Influence of Nitrogen Supply on the Susceptibility of Wheat to Septoria tritici

Nitrogen (N) fertilization is required for achieving high yields in wheat (Triticum aestivum L.) but may enhance the development of Septoria tritici Rob. ex Desm. A study was undertaken to investigate how N supply influences the area under disease progress curve (AUDPC). Two field experiments were carried out in a typical argiudol in 1996 and 1997. Six cultivars were grown at two N treatments (0 and 100–150 kg N/ha) in a split plot design. Percentage of necrosis (severity) of the two upper leaves of each treatment was evaluated three times during the growing season. Height and heading date were also scored and related to the resistance level. In 1996, with weather conditions conducive to the disease, AUDPC values were higher in the fertilized treatment. In 1997, with insufficient rain immediately after inoculation, the disease only progressed faster under N fertilization in the flag leaf, which was exposed to conducive environmental conditions from its appearance. The effect of N fertilization was influenced by the cultivar characteristics, climatic and agronomic conditions. Knowledge that N fertilization promotes the development of S. tritici in conducive conditions will be useful for deciding management strategies of the cultivars and for optimizing conditions for the selection in breeding programmes.     

Changes in Wheat Leaf Extracellular Proteolytic Activity after Infection with Septoria tritici

The proteolytic activity of the leaf extracellular space of wheat cultivars Pigüé and Isla Verde was estimated after inoculation of either detached leaves or plants with the fungus Septoria tritici. Pigüé is resistant, whereas Isla Verde is susceptible to the disease caused by S. tritici. Viable conidiospores of the fungus caused similar increases in both hydrogen peroxide production and chitinase activity of the cultivars studied. In contrast, they caused a decrease in the extracellular serine proteinase activity of Isla Verde and a significant increase in that of Pigüé. Independently of the cultivar from which it was extracted, the extracellular serine proteinase inhibited the germination of Septoria tritici conidiospores. These results suggest that the proteolytic activity of the leaf extracellular space can participate in the defence of wheat plants against Septoria tritici. Its regulation may be controlled by specific defence components of each cultivar.     

Stability of Adult‐plant Resistance to Septoria tritici blotch in 24 European Winter Wheat Varieties Across Nine Field Environments

Septoria tritici blotch (STB) is one of the most important leaf diseases in wheat worldwide. Objectives of this study were (i) to compare inoculation and natural infection; (ii) to evaluate the level of adult‐plant resistance to STB using four isolates; and (iii) to analyse environmental stability of 24 winter wheat (Triticum aestivum L.) varieties in inoculated vs. non‐inoculated field trials across 3 years including nine environments (location × year combinations). Field trials were sown in split‐plot design inoculated with four aggressive isolates of S. tritici plus one non‐inoculated variant as main factor and 24 wheat varieties as subfactor. Septoria tritici blotch severity was visually scored as percentage flag leaves covered with lesions bearing pycnidia. Overall STB rating ranged from 8% (Solitär) to 63% (Rubens) flag leaf area affected, resulting in significant (P < 0.01) genotypic variance. Variance of genotype × environment interaction amounted to approximately 50% of the genotypic variance. Genotype × isolate interaction variance was significant too (P < 0.01) but of minor importance. Therefore, environmental stability of varieties should be a major breeding goal. The varieties Solitär, History and Florett were most resistant and stable as revealed by a regression approach, and the susceptible varieties were generally unstable. Hence, STB resistance and stability are correlated (P < 0.01), but there were some exceptions (Tuareg, Ambition). Promising candidates for an environmentally stable, effective adult‐plant resistance have been identified.     

Role of hydrogen peroxide during the interaction between the hemibiotrophic fungal pathogen Septoria tritici and wheat

Hydrogen peroxide (H(2)O(2)) is reported to inhibit biotrophic but benefit necrotrophic pathogens. Infection by necrotrophs can result in a massive accumulation of H(2)O(2) in hosts. Little is known of how pathogens with both growth types are affected (hemibiotrophs). The hemibiotroph, Septoria tritici, infecting wheat (Triticum aestivum) is inhibited by H(2)O(2) during the biotrophic phase, but a large H(2)O(2) accumulation occurs in the host during reproduction. Here, we infiltrated catalase, H(2)O(2) or water into wheat during the biotrophic or the necrotrophic phase of S. tritici and studied the effect of infection on host physiology to get an understanding of the survival strategy of the pathogen. H(2)O(2) removal by catalase at both early and late stages made plants more susceptible, whereas H(2)O(2) made them more resistant. H(2)O(2) is harmful to S. tritici throughout its life cycle, but it can be tolerated. The late accumulation of H(2)O(2) is unlikely to result from down-regulation of photosynthesis, but probably originates from damage to the peroxisomes during the general tissue collapse, which is accompanied by release of soluble sugars in a susceptible cultivar.     

Sources of resistance and susceptibility to Septoria tritici blotch of wheat

An association genetics analysis was conducted to investigate the genetics of resistance to Septoria tritici blotch, caused by the fungus Zymoseptoria tritici (alternatively Mycosphaerella graminicola), in cultivars and breeding lines of wheat (Triticum aestivum) used in the UK between 1860 and 2000. The population was tested with Diversity Array Technology (DArT) and simple‐sequence repeat (SSR or microsatellite) markers. The lines formed a single population with no evidence for subdivision, because there were several common ancestors of large parts of the pedigree. Quantitative trait loci (QTLs) controlling Septoria resistance were postulated on 11 chromosomes, but 38% of variation was not explained by the identified QTLs. Calculation of best linear unbiased predictions (BLUPs) identified lineages of spring and winter wheat carrying different alleles for resistance and susceptibility. Abundant variation in Septoria resistance may be exploited by crossing well‐adapted cultivars in different lineages to achieve transgressive segregation and thus breed for potentially durable quantitative resistance, whereas phenotypic selection for polygenic quantitative resistance should be effective in breeding cultivars with increased resistance. The most potent allele reducing susceptibility to Septoria, on chromosome arm 6AL, was associated with reduced leaf size. Genes which increase susceptibility to Septoria may have been introduced inadvertently into UK wheat breeding programmes from cultivars used to increase yield, rust resistance and eyespot resistance between the 1950s and 1980s. This indicates the need to consider trade‐offs in plant breeding when numerous traits are important and to be cautious about the use of non‐adapted germplasm.     

Wheat Resistance to Spot Blotch Potentiated by Silicon

Spot blotch, caused by the fungus Bipolaris sorokiniana, is one of the most important diseases on wheat. The effects of silicon (Si) on this wheat disease were studied. Plants of wheat cultivars BR‐18 and BRS‐208 were grown in plastic pots containing Si‐deficient soil amended with either calcium silicate (+Si) or calcium carbonate (?Si). The content of Si in leaf tissue was significantly increased by 90.5% for the +Si treatment. There was no significant difference between Si treatments for calcium content, so variations in Si accounted for differences in the level of resistance to spot blotch. The incubation period was significantly increased by 40% for the +Si treatment. The area under spot blotch progress curve, number of lesions per cm² of leaf area, and real disease severity significantly decreased by 62, 36 and 43.5% in +Si treatment. There was no significant effect of Si on lesion size. The role played by total soluble phenolics in the increased resistance to spot blotch of plants from both cultivars supplied with Si was not clear. Plants from cultivar BR‐18 supplied with Si showed the highest values for concentration of lignin‐thioglycolic acid derivatives during the most advanced stages of fungus infection. Chitinase activity was high at the most advanced stages of fungus infection on leaves from both cultivars supplied with Si and may have had an effect on fungus growth based on the reduction of the components of resistance evaluated. Peroxidase activity was found to be high only at 96 h after inoculation of both cultivars supplied with Si. Polyphenoloxidase activity had no apparent effect on resistance regardless of Si treatments. Results revealed that supplying Si to wheat plants can increase resistance against spot blotch.     

Progress in Wheat Resistance to Spot Blotch in Bangladesh

Spot blotch, caused by Cochliobolus sativus, is considered one of the most destructive diseases of wheat (Triticum aestivum) in the warm areas of South Asia. Over the past 20 years, wheat breeding efforts in the region have improved spot blotch resistance in susceptible commercial cultivars. This study assessed resistance and spot blotch‐induced yield losses in newly released wheat cultivars developed in Bangladesh since the release of the landmark wheat variety ‘Kanchan’. Replicated field studies were conducted during the 2003 and 2004 wheat seasons at two sites: a farmer's field and a research station in a warm region of Bangladesh where spot blotch has been a serious problem. Spot blotch affected 60% of the crop and caused yield losses of from 2% to 22%. Disease severity and disease‐induced grain yield reductions were less in wheat genotypes developed since 1983, with a corresponding trend towards higher yield in newly developed varieties. The level of resistance to spot blotch in the new cultivars and advanced breeding lines represents considerable progress in breeding for resistance over the past two decades.     

Rapidly Evolving Genes Are Key Players in Host Specialization and Virulence of the Fungal Wheat Pathogen Zymoseptoria tritici (Mycosphaerella graminicola)

The speciation of pathogens can be driven by divergent host specialization. Specialization to a new host is possible via the acquisition of advantageous mutations fixed by positive selection. Comparative genome analyses of closely related species allows for the identification of such key substitutions via inference of genome-wide signatures of positive selection. We previously used a comparative genomics framework to identify genes that have evolved under positive selection during speciation of the prominent wheat pathogen Zymoseptoria tritici (synonym Mycosphaerella graminicola). In this study, we conducted functional analyses of four genes exhibiting strong signatures of positive selection in Z. tritici. We deleted the four genes in Z. tritici and confirm a virulence-related role of three of the four genes ΔZt80707, ΔZt89160 and ΔZt103264. The two mutants ΔZt80707 and ΔZt103264 show a significant reduction in virulence during infection of wheat; the ΔZt89160 mutant causes a hypervirulent phenotype in wheat. Mutant phenotypes of ΔZt80707, ΔZt89160 and ΔZt103264 can be restored by insertion of the wild-type genes. However, the insertion of the Zt80707 and Zt89160 orthologs from Z. pseudotritici and Z. ardabiliae do not restore wild-type levels of virulence, suggesting that positively selected substitutions in Z. tritici may relate to divergent host specialization. Interestingly, the gene Zt80707 encodes also a secretion signal that targets the protein for cell secretion. This secretion signal is however only transcribed in Z. tritici, suggesting that Z. tritici-specific substitutions relate to a new function of the protein in the extracellular space of the wheat-Z. tritici interaction. Together, the results presented here highlight that Zt80707, Zt103264 and Zt89160 represent key genes involved in virulence and host-specific disease development of Z. tritici. Our findings illustrate that evolutionary predictions provide a powerful tool for the identification of novel traits crucial for host adaptation and pathogen evolution.     

Chromosomal location of a gene for resistance to septoria tritici blotch (Mycosphaerella graminicola)in the hexaploid wheat ’Synthetic 6x’

Septoria tritici blotch, caused by the fungus Mycosphaerella graminicola,is currently the major foliar disease of wheat world-wide, and new sources of resistance and knowledge about the genetics of resistance are needed to improve breeding for resistance to this disease. Sears’s ’Synthetic 6x’ hexaploid wheat, derived from a hybrid of Triticum dicoccoides and Triticum tauschii, was resistant to 12 of 13 isolates of M. graminicola tested. Chromosome 7D of ’Synthetic 6x’ was identified as carrying resistance to all 12 isolates in tests of seedlings of inter-varietal chromosome substitution lines of ’Synthetic 6x’ into ’Chinese Spring’ and to two isolates in tests of adult plants. A septoria tritici blotch resistance gene, named Stb5, was identified using the M. graminicola isolate IPO94269 and mapped on the short arm of chromosome 7D, near the centromere, in a population of single homozygous chromosome-recombinant lines for the 7D chromosome. Received: 1 February 2001 / Accepted: 17 April 2001     

Study of Genetic Variability Among Monopycnidial and Monopycnidiospore Isolates Derived from Single Pycnidia of Stagonospora ssp. and Septoria tritici with the use of RAPD-PCR, MP-PCR and rep-PCR Techniques

Thirty-six isolates of Stagonospora avenae f. sp. triticea, S. nodorum and Septoria tritici recovered from asexual fruiting bodies - pycnidia and their spores were assessed for DNA polymorphism with the use of three molecular techniques: microsatellite-primed polymerase chain reaction (MP-PCR), primers correspond to dispersed repetitive elements (rep-PCR) and random amplified polymorphic DNA (RAPD-PCR). These polymerase chain reaction (PCR)-based techniques were simultaneously evaluated for their capacity to detect genetic variation at DNA level. The most polymorphic DNA profiles of monopycnidial and monopycnidiospore isolates were detected with diverse microsatellite motifs used for PCR priming. The lowest similarity values 0.86, 0.76 and 0.84 were identified among monopycnidiospore isolates derived from the same pycnidium of S. avenae f. sp. triticea, S. nodorum and S. tritici, respectively. The above, rather low similarities, found for isolates recovered from single pycnidia, supported a hypothesis that heterokaryosis resulted from high mutation rate of microsatellites and transposons activity. This would have fundamental consequences for the genetic status of asexual populations of Stagonospora spp. and S. tritici. The data produced by this study indicate that more attention should be paid to asexual reproduction as a possible source of genetic variability among populations of the pathogens.     

Histopathology of Leaf Rust Infection and Development in Wheat Genotypes containing Lr12 and Lr13

Evidence exists that certain genes for resistance to leaf rust in wheat, e.g. Lr13 and Lr34 , may interact with other genes to condition higher levels of resistance than that conferred by each gene individually. In this study, the hypothesis that Lr12 and Lr13 , both genes for adult plant resistance to Puccinia recondita Roberge ex. Desmaz f. sp. tritici Eriks. and Henn., interact to confer an improved level of resistance, was investigated using fluorescence and phase-contrast microscopy. Flag leaf segments of monogenic and digenic Thatcher lines, sampled 64 and 240 h post-inoculation, were stained with Uvitex 2B and screened, using fluorescence microscopy, for development of infection structures or host response. To study cell wall appositions, specimens were stained with trypan blue and a solution of picric acid in methyl salicylate. Aborted penetration, consisting of nonpenetrating appressoria and aborted substomatal vesicles, showed that inhibition of fungal growth in wheat lines containing Lr12 and/or Lr13 was activated, to a certain degree, before haustoria were formed. At 240 h after inoculation colony size indicated that fungal colonies in the Lr gene combination lines were generally smaller than in the parents, but not necessarily smaller than those in a line with Lr13 only. Host cell necrosis was more frequently associated with infection sites, specifically of pathotype UVPrt2, in the combination lines than in the parents. The morphology of cell wall appositions varied considerably from a narrow, luminous zone slightly wider in the centre, to a thick central part opposite the haustorium mother cell, sharply decreasing towards both ends. Histological assessments could, however, not conclusively prove pronounced resistance enhancement or unconventional resistance mechanisms due to combining the genes Lr12 and Lr13 .     

Remarkable recent changes in the genetic diversity of the avirulence gene AvrStb6 in global populations of the wheat pathogen Zymoseptoria tritici

Septoria tritici blotch (STB), caused by the fungus Zymoseptoria tritici, is one of the most economically important diseases of wheat. Recently, both factors of a gene-for-gene interaction between Z. tritici and wheat, the wheat receptor-like kinase Stb6 and the Z. tritici secreted effector protein AvrStb6, have been identified. Previous analyses revealed a high diversity of AvrStb6 haplotypes present in earlier Z. tritici isolate collections, with up to c.18% of analysed isolates possessing the avirulence isoform of AvrStb6 identical to that originally identified in the reference isolate IPO323. With Stb6 present in many commercial wheat cultivars globally, we aimed to assess potential changes in AvrStb6 genetic diversity and the incidence of haplotypes allowing evasion of Stb6-mediated resistance in more recent Z. tritici populations. Here we show, using targeted resequencing of AvrStb6, that this gene is universally present in field isolates sampled from major wheat-growing regions of the world in 2013–2017. However, in contrast to the data from previous AvrStb6 population studies, we report a complete absence of the originally described avirulence isoform of AvrStb6 amongst modern Z. tritici isolates. Moreover, a remarkably small number of haplotypes, each encoding AvrStb6 protein isoforms conditioning virulence on Stb6-containing wheat, were found to predominate among modern Z. tritici isolates. A single virulence isoform of AvrStb6 was found to be particularly abundant throughout the global population. These findings indicate that, despite the ability of Z. tritici to sexually reproduce on resistant hosts, AvrStb6 avirulence haplotypes tend to be eliminated in subsequent populations.     

Identification of New Sources of Resistance to Septoria Tritici Blotch Caused by Zymoseptoria tritici

Twenty‐nine synthetic hexaploid wheats (SHWs) were evaluated for resistance to five isolates of Zymoseptoria tritici, a devastating wheat pathogen worldwide. The five Z. tritici isolates varied in their virulence spectra towards wheat genotypes, indicating that they have distinct set of avirulence genes. New isolate‐specific resistances were identified that could be used in wheat breeding programmes. Comparing with the previous studies, the number of specific resistances identified in this study is considerable. Among 150 interactions, 78 isolate‐specific resistances were identified. Interestingly, 21 wheat genotypes showed specific responses to one or more isolates tested. Of these, 12 genotypes were highly resistant to all isolates, indicating that they possess known or novel effective resistance genes. The Stb15 and Stb16/Stb17 are effective resistance genes towards isolates used in this study, indicating that the conferred resistance in these genotypes is due to the presence of either of these genes in combination or individually. Alternatively, they may carry novel broad‐spectrum resistance gene(s) that their identification is of interest. Our data suggest that the presence of complete resistance to various Z. tritici isolates in SHWs justifies the need for more in‐depth research to characterize the likely novel genes.     

Phenotypic and genetic analysis of the Triticum monococcum-Mycosphaerella graminicola interaction

Here, the aim was to understand the cellular and genetic basis of the Triticum monococcum-Mycosphaerella graminicola interaction. Testing for 5 yr under UK field conditions revealed that all 24 T. monococcum accessions exposed to a high level of natural inocula were fully resistant to M. graminicola. When the accessions were individually inoculated in the glasshouse using an attached leaf seeding assay and nine previously characterized M. graminicola isolates, fungal sporulation was observed in only three of the 216 interactions examined. Microscopic analyses revealed that M. graminicola infection was arrested at four different stages post-stomatal entry. When the inoculated leaves were detached 30 d post inoculation and incubated at 100% humidity, abundant asexual sporulation occurred within 5 d in a further 61 interactions. An F(2) mapping population generated from a cross between T. monococcum accession MDR002 (susceptible) and MDR043 (resistant) was inoculated with the M. graminicola isolate IPO323. Both resistance and in planta fungal growth were found to be controlled by a single genetic locus designated as TmStb1 which was linked to the microsatellite locus Xbarc174 on chromosome 7A(m). Exploitation of T. monococcum may provide new sources of resistance to septoria tritici blotch disease.     

Inheritance of resistance to leaf and glume blotch caused by Septoria nodorum Berk. in winter wheat

Sixteen crosses between eight winter wheat cultivars were screened for resistance to Septoria nodorum leaf and glume blotch in the F₁ and F₄ generations using artificial inoculation in the field. The F₁ of most crosses showed dominance for susceptibility on both ear and leaf. The effects of general combining ability were of similar magnitude as the effects for specific combining ability. On the basis of the phenotypic difference of the parents, no prediction was possible about the amount and the direction of genetic variance in the segregating populations. The variation observed in this study both within and among the segregating populations suggests a quantitative inheritance pattern influencing the expression of the two traits. The components of variance between F₂ families within a population were as high as (for S. nodorum blotch on the ear) or higher (for S. nodorum blotch on the leaf) than those between populations. Therefore, strong selection within a few populations may be as effective to obtain new resistant genotypes as selection in a large number of populations. In almost all crosses, progenies were found that were more resistant than the better parent. Thus transgression breeding may be a tool to breed for higher levels of resistance to S. nodorum blotch. Highly resistant genotypes were found even in combination with two susceptible parents. The genetic source for Septoria resistance is probably broader than is generally assumed and could be used to improve S. nodorum resistance by combination breeding followed by strong selection in large populations. Received: 18 January / Accepted: 30 April 1999     

The effect of Trichoderma harzianum and T. koningii on the control of tan spot (Pyrenophora tritici-repentis) and leaf blotch (Mycosphaerella graminicola) of wheat under field conditions in Argentina

The effect of six isolates of Trichoderma harzianum and one isolate of T. koningii on the incidence and severity of tan spot (Pyrenophora tritici-repentis) and leaf blotch of wheat (Mycosphaerella graminicola) was evaluated under field conditions. Significant differences between wheat cultivars, inoculum types and growth stages were found. Three of the isolates tested (T2 for M. graminicola, T7 for P. tritici-repentis and T5 for both of them) showed the best performance in controlling leaf blotch and tan spot when coated onto seed or sprayed onto wheat leaves at different growth stages, with significant severity reduction up to 56%. At tillering, six of the isolates reduced the severity of P. tritici-repentis and M. graminicola compared to the control by up to 39% and 12-53%, respectively. In some experiments, the biocontrol preparation (T2 and T5) gave a level of disease control similar to that obtained with Tebuconazole (70 and 48%, respectively). The effect of Trichoderma against P. tritici-repentis was also observed at the heading stage, when six of the treatments reduced disease severity by 16-35%. This is the first report on the efficacy of Trichoderma spp. against wheat necrotrophic pathogens under field conditions in Argentina.     

Comparing single- vs. mixed-genotype infections of Mycosphaerella graminicola on wheat: effects on pathogen virulence and host tolerance

Mixed-genotype infections (infections of a host by more than one pathogen genotype) are common in plant-pathogen systems. However their impact on the course of the infection and especially on pathogen virulence and host response to infection is poorly understood. We investigated the effects of mixed-genotype infections on several parameters: host resistance and tolerance, as well as pathogen aggressiveness and virulence. For these purposes, we inoculated three wheat lines with three Mycosphaerella graminicola genotypes, alone or in mixtures, in a greenhouse experiment. For some of the mixtures, disease severity and virulence were lower than expected from infection by the same genotypes alone, suggesting that competition between genotypes was reducing their aggressiveness and virulence. One host line was fully resistant, but there were differences in resistance in the other lines. The two host lines that became infected differed slightly in tolerance, but mixed-genotype infections had no effect on host tolerance.     

小麦PAL基因的克隆及赤霉菌诱导下的表达分析

利用苯丙氨酸解氨酶(PAL,phenylalanine ammonia-lyase)基因保守区域从小麦抗赤霉病材料苏麦3号中克隆获得4个PAL基因,分别命名为Ta PAL1、Ta PAL2、Ta PAL3、Ta PAL4。4个基因的开放阅读框(ORF,open reading frame)长度分别为2142 bp、2016 bp、2118 bp和2139 bp,分别编码714个、672个、706个和713个氨基酸。基因序列比对发现其相似性达到88.35%,所编码的氨基酸相似性为91.92%,氨基酸序列分析表明4个基因都包含HAL-PAL结构域及PAL结构域。通过接种禾谷镰刀菌,利用荧光定量PCR对PAL基因进行表达分析发现,4个PAL基因全部为上调表达,其中Ta PAL2、Ta PAL3和Ta PAL4最为明显。PAL基因的上调表达,说明PAL基因在小麦抵抗赤霉病菌侵染的机制中可能起着重要作用。     

Role of Antioxidant Systems in Wheat Genotypes Tolerance to Water Stress

The role of plant antioxidant systems in stress tolerance was studied in leaves of three contrasting wheat genotypes. Drought imposed at two different stages after anthesis resulted in an increase in H2O2 accumulation and lipid peroxidation and decrease in ascorbic acid content. Antioxidant enzymes like superoxide dismutase, ascorbate peroxidase and catalase significantly increased under water stress. Drought tolerant genotype C 306 which had highest ascorbate peroxidase and catalase activity and ascorbic acid content also showed lowest H2O2 accumulation and lipid peroxidation (malondialdehyde content) under water stress in comparison to susceptible genotype HD 2329 which showed lowest antioxidant enzyme activity and ascorbic acid content and highest H2O2 content and lipid peroxidation. HD 2285 which is tolerant to high temperature during grain filling period showed intermediate behaviour. Superoxide dismutase activity, however, did not show significant differences among the genotypes under irrigated as well as water stress condition. It seems that H2O2 scavenging systems as represented by ascorbate peroxidase and catalase are more important in imparting tolerance against drought induced oxidative stress than superoxide dismutase alone. This revised version was published online in July 2006 with corrections to the Cover Date.