Sporulation of Septoria nodorurn (and S. tritici) on spring wheat cvs Kolibri and Maris Butler in relation to growth stage, plant part and time of season

Sporulation of Septoria nodorum was assessed on inoculated spring wheat cvs Kolibri and Maris Butler. The time from inoculation to first production of spores (latent period) was similar over a range of constant temperatures (12–18°C) but was shorter for Kolibri at 6°C and at 24°C. Spore production was greatest during the period of stem extension and, for equal amounts of disease, was twice as great on Kolibri as on Maris Butler. Spore production on leaves was much more intense with Septoria tritici than S. nodorum for equal amounts of disease, and was less intense on heads than on leaves for both pathogens. Inoculation of plants resulted in significant losses in yield. Mans Butler consistently out-performed Kolibri on each component of yield. Kolibri was particular affected: 1000-grain weights were reduced at each growth stage tested especially by S. nodorum, grain numbers and yield/head were reduced particularly following inoculation at heading. Spore production of S. nodorum during the period of stem extension increased to a peak in early July and then declined but no reliable relationship with monitored weather accounted for this seasonal trend.     

Effect of pH and oxalic acid on the reduction of Fe3+ by a biomimetic chelator and on Fe3+ desorption/adsorption onto wood: Implications for brown-rot decay

Fenton reaction is thought to play an important role in wood degradation by brown-rot fungi. In this context, the effect of oxalic acid and pH on iron reduction by a biomimetic fungal chelator and on the adsorption/desorption of iron to/from wood was investigated. The results presented in this work indicate that at pH 2.0 and 4.5 and in the presence of oxalic acid, the phenolate chelator 2,3-dihydroxybenzoic acid (2,3-DHBA) is capable of reducing ferric iron only when the iron is complexed with oxalate to form Fe³⁺-mono-oxalate (Fe(C₂O₄)⁺). Within the pH range tested in this work, this complex formation occurs when the oxalate:Fe³⁺ molar ratio is less than 20 (pH 2.0) or less than 10 (pH 4.5). When aqueous ferric iron was passed through a column packed with milled red spruce (Picea rubens) wood equilibrated at pH 2.0 and 4.5, it was observed that ferric iron binds to wood at pH 4.5 but not at pH 2.0, and the bound iron could then be released by application of oxalic acid at pH 4.5. The release of bound iron was dependent on the amount of oxalic acid applied in the column. When the amount of oxalate was at least 20-fold greater than the amount of iron bound to the wood, all bound iron was released. When Fe–oxalate complexes were applied to the milled wood column equilibrated in the pH range of 2–4.5, iron from Fe–oxalate complexes was bound to the wood only when the pH was 3.6 or higher and the oxalate:Fe³⁺ molar ratio was less than 10. When 2,3-DHBA was evaluated for its ability to release iron bound to the milled wood, it was found that 2,3-DHBA possessed a greater affinity for ferric iron than the wood as 2,3-DHBA was capable of releasing the ferric iron bound to the wood in the pH range 3.6–5.5. These results further the understanding of the mechanisms employed by brown-rot fungi in wood biodegradation processes.     

Endophytic Fungi in Four Winter Wheat Cultivars (Triticum aestivum L.) Differing in Resistance Against Stagonospora nodorum (Berk.) Cast. & Germ. =Septoria nodorum (Berk.) Berk.

Four winter wheat cultivars with different levels of resistance to Septoria nodorum were investigated at four locations during two vegetation periods. Forty plants per cultivar and site were collected at random at seven defined growth stages from crop emergence to harvest. Samples from roots, culms, leaves, glumes and kernels were examined for the occurrence of endophytic fungi after surface sterilization. 83% of the 26944 isolates sporulated and were assigned to 213 species. The most frequent were: Septoria nodorum (20.1%), Alternaria tenuissima (9.8%), Epicoccum purpurascens (9.1%), Idriella bolleyi (6.9%), Fusarium graminearum (5.3%), Fusarium culmorum (4.0%), Cladosporium oxysporum (3.7%), Didymella exitialis (3.1%), Fusarium nivale (2.8%) and Rhizoctonia solani (2.1%). Each species occurred preferentially in one or more plant organs. A factorial analysis of variance showed that plant organ, sampling site, vegetation, period and cultivar in decreasing order of importance influenced the quantitative and qualitative composition of the fungal populations. No relationship between endophytic fungi was found to be constantly antagonistic or mutualistic. Septoria nodorum was isolated mainly from culms. The number of S. nodorum isolates differed significantly between cultivars in culms and glumes but not in flag leaves. The results are discussed in relation to resistance breeding and the effect endophytic fungi, might have on yield.     

Hydrogen peroxide production in wheat leaves infected with the fungus Septoria nodorum Berk. Strains with different virulence

The effect of two strains of the phytopathogenic fungus Septoria nodorum Berk. of different virulence on the intensity of local generation of hydrogen peroxide in common wheat leaves and the role of oxidoreductases in this process was studied. Differences in the pattern of hydrogen peroxide production in wheat plants infected with high- and low-virulence pathogen strains have been found. The low-virulent S. nodorum strain caused a long-term hydrogen peroxide (H₂O₂) generation in the infection zone, whereas the inoculation of leaves with the highly virulent strain resulted in a transient short-term increase in the H₂O₂ concentration at the initial moment of contact between the plant and the fungus. It was shown that the low level of H₂O₂ production by plant cells at the initial stages of pathogenesis facilitates S. nodorum growth and development. The decrease in the H₂O₂ concentration induced by the highly virulent S. nodorum strain is determined by inhibition of the oxalate oxidase activity in plant tissues and by the ability of the fungus to actively synthesize an extracellular catalase. The pattern of hydrogen peroxide generation at the initial stages of septoriosis may serve as an index of virulence of S. nodorum population.     

Comparative studies on the activities of chitinase, β-1,3-glucanase, peroxidase and phenylalanine ammonia lyase in the leaves of triticale and wheat infected with Stagonospora nodorum

The activities of chitinase, β-1,3-glucanase, peroxidase and phenylalanine ammonia lyase, constitutive and induced by Stagonospora nodorum were examined in the 10 – 14 day old seedlings of three triticale and two wheat cultivars under controlled environmental conditions and in flag leaves of two triticale cultivars in the field. Two S. nodorum isolates of different virulence were used. Both the constitutive and induced activities in triticale and wheat depended on genotype and in triticale the effect of growth conditions was also evidenced. The constitutive activities of chitinase, β-1,3-glucanase and peroxidase were several fold lower in flag triticale leaves in plants from the field than in the seedlings, growing under controlled conditions, but induction in the infected flag leaves was significantly more pronounced. In triticale genotypic differences in the response to infection were revealed only upon inoculation by S. nodorum isolate of higher virulence. The enzymatic activities increased several fold during successive days after the infection except for phenylalanine ammonia lyase. Induction of this enzyme was only transient and the activity decreased 48 or 96 h after infection when the activities of other enzymes were rising. In flag leaves in the field this activity was differentiated only after infection with more a virulent strain. A tendency appeared in triticale seedlings for association of the resistance to the pathogen with lower enzymatic constitutive activities. This relationship became more evident in triticale infected by S. nodorum and may imply that although the investigated enzymes are certainly involved in general, non-specific defense mechanism, they do not decide on the resistance to pathogen at least in the early stages of infection and cooperate with other factors in the complex pathogen-plant interaction. One can also assume that the enzymatic activities are associated with severity of infection rather than resistance to pathogen.     

Jasmonic Acid and Salicylic Acid Induce Accumulation of β-1,3-Glucanase and Thaumatin-Like Proteins in Wheat and Enhance Resistance Against Stagonospora nodorum

The effect of application of jasmonic acid (JA) and salicylic acid (SA) on the induction of resistance in wheat to Stagonospora nodorum and on the induction of -1,3-glucanase and thaumatin-like proteins (TLPs) was studied. Western blot analysis revealed that two -1,3-glucanases with apparent molecular masses of 31 and 33 kDa that cross-reacted with a barley glucanase antiserum were induced in wheat leaves after treatment with JA and SA. When wheat plants were treated with SA and JA, a TLP with an apparent molecular mass of 25 kDa and several other isoforms of TLP were induced. Pre-treatment of wheat plants with SA and JA significantly reduced (up to 56 %) the incidence of leaf blotch disease incited by S. nodorum compared with untreated control plants.     

Ferrous iron dependent nitric oxide production in nitrate reducing cultures of Escherichia coli

l-Lactate-driven ferric and nitrate reduction was studied in Escherichia coli E4. Ferric iron reduction activity in E. coli E4 was found to be constitutive. Contrary to nitrate, ferric iron could not be used as electron acceptor for growth. Ferric iron reductase activity of 9 nmol Fe²⁺ mg^-1 protein min^-1 could not be inhibited by inhibitors for the respiratory chain, like Rotenone, quinacrine, Actinomycin A, or potassium cyanide. Active cells and l-lactate-driven nitrate respiration in E. coli E4 leading to the production of nitrite, was reduced to about 20% of its maximum activity with 5 mM ferric iron, or to about 50% in presence of 5 mM ferrous iron. The inhibition was caused by nitric oxide formed by a purely chemical reduction of nitrite by ferrous iron. Nitric oxide was further chemically reduced by ferrous iron to nitrous oxide. With electron paramagnetic resonance spectroscopy, the presence of a free [Fe²⁺-NO] complex was shown. In presence of ferrous or ferric iron and l-lactate, nitrate was anaerobically converted to nitric oxide and nitrous oxide by the combined action of E. coli E4 and chemical reduction reactions (chemodenitrification).     

Predisposition to Septoria nodorum as a result of take-all (Gaeumannomyces graminis) infection of wheat

In a series of experiments excised leaves from take-all infected wheat plants and from control plants were inoculated with Septoria nodorum. Larger lesions, more lesions/leaf and more pycnidia/unit area of lesion were produced from take-all plants. Significant effects of predisposition were demonstrated when only 3% of the area of the total root system was infected by take-all. Microscopical investigations revealed that germ-tubes of S. nodorum grew more rapidly on leaves from take-all plants, but the time of penetration was not affected. It is proposed that the observed effects of predisposition arose because more germ-tubes produced successful infections and host tissue was more rapidly colonised. The importance of these results for the epidemiology of glume blotch is discussed.     

Spores of the mycorrhizal fungus <Emphasis Type="Italic">Glomus mosseae</Emphasis> host yeasts that solubilize phosphate and accumulate polyphosphates

The present paper reports the presence of bacteria and yeasts tightly associated with spores of an isolate of Glomus mosseae. Healthy spores were surface disinfected by combining chloramine-T 5%, Tween-40, and cephalexin 2.5 g L⁻¹ (CTCf). Macerates of these spores were incubated on agar media, microorganisms were isolated, and two yeasts were characterized (EndoGm1, EndoGm11). Both yeasts were able to solubilize low-soluble P sources (Ca and Fe phosphates) and accumulate polyphosphates (polyPs). Sequence analysis of 18S ribosomal deoxyribonucleic acid showed that the yeasts belong to the genera Rhodotorula or Rhodosporidium (EndoGm1) and Cryptococcus (EndoGm11). Results from inoculation experiments showed an effect of the spore-associated yeasts on the root growth of rice, suggesting potential tripartite interactions with mycorrhizal fungi and plants.     

Change of virulence of Septoria nodorurn during passage through barley and wheat

The barley biotype of Septoria nodorum was recovered from isolates of the wheat biotype after 2–6 passages through barley. Shifts in the population of the two biotypes during host passage were determined by isolating single spores after passage and observing changes in symptom expression on both hosts. The number of passages required before the barley biotype was recovered varied with the fungus isolate and barley cultivar. Attempts to recover the wheat biotype from isolates of the barley biotype during several passages through wheat were unsuccessful when 10⁵ to 10⁶ conidia m1^-1 were used as inoculum. However, the wheat biotype was recovered from two isolates of the barley biotype during a single passage when a very high concentration of conidia (>10¹² ml^-1) were used as inoculum. From the results of this study and a previous report by the author, the biotype of S. nodorum on barley which occurs in the southeastern United States appears to be largely restricted to barley.     

Impact of PGPR inoculation on growth and antioxidant status of wheat under saline conditions

Two plant growth‐promoting rhizobacterial (PGPR) strains, Bacillus subtilis SU47 and Arthrobacter sp. SU18, were found to tolerate 8% NaCl. Wheat co‐inoculated with these two PGPR strains, and grown under different salinity regimes (2–6 dS m^?1), showed an increase in dry biomass, total soluble sugars and proline content. Wheat sodium content was reduced under co‐inoculated conditions but not after single inoculation with either strain or in the control. The activity of antioxidant enzymes in wheat leaves decreased under salinity stress after PGPR co‐inoculation, suggesting these PGPR species could be used for amelioration of stress in wheat plants. Activity of three antioxidant enzymes in wheat grown with both PGPR strains was reduced, most notably that of catalase activity at a salinity of 6 dS m^?1, when compared with the control. The results indicate that co‐inoculation with B. subtilis and Arthrobacter sp. could alleviate the adverse effects of soil salinity on wheat growth.     

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.     

Components of partial resistance as criteria for identifying resistance

The components of partial resistance, incubation period, lesion area, latent period and sporulation were recorded on plants of six winter and two spring wheat cultivars which had been artificially inoculated with Septoria nodorum spores. Incubation period gave a guide as to how the cultivars would respond in the field to Septoria nodorum but statistical analysis showed that it could not be used alone to predict accurately the resistance of each cultivar to the pathogen. Average sporulation, however, could be used with more confidence for predicting the field resistance of the cultivars. From a regression analysis of NIAB rating versus incubation period, lesion area, latent period and sporulation, an equation was devised to obtain resistance indices for each cultivar. These resistance indices clearly reflected the NIAB ratings for the cultivars. It would therefore appear that resistance indices could be used as a pre-field evaluation method for identifying resistance to Septoria nodorum and thus be a valuable technique in breeding programmes.     

Application of an M13 bacteriophage displaying tyrosine on the surface for detection of Fe3+ and Fe2+ ions

Ferric and ferrous ion plays critical roles in bioprocesses,their influences in many fields have not been fully explored due to the lack of methods for quantification of ferric and ferrous ions in biological system or complex matrix.In this study,an M13 bacteriophage(phage) was engineered for use as a sensor for ferric and ferrous ions via the display of a tyrosine residue on the P8 coat protein.The interaction between the specific phenol group of tyrosine and Fe~(3+)./ Fe~(2+).was used as the sensor.Transmission electron microscopy showed aggregation of the tyrosine-displaying phages after incubation with Fe~(3+) and Fe~(2+).The aggregated phages infected the host bacterium inefficiently.This phenomenon could be utilized for detection of ferric and ferrous ions.For ferric ions,a calibration curve ranging from 200 nmol/L to 8 μmol/L with a detection limit of 58 nmol/L was acquired.For ferrous ions,a calibration curve ranging from 800 nmol/L to 8μmol/L with a detection limit of 641.7 nmol/L was acquired.The assay was specific for Fe~((3+)) and Fe~((2+)) when tested against Ni~(2+),Pb~(2+),Zn~(2+),Mn~(2+),Co~(2+),Ca~(2+),Cu~(2+),Cr~(3+),Ba~(2+),and K~+.The tyrosine displaying phage to Fe~(3+) and Fe~(2+) interaction would have plenty of room in application to biomatenals and bionanotechnology.     

Effect of Four Fungicides on Barley Net Blotch Caused by Drechslera teres

The effect of four fungicides (benomyl, carbendazim, triadimefon and sulp hur), used in intensive barley cropping on the epidemiology of Drechslera teres f. maculata and f. teres, was investigated under a controlled environment. Two fungicides (benomyl and carbendazim) increased disease development, especially at the recommended field rate, sulphur had the same effect at lower concentrations and triadimefon promoted net blotch caused by D. teres f. teres. At the recommended field rate, carbendazim enhanced sporulation in the two pathogen forms, whereas benomyl and sulphur enhanced 14 sporulation only on the maculata form. The results showed that, under controlled conditions, most of the fungicides promotednet blotch development and that sporulation increased either as a result of the pathogen form or of the concentration of the active ingredient.     

The role of iron uptake in pathogenicity and symbiosis in <Emphasis Type="Italic">Photorhabdus luminescens</Emphasis> TT01

Background  

Photorhabdus are Gram negative bacteria that are pathogenic to insect larvae whilst also having a mutualistic interaction with nematodes from the family Heterorhabditis. Iron is an essential nutrient and bacteria have different mechanisms for obtaining both the ferrous (Fe²⁺) and ferric (Fe³⁺) forms of this metal from their environments. In this study we were interested in analyzing the role of Fe³⁺ and Fe²⁺ iron uptake systems in the ability of Photorhabdus to interact with its invertebrate hosts.     

Disease progress of non-specialised fungal pathogens in intraspecific mixed stands of cereal cultivars. II. Field experiments

Epidemic development of Septoria nodorum was examined in pure stands and mixtures of two spring wheat cultivars, Kolibri and Maris Butler. In mixtures, disease development was reduced almost to that of the more resistant pure stand (Maris Butler). Disease levels, however, were low and no significant differences in grain yield per head were demonstrated. Disease development was further investigated for the host-pathogen combinations winter wheat/S. nodorum and winter barley/Rhynchosporium secalis. With both combinations the mixed stands again reduced disease levels to almost that of the more resistant pure stands. In general, mixed stands were more effective against R. secalis, although, with S. nodorum, disease levels were low. The yield response in mixed stands differed for the host cultivars and was not significantly changed by the presence of disease. The complexity of analysing such situations and the implications of these findings for mixtures of cultivars differing in resistance to non-specialised pathogens are discussed.     

Salicylic and Jasmonic acids in regulation of the proantioxidant state in wheat leaves infected by <Emphasis Type="Italic">Septoria nodorum</Emphasis> Berk

Influence of mediators of the signal systems of salicylic (SA) and jasmonic (JA) acids and their mixture on reactive oxygen species’ (ROS) (superoxide radical and O₂^·− H₂O₂) generation and activity of oxidoreductases (oxalate oxidase, peroxidase and catalase) in leaves of wheat Triticum aestivum L. infected by Septoria leaf blotch pathogen Septoria nodorum Berk has been studied. Presowing treatment of seeds by SA and JA decreased the development rate of fungus on wheat leaves. SA provided earlier inductive effect on production of O₂^·− and H₂O₂ compared with JA. The protective effect of the salicylic and jasmonic acids against Septoria leaf blotch pathogen was caused by activation of oxalate oxidase, induction of anion and cation peroxidases, and decrease of catalase activity. Ability of compounds to stimulate ROS in the plant tissues can be used as criteria for evaluation of immune-modulating activity of new substances for protection of the plants.     

Response of Arbuscular Mycorrhizal Fungi on Wheat (Triticum aestivum L.) Grown Conventionally and on Beds in a Sandy Loam Soil

The present study was undertaken to assess the benefit and compare the functioning of AM fungi on wheat grown conventionally and on beds. Ten treatment combinations were used, treatments 1 and 2: no fertilizers with and without arbuscular mycorrhizal (AM) fungi (In vitro produced Glomus intraradices); 3:100% of recommended NPK: (120 kg ha⁻¹ N; 60 kg ha⁻¹ P; 50 kg ha⁻¹ K), and 4 and 5: 75% of recommended NPK dose with and without AM inoculation in a 5 × 2 split-plot design on wheat using conventional/flat system and elevated/raised bed system. The maximum grain yield (3.84 t ha⁻¹) was obtained in AM fungi inoculated plots of raised bed system applied with 75% NPK and was found higher (although non- significant) than the conventional (3.73 t ha⁻¹) system. The AM inoculation at 75% fertilizer application can save 8.47, 5.38 kg P and 16.95, 10.75 kg N ha⁻¹, respectively, in bed and conventional system. While comparing the yield response with 100% fertilizer application alone, AM inoculation was found to save 20.30, 15.79 kg P and 40.60, 31.59 kg N ha⁻¹, respectively, in beds and conventional system. Mycorrhizal inoculation at 75% NPK application particularly in raised bed system seems to be more efficient in saving fertilizer inputs and utilizing P for producing higher yield and growth unlike non-mycorrhizal plants of 100% P. Besides the yield, mycorrhizal plants grown on beds had higher AM root colonization, soil dehydrogenases activity, and P-uptake. The present study indicates that the inoculation of AM fungi to wheat under raised beds is better response (although non-significantly higher) to conventional system and could be adopted for achieving higher yield of wheat at reduced fertilizer inputs after field validation.     

Study of the Protective Activity of Chitosan Hydrolyzate Against Septoria Leaf Blotch of Wheat and Brown Spot of Tobacco

Chitosan hydrolyzate containing low-molecular-weight chitosan (≤24 kDa) and its oligomers (≤1.2 kDa) has been obtained via chemical depolimerization of high-molecular-weight chitosan by nitric acid. The fractions of the obtained chitosan hydrolyzate have been characterized by high performance gel permeation chromatography and proton magnetic resonance. The test performed on detached leaves of wheat has shown that the hydrolyzate completely inhibits the development of Stagonospora nodorum, a casual agent of the septoria leaf blotch, at a concentration of 200 μg/mL. A similar test with detached tobacco leaves has shown that the hydrolyzate at a concentration of 100 μg/mL also inhibits the development of Alternaria longipes, which causes brown spot of tobacco, by 75%.