Hyphal density as a measure of suppression of Gaeumannomyces graminis var. tritici on wheat roots

Runner hyphae of Gaeumannomyces graminis (Sacc.) Arx & Olivier var. tritici Walker on seminal roots of wheat seedlings were photographed and their length measured. As well, their length was estimated using the line-intercept method. The correlation of 0.904 between measured and estimated lengths of hyphae was highly significant. This line intercept method was used to estimate the density (length/unit area) of hyphae on roots of plants growing in the presence and absence of a soil suppressive to G. graminis var. tritici. Estimations were made eight times during 28 days growth at 15°C. In fumigated soil (non-suppressive) inoculated with 0.1% ground oat grain infested with G. graminis var. tritici, the density of hyphae on roots started to increase at five days compared with 15 days when soil there was a 10.8% cover of the root surface after 15 days when the hyphae had reached maximum density. Suppression to G. graminis var. tritici is normally detected by a difference in disease rating of roots at 28 days but this study has shown that suppression can be demonstrated by the difference in the density of hyphae if roots are examined between seven and 19 days.     

Cross-protection against the wheat and oat take-all fungi by Gaeumannomyces graminis var. graminis

Glasshouse experiments have shown that the prior colonisation of wheat roots by Gaeumannomyces graminis var. graminis, a fungus closely related to the wheat and oat take-all fungi but non-pathogenic to temperate cereals, reduced take-all infection along the roots. Cross-protected wheat plants produced grain yields significantly greater than those of unprotected plants but not significantly different to those of healthy wheat plants. A Phialophora-like fungus from grass roots did not confer the same degree of protection. There is some evidence that the cross-protection mechanism may be a specific host response nduced by var. graminis. The possible use of var. graminis in the biological control of take-all is discussed.     

Dynamics of root colonization by the take-all fungus, Gaeumannomyces graminis

The progressive colonization of wheat seminal roots by Gaeumannomyces graminis var. tritici was monitored following inoculation by single inoculum units. G. graminis grew equally well above and below inoculation sites prior to blockage of the stele but after this growth was favoured up roots, above inoculation sites, rather than down, resulting in an asymmetrical pattern of root colonization. This asymmetrical pattern was common to superficial and cortical runner hyphae. It is suggested that cessation of host assimilate supply to the distal portion of infected roots inhibited further extensive growth of G. graminis. This hypothesis was tested by comparing extents of colonization by G. graminis on seminal roots of wheat with normal, enhanced and diminished assimilate supplies. A diminished assimilate supply to infected roots retarded the extent of pathogen colonization.     

Soils suppressive to Gaeumannomyces graminis var. tritici: Effects on other fungi

The effect of soils suppressive to Gaumannomyces graminis var. tritici (Ggt) on the severity of root and crown rots caused by Rhizoctonia solani, Gibberella zeae, Pythium irregulare, Cochliobolus sativus and Fusarium culmorum was tested in pot bioassays. An induced suppressive soil was obtained from the rhizosphere of wheat plants grown at 15°C for 28 days in fumigated soil inoculated with live inoculum (colonized oat grain) of Ggt.Root rot caused by R. solani was significantly less in soil amended with either induced or naturally suppressive soil. Disease caused by the other pathogens was also reduced by the induced suppressive soil, with the least reduction occurring with F. culmorum.Colonization of the surfaces of seminal roots of wheat plants by Gaeumannomyces graminis var. graminis (Ggg) and a Phialophora-like fungus (Plf 119) was also studied using the line-intercept method. In non-suppressive soil the maximum area of the primary seminal root colonized by Ggg was 7.4 per cent and by Plf 119 was 3.3 per cent. Colonization of roots by Ggg and Plf 119 was reduced substantially by the addition of induced suppressive soil.     

Soils suppressive to Gaeumannomyces graminis var. tritici: Induction by other fungi

Rhizosphere soils were obtained from wheat plants growing in fumigated soil inoculated with one of the pathogens, Rhizoctonia solani, Gibberella zeae, Fusarium culmorum, Cochliobolus sativus and Pythium irregulare or one of the non-pathogenic fungi, Gaeumannomyces graminis var. graminis (Ggg) or a Phialophora-like fungus (Plf). Using a pot bioassay, these soils were tested for suppression of Gaeumanomyces graminis var. tritici (Ggt) and the fungus involved in the initial induction. G. zeae was the only fungus that induced suppression to Ggt and to itself. Ggg but not Plf induced suppression to Ggt although both fungi induced suppression to themselves.Fungi capable of inducing suppression of take-all have two characteristics in common, they induce suppression of themselves and their saprophytic survival is restricted to organic matter.     

Age of wheat and barley roots and infection by gaeumannomyces graminis var tritici

Seminal roots of wheat and barley seedlings were inoculated with G. graminis var tritici on regions 0, 5- and 15-days old, and assessed for intensity and extent of infection after standard times. Wheat roots were most heavily infected on young regions, whereas barley roots were most heavily infected on old ones. The effect of root age in wheat was similar in both unsterile and aseptic conditions, so it could not be ascribed to saprophytic rhizosphere micro-organisms interacting with G. graminis.The contrasting results for wheat and barley are explained by a single hypothesis, based on decreasing host-resistance in the root cortex but increasing resistance at or near the endodermis as the roots age. It is suggested that, under some conditions, even small amounts of non-pathogenic root cortex death can enhance infection by G. graiminis. This interpretation may explain several aspects of take-all and its biological control by other dark mycelial parasites.     

Growth of Gaeumannomyces graminis var. tritici in soil: Effects of temperature and water potential

The effects of temperature and water on the growth of the take-all fungus, Gaeumannomyces graminis var. tritici (Ggt), were examined in two factorial experiments. The first examined the effects of temperature and water potential on the growth of two isolates of Ggt on agar media, using osmotically-adjusted water potentials. The second experiment was concerned with the growth of the Ggt isolates in one sterile and two natural soils at two water regimes in the absence of a living host. Three temperatures (10, 18 and 26°C) were used in these experiments. A third experiment determined growth through soil.Growth was greatest at high temperatures and low water potential in axenic culture, but in unsterile soil growth at different temperatures and water potentials was strongly influenced by competition from the soil biota. The best temperature for growth in unsterile soil was 18°C. Growth at 26°C in unsterile soil was greatly reduced, this being attributed to more intense microbial competition. In sterile soil Ggt grew equally well at 18 and 26°C. At 10°C, both isolates of Ggt grew better in unsterile soil than in sterile soil.Under suitable conditions Ggt grew out readily from infected straw into unsterile soil (up to 5 cm in 10 days) in the absence of a host plant, forming melanized, hyaline and branched hyphae. These hyphae were infectious after dry storage for 5 months in the laboratory. Ggt thus appears to be a more successful soil inhabitant than is widely believed. Our experiments could explain many of the host-based concepts related to field expression of disease.The technique presented here could be of value for testing the suppressiveness or conduciveness of soils by measuring fungal growth in soil.     

Colonisation of seminal roots of wheat and barley by egg-parasitic nematophagous fungi and their effects on Gaeumannomyces graminis var. tritici and development of root-rot

This study provides evidence that egg-parasitic nematophagous fungi, Pochonia chlamydosporia, Pochonia rubescens and Lecanicillium lecanii, can also reduce root colonisation and root damage by a fungal pathogen. Interactions of nematophagous fungi with the take-all fungus, Gaeumannomyces graminis var. tritici (Ggt), and their influence on severity of the root disease it causes were studied in laboratory and pot experiments. In Petri dish experiments the three nematophagous fungi reduced colonisation of barley roots by Ggt and also reduced necrotic symptoms. On the contrary, root colonisation by nematophagous fungi was unaffected by Ggt. In growth tube experiments, the three nematophagous fungi again reduced Ggt root colonisation and increased effective root length of barley seedlings. This was true for both simultaneous and sequential inoculation of nematophagous fungi versus Ggt. In the pot experiments the inoculum of the tested fungi in soil was applied in the same pot, as a mixture or in layers, or in coupled pots used for wheat grown with a split-root system. The nematophagous fungi P. chlamydosporia (isolate 4624) and L. lecanii (isolate 4629), mixed with Ggt or in split root systems with the pathogen, promoted growth of wheat (i.e. increased shoot weight), although no disease reduction was found. In split root systems, lower levels of peroxidase activity were found in seedlings inoculated with Ggt in combination with the nematophagous isolates 4624 and 4629 than when the take-all fungus was applied alone.Our results show that nematophagous fungi reduce root colonisation by Ggt, root damage and stress induced senescence in Ggt-inoculated plants.     

The incidence of Gaeumannomyces graminis var. Tritici and Phialophora radicicola var. Graminicola on wheat grown after different cropping sequences

The effects of three treatment cropping sequences (fallow, lucerne or grass-clover ley) on the incidence of Gaeumannomyces graminis var. tritici and Phialophora radicicola var. graminicola were measured in a field experiment. Increases in G. g. tritici population in the soils of the first wheat crop and the incidence of take-all in the second wheat crop were greater after fallow or lucerne than after grass-clover. These differential increases were not associated with differences in survival of G. g. tritici during the treatment cropping but were correlated negatively with the population of P.r. graminicola in the soil. After the third wheat crop the P. r. graminicola population after grass-clover had decreased and take-all was as prevalent as after fallow or lucerne.     

Biological control of the take-all fungus, Gaeumannomyces graminis, by Phialophora radicicola and similar fungi

Phialophora radicicola is an avirulent fungal root-parasite of grasses and cereals, with runner hyphae like those of Gaeumannomyces graminis. Weakly and non-pathogenic varieties of these fungi control the pathogens, G. graminis vars. tritici and avenue. Biology of these fungi is considered and the evidence for biological control and possible mechanisms reviewed; control is probably widespread in natural plant communities, and host-mediated, perhaps by induction of plant resistance mechanisms.Prospects for application of biological control seem best for P. radicicola var. graminicola established on grass crops, as this is already exploited in British agriculture. New evidence is presented on the effects of grassland factors on this fungus, especially sward composition, age, mineral nutrition and management practices: its population might often be limited by the rate of new root production to replace those with cortices already colonized. Prospects for control by seed inoculation with P. radicicola var. radicicola and G. graminis var. graminis also seem good, but possible dangers of introducing them into cereal cropping are emphasized. The weak pathogens might be used also for indirect control by establishing hyper-parasites or inducing disease suppression (like take-all decline) in soils, but there is no evidence for ‘Phialophora decline’, at least in well-managed grasslands. Finally, different biocontrols of take-all might be combined, and biological with chemical ones for ophiobolus patch disease of turf.P. radicicola var. graminicola has a slight beneficial effect on grass yield, even when the pathogens, G. graminis vars. tritici and arenae arc absent; this probably contributes to its abundance in natural grasslands in Britain. The scale of biological control by this and similar fungi might explain why, in their absence, effective plant resistance to G. graminis is uncommon in the Gramineae.     

Infection of wheat seminal roots by varieties of Phialophora radicicola and Gaeumannomyces graminis

The growth of isolates of Phialophora radicicola var. radicicola, P. radicicola var. graminicola, Gaeumannomyces graminis var. graminis, G. graminis var. tritici and Leptosphaeria narmari was compared on the coleoptiles and roots of wheat seedlings. Fungal growth was measured as the extent and density of dark runner hyphae. All except P. radicicola var. graminicola grew on coleoptiles and all grew on roots although only G. graminis var. tritici extensively colonized the root stele. Growth rate on roots was positively correlated with that on agar, P. radicicola var. graminicola and L. narmari growing at about half the rate of the other fungi; hyphal density was high for P. radicicola var. graminicola but relatively low for the other fungi. For P. radicicola var. radicicola, P. radicicola var. graminicola and G. graminis var. tritici growing from buried inocula, the extent and density of hyphae up roots towards the seed was similar to that down, but G. graminis var. tritici caused chocolate-brown stelar discoloration up roots only.Root invasion by P. radicicola var. radicicola, P. radicicola var. graminicola and G. graminis var. tritici was described from sections. Each gave a different pattern of hyphae and host response within an inoculum layer, and progressive changes occurred away from the inoculum. Studies of the rate of penetration by each fungus and the rate and pattern of death of cortical cells explained the differences between fungi. G. graminis var. tritici penetrated living cells in advance of other soil micro-organisms, and hence by hyaline hyphae inducing much lignituber formation as a host resistance reaction. P. radicicola var. graminicola penetrated only senescent or dead cells in association with other soil microorganisms, and hence by dark hyphae, inducing little lignituber formation. P. radicicola var. radicicola was intermediate in all these respects. The high hyphal density of P. radicicola var. graminicola was due to the colonization of cortical cells and spaces by dark, clearly visible, rather than hyaline hyphae, which are invisible in unstained roots. Cell death in the outer cortex explained the observed progressive restriction of growth by all fungi to the inner cortex with increasing distance from the inoculum. Spread by G. graminis var. tritici up roots was ectotrophic relative to the stele but down roots hyphae spread rapidly within the stele. Stelar reactions suggested as resistance mechanisms occurred up roots only. Their absence down roots is attributed to infection disrupting stelar transport.     

The bioprotective effect of AM root colonization against the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici in barley depends on the barley variety

The systemic effect of root colonization by the arbuscular mycorrhizal fungus (AMF) Glomus mosseae on the susceptibility of old and modern barley varieties to the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici (Ggt) was studied in a split-root system. Plants were precolonized on one side of the split-root system with the AMF and thereafter the other side of the split-root system was inoculated with the pathogen. At the end of the experiment the level of bioprotection was estimated by quantifying lesioned roots and the determination of the root fresh weight. AM root colonization provided protection in some of the barley genotypes tested, but not in others. This protective effect seemed to vary in the oldest and the most modern barley variety tested.     

Colonization of wheat roots by Gaeumannomyces graminis inhibited by specific soils,microorganisms and ammonium-nitrogen

Lineal extension of Gaeumannomyces graminis var. tritici hyphae along roots of intact wheat plants growing in soils was measured. Hyphal growth rates were lower in soils treated with NH₄⁺-N than with NO₃^?-N. In a soil that is suppressive to the take-all disease, the controlling influence of NH₄⁺-N was eliminated by soil fumigation (methyl bromide), and reintroduced to fumigated soil by additions of 1% nonsterile soil. Effects of fumigation on hyphal growth were absent in a nonsuppressive soil, and in NO₃^?-treatments of the suppressive soil. When inocula of selected groups of wheat rhizoplane microflora were reintroduced into a fumigated or a soil-reinoculated soil via a root-food base, the Pseudomonas spp. consistently appeared more suppressive in NH₄⁺-N treatments than the general bacterial flora, Bacillus spp. spores, streptomycetes, and fungi.     

Effects of wheatfield soil on inocula of gaeumannomyces graminis (Sacc.) Arx & Olivier var. tritici J. Walker in relation to take-all decline

Organic debris extracted from wheatfield soils was found to be significantly more infective following 2–3 yr continuous wheat than after the fifth successive crop. Plots with added nitrogen yielded more infective debris than those without. Short periods of contact with decline soil or its suspension, reduced infection of wheat roots by Gaeumannomyces when they were subsequently grown in non-decline soil. Hyphae emerging from previously infected root pieces or culture inocula showed a positive growth response to wheat roots or their exudates. This response was negatively correlated to the distance between root and inoculum. In the presence of decline soil, hyphal response was significantly reduced. This reduction can be related to the decreased infectivity of decline soil and may help to explain take-all decline.     

Soil moisture affects the interaction between Gaeumannomyces graminis var. Tritici and antagonistic bacteria

Reduction in the soil moisture from a potential of ?10 to?108 kPa reduced the growth of Gaeumannomyces graminis. The antagonist Bacillus cereus only spread through the soil or sand at potentials between ?10 and ?60 kPa. The spread of B. pumilus was not affected by the water potential within the range tested. Both B. cereus and B. pumilus were less antagonistic in the drier sand and soil. When the bacteria and the fungus were grown together the maximum growth of the fungus occurred at those potentials when bacterial antagonism was reduced but the fungus was still able to grow.     

Infectivity of lesioned wheat root tissue related to the presence of dehydrogenase enzymes in hyphae of Gaeumannomyces graminis var. Tritici in the lesions

Transverse sections of lesioned tissue taken from wheat roots grown in soil naturally infested with Gaeumannomyces graminis var, Tritici were stained with trypan blue and the area of stele occupied by hyphae or by brown host deposits was measured. The area of mycelium in lesioned pieces taken from seedling or tillering plants and used as inoculum in host infectivity tests was positively correlated with the disease produced and the area of brown deposits in lesioned pieces taken from tillering or mature plants was negatively correlated. Whole pieces of lesioned tissue were examined cytochemically for glutamic and succinic dehydrogenases in the invading hyphae. Groups of host cells in the endodermal region were filled with hyphae showing positive reactions for both dehydrogenases (active) and separated by areas of brown discoloured host tissue containing few active hyphae. Less than half the discoloured lesion was occupied by active hyphae. The area of lesion containing hyphae with dehydrogenase enzymes was positively correlated with the measure of disease severity of the roots and with the infectiveness of the lesioned tissue when inoculated on to axenic wheat seedlings. The progress of infection in axenie seedlings inoculated at 3 or 8 cm from the seed differed with the two placements, notably in the host response and the growth of active hyphae in the cortex.     

Antagonists of Gaeumannomyces graminis from the rhizoplane of wheat in soils fertilized with ammonium- or nitrate-nitrogen

Take-all of wheat caused by Gaeumannomyces graminis var. tritici was less when soils in glasshouse pots were fertilized with NH₄⁺-N than with NO₃^?-N. The form of N did not alter countable populations of microorganisms in the rhizosphere or rhizoplane, but altered the numbers of bacteria and streptomycetes that inhibited the pathogen's growth in vitro. The pH of the medium used to isolate these microorganisms, whether similar or dissimilar to the pH of the rhizosphere, had some influence both upon countable populations and upon the proportions of antagonists. Highest counts of the rhizoplane microflora were on agar media with a pH similar to that of the soil. Most antagonists were isolated from a soil that is physically and chemically conducive to parasitism of wheat roots by Gaeumannomyces, but which contains a microflora suppressive toward the parasitic colonization of the roots. Isolates of the general bacterial flora, of Pseudomonas spp. and of streptomycetes, but not of Bacillus spp. inhibited the in vitro growth of G. graminis.     

Pathozones of genetic subtypes of Gaeumannomyces graminis in cereals

The extent of damage to the host plant caused by Gaeumannomyces graminis var. tritici (Ggt) and var. graminis (Ggg) is a result of a net effect of host susceptibility and mycelium infectivity. The disease severity on cereal roots caused by G. graminis (Gg) fungi varies considerably depending on the genetic subtypes. Results of our rhizobox placement experiments additionally showed a subtype-specific effect of the spatial distance between host and fungus on the infection. The highest pathogenicity of each subtype was found in different zones of the root system: pathozones of different subtypes alternated along the root. The extent of the pathozone profiles did not depend on the infectivity of the inoculum and plant age. However, disease severity was shown to be affected by defence reactions of the host plant. An attack of a fungal subtype that is easily recognized by the host plant leads to defence reactions like increased root growth, thus minimizing the damage to the shoot. Detailed analysis showed that a Ggt subtype had a high potential for colonizing root laterals. It formed concentric zones of high colonization efficiency at a distance of ca. 5 cm around the shoot.     

Saprophytic survival of the take-all fungus and its antagonist, Gaeumannomyces graminis var. graminis under conventional and no-tillage

The saprophytic survival of the take-all fungus, Gaeumannomyces graminis var. tritici, and its antagonist, G. graminis var. graminis, was studied over 9-month periods in 1982 and 1983 under conventional and no-tillage cultivation. In 1982, when the rainfall for the 9-month period was 30.4% below the long-term average, the survival of the fungi was comparable under the two tillage systems. However, the survival of G. graminis var. graminis was significantly greater (P 0.05) than that of G. graminis var. tritici at each sampling time. In 1983, when the rainfall for the 9-month period was 25.1% above the long-term average, the survival of both fungi was significantly greater under no-tillage than under conventional tillage. The survival of the two isolates of G. graminis var. graminis after 3 months was significantly greater (P 0.05) than that of the two isolates of the pathogen. The implications for the greater survival of the fungal antagonist compared to the take-all fungus under no-tillage are discussed in relation to the biological control of take-all.     

The role of bacteria in the biological control of Gaeumannomyces graminis by suppressive soils

The suppression of Gaeumannomyces graminis var. tritici by certain soils or following certain soil treatments is considered to be an expression of either specific or general antagonism sensu Gerlagh (1968). Specific antagonism is effective in dilutions as high as 1 in 1,000, can be transferred from soil to soil, operates near or on wheat roots, is destroyed by 60°C moist heat for 30 min. or desiccation, is fostered by wheat monoculture but may be lost from a soil by fallow or rotation with certain crops, especially legume hay or pasture crops. Strains of Pseudomonas fluorescens may be involved. General antagonism is a soil property which cannot be transferred and is resistant to 80°C moist heat for 30 min, to methyl bromide and chloropicrin, but not to autoclaving. Take-all control by organic amendments, minimum tillage, or a soil temperature of 28°C may be expressions of increased general antagonism.In much of the southern Australian wheat belt, where take-all can cause heavy crop losses, some general but rarely specific antagonism is apparently operative. Both types of antagonism are probably operative in long-term wheat growing areas of the Pacific Northwest U.S.A. where take-all is virtually nonexistent.