene were produced and analysed. The fungal xylanase was produced in the developing endosperm under the control of either the rice glutelin B-1 (GluB-1) or barley B1 hordein (Hor2-4) promoter. The rice GluB-1 promoter provided an apparently higher expression level of recombinant proteins in barley grain than the barley Hor2-4 promoter in both transient and stable expression experiments. In particular, the mean value for the fungal xylanase activity driven by the GluB-1 promoter in the mature grains of transgenic barley was more than twice that with the Hor2-4 promoter. Expression of the xylanase transgene under these endosperm-specific promoters was not observed in the leaf, stem and root tissues. Accumulation of the fungal xylanase in the developing grains of transgenic barley followed the pattern of storage protein deposition. The xylanase was stably maintained in the grain during grain maturation and desiccation and post-harvest storage. These results indicate that the cereal grain expression system may provide an economic means for large scale production of feed enzymes in the future.     

Fusarium culmorum Infection of Barley Seedlings: Correlation between Aggressiveness and Deoxynivalenol Content

Fusarium culmorum is a serious plant pathogen, especially on cereals. The production of deoxynivalenol (DON) by F. culmorum is believed to play a role in pathogenesis. This relationship has been almost exclusively studied in connection with head blight. The present paper reports the first finding of DON in cereal seedlings infected with F. culmorum . A pathogenicity test was performed, including 70 isolates of this pathogen from different sites within northern and central Europe. All isolates caused disease on barley seedlings. For 15 isolates with varying aggressiveness, the DON content in the 19-day-old-barley seedlings was determined. There was a significant correlation between DON concentration and disease index. The aggressiveness of two outlying isolates with very low DON production is discussed. The results indicate that for F. culmorum isolates of the DON chemotype, production of this toxin influences the aggressiveness of the isolates towards barley seedlings.     

Intra- and inter-species variability of the aggressiveness in four Fusarium head blight species on durum wheat plants detected in an in vitro Petri-dish assay

Abstract

Aggressiveness assessment of Fusarium head blight (FHB) is crucial for explaining the interaction in FHB–wheat association. Currently, in vitro tools have proved to be very useful in identifying disease responses in wheat to FHB infection. To update our knowledge, intra- and inter-species variability of the aggressiveness was studied in 16 isolates of four FHB species (F. culmorum, F. solani, F. verticillioides and F. equiseti) using an in vitro Petri-dish test. Three aggressiveness criteria, germination rate reduction, standardised area under disease progress curve (AUDPC_standard) and coleoptile length reduction, were evaluated on a durum wheat cultivar showing a moderate level of quantitative resistance. Regarding AUDPC_standard, intra- and inter-species variability was detected. The other two aggressiveness criteria did not distinguish isolates within and among species. The three aggressiveness criteria were not significantly correlated. However, it was not possible to cluster the isolates based on their species origins because of similarity in pathogenic level among the 16 fungal isolates. The results provide evidence that there is a differential interaction between quantitative resistance genes in wheat plants and the 16 tested FHB isolates. The values of AUDPC_standard showed significant correlation with both: disease incidence and disease severity obtained under controlled conditions (r?=?0.627** and r?=?0.539*, respectively), and disease incidence generated by field conditions (r?=?0.525*). AUDPC_standard could be of potential use in evaluating the aggressiveness of FHB in adult wheat plants under controlled and field conditions.     

Identification of 50 K Illumina-chip SNPs associated with resistance to spot blotch in barley

Background

Spot blotch, caused by Cochliobolus sativus, is one of the most widespread and harmful diseases in barley. Identification of genetic loci associated with resistance to C. sativus is of importance for future marker-assisted selection. The goal of the current study was to identify loci conferring seedling resistance to two different pathotypes of C. sativus in the Siberian spring barley core collection.

Results

A total of 96 spring barley cultivars and lines were phenotyped at the seedling stage with two C. sativus isolates (Kr2 and Ch3). According to the Fetch-Steffenson rating scale 16%/17% of genotypes were resistant and 26%/30% were moderate-resistant to the Kr2/Ch3 isolates respectively. A total of 94 genotypes were analyzed with the barley 50 K Illumina Infinium iSELECT assay. From 44,040 SNPs, 40,703 were scorable, from which 39,140 were polymorphic. 27,319 SNPs passed filtering threshold and were used for association mapping. Data analysis by GLM revealed 48 and 41 SNPs for Kr2 and Ch3 isolates, respectively. After application of 5% Bonferroni multiple test correction, only 3 and 27 SNPs were identified, respectively. A total of three genomic regions were associated with the resistance. The region on chromosome 3H associated with Ch3-resistance was expanded between markers SCRI_RS_97417 and JHI-Hv50k-2016-158003 and included 11 SNPs, from which JHI-Hv50k-2016-157070, JHI-Hv50k-2016-156842 had the lowest p-values. These two SNPs were also significant in case of Kr2 isolate. The region on chromosome 2H included 16 loci (7 of them with the lowest p-values were tightly linked to BOPA2_12_11504). Three loci corresponding to this region had suggestive p-values in case of Kr2 tests, so the locus on chromosome 2H may also contribute to resistance to Kr2 isolate. The third region with significant p-value in case of Kr2 tests was identified on chromosome 1H at the locus JHI-Hv50k-2016-33568.

Conclusions

Three genomic regions associated with the resistance to one or both isolates of C. sativus were identified via screening of the Siberian spring barley core collection. Comparison of their location with QTLs revealed previously either with biparental mapping populations studies or with GWAS of distinct germplasm and other isolates, demonstrated that resistance to isolates Kr2 and Ch3 is conferred by known spot blotch resistance loci. Information on SNPs related can be used further for development of DNA-markers convenient for diagnostics of resistance-associated alleles in barley breeding programs.

     

Production of Wood Decay Enzymes,Loss of Mass,and Lignin Solubilization in Wood by Diverse Tropical Freshwater Fungi

In vitro production of cellulase and xylanase was common among diverse freshwater ascomycetes and their hyphomycetous anamorphs. Production of enzymes involved in lignin degradation was rare. Most isolates were capable of causing mass loss in angiosperm wood, although values were low, at ~10% during a 24-week period. A few isolates caused higher mass loss of up to 26.5%, and five of these were shown to solubilize significant amounts of lignin. This is the first report of lignin solubilization by freshwater fungi. Torula herbarum (hyphomycete) and Ophioceras dolichostomum (ascomycete) produced indices of lignin solubilization equivalent to those of terrestrial white-rot basidiomycetes. In all cases wood decay was 2.2- to 3-fold higher in exposed rather than submerged conditions.     

Cultural and Aggressiveness Variability of Cercospora coffeicola

Although brown eye spot of coffee, caused by Cerco‐spora coffeicola, is important for coffee production in Brazil, there is a general lack of knowledge regarding the disease. In this study, we evaluated the variability of both the cultural and aggressiveness traits of 60 isolates from coffee plants grown under conventional and organic systems in three regions of Minas Gerais State, Brazil. Variability among the isolates was detected with regard to all of the traits and was unrelated to an effect of either the region or cropping system. Mycelial growth, cercosporin production and sporulation were assessed in the laboratory. Of the 60 isolates, 27 did not sporulate at 25°C; the mycelial growth of all of the isolates and cercosporin production by 18 of the isolates linearly increased as the temperature rose from 18 to 26°C. We inoculated six selected isolates on plants of two coffee cultivars (‘Catuaí Vermelho IAC44’ and ‘Catucaí Vermelho 785‐15’) and evaluated the incubation period (IP), latent period (LP) and disease severity. All three of these traits were affected by temperature postinoculation and KCl amendment. The significant correlations were as follows: IP and LP in both cultivars; severity and leaf fall in both cultivars; and cercosporin production in vitro and severity values in ‘Catucaí Vermelho 785‐15’. In conclusion, we found that (i) C. coffeicola is highly variable for both cultural and aggressiveness traits; (ii) laboratory and glasshouse experiments were suitable to assess the pathogen variability; (iii) research protocols should account for the effect of environmental factors, such as temperature and KCl, on the traits evaluated; and (iv) these protocols should include the assessment of the IP instead of the LP, as both are correlated, and the IP is easier to evaluate.     

Role of Melanin in Release of Extracellular Enzymes and Selection of Aggressive Isolates of Bipolaris sorokiniana in Barley

Eighteen barley isolates of Bipolaris sorokiniana belonging to wild and clonal type of black, mixed and white subpopulations were quantitatively assayed for their melanin content and aggressiveness with respect to production of some of the extracellular enzymes such as cellulase, pectinase, amylase and protease. Cellulase and pectinase constituted major portion of the enzymes recovered from the black, mixed and white isolates. Enzyme production and aggressiveness were relatively higher in melanin devoid or low melanin isolates. The melanin deficient isolates were also differentiated from black and mixed isolates on the basis of variation in internal transcribed spacer region of the ribosomal DNA. Higher enzyme productions positively correlated with area under disease progress curve (AUDPC) and lesion development. Melanin content was negatively correlated with extracellular enzymes and aggressiveness of the isolates. Based on melanin content, lesion size, AUDPC and extracellular enzymes, the isolates were grouped in two major clusters (I and II) with further division of cluster II into two sub-clusters (II-A and II-B). The results appears to indicate a possible role of melanin in release of extracellular enzymes and hence in evolution and selection of aggressive isolates of B. sorokiniana in barley.     

Effect of the incubation conditions on the production of patulin by Penicillium griseofulvum isolated from wheat

A total of 290 Candida isolates from patients were investigated for in vitro proteinase production. Overall, sixty percent of these strains were found to be proteinase producers. Of the C. albicans strains, 81.4% of the significant isolates in contrast to 19.7% of nonsignificant isolates were proteinase producers, the difference being statistically significant (P<0.001). Amongst the different Candida species, the proteinase production was found not only in Candida albicans, but also in C. tropicalis, C. parapsilosis and C. glabrata. Thus this in vitro method of demonstration of proteinase may be a good adjunct to smear and culture examination in identifying pathogenic Candida species from anatomical sites where they can also be present as commensals.     

Aggressiveness and diversity of Phytophthora capsici on vegetable crops in Georgia

Phytophthora blight induced by Phytophthora capsici causes significant yield loss in a number of vegetable crops. It is imperative to understand the diversity and aggressiveness of the pathogen to design more efficient disease management programs. A collection of P. capsici strains isolated from different vegetable crops in Georgia, USA, were characterised in this study. Of the 49 isolates tested, 24 were A1 and 25 were A2 mating type, respectively, with both mating types found in the same fields. Variability of the isolates was assessed in terms of their aggressiveness on six pepper genotypes. The isolates differed in their aggressiveness on different pepper cultivars with 10 pathotypes identified. No correlation between aggressiveness of the isolates and their host origin or geographical location of isolation was observed. Randomly amplified polymorphic DNA (RAPD) analysis was used to evaluate genetic variability among P. capsici populations. RAPD analysis using 15 random primers resulted in 133 reproducible bands and cluster analysis separated the isolates into 5 groups. Analysis of molecular variance showed that there was moderate genetic differentiation associated with host origin and geographical location of the isolates. No correlation was found between RAPD groups and pathotypes or mating types. These results indicate that P. capsici populations infecting vegetable crops in Georgia were genetically diverse, which should be taken into account in developing resistant cultivars or other disease management programmes.     

Effect of Carbon Source on Production of Lytic Enzymes by the Sclerotial Parasites Trichoderma atroviride and Coniothyrium minitans

Three isolates of Trichoderma atroviride and two isolates of Coniothyrium minitans known to efficiently degrade sclerotia of Sclerotinia sclerotiorum were cultured on minimal medium with sucrose, carboxymethyl cellulose (CMC), xylan, laminarin, colloidal chitin or powdered sclerotia as carbon source. The activity of endochitinase, endo‐β‐1,3‐glucanase, endoxylanase and endocellulase in culture filtrates was determined after 7 and 15 days of culture using dye‐labelled substrates. The strongest inducers of chitinase were colloidal chitin and sclerotia powder. Chitinase activity appeared to be faster induced in the isolates of T. atroviride than in the isolates of C. minitans, but the maximum level of activity present in culture filtrates of the two species was similar. With CMC and xylan as carbon source, concurrent production of the corresponding enzymes was observed for the Trichoderma isolates. The isolates of C. minitans produced cellulase on xylan but not on CMC, whereas xylanase was produced on both carbon sources. Laminarin induced the formation of glucanases in the three isolates of T. atroviride but not the isolates of C. minitans. However, in the sclerotia‐containing cultures of C. minitans glucanase activity was even higher than in the respective cultures of the Trichoderma isolates. During the 31‐day study period, the pattern of enzyme production in shake cultures containing sclerotia powder was very similar for the isolates of T. atroviride and C. minitans. Glucanase activity generally reached a peak 24 days after inoculation of the flasks, whereas the activity of chitinase, cellulase and xylanase remained fairly constant throughout the experiment.     

Heterogeneity in the ITS of the ribosomal DNA of <Emphasis Type="Italic">Pyrenophora graminea</Emphasis> isolates differing in xylanase and amylase production

Xylanase and amylase have gained increasing interest because of their various biotechnology applications. In this research, the restriction of PCR-amplified internal transcribed spacers (ITS) of ribosomal DNA (rDNA) was used to confirm the genetic variation among 22 isolates of Pyrenophora graminea differing in their xylanase and amylase production. The fingerprints generated from the six restriction digestions of the rDNA ITS region showed high levels of intraspecific variation within the P. graminea population. Neighbour-Joining diagram, based on Nei’s genetic distances, showed that isolates formed two phylogenetic groups. No apparent association could be observed between xylanase and amylase production and genetic diversity among the twenty-two isolates.     

Pyrenophora teres: Population structure,virulence and aggressiveness in Southern Russia

The barley net blotch agent Pyrenophora teres (Died) Drechs. is one of the dominant fungal pathogens in agricultural crops worldwide. Here we aim to study the aggressiveness and virulence of P. teres populations collected at different ontogenesis stages (BBCH 30 and BBCH 47) from winter barley cultivars of various resistance types: moderately resistant, moderately susceptible and highly susceptible. We observed a direct proportional relationship between cultivar resistance and the aggressiveness of P. teres populations collected in both growth phases of the host plant. The isolates collected at an early stage of host plant development have a large difference in aggressiveness criteria: colony growth rate, sporulation intensity, latency period, plant damage degree, and the number of identified races. At the BBCH 30 growth stage, the growth rate of fungus colonies selected from a resistant cultivar is 1.2 times higher than that of a susceptible cultivar. The growth rate of colonies selected from resistant and susceptible cultivars in the earlier BBCH 30 stage is 1.04 times higher than the growth rate of colonies selected from the later phase. The sporulation intensity of fungal populations selected from a resistant cultivar is higher than that of populations selected from a susceptible cultivar (for BBCH 30–5.4 times, for BBCH 47–4.0 times); and it is 1.3 times higher in an earlier phase of plant development. Correlation between colony growth rate and spore formation rate in the BBCH 30 is r = 0.4. A high correlation level (r = 0.9) and notable difference between the variants were revealed when studying the duration of the latent period. The average value of plant damage by the P. teres from resistant cultivar is 4 times higher than from the susceptible cultivar in the BBCH 30 stage; and 12 times – in the BBCH 47 stage. There is a moderate negative correlation between the plant damage degree and the number of races identified from the fungal population, r = ?0.59 for the BBCH 30, r = ?0.8 for the BBCH 47. The number of races identified from P. teres populations collected in the late phase of plant growth was one third less. Our study helped to acquire new knowledge about intrapopulation processes under the influence of various factors – plant growth stage and cultivar genotype. The results obtained are the basis for the development of adaptive-integrated techniques for managing populations of the hemibiotrophic pathogen, barley net blotch.