The invasion and development of the cereal cyst-nematode, Heterodera avenae in the roots of autumn-and spring-sown cereals

Observations were made at 2 or 4 wk intervals from December to harvest on all stages of Heterodera avenae in winter oats growing on infested land. Second-stage larvae were present in all soil samples except on 5 and 20 July. Invasion and development of larvae was slow during winter. The nodal and seminal roots of winter oats were both heavily invaded by the nematode; larvae which invaded seminal roots tended to become male whereas those in nodal roots tended to become female. There was a small second invasion in August. Females were first observed on the roots of winter oats on 17 May, 214 days after the crop was sown and 62 days after the first fourth-stage larva was observed. The nodal roots of spring barley contained few H. avenae larvae whereas these roots were heavily invaded in winter wheat and oats. In spring barley the nodal roots were developing in June and July when few second-stage larvae were in the soil whereas in winter oats and wheat the nodal roots were growing rapidly in April when larvae were most numerous, and so were heavily invaded.     

Control of the take-all fungus byMicrodochium bolleyi,and interactions involvingM. bolleyi,Phialophora graminicola andPericonia macrospinosa on cereal roots

Summary In glasshouse experiments,Microdochium bolleyi (Mb) significantly reduced infection of wheat roots by the take-all fungus,Gaeumannomyces graminis vartritici (Ggt), when inocula were dispersed in soil at ratios of 10∶1 (Mb:Ggt) or more. Spread of take-all lesions up roots from a layer of inoculum also was reduced when Mb was inoculated immediately below the crown. In contrast,Periconia macrospinosa did not control take-all even at an inoculum ratio of 100∶1. M. bolleyi interfered with growth on roots byPhialophora graminicola, a known biocontrol agent of take-all. It is suggested that this phenomenon and control of take-all by these fungi occur by competition for cortical cells that senesce in the normal course of root development.     

The effect of minimum cultivation on the incidence of take-all down the root profile of winter wheat

The effects of direct drilling, shallow cultivation and ploughing on the infection of winter wheat roots by the take-all fungus (Gaeumannomyces graminis var. tritici) were studied on three field sites over a number of years. All three soil types were categorised by Cannell, Davies, Mackney & Pidgeon (1978) as suitable for sequential direct drilling. The results show that a smaller proportion of roots was infected at depth in the direct-drilled plots in May/June. However by July these differences had all but disappeared and an estimate of infection in the top 7 cm of the roots (approximately equivalent to traditional hand sampling for take-all) gave a reliable comparison of the total take-all on plants grown under these different cultivation systems.     

The effect of Heterodera avenae, on the root system of susceptible and resistant oat seedlings

The effects of invasion by the cereal cyst nematode, Heterodera avenae, on the root growth of susceptible and resistant oat seedlings were similar. Nematode invasion directly inhibited root extension and lateral root production, which was dependent upon the developmental status of the invaded root tissue and the nematode density cm-¹ root. Both responses are caused by mechanical damage due to nematode invasion leading to decreased top growth in the host. Reduced root development, decreased efficiency of water and nutrient uptake in invaded plants being indirect effects of nematode invasion.     

Are Pathogenesis-Related Proteins Induced by Meloidogne javanica or Heterodera avenae lnvasion?

Changes in root- and leaf-soluble proteins were investigated in tomato after invasion by the root-knot nematode Meloidogyne javanica, or in barley and wheat after invasion by the cereal cyst nematode Heterodera avenae. Infection of susceptible tomato plants by M. javanica did not cause any change in the soluble-protein composition of leaves or roots compared with uninoculated plants at an early infection stage. No pathogenesis-related proteins (chitinase, glucanase, or P-14) were induced in the leaf apoplast. Changes in leaf proteins were not observed after invasion of wheat cultivars by H. avenae, whereas, in barley, a few changes in intercellular leaf proteins were recorded in resistant cultivars. These changes, however, were not the same among different H. avenae-resistant cultivars. Protein changes were found at an early stage of infection in barley and wheat roots infected with H. avenae, but no difference was found between resistant and susceptible cultivars.     

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.     

Manganese nutrition and accumulation of phenolics and lignin as related to differential resistance of wheat genotypes to the take-all fungus

Differential resistance of four Triticum aestivum L. genotypes to isolates of take-all fungus (Gaeuman-nomyces graminis var. ritici Walker) was tested in a complete factorial experiment set up in a growth chamber using Mn-deficient Wangary sand amended with four rates of Mn. Mn-efficient cultivars produced more dry matter at low supply of Mn. Fertilization with Mn significantly increased its accumulation in roots and shoots. The most sensitive measure of take-all infection was the total length of root stellar lesions; these lesions were reduced by Mn fertilization and were shorter in Mn-efficient genotypes. The resistance-enhancing effect of Mn was the most obvious in the Mn-inefficient genotype (Bayonet) and the least obvious in the Mn-efficient one (C8MM). Phenolics biosynthesis in roots was clicited by fungal infection, especially in the case of the highly virulent isolate. The weakly virulent isolate increased phenolics concentration in roots much more if no Mn was added, indicating that the resistance-enhancing effect of Mn may not be directly exerted through the effects on phenolics biosynthesis. Lignin concentration in roots decreased due to Mn fertilization, while no effect of take-all infection was noted. It appears that biosynthesis of phenolics and lignin in wheat roots has a low Mn requirement which can be satisfied at environmental Mn concentrations below those necessary for optimum plant growth. ei]Section editor: A C Borstlap ei]Section editor: H Lambers     

Studies of the cereal cyst-nematode, Heterodera avenae under continuous cereals, 1975–1978. II. Fungal parasitism of nematode females and eggs

The development and fungal parasitism of Heterodera avenae females and eggs on susceptible cereals was studied from 1975 to 1978 in a sandy loam soil. Despite the production of many females on the roots nematode numbers declined in two years (1975 and 1978), and it is female survival and fecundity and not female numbers which often limit H. avenae multiplication. Fungal parasites may totally destroy females on roots or result in the formation of small cysts which are often empty. Fecundity is reduced and many eggs may became infected. Parasitism of females and eggs was decreased and nematode multiplication was increased in soil treated with formalin (38% formaldehyde) at 3000 litres/ha, but because it is nematicidal and fungicidal interpretation of the effects of the sterilant are difficult. Formalin has a greater effect on H. avenae multiplication in wet summers when fungal parasites, particularly Nematophthora gynophila are more active. Parasitised females which may be destroyed in about 7 days are fragile and difficult to extract from soil. Rates of parasitism tend to be underestimated. Approximately 60% of the females which failed to form cysts containing eggs can be accounted for by N. gynophila and Verticillium chlamydosporium. Fungal parasitism is therefore the major factor in limiting the multiplication of H. avenae.     

Use of a root assessment tray for the detection of take-all on lateral roots of wheat seedlings

A root assessment tray was designed for the meticulous assessment of take-all on wheat seedling roots from soil bioassays. Subsequently, the detection of lateral root infections (in addition to the more obvious infections on main axes of seminal roots) resulted in increased estimates of propagule numbers of the take-all fungus (Gaeumannomyces graminis var.tritici) for 196 of the 368 soil samples bioassayed in a field study conducted in Western Australia between 1984 and 1986.     

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.     

Plant genotype, micronutrient fertilization and take-all infection influence bacterial populations in the rhizosphere of wheat

The relationship between micronutrient efficiency of four wheat (Triticum aestivum L.) genotypes, tolerance to take-all disease (caused by Gaeumannomyces graminis (Sacc.) Arx and Olivier var. tritici Walker), and bacterial populations in the rhizosphere was tested in soil fertilized differentially with Zn and Mn. Plant growth was reduced by Mn or Zn deficiency and also by take-all. There was an inverse relationship between micronutrient efficiency of wheat genotypes when grown in deficient soils and the length of take-all lesions on roots (efficient genotypes had shorter lesions than inefficient ones). In comparison to the rhizosphere of control plants of genotypes Aroona and C8MM receiving sufficient Mn and Zn, the total numbers of bacterial cfu (colony forming units) were greater in the rhizosphere of Zn-efficient genotype Aroona under Zn deficiency and in Mn-efficient genotype C8MM under Mn deficiency. These effects were not observed in other genotypes. Take-all decreased the number of bacterial cfu in the rhizosphere of fully-fertilized plants but not of those subjected to either Mn or Zn deficiency. In contrast, the Zn deficiency treatment acted synergistically with take-all to increase the number of fluorescent pseudomonads in the rhizosphere. Although numbers of Mn-oxidising and Mn-reducing bacteria were generally low, take-all disease increased the number of Mn reducers in the rhizosphere of Mn-efficient genotypes Aroona and C8MM. Under Mn-deficiency conditions, the number of Mn reducers in the rhizosphere increased in Aroona but not in C8MM wheat. The results suggest that bacterial microflora may play a role in the expression of Mn and Zn efficiency and tolerance to take-all in some wheat genotypes.     

Cereal Cyst Nematode (Heterodera avenae) on Oats. II. Early Root Development and Nematode Tolerance

The effect of Heterodera avenae infestation on early seminal and lateral root growth was examined in four oat genotypes differing in tolerance to H. avenae. Recently emerged seminal roots were inoculated with a range of H. avenae larval densities, then transferred a hydroponic system to remove the effect of later nematode penetration on root development. Intolerance to H. avenae was assessed in terms of impairment of seminal root extension resulting in fewer primary lateral roots emerging from the seminal root below the zone of juvenile penetration. Tolerant plants infested with H. avenae had longer lateral root systems than infested intolerant plants. The decline in lateral root growth below the penetration zone was partly offset by increased growth above. This did not contribute to tolerance, however, as there were no differences between cultivars for this feature. Nematodes induced earlier nodal root emergence in all cultivars. Nodal root development was most advanced on the most tolerant cultivar.     

Impact of intercropping aphid-resistant wheat cultivars withoilseed rape on wheat aphid (Sitobion avenae) and its natural enemies

The effects of intercropping of wheat cultivars and oilseed rape on the densities of wheat aphid, Sitobion avenae, and their arthropod natural enemies were evaluated. Three winter wheat cultivars with different resistant levels to S. avenae were used: ‘KOK’ (high resistance), ‘Xiaobaidongmai’ (low resistance) and ‘Hongmanghong’ (susceptible). The results showed that the densities of S. avenae were significantly higher on the monoculture pattern than on either the 8-2 intercropping pattern (eight rows of wheat with two rows of oilseed rape) or the 8-4 intercropping pattern (eight rows of wheat with four rows of oilseed rape). The mean number of predators and the mummy rates of S. avenae were significantly higher in two intercropping patterns than those in the monoculture pattern. The densities of S. avenae, ladybeetles, and mummy rate of S. avenae were significantly different among different wheat cultivars. The highest densities of S. avenae and ladybeetles were found on wheat cultivar Hongmanghong. The lowest densities of S. avenae associated with high mummy rate of S. avenae were found on wheat cultivar Xiaobaidongmai. The results showed that wheat-oilseed rape intercropping conserved more predators and parasitoids than in wheat monoculture fields, and partial resistance of wheat cultivar Xiaobaidongmai had complementary or even synergistic effects on parasitoid of S. avenae.     

Biological control of take-all in wheat by endophytic Bacillus subtilis E1R-j and potential mode of action

The bacterial strain E1R-j, isolated as an endophyte from wheat roots, exhibited high antifungal activity to Gaeumannomyces graminis var. tritici (Ggt). Strain E1R-j was identified as Bacillus subtilis based on morphological, physiological and biochemical methods as well as on 16S rDNA analysis. This strain inhibited mycelium growth in vitro of numerous plant pathogenic fungi, especially of Ggt, Coniothyrium diplodiella, Phomopsis sp. and Sclerotinia sclerotiorum. In greenhouse experiments, soil drenches with cell densities of 10⁶, 10⁹ and 10¹² CFU ml⁻¹ E1R-j reduced significantly take-all disease, caused by Ggt, in wheat seedling by 62.6%, 68.6% and 70.7%, respectively, compared to the inoculated control, 4 weeks after sowing. Growth parameters such as lengths and fresh weights of roots and shoots of Ggt-inoculated control plants were significantly lower compared to Ggt-inoculated and E1R-j treated plants. Field experiments in the season 2006/2007, heights of wheat plants in the Ggt inoculated plots were significantly reduced compared to the non inoculated treatments. Yield parameters such as kernels per head and thousand kernel weight (TKW) in inoculated control plants were lower compared to the other treatments. In the experimental year 2007/2008, independent treatments with the bacterial strain E1R-j and the fungicide Triadimefon reduced take-all disease in wheat roots by 55.3% and 61.9%, compared to the inoculated control plants. In this season plant height in inoculated control was significantly lower and also the yield parameters seeds per head and especially TKW were drastically reduced compared to the other treatments. E1R-j treatment alleviated the detrimental effects of take-all on grain yield parameters to a similar extent as Triadimefon application. SEM studies revealed that in the presence of E1R-j, hyphae of Ggt showed leakage, appeared ruptured, swollen and shriveled. Following root drench, strain E1R-j was able to colonize endophytically roots and leaves of wheat seedlings. While the population of the bacterial strain in wheat roots steadily increased from the second to the fourth leaf stage, in the leaf tissue the population of the strain rapidly declined. TEM studies also showed that cells of E1R-j were present in roots of wheat seedlings and effectively retarded infection and colonization of Ggt in root tissue; suppression of Ggt by E1R-j was accompanied by disintegration of hyphal cytoplasm. In addition, in the presence of E1R-j cells in Ggt-infected root tissue morphological defense reactions were triggered such as formation of wall appositions and papillae. The results presented indicate that the endophytic strain E1R-j of B. subtilis meets demands required for biocontrol of take-all.     

Studies on virulence of the take-all fungus, Gaeumannomyces graminis, with reference to methology

Current methods for take-all assessment in laboratory experiment were examined; it was shown that the extent of vascular discoloration may not reflect virulence of a fungal isolate or host resistance to the pathogen under some experimental conditions. A new assessment method for take-all is described, based on the ability of transport eosin past infection sites. It enables hosts or isolates to be compared by ET₅₀ values, the times from inoculation when 50% of plants fail in eosin-uptake through the three oldest seminal roots. Use of this technique suggested that barley roots were less affected than were wheat roots by Gaeumannomyces graminis var. tritici. Further experimental results showed that an isolate of G. graminis that had lost part of its virulence in culture yielded some single-conidium progeny more virulent than itself. When single-condium isolates or a mycelial isolate and its single-conidium progeny were jointly inoculated on wheat, the amount of disease was less than that caused by the more virulent isolate alone.     

Identity and frequency of occurrence ofTrichoderma spp. in roots of wheat and rye-grass in Western Australia and their effect on root rot caused byGaeumannomyces graminis var.tritici

Trichoderma hamatum, T. harzianum andT. koningii were isolated from wheat and rye-grass roots from a field in Western Australia. Frequency of occurrence ofTrichoderma spp. was higher on roots subjected to washing only, for both wheat and rye-grass than the roots which were surface-sterilized with 0.6% or 1.25% NaOCl.Trichoderma spp. were recovered at a higher frequency on PDA amended with lactic acid (pH 4.5) than on PDA alone (pH 5.6) or PDA with streptomycin. In general,Trichoderma spp. were isolated at a higher frequency from roots of wheat than that of rye-grass.T. hamatum occurred at a higher frequency in rye-grass roots than in wheat, whereasT. harzianum was more common in roots of wheat than in rye-grass, especially in seedling and milky ripe stages.T. koningii was recovered at a higher frequency from roots at seedling stage of rye-grass than wheat, the reverse being true at tillering stage.T. koningii was not recovered from roots of either host in any sampling when they were surface sterilized with 1.25% NaOCl.The take-all fungus was isolated from wheat and rye-grass roots more frequently at tillering and stem extension stages than others. It was severely pathogenic to both hosts in sterilized and non-sterilized soil.Addition of lactic acid, HCl or streptomycin to PDA did not affect the growth of theTrichoderma spp. tested, but the growth was slower on Martin's medium than on other media. In generalT. harzianum andT. koningii grow faster thanT. hamatum. The growth of the three species were not different at 20 and 25°C, but at 15°c growing of all species was significantly reduced.Incorporation of lactic acid into PDA prevented the bacterial growth in all treatments. Streptomycin too reduced but to a lesser degree than lactic acid. Surface sterilization with NaOCl decreased the recovery of both bacteria and fungi. T. hamatum andT. koningii reduced the mortality of wheat and rye-grass plants inoculated with the take-all fungus in sterilized and non-sterilized soil, whereT. harzianum did not protect wheat or rye-grass from infection by the take-all fungus.     

A cereal cyst nematode (Heterodera avenae Woll.) resistance gene transferred from Aegilops triuncialis to hexaploid wheat

 The cereal cyst nematode (Heterodera avenae) is an important root parasite of common wheat. A high level of resistance was transferred to wheat from Aegilops triuncialis (TR lines) using the cross [(T. turgidum×Ae. triuncialis)×T. aestivum]. Low fertility (3–5 viable kernels per plant) was observed during the process but the surviving hybrid plants were highly vigorous. To obtain stable resistant lines further crosses to T. aestivum were performed. The resistance in TR lines seems to be transferred from the C genome of Ae. triuncialis (genomes CCUU). Ae. triuncialis was highly resistant to the two Spanish populations of H. avenae tested, as well as to four French races and two Swedish populations. The histological analysis showed a hypersensitive reaction in the roots of a resistant TR line inoculated with the Ha71 pathotype of H. avenae, whereas well-formed syncytia were observed in the roots of the susceptible control. Resistance to the H. avenae Ha71 pathotype seemed to be inherited as determined by a single dominant factor in the crosses between resistant TR lines and susceptible cultivars. Received: 11 November 1997 / Accepted: 9 December 1997     

Interactions between Heterodera avenae and Pratylenchus neglectus on Wheat

In a long-term field experiment, differential population densities of Heterodera avenae were produced by frequent cropping with resistant (cv. Panema) or susceptible (cv. Peniarth) oat. The two oat cultivars were equally good hosts of Pratylenchus neglectus in a glass house experiment with field soil. On wheat crops grown after oats in field experiments, P. neglectus population densities in roots were higher in plots where H. avenae had been controlled than in plots with moderate infestations (40 H. avenae eggs/g soil). The field observations indicated that the reduction in population densities of P. neglectus coincided with the development in roots of sedentary stages of the cyst nematode. Evidence for an indirect effect of H. avenae on P. neglectus was found in vitro in a split-root experiment. In the same field, grain yields of two wheat cultivars susceptible or resistant to H. avenae, but both susceptible to P. neglectus, was not reduced by P. neglectus. Alternation of H. avenae resistant and susceptible cultivars is a possible way of exploiting the inverse relationship between these nematodes, whilst controlling cyst nematode -populations in intensive cereal production systems.     

Studies of the cereal cyst-nematode, Heterodera avenae under continuous cereals, 1974–1978. I. Plant growth and nematode multiplication

Population changes of Heterodera avenae and crop growth in a sandy loam soil were studied from 1974 until 1978; the nematode decreased plant growth but failed in two of the years to multiply on susceptible hosts. Spring oats were the most heavily invaded cereal and produced the smallest shoots. Second-stage juveniles invaded cereal roots in decreasing numbers: spring oats > autumn oats > spring barley > spring wheat > autumn barley > autumn wheat. Numbers of females developing on the different cultivars were in a similar order. Most females developed on roots in 1976 despite poor crop growth in the severe drought. Numbers of H. avenae in soil treated with oxamyl (Vydate) at 8.8 kg/ha a. i. were less in all years except 1975. In the dry winter and spring of 1975/76 nematode multiplication was prevented in soil treated with oxamyl before drilling in the autumn. In all years large numbers of females were produced on the roots of all cultivars but in 1975 and 1978 nematode populations declined because few females survived to form cysts containing eggs and their fecundity was reduced. Numbers of cysts after harvest were not affected by formalin (38% formaldehyde) applied as a drench at 3000 litres/ha in 1977 but fecundity doubled in treated soil, and nematode multiplication increased from 3.8 × in untreated plots to 18.6 ×. When the plots were irrigated in 1978 numbers of cysts and fecundity increased in formalin treated soil resulting in an increase in multiplication from 0.3 × to 14.6 ×. Fungal parasites attacking H. avenae females and eggs are considered responsible for the poor multiplication of the nematode.     

Influence of wheat and oat cultivars on the development of the cereal aphid parasitoid Aphidius rhopalosiphi and the generalist aphid parasitoid Ephedrus plagiator

The effects of three wheat cultivars and two oat cultivars on the development of the cereal aphid parasitoid Aphidius rhopalosiphi De Steph. and the generalist aphid parasitoid Ephedrus plagiator (Nees) (Hymenoptera: Braconidae) were evaluated in the laboratory. The level of hydroxamic acids, a family of secondary metabolites that can affect the mean relative growth rate of cereal aphids in cereals, were measured in the different cultivars. The parasitoids were reared in Sitobion avenae (F.) (Homoptera: Aphididae), using plants grown under greenhouse conditions. A. rhopalosiphi showed a longer developmental time on wheat relative to oat cultivars. This effect was accounted for by a significant increase in the time from oviposition to pupation (mummy formation), while the duration of the pupal stage remained constant between treatments. No further effects were observed in other variables evaluating A. rhopalosiphi performance, such as adult longevity, adult body weight and secondary sex ratio. The generalist E. plagiator did not show significant differences in any of the variables analysed, both between cultivars and cereal species. Hydroxamic acids levels correlated negatively with mean relative growth rates of S. avenae, but positively with the observed developmental time of A. rhopalosiphi. The results are discussed in terms of tritrophic effects and the development of breeding programmes trying to improve plant resistance to aphids.