Linear relationship between Gaeumannomyces graminis var. tritici (Ggt) genotypic frequencies and disease severity on wheat roots in the field

In order to investigate potential links existing between Gaeumannomyces graminis var. tritici (Ggt) population structure and disease development during polyetic take-all epidemics in sequences of Ggt host cereals, seven epidemics in fields with different cropping histories were monitored during the seasons 2001/2002 (two fields), 2002/2003 (two fields) and 2003/2004 (three fields). Take-all incidence and severity were measured at stem elongation and Ggt populations were characterized. The 73 isolates collected in the two fields in 2001/2002 were distributed into two multilocus genotypes, G1 and G2 according to amplified fragment length polymorphism analysis. A monolocus molecular marker amplified by F-12 random amplification polymorphism DNA primer sizing between 1.9 and 2.0 kb that gave strictly the same distinction between the two multilocus genotypes was further applied to measure G1/G2 frequencies among Ggt populations in all fields (266 isolates). The ratios of G1 to G2 differed between fields with different cropping histories. A linear relationship between G2 frequency among Ggt populations and disease severity at stem elongation was measured during the three cropping seasons. When take-all decline was observed, G2 frequencies were low in first wheat crops, highest in short-term sequences and intermediate in longer sequences of consecutive crops of Ggt host cereals. This pattern could be the result of population selection by environmental conditions, in particular by microbial antagonism during the parasitic phase of the fungus. In order to better understand take-all epidemic dynamics, the distinction between these two genotypes could be a basis to develop models that link approaches of quantitative epidemiology and advances in population genetics of Ggt.     

Control of Mn status and infection rate by genotype of both host and pathogen in the wheat take-all interaction

Take-all is a world-wide root-rotting disease of cereals. The causal organism of take-all of wheat is the soil-borne fungus Gaeumannomyces graminis var tritici (Ggt). No resistance to take-all, worthy of inclusion in a plant breeding programme, has been discovered in wheat but the severity of take-all is increased in host plants whose tissues are deficient for manganese (Mn). Take-all of wheat will be decreased by all techniques which lift Mn concentrations in shoots and roots of Mn-deficient hosts to adequate levels. Wheat seedlings were grown in a Mn-deficient calcareous sand in small pots and inoculated with four field isolates of Ggt. Infection by three virulent isolates was increased under conditions which were Mn deficient for the wheat host but infection by a weakly virulent isolate, already low, was further decreased. Only the three virulent isolates caused visible oxidation of Mn in vitro. The sensitivity of Ggt isolates to manganous ions in vitro did not explain the extent of infection they caused on wheat hosts. In a similar experiment four Australian wheat genotypes were grown in the same Mn-deficient calcareous sand and inoculated with one virulent isolate of Ggt. Two genotypes were inefficient at taking up manganese and were very susceptible to take-all, one was very efficient at taking up manganese and was resistant to take-all, and the fourth genotype was intermediate for both characters. All genotypes were equally resistant under Mn-adequate conditions.     

Occurrence of Phialophora radicicola var. graminicola and Gaeumannomyces graminis var. tritici on roots of wheat in field crops

Assessments of Phialophora radicicola var. graminicola (PRG) and Gaeumannomyces graminis var. tritici (GGT) were made by culturing and by direct microscopic examination of pieces of seminal roots from 16 winter wheat crops grown in different cropping sequences and with different phosphate manuring. PRG occurred on all wheat crops, but was abundant only on wheat after grass, where it seemed to delay the onset of damaging take-all by 1 yr. Delayed occurrence of take-all by phosphate fertiliser was not related to differences in populations of PRG. Wheat grown in ‘take-all decline’ soils had only small amounts of PRG, indicating that the development and the decline of take-all epidemics may be influenced by different biological control mechanisms; breaking sequences of wheat crops by 1 yr grass leys might harness the advantages of both mechanisms.     

Grain yield and incidence of take-all (Ophiobolus graminis Sacc.) in wheat grown in different crop sequences

The yield of wheat and the incidence of take-all were measured in crops grown in six different 4-year sequences, repeated in 3 successive years. The first crop of winter wheat grown after oats or beans yielded 13–23 cwt/acre (1632–2887 kg/ha) more grain than wheat after wheat or barley. Spring wheat after oats yielded 2–5 cwt/acre (250–625 kg/ha) more than spring wheat after wheat. The smaller yields of wheat after wheat or barley were caused mostly by greater prevalence of take-all. Regression analysis indicates that each 1 % increase in straws with take-all decreased yield of winter wheat by 0·6%. Take-all was more prevalent in the second and third successive wheat crops after oats than in the fourth crop.     

Gluconic acid: an antifungal agent produced by Pseudomonas species in biological control of take-all

Pseudomonas strain AN5 (Ps. str. AN5), a non-fluorescent Australian bacterial isolate, is an effective biological control (biocontrol) agent of the take-all disease of wheat caused by the fungus Gaeumannomyces graminis var. tritici (Ggt). Ps. str. AN5 controls Ggt by producing an antifungal compound which was purified by thin layer and column chromatography, and identified by NMR and mass spectroscopic analysis to be d-gluconic acid. Commercially bought pure gluconic acid strongly inhibited Ggt. Two different transposon mutants of Ps. str. AN5 which had lost take-all biocontrol did not produce d-gluconic acid. Gluconic acid production was restored, along with take-all biocontrol, when one of these transposon mutants was complemented with the corresponding open reading frame from wild-type genomic DNA. Gluconic acid was detected in the rhizosphere of wheat roots treated with the wild-type Ps. str. AN5, but not in untreated wheat or wheat treated with a transposon mutant strain which had lost biocontrol. The antifungal compounds phenazine-1-carboxylic acid and 2,4-diacetylphloroglucinol, produced by other Pseudomonads and previously shown to be effective in suppressing the take-all disease, were not detected in Ps. str. AN5 extracts. These results suggest that d-gluconic acid is the most significant antifungal agent produced by Ps. str. AN5 in biocontrol of take-all on wheat roots.     

Comparison of natural and artificial epidemics of take-all in sequences of winter wheat crops

Winter wheat was grown for six successive years (Expt 1) and for three successive years (Expt 2) in field experiments on different soil types. Artificial inoculum of the take-all fungus (Gaeumannomyces graminis var. tritici cultured on autoclaved oat grains) was incorporated in the soil of some of the plots just before, or at, sowing of the first winter wheat crop. Expt 1 tested the incorporation of similar amounts of inoculum (212 kg ha^-1) at different depths. Expt 2 tested different amounts of inoculum at the same, shallow depth. Early sowing (September), late sowing (October) and spring inoculation were additional treatments, applied to the first crop only, in Expt 2. Seasonal factors apart, the disease outcome in the first year after inoculation depended on amounts and placement of applied inoculum, as well as date of sowing. Deeper inoculum resulted in less disease (Expt 1). Severe take-all was produced in Expt 2 by incorporating inoculum shallowly in sufficient quantities (400 kg ha^-1 or more). Less inoculum (200 kg ha^-1) generated less disease, especially in earlier-sown plots. Differences in disease amongst inoculum treatments were greatest in the first year and diminished subsequently, particularly where sowing had been early in the first year. In Expt 1, where first crops exposed to artificial inoculum developed moderate-to-severe disease, disease in subsequent second and/or third crops was less. In the fourth crop a second peak of disease occurred, coinciding with a first peak in sequences without added inoculum. Take-all decline (TAD) appeared to be expressed in all sequences thereafter. In Expt 2 in sequences without added inoculum, TAD occurred after a peak of disease in the second crop. Where 400 kg ha^-1 or more of inoculum were added, disease was severe in the first year and decreased progressively in successive years. Disease was less patchy in plots that received artificial inoculum. However, it remains uncertain mat severe disease caused by artificial inoculation achieved an early onset of true TAD. The infectivity of the top 12 cm of soil in the first 3 yr of Expt 1, determined by bioassay, depended on the depth of added inoculum and amount of disease in subsequent crops. However, at the time of the naturally occurring peak of disease severity (in either inoculated or non-inoculated plots) it did not predict either disease or TAD. Differences and similarities amongst epidemics developing naturally and those developing from different amounts and placement of applied inoculum have been revealed. The epidemiological implications of adding inoculum and the potential value of artificially-created epidemics of take-all in field trials are discussed.     

Enrichment and genotypic diversity of phlD-containing fluorescent Pseudomonas spp. in two soils after a century of wheat and flax monoculture

Fluorescent Pseudomonas spp. producing the antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) play a key role in the suppressiveness of some soils to take-all of wheat and other diseases caused by soilborne pathogens. Soils from side-by-side fields on the campus of North Dakota State University, Fargo, USA, which have undergone continuous wheat, continuous flax or crop rotation for over 100 years, were assayed for the presence of 2,4-DAPG producers. Flax and wheat monoculture, but not crop rotation, enriched for 2,4-DAPG producers, and population sizes of log 5.0 CFU g root(-1) or higher were detected in the rhizospheres of wheat and flax grown in the two monoculture soils. The composition of the genotypes enriched by the two crops differed. Four BOX-PCR genotypes (D, F, G, and J) and a new genotype (T) were detected among the 2,4-DAPG producers in the continuous flax soil, with F- and J-genotype isolates dominating (41 and 39% of the total, respectively). In contrast, two genotypes (D and I) were detected in the soil with continuous wheat, with D-genotype isolates comprising 77% of the total. In the crop-rotation soil, populations of 2,4-DAPG producers generally were below the detection limit, and only one genotype (J) was detected. Under growth-chamber and field conditions, D and I genotypes (enriched by wheat monoculture) colonized the wheat rhizosphere significantly better than isolates of other genotypes, while a J-genotype isolate colonized wheat and flax rhizospheres to the same extent. This study suggests that, over many years of monoculture, the crop species grown in a field enriches for genotypes of 2,4-DAPG producers from the reservoir of genotypes naturally present in the soil that are especially adapted to colonizing the rhizosphere of the crop grown.     

Evaluation of RAPD, ISSR and AFLP Markers for Characterization of the Loose Smut Fungus Ustilago tritici

Random Amplified Polymorphic DNA (RAPD), inter simple sequence repeat (ISSR) and Amplified Fragment Length Polymorphism (AFLP) profiling were evaluated for assessing the extent of genetic variation among the isolates of Ustilago tritici (Pers.) Rostr., which causes the loose smut disease of wheat.Thirty random decamer primers, six random primer pairs, four SSR primers such as (GACA)₄, (GATA)₄, (CAA)₅ and (GTG)₅ and nine combinations of AFLP selective primers were used to characterize nine isolates of the fungus. These isolates were collected from infected earheads of seven commercial wheat cultivars grown at eight different locations in Haryana, which is a major wheat growing state in the North-West Plain Zone of India. The RAPD and ISSR primers generated 21 0 scorable amplified fragments, all of which were monomorphic among the isolates.The AFLP primer combinations generated 239 fragments out of which 193 were polymorphic. All the isolates could be precisely differentiated from each other employing AFLP and grouped into two distinct clusters.The molecular classification partly corresponded with geographic distribution and host origin of the isolates. AFLP profiling was found superior to RAPD and ISSR and can be effectively utilized for further characterization of loose smut pathogen.     

II. Cereal disease: the interplay between resistance and pathogenicity: Inheritance of pathogenicity in Gaeumannomyces graminis var. tritici

Variation in host response of isolates of the eyespot pathogen from different sources was examined over a number of years. Pathogen types were found in intensively-cropped couch-infested cereal sites that were almost as virulent on Agropyron repens (couch) as on wheat or barley. The commonly occurring wheat (W) type isolates from couch-free cereal crops were virulent on wheat and barley but avirulent on couch. Couch (C) types were isolated not only from couch but also from wheat, barley and oat crops with couch infestation. In pathogenicity tests on rye, C. types did not differ in virulence from the more commonly occurring W types. Aegilops ventricosa was equally resistant to both types. W type isolates from wheat and barley were examined to assess differential pathogenicity on wheat and barley. Sequential cropping with single cereal crops was used to separate out possible specific types. Isolates from fourth wheat and fourth barley crops were more pathogenic on the original than on the alternative host. When comparisons were made between isolates from third and fifth consecutive wheat and barley crops only those from barley showed a preference for the original host. An experiment comparing isolates from third and seventh consecutive wheat and barley crops showed a decline in virulence from the short to the longer sequences on the alternative but not on the original host.     

Effect of previous crops on the incidence of eyespot on winter wheat

Surveys of winter wheat from 1939 to 1946 show that eyespot (Cercosporella herpotrichoides Fron.) occurs throughout Britain and that its incidence depends largely on previous cropping and on weather. Examination of 551 crops on land whose cropping for the previous 4 years was known showed that the incidence rose steadily with increasing numbers of preceding wheat and barley crops: where neither crop had been taken for 4 years the proportion of crops with more than 70% infected straws was 2%, rising to 45% where three or four such crops had been taken and the average straws infected rose from 6 to 55%.
The percentage infection to be expected in various groups of crops was calculated from previous cropping; it was compared with the actual infection and so used to assess the importance of other factors in determining the incidence of eyespot. High spring rainfall, early sowing and a dense plant increased incidence and low spring rainfall, late sowing and a thin plant reduced it.
Eyespot was not usually severe on newly ploughed grassland until the third or fourth crop of wheat, but under very wet conditions it was sometimes severe in the second crop.
Oats is much less susceptible than wheat or barley, but some crops were found with a third of their straws infected.
A brief survey of winter wheat in Holland suggested possible causes for the rise and fall of eyespot in recent years and for its present lower incidence there as compared with East Anglia.     

Genetic variation, pathogenicity and mycelial growth rate differentiation between Gaeumannomyces graminis var. tritici isolates derived from winter and spring wheat

The variability of Gaeumannomyces graminis var. tritici ( Ggt ) isolates was evaluated at an intravarietal level using non-molecular and molecular methods. Pathogenicity and linear growth rate of the pathogen were estimated. Very high pathogenicity was found in 44% of the isolates, medium in 20% and low only in 8%. Significant differences in mycelial growth rate were observed. The quickest linear growth rate of Ggt mycelium was observed at 25°C. Isolates derived from winter wheat grew faster than those obtained from spring wheat. The correlation between growth rates and pathogenicity was not significant. DNA polymorphism determined by random amplified polymorphic DNA (RAPD)–PCR was used to assess genetic variation among isolates. Thirty-two RAPD markers revealed DNA polymorphism suitable for assessing variability among isolates examined. Cluster analysis of RAPD data identified a few groups of isolates. RAPD markers associated with pathogenicity as well as mycelium growth rate were found.     

EFFECTS OF CULTURAL TREATMENTS ON WHEAT AND ON THE INCIDENCE OF EYESPOT, LODGING, TAKE-ALL AND WEEDS. Field Experiments 1945–8

An experiment was made on the fourth, fifth and sixth successive crops of winter wheat to determine the effects of various treatments on the troubles which result from close cereal cropping. Eyespot and lodging were prevalent in the first year (1946); weeds in the second; eyespot, lodging, take-all and weeds in the third.
Spraying with H₂SO₄ reduced the incidence of eyespot, lodging and weeds, and increased yield of grain on plots which received sulphate of ammonia (by 2.7, 2.2 and 10.0 cwt./acre in successive years).
Sulphate of ammonia increased the incidence at harvest of eyespot and lodging, reduced take-all and consistently increased yield of straw. Eyespot and lodging reduced the effect of the fertilizer on yield of grain, take-all increased it.
Increase in seed rate increased the incidence of severe eyespot and of take-all; it increased lodging except when plants were dwarfed by take-all.
Weight of straw and percentage straws with severe eyespot lesions independently affected lodging, together accounting for 51% of the variance in percentage area lodged at harvest and 64 % of that lodged 33 days earlier.
Mean yields of grain on untreated plots sown with 3.3 1/2 bushels seed/acre fell from 26.0 to 22.5 to 11.7 cwt./acre in successive years, whereas yields of 28.4, 29.9 and 29.1 cwt./acre were obtained on sprayed plots sown with 1 1/2.2 bushels seed/acre which received 4 cwt./acre sulphate of ammonia, showing that high yields were maintained when eyespot, lodging, take-all and weeds were controlled.
By 1948 yields of grain on unsprayed plots had fallen to the level of those on similarly manured plots on the continuous wheat experiment on Broadbalk field. Spraying increased grain by amounts similar to those resulting from one year's fallow on Broadbalk; but fallow had its greatest effects on plots with low nitrogen, spraying on those with high nitrogen.     

Exploiting genotypic diversity of 2,4-diacetylphloroglucinol-producing Pseudomonas spp.: characterization of superior root-colonizing P. fluorescens strain Q8r1-96

The genotypic diversity that occurs in natural populations of antagonistic microorganisms provides an enormous resource for improving biological control of plant diseases. In this study, we determined the diversity of indigenous 2,4-diacetylphloroglucinol (DAPG)-producing Pseudomonas spp. occurring on roots of wheat grown in a soil naturally suppressive to take-all disease of wheat. Among 101 isolates, 16 different groups were identified by random amplified polymorphic DNA (RAPD) analysis. One RAPD group made up 50% of the total population of DAPG-producing Pseudomonas spp. Both short- and long-term studies indicated that this dominant genotype, exemplified by P. fluorescens Q8r1-96, is highly adapted to the wheat rhizosphere. Q8r1-96 requires a much lower dose (only 10 to 100 CFU seed(-1) or soil(-1)) to establish high rhizosphere population densities (10(7) CFU g of root(-1)) than Q2-87 and 1M1-96, two genotypically different, DAPG-producing P. fluorescens strains. Q8r1-96 maintained a rhizosphere population density of approximately 10(5) CFU g of root(-1) after eight successive growth cycles of wheat in three different, raw virgin soils, whereas populations of Q2-87 and 1M1-96 dropped relatively quickly after five cycles and were not detectable after seven cycles. In short-term studies, strains Q8r1-96, Q2-87, and 1M1-96 did not differ in their ability to suppress take-all. After eight successive growth cycles, however, Q8r1-96 still provided control of take-all to the same level as obtained in the take-all suppressive soil, whereas Q2-87 and 1M1-96 gave no control anymore. Biochemical analyses indicated that the superior rhizosphere competence of Q8r1-96 is not related to in situ DAPG production levels. We postulate that certain rhizobacterial genotypes have evolved a preference for colonization of specific crops. By exploiting diversity of antagonistic rhizobacteria that share a common trait, biological control can be improved significantly.     

Soil-applied fungicides for controlling take-all in field experiments with winter wheat

Soil treatment fungicides were tested against take-all (Gaeumannomyces graminis var. tritici) in three field experiments with winter wheat. Fungicides were applied as drenches either before sowing in autumn, and incorporated by rotary harrowing, or to the crop in spring. The most effective treatments were autumn applied benomyl (20 kg/ha) and nuarimol (0·55-4·4 kg/ha). However, the highest nuarimol concentration depressed yield. Benomyl sometimes induced a resurgence of take-all in the second wheat crop after treatment. Nuarimol had no adverse effects in subsequent crops, and neither fungicide hindered the onset of take-all decline in a third crop after treatment. The possible value of soil treatment in future control strategies is discussed.     

EFFECTS OF PREVIOUS WHEAT CROPS, SEED-RATE AND NITROGEN ON EYESPOT, TAKE-ALL, WEEDS AND YIELDS OF TWO VARIETIES OF WINTER WHEAT: FIELD EXPERIMENT 1954-56

In a replicated field experiment mean yields of wheat from plots that, in the preceding 2 years, had carried oats, beans or potatoes were 39.2 and 42.6 cwt. per acre in 1954 for Holdfast and Cappelle, respectively; 42.8 and 55.8 in 1955 and 34.9 and 49.6 in 1956. Previous wheat crops had more effect than any other treatment in increasing the incidence of eyespot, take-all and weeds and in decreasing the number of ears per unit area and the yield of grain. In 1956 on plots carrying the first, second and third successive wheat crops the percentages of straws with eyespot were respectively 12, 54 and 42 and with take-all 0.1, 1 and 16. Cappelle was less severely infected by eyespot than Holdfast. The second and third successive wheat crops yielded an average of 23.3 cwt./acre less than the first wheat crop. Cappelle consistently yielded more than Holdfast, the mean difference being 13.8 cwt./acre after potatoes but only 3.8 cwt./acre after two wheat crops. The higher seed-rate gave an average increase in grain yield of 3.3 cwt./acre; but where eyespot and take-all were both severe the lower seed-rate yielded as much total and more dressed grain than the higher. Wheat given a spring top dressing of 6 cwt./acre Nitro-Chalk yielded an average of 4 cwt./acre more grain than wheat given 3 cwt./acre.     

AFLP和RAPD标记技术在栉孔扇贝遗传多样性研究中的应用比较

AFLP和RAPD标记技术是近年来发展最快的基于PCR基础上的两种DNA标记技术,本文比较了两种标记技术在我国栉孔扇贝群体遗传多样性研究中的应用。共筛选20个RAPD引物和7个AFLP引物组合,检测到AFLP标记的有效等位基因数和平均多态信息量稍低于RAPD标记,但AFLP标记在每单位分析中扩增到的野生和养殖群体的多态性条带数(23．8,24．8)分别高于RAPD标记(5．6,5．6),AFLP多态性检测效率显著高于RAPD标记。AFLP和RAPD两种标记技术所揭示的野生种群与养殖群体间的近交系数、遗传距离两项指标均表明,我国栉孔扇贝养殖群体和野生种群之间尚未出现明显的遗传分化。研究结果表明：RAPD和AFLP这两种标记技术均可用于栉孔扇贝遗传多样性的分析,其分析结果是一致的。     

Natural suppression of take-all disease of wheat in Montana soils

This research was initiated to determine whether soils suppressive to take-all of wheat caused by Gaeumannomyces graminis var. tritici (Ggt) occur in Montana, and to identify the organisms most likely involved in this suppression. From an initial screening of eight soils collected from different wheat growing areas of Montana, two were highly suppressive to take-all. Microbial characterization of these soils indicated that different mechanisms were involved in the suppression. In Larslan soil, mycoparasitism appeared to be the main mechanism. Two different fungi with exceptional ability to reduce the severity of take-all were isolated from this soil. One of these fungi could parasitize the hyphae of Ggt. Field tests with these fungi in Ggt infested soil showed increases of over 100% in both harvestble tillers and grain yield as compared to treatments without these two fungi. In tests with 48 different bacteria and 10 actinomycetes from Larslan soil, none were able to consistently reduce severity of take-all alone, or in mixtures. In Toston soil, antibiosis by actinomycetes and perhaps the involvement of Pseudomonas spp. in production of antibiotics and/or siderophores appeared to be the most likely mechanisms involved in take-all suppression. Increases in shoot dry weight over that in the Ggt infested control using mixtures of pseudomonads and actinomycetes ranged from 25% to 87%. Actinomycetes added individually or in mixtures to soil infested with Ggt consistently reduced the severity of the disease to a greater extent than did mixtures of Pseudomonas spp.     

An experimental design and procedures for testing putative controls against naturally-occurring take-all in the field

In experiments during 1983–86 take-all was more severe and eyespot and sharp eyespot less frequent in 2nd-4th crops of winter wheat at Woburn (Beds.) than at Rothamsted (Herts.). Third crops had most take-all and yielded least grain. Against this background, small plots, 37 cm × 31 cm, in which all plants were sampled, were tried as a means of increasing experimental precision. They were arranged in fours in incomplete blocks and blocks with complementary treatments (putative controls of take-all) were paired. Thirty of these block-pairs were distributed throughout each experimental site in each year to provide one replicate of the design for each of three sampling times: April, June and August. Unattributed variation in disease and plant growth for plots within blocks was compared to that in other strata (block-pairs and blocks within block-pairs) of the experiment. The variability amongst block-pairs scattered throughout the site was nearly always greater than that for blocks within block-pairs (98% of take-all assessments, 71% of soil infectivity estimates, 94% of eyespot and sharp eyespot assessments and 86% of all plant measurements). The variability of blocks within block-pairs exceeded that of plots much less frequently (56% and 69% of take-all assessments, 33% and 25% of soil infectivity measurements, 63% and 56% of eyespot and sharp eyespot assessments and 50% and 63% of plant measurements; Rothamsted and Woburn, respectively). Small plots were judged mostly on this last comparison, where a variance ratio in excess of 1 indicated that the small plots had decreased variability and increased precision. Variance ratios for different assessments of take-all indicates that small plots: i) most consistently decreased disease variability during the years of maximum disease, ii) were slightly less effective at Rothamsted than at Woburn, and iii) were usually less effective in fourth crops than in previous crops. Soil infectivity was most uniform after crops with most disease and blocks were rarely more variable than plots. hxcept when disease was severe, soil infectivity in August tended to be positively associated with the yield of the crop just harvested. These findings reveal changes in the scale of disease patterns, both during the crop sequence and within individual crops, and suggest more than one scale of pattern in take-all-infested fields. This is discussed in relation to field experimentation and take-all decline.     

Genetic variability in Puccinia striiformis f. sp. tritici populations sampled on a local scale during natural epidemics

This study investigated genetic polymorphism on a local scale in Puccinia striiformis f. sp. tritici populations during natural epidemics, in four fields located in northern France and sampled in 1998 or 1999. Two hundred and forty-seven isolates were analyzed for their amplified fragment length polymorphism (AFLP) pattern through four primer combinations, and 194 of them were also tested for their virulence factors. Only one or two pathotypes were found in each field, and all isolates had virulence v17, matching the recently introduced Yr17 resistance gene. Polymorphism on a field scale was low. Although 67 loci were polymorphic, 77% of the isolates had the same AFLP pattern, all other patterns being rare or unique. Analyses of the genetic distance between AFLP patterns based on the Jaccard index allowed us to define 12 groups, but a bootstrap analysis showed that all isolates could be assigned to a single clonal lineage. This leads us to conclude that P. striiformis f. sp. tritici populations are clonal on a field scale in northern France.     

Soil conduciveness to take-all of wheat: Influence of the nitrogen fertilizers on the structure of populations of fluorescent pseudomonads

In a field cropped with wheat, a high and low level of soil conduciveness to take-all were induced by applying a nitrogen fertilizer with either calcium nitrate or ammonium sulphate. From these two soils, two representative populations of fluorescent pseudomonads were tested for their in situ behaviour. Take-all index and root dry weight were assessed on plants cropped in soils infested with Gaeumannomyces graminis var tritici (Ggt) and each bacterized with one of the isolates of fluorescent pseudomonads. The bacteria tested can be split into three groups: antagonists which reduce take-all, deleterious isolates which aggravate the disease and neutral without evident effect on the disease. The predominance of antagonistic fluorescent pseudomonads in the NH₄-treated soil and the predominance of deleterious ones in the NO₃-treated soil was confirmed after statistical analysis. The microbial impact on take-all must be more considered as the resulting effect of divergent activities of both rhizobacteria types than the only consequences of the presence of antagonistic pseudomonads. All the high cyanogenic pseudomonads were antagonists in situ and were more numerous in the NH₄-treated soil than in the NO₃-treated soil.