Aubian wheat mosaic virus, a new soil-borne wheat virus emerging in France

The properties of Aubian wheat mosaic virus (AWMV), a new soil-borne wheat virus in France, were investigated. Symptoms include foliar mosaic and severe stunting of winter wheat. The vector of the disease is unknown but the plants infected carry Polymyxa graminis in the roots. AWMV was transmitted mechanically to wheat and to two dicotyledoneous species: Lactuca sativa and Vicia faba. This virus was transmitted by seed to three winter wheat cultivars tested. Purified preparations contained rod-shaped particles with a variable length of 150–700 nm. Certain particles are very long and appear flexible. Antiserum raised against AWMV reacted specifically with AWMV in both indirect and direct enzyme-linked immunosorbent assays (ELISA). The incidence of AWMV in 26 winter wheat cultivars was investigated in the field during the growing season of 1999–2000. AWMV was detected in roots and shoots of all cultivars regardless of the symptoms. Twelve virus species belonging to the genera Benyvirus, Bymovirus, Furovirus, Pecluvirus and Pomovirus did not react with the AWMV antisera. A new tubular virus described in winter wheat in Bedfordshire in England reacted strongly with AWMV in ELISA. It is concluded that AWMV and probably the Bedford-virus constitute a previously undescribed tubular virus biologically and serologically distinct from other soil-borne viruses of wheat.     

Assessing Four-Way Mixtures of Winter Wheat Cultivars from the Performances of their Two-Way and Individual Components

Seven four-way bread wheat mixtures were compared to their components (individual cultivars and two-way mixtures) for septoria tritici blotch severity, grain protein content, and grain yield. Four-way mixtures and two-way mixtures performed better than the average of individual pure line components. Disease severity and protein content were most influenced by mixtures, with mixture efficiencies being greater in the four-way than in the two-way mixtures. Performances of four-way mixtures in terms of diseased leaf area reduction, grain yield, 1000-grain weight, and grain protein content were better predicted by performances of two-way mixtures than by those of pure lines. Our results suggest that two-way mixtures should be screened to remove unfavourable cultivar pairs and those binary mixtures that show complementarity should be selected in order to construct four-way mixtures.     

Natural Infection of Wheat by the Type Strain of Soil-borne Wheat Mosaic Virus in a Field in Southern Germany

Molecular analyses revealed that a virus causing a severe disease of wheat in one field in Southern Germany is closely related to the Nebraska type strain of Soil-borne wheat mosaic virus (SBWMV) and only distantly related to Soil-borne cereal mosaic virus that is widely distributed in Europe. The latter virus was not found in the SBWMV-containing leaf samples. This is the first report of the occurrence of SBWMV in Germany, and perhaps in all of Europe, which has been confirmed on the molecular level.     

Temperature Dependent Seed Transmission of Stagonospora nodorum in Wheat

The transmission of Stagonospora nodorum from four naturally infected winter wheat seedlots was quantified in controlled environment germination chambers at 9, 13, 17, 21, and 25 °C. Seedlings were harvested when the second leaf began to emerge. Coleoptiles and first seedling leaves were examined for the presence of lesions caused by S. nodorum. First leaves were incubated on Bannon's medium for 2 weeks, after which they were examined for pycnidia of S. nodorum. Transmission to the coleoptile occurred at all temperatures, but decreased from 100% to 72% as temperature increased from 9 to 25 °C. Transmission to the first leaf was less, dropping from 37% to 2% as temperature increased from 9 to 25 °C. At least 44% of infected first leaves were symptomless at all temperatures, with 96% of infected leaves showing no symptoms at 25 °C. Transmission to seedling leaves occurred over a broad temperature range. Under the high densities at which wheat is sown, a significant number of infected seedlings per unit area may originate from relatively low initial seed infection levels and transmission efficiencies.     

Molecular characterisation of <Emphasis Type="Italic">Rice stripe necrosis virus</Emphasis> as a new species of the genus <Emphasis Type="Italic">Benyvirus</Emphasis>

The complete nucleotide sequences of RNAs 1 and 2 of Rice stripe necrosis virus (RSNV) were determined and compared to the corresponding genomes of all sequenced, rod-shaped plant viruses. The genome organisation of RSNV RNA1 and RNA2 is nearly identical to that of Beet necrotic yellow vein virus (BNYVV) and Beet soil-borne mosaic virus (BSBMV), definitive species of the genus Benyvirus. As demonstrated for BNYVV and BSBMV, the RNA1 of RSNV also encodes a single ORF with putative replicase-associated motifs, which distinguishes benyviruses from all other viruses possessing rod-shaped particles. As described for BNYVV, RNSV RNA-2 also contains six ORFs: the capsid protein gene, the read-through protein gene, a triple gene block gene that codes for three different proteins, and a 17 kDa cysteine-rich protein. RNAs 3 and 4 (or 5 in the case of BNYVV), identified in natural infections of BNYVV and BSBMV, were not detected in any of the 44 RSNV cDNA clones obtained in this investigation. Nevertheless, phylogenetic and amino comparative acid sequence analyses demonstrated that RSNV is more closely related to BNYVV and BSBMV than to any other rod-shaped plant virus characterised to date.     

Surveys of cereal diseases and pests in the Netherlands. 4. Occurrence of powdery mildew and rusts in winter wheat

Results of annual surveys of winter wheat fields from 1974 to 1986 were compiled to describe epidemics of powdery mildew and rusts in relation to weather and cultivar resistance.An average of 29 and 70% of fields were infected by powdery mildew in May and July, respectively. Mildew prevalence in May was positively correlated with average temperature in October and with average temperature over the months December, January, February and March. In addition, it was correlated negatively with the average grade of mildew resistance of the cultivars sown each year. Prevalence of mildew in July did not show consistent correlations with weather characteristics nor with mildew prevalence in May.Yellow rust was usually not detected in May and on average 18% of the fields was infected in July. The occurrence of yellow rust decreased after 1977, when the farmers adopted cultivars resistant or moderately resistant to yellow rust.Brown rust was usually not detected in May, while in July on average 48% of the fields was infected. Brown rust intensity in July was high in years with a high March temperature and high precipitation during April and May.Black rust was rare in the Netherlands, with 3 and 1% of the fields infected in July 1977 and 1981, respectively.     

Surveys of cereal diseases and pests in the Netherlands. 2. Stem-base diseases of winter wheat

In the period 1974–1986, ca. 100 commercial winter wheat fields were surveyed annually for stembase diseases. In these years, on average 6% of the tillers was infected with eyespot in spring at the first-second, node stage. Eyespot intensity in spring was high in years with high temperature during winter. Eyespot, intensity in the milky-ripe stage in July, averaged 13% of the culms and was high in years with high temperature in April and high precipitation in March, April and May. These data may improve forecasts.In the milky-ripe stage, 2% of the culms were infected with sharp eyespot, but its intensity has gradually increased during the survey years. No significant correlation of sharp eyespot intensity with dry periods in autumn, spring or summer was found.Symptoms of take-all were found on 0.2% of the stem-bases during ripening. Its intensity was low in years with high precipitation in March, April, May and June and high temperature in May and June.Samenvatting Een honderdtal percelen wintertarwe werd in 1974–86 jaarlijks op voetziekten geïnventariseerd. In het voorjaar, tijdens het, eerste en tweede knoopstadium van het gewas, was gemiddeld 6% van de spruiten aangetast door oogvlekkenziekte. De intensiteit van de ziekte was hoog in jaren met een hoge gemiddelde wintertemperatuur. Tijdens het melkrijpe stadium, in juli was gemiddeld 13% van de halmen aangetast door oogvlekkenziekte. De aantasting in juli was hoog in jaren met een hoge temperatuur in april en veel neerslag in de maanden maart, april en mei. Met deze gegevens kunnen adviessystemen worden verbeterd.Scherpe oogvlekkenziekte was op gemiddeld 2% van de halmen in juli aanwezig. De ziekte nam geleidelijk met de jaren toe. De jaarlijkse intensiteit was niet gecorreleerd met droge perioden in de herfst, voorjaar of zomer.Symptomen van halmdoder waren op gemiddeld 0.2% van de halmen aanwezig. De intensiteit van de ziekte was hoog in jaren met weinig neerslag in maart, april, mei en juni en met een lage temperatuur in mei en juni.     

Comparison and differentiation of Wheat Yellow Mosaic Virus(WYMV), Wheat Spindle Streak Mosaic Virus (WSSMV) and Barley Yellow Mosaic Virus (BaYMV) isolates using WYMV monoclonal antibodies

Twelve monoclonal antibodies (MAbs) were obtained by immunizing mice with a French isolate (F1) of wheat yellow mosaic virus (WYMV). Three of these (3D12, 2C1, 6C3) belong to the IgM class and the nine others to the IgG class (3D8, 3H1, 2B8, 1F2, 3C10, 4F12, 3H9, 1G5, 54). In antigen-coated plate (ACP) ELISA and indirect double antibody sandwich (IDAS) ELISA, all MAbs recognize the WYMV (F1) both in the form of purified particles and in wheat leaf extract. The analysis of numerous French isolates of WYMV shows a variable reactivity with MAbs 3D8, 3H1, 2B8, 3C10, 3H9 and 1G5 in IDAS — and ACP-ELISA. The Japanese isolate of WYMV and United States isolates of wheat spindle streak mosaic virus (WSSMV) were detected in IDAS- and ACP-ELISA by ten of the MAbs tested showing that the wheat bymoviruses originating from the three locations share a high epitopic homology. French isolates of barley yellow mosaic virus (BaYMV; pathotypes 1 and 2) were only detected in ACP-ELISA with MAbs 6C3, 3D8, 3H1 and 2B8 whereas the two Japanese strains (I-1, II-1) of MaYMV were recognized with these and also with that of 3C10. In IDAS-ELISA, the two Japanese strains were clearly detected by MAbs, 6C3, 3D8, 3H1, 1F2, 3C10 and 1G5 and the British and Belgian (pathotype 2) isolates only by that of 6C3. Only the Japanese strain of BaYMV, 1-1 could be detected with MAb 3H9 in this ELISA system.     

Measures of disease intensity in powdery mildew (Erysiphe graminis) of winter wheat. 1. Errors in estimating pustule number

Assessments of pustule number and severity of powdery mildew on winter wheat in the Netherlands were made in commercial fields and in experimental plots. The sample variance (s²) of the number of pustules per leaf (m) was fairly constant over years, varieties, growth stages and leaf postitions, but depended strongly on the average pustule number: . The effect of sample size on the precision of the estimate is discussed and it is concluded that it is difficult to estimate low disease intensities accurately. Estimates are given for the detection level of pustule counts in relation to sample size.Mildew intensity on the lower surface of leaves can be estimated from the intensity on the upper surface. This method reduces the duration of the observation, but introduces an additional error. At low disease intensities and small sample sizes this method is more efficient than sampling mildew on both surfaces of leaves. The common practice of assessments of the upper surface of leaves only may not be the most efficient method.Samenvatting Aantallen puistjes meeldauw per blad werden geteld in praktijkpercelen en veldproeven met wintertarwe. De steekproefvariantie van het aantal puistjes was tamelijk constant in de jaren, rassen, gewasstadia en bladposities, maar was sterk afhankelijk van het gemiddeld aantal puistjes . Het effect van de steekproefgrootte op de nauwkeurigheid van de schatting wordt besproken en het blijkt dat het moeilijk is om lichte aantastingen nauwkeurig te schatten. Er worden schattingen gegeven van de detectiegrens in afhankelijkheid van de steekproefgrootte.Meeldauwaantastingen aan de onderkant van het blad, kunnen worden geschat uit de aantasting op de bovenkant van het blad. Deze methode levert een tijdsbesparing op, maar ook een extra onnauwkeurigheid. Alleen bij lichte aantastingen en kleine steekproeven is deze methode efficiënter dan een directe tweezijdige bemonstering. Het schatten van meeldauw op de bovenkant van bladeren is, hoewel algemeen gebruikelijk, waarschijnlijk niet de meest efficiënte methode.     

Yield Reduction in Wheat in Relation to Leaf Disease From Yellow (tan) Spot and Septoria Nodorum Blotch

Yellow or tan spot (caused by Pyrenophora tritici-repentis) and septoria nodorum blotch (caused by Phaeosphaeria nodorum) occur together and are a constraint to wheat yields in Australia. Recently, higher crop yields and lower fungicide costs have made fungicides an attractive management tool against these diseases. Yield-loss under different rates of progress of yellow spot and septoria nodorum blotch was examined in four experiments over three years to define the relationship between disease severity and yield. In these experiments, differences in disease were first promoted by inoculations either with P. tritici-repentis-infected stubble or aqueous spore suspensions of P. nodorum. Disease progress was further manipulated with foliar application of fungicide. The pattern of disease development varied in each year under the influence of different rainfall patterns. The inoculation and fungicide treatments produced differences in disease levels after flag leaf emergence. The infection of yellow spot or septoria nodorum blotch caused similar losses in grain yield, ranging from 18% to 31%. The infection by either disease on the flag or penultimate leaf provided a good indication of yield-loss. Disease severity on flag leaves during the milk stage of the crop or an integration of disease as area under the disease progress curve on the flag leaves based on thermal time explained more than 80% variance in yield in a simple regression model. The data provided information towards the development of disease management strategies for the control of septoria nodorum blotch and yellow spot.     

Studies on the Interaction Between Septoria tritici and Stagonospora nodorum in Wheat

Interactions between Stagonospora nodorum and Septoria tritici were studied. Results from a detached glume experiment indicated that the interaction may be isolate-dependent, as it was shown that the interaction between the two pathogens may be beneficial or antagonistic depending on the isolate of each pathogen present. The number of spores produced by both pathogens was significantly greater when an aggressive isolate of S. tritici was mixed with a non-aggressive isolate of S. nodorum, whereas the number of spores produced by both pathogens was significantly less when two non-aggressive isolates were mixed. There was a significant reduction in disease level when S. tritici was applied prior to S. nodorum, compared to vice versa in the growth chamber. Results from growth chamber and field studies showed that S. nodorum produced significantly more spores when both pathogens were present together. It is concluded that S. tritici has a stimulatory effect on spore production by S. nodorum. However, there was a reduction of S. tritici spores observed in the dual inoculation treatments, suggesting that S. nodorum inhibits S. tritici.     

Uromyces Viciae-fabae Haustorium Formation in Susceptible and Resistant Faba Bean Lines

Haustorium formation by the faba bean rust (Uromyces viciae-fabae) was studied on susceptible and resistant faba bean lines. The resistant lines showed incomplete resistance, based on late acting hypersensitivity or on non-hypersensitive resistance acting before haustorium formation. Histological observations on infected leaves showed that both the number of haustoria per infection unit and their developmental stage was reduced in both resistant lines. Isolation of haustoria confirmed that both the number and the size of haustoria were reduced in resistant lines, irrespective of whether the resistance was associated with hypersensitivity. Plant age had no detectable effect on both parameters.     

Molecular Identification of Oat Mosaic Virus as a Bymovirus

A partial sequence of Oat mosaic virus (OMV) has been obtained for four isolates of the virus from four European countries. This represents the first available sequence data for this important disease of winter-sown oats. The longest clone of 1699 nucleotides was obtained from infected English oats using a degenerate primer, designed to members of the Potyviridae family. Alignment of the predicted amino acid sequence with members of the Potyviridae showed closest identity with viruses of the Bymovirus genus. The predicted amino acid sequence has one open reading frame corresponding to part of the NIb and capsid protein, with a 3 untranslated region of 351 nucleotides, followed by a poly(A) tail. PCR primers were designed to the coat protein and NIb gene of members of the Bymovirus genus and used to obtain partial sequences of 1441 nucleotides at the 3 end of infected oats from both Wales and France. A specific primer set designed to the English isolate was used to generate a product of 701 nucleotides from OMV-infected oat leaves from Ireland. All four isolates are highly conserved at the amino acid level.The first two authors contributed equally to the work     

Components of Partial Resistance to Powdery Mildew in Wheat Mutants

Components of resistance to powdery mildew (latent period, pustule density and conidium production) were analysed on glasshouse-grown artificially inoculated plants of wheat mutants (n = 11) derived from either induced mutagenesis or adventitious regeneration of cv. Guardian. These mutants had previously been shown to be partially resistant to the disease in the field over a two-year period. Analyses were carried out at three stages of development: seedling, tillering and heading. None of the mutants exhibited a latent period significantly different from that of the susceptible parent Guardian at any stage of development tested. With respect to pustule density, one mutant (M61) was significantly more resistant than Guardian at all stages of development, two mutants (M19 and SC100) were more resistant at tillering and heading, and two (M156 and SC267) were more resistant only at heading. Significantly reduced conidium production per pustule (as a measure of pustule size) over a 24h period was observed on three mutants (M61, SC100 and SC231) at tillering and heading, while mutant SC68 exhibited this trait at heading only. On the basis of resistance components and relative grain yield in the presence of mildew, the eleven mutants were categorised into five categories. Tests for associations between field resistance score and components of resistance measured in the glasshouse, both measured at heading, revealed a significant positive correlation only between pustule density and field score. To assess the effects of combining different components (i.e. pyramiding different partial resistance genes), mutants were crossed. Transgressive segregation for at least one component was observed in the F₂ generation in crosses between mutants with complementary modes of action (i.e. involving different components of partial resistance), but not in cases where the parents exhibited the same component of resistance.     

Effects of powdery mildew and weather on winter wheat yield. 2. Effects of mildew epidemics

Wheat yield losses caused by powdery mildew were computed based on effects of the disease on leaf photosynthesis. Powdery mildew was introduced in a crop model of wheat by quantification of five parameters, taking the vertical and horizontal distribution of mildew in the crop into account. The most important parameters were those of the mildew intensity, the distribution of mildew in the crop, and the effect of mildew on assimilation at light saturation. Measured mildew epidemics in field experiments in three different years, were used to compute yield losses. Computed losses were compared to measured losses. On average, computed yield loss approached measured, but measured yield loss was underestimated, especially in early mildew epidemics due to the computation of partitioning and reallocation of assimilates. Other processes which may cause an underestimation are described. The use of crop models as a method to upgrade disease management systems is discussed.Samenvatting Opbrengstderving van wintertarwe werd berekend aan de hand van het effect dat meeldauw heeft op de blad-fotosynthese. Een rekenmodel voor de gewasgroei van tarwe werd uitgebreid met meeldauw. Met inachtneming van de vertikale en horizontale verdeling van meeldauw in het gewas, werd meeldauw in het model gekwantificeerd door vijf parameters. De belangrijkste parameters waren die van de meeldauwintensiteit, de verdeling van meeldauw in het gewas en het effect van meeldauw op de assimilatie bij een overvloed aan licht. Epidemieën van meeldauw, gemeten in veldproeven in drie verschillende jaren, werden gebruikt om opbrengstdervingen te berekenen. Gemiddeld kwam deze redelijk overeen met de in de veldproeven gemeten opbrengstderving. De gemeten opbrengstderving werd echter onderschat, vooral bij vroege epidemieën van meeldauw door de wijze waarop de (her)verdeling van assimilaten wordt berekend. Andere mechanismen, die een onderschatting van opbrengstderving kunnen veroorzaken worden besproken. Of deze modellen als methode gebruikt kunnen worden om systemen voor de geleide bestrijding van ziekten te verbeteren wordt bediscussieerd.     

Requirement of Host Signaling Mechanisms for the Action of Ptr ToxA in Wheat

Ptr ToxA, the host-selective toxin produced by Pyrenophora tritici-repentis, is genetically associated with the development of tan spot disease of wheat. The toxin was shown previously to cause a programmed cell death in the host that requires de novo mRNA and protein synthesis. In the present study, inhibitors of plant signaling mechanisms protected wheat leaves from toxin action, as determined by electrolyte leakage bioassays, when applied to leaves with toxin. Okadaic acid, calyculin A and phenylarsine oxide, all inhibitors of protein phosphatase activity, reduced toxin-induced electrolyte leakage by more than 90%. Inorganic calcium channel blockers (LaCl₃ and CoCl₂ reduced toxin-induced electrolyte leakage by 78–95%, depending on inhibitor and time of measurement. By comparison, about 50% protection was achieved by the application of the protein kinase inhibitors staurosporine and K-252A. Nonetheless, the reduction in toxin-induced electrolyte leakage by protein kinase inhibitors was reproduced in multiple trials and was statistically significant. The data indicate that host signaling mechanisms, including calcium fluxes and a protein phosphorylation cascade, are required for the Ptr ToxA-induced cell death in wheat. Our current model holds that the signaling events occur between toxin perception by the cell and the toxin-directed gene expression in the host associated with cell death. As an alternative, the toxin-induced mRNA synthesis required for cell death may be for protein phosphatase and/or protein kinase genes. Additional work is required to resolve these possibilities.     

Effect of Juglone on Active Oxygen Species and Antioxidant Enzymes in Susceptible and Partially Resistant Banana Cultivars to Black Leaf Streak Disease

The black leaf streak disease (BLSD), caused by Mycosphaerella fijiensis, is the most destructive disease of bananas and plantains around the world. Breeding for resistance is the most promising strategy to fight this disease especially in small farmer plantations. Mycosphaerella fijiensis produces many phytotoxins such as juglone, which can be used, jointly with field and inoculations under controlled conditions, for screening banana cultivars for BLSD-resistance. This non-host specific phytotoxin has been shown to act on chloroplasts and disturbs the proton electrochemical gradient across the plasmalemma membrane. Moreover, an involvement of the oxidative burst during the interaction has been suggested. The present study was carried out using two cultivars that differed for either their juglone-responses or their resistance to BLSD (cv. Grande Naine susceptible to BLSD and juglone and cv. Fougamou partially resistant to BLSD and highly tolerant to juglone). The production of active oxygen species (AOS) and the enhancement of the enzymatic and/or non-enzymatic AOS-scavenging systems were investigated after treatment of the two cultivars with juglone. The time-course of AOS-production and AOS-scavenging was shown to be the key difference between these two tested cultivars after treatment with juglone. Thus, an early release of AOS (O₂⁻ radical and H₂O₂) and a quick stimulation of a preferment anti-oxidant system (superoxide dismutases, catalases, and peroxidases) was observed for cv. Fougamou as compared to cv. Grande Naine for which a late and weak generation of AOS accompanied by a late stimulation of the anti-oxidant systems were detected.