Invasion and development of the cereal cyst-nematode Heterodera avenae in roots of barley infected by the take-all fungus Gaeumannomyces graminis

Observations on the invasion and development of Heterodera avenae in relation to take-all infection of barley roots were made in three pot experiments. Post-cropping H. avenae populations were lower on fungus-infected plants, as a result of the production of fewer, smaller and less fecund cysts. The reduction in cyst numbers was associated with reduced larval invasion of take-all infected roots and with the effects of the unfavourable environment of these roots on female development, possibly through its effects on sex determination and survival of the larvae.     

Barley segregates resistant and susceptible to the cereal cyst-nematode (Heterodera avenae Woll.)

In the first year of this experiment both nematode-resistant and susceptible segregates of spring barley were equally invaded by Heterodera avenae larvae, but few females developed in the roots of the resistant plants and numbers of eggs in the soil were fewer after the resistant crop. In the second year, a commercial, nematode-susceptible barley crop yielded significantly more grain following the resistant segregate. The resistant segregate, derived from Hordeum pallidum var 191, suppresses the development of Heterodera avenae pathotypes 1 and 2 [British nomenclature] and should be useful in areas where these pathotypes predominate.     

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.     

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.     

The invasion of root tips of cereals by the cereal cyst nematode Heterodera avenue

Newly germinated seedlings of susceptible cultivars of oats, wheat, barley and rye were inoculated with second-stage juveniles of Heterodera avenae in pots of sand. Subsequent examination showed oat root tips to be more commonly invaded, and by a greater range of nematode numbers than the other cereals. A comparison of oats and barley showed that lower nematode numbers in barley were not due to a higher emigration from barley; invasion, establishment and emigration by nematodes all being greater in oats. Second-stage juveniles were more likely to migrate prior to establishment in barley than in oats.     

The distribution of pathotypes of the cereal cyst-eelworm Heterodera avenae in England and Wales

Two pathotypes of the cereal cyst-eelworm (Heterodera avenae), occurred commonly in England and Wales and a third pathotype was apparently present in low numbers in a few areas. Pathotype 1, which can breed on most cultivars of wheat, barley and oats, but not on Drost barley nor barley cultivars containing the gene for resistance from barley No. 191, was most common in the south west of England. Pathotype 2, which can breed on most cereals, including Drost barley but not No. 191, was common throughout England and Wales and mixtures of these pathotypes occurred in many places. Pathotype 3, which can breed on barley No. 191 and probably on Drost and most cultivars of cereals, was rare and was found only in low numbers together with either or both Pathotypes 1 and 2. In the initial tests it was present or suspected in only nine fields (6%), but when these soils were retested in more detail the following year, Pathotype 3 was confirmed at only one site.     

The effects of soil sterilants on the cereal cyst-nematode (Heterodera avenae Woll.), take-all (Ophiobolus graminis Sacc.) and yields of spring wheat and barley

In a 3-year field experiment on sandy loam at Woburn, methyl bromide, chloropicrin, D-D mixture, dazomet, formalin and mercury salts were applied in the first year only, or in the first and second years, before drilling spring wheat. In the third year, their residual effects on spring barley were measured. All sterilants except mercury decreased cereal cyst-nematode ( Heterodera avenae ) numbers and take-all ( Ophiobolus graminis ) incidence, and increased crop yields in the year they were applied. Dazomet gave the best control of H. avenae in the first year and controlled O. graminis best in both years of application. In the third year, O. graminis increased in all plots previously treated (except after two successive D-D treatments), but H. avenae increased only after formalin. Two formalin applications more than trebled the H. avenae egg count by the end of the third year and depressed the yield of barley. Two successive applications of chloropicrin gave the best nematode control.
Other fungus diseases, Fusarium foot rot, Pythium root rot, eye-spot ( Cercosporella herpotrichoides Fron.) and sharp eye spot ( Rhizoctonia solani Kühn), were uncommon and sterilants had no detectable effects on them. The largest aggregate straw yield (3-year total) was obtained from two applications of dazomet, but the best yields after treatment in the first year only were given by D-D and methyl bromide.     

Some effects of differently-acting nematicides on the cereal cyst-nematode (Heterodera avenae) and on the appearance of ''scorch'' in spring wheat on light loamy sand

In fungitoxicity tests against Phytophthora cinnamomi on Chamaecyparis lawsoniana cv. Ellwoodii, a drench of furalaxyl (1000 mg a.i./l) applied to the compost in which 1-yr-old plants were growing, 1 wk before they were inoculated with 650 000 zoospores, controlled disease for at least 12 months. With an inoculum dose of 650 zoospores/plant, furalaxyl at 500 mg a.i./l controlled disease even when inoculation was 12 wk after fungicide treatment. Aluminium tris (ethyl phosphonate) (2000 mg a.i./l) applied as a drench 1 wk before inoculation with 650 000 zoospores/plant did not prevent root infection but delayed foliar symptoms for 9 months: the same treatment, using etridiazole (500 mg a.i./l) only slightly reduced disease incidence. When applied as a single drench 2 days before inoculation, prothiocarb (2000 mg a.i./l) and cuprammonium compounds (200 mg a.i./l) were much less effective than furalaxyl (1200 mg a.i./l), sodium ethyl phosphonate (1500 mg a.i./l), aluminium tris (ethyl phosphonate) (1500 mg a.i./l) or etridiazole (500 mg a.i./l). However, a drench of furalaxyl at 1000 mg a.i./l, aluminium tris (ethyl phosphonate) at 2000 mg a.i./l or etridiazole at 500 mg'a.i./l did not eradicate P. cinnamomi from compost containing infected root debris. Pre-planting drenching of the compost was ineffective. All fungicide treatments were non-phototoxic to 1-yr-old C. lawsoniana cv. Ellwoodii. These results are of special relevance to the control of P. cinnamomi on container-grown woody ornamentals.     

Effect of some fertilizers on hatching of cereal cysts nematode,Heterodera avenae

Experiments were conducted in laboratory and pot conditions to determine the effects of urea, di-ammonium phosphate (DAP), single super phosphate (SSP), muriate of potash (MOP) and zinc sulphate (ZnSO₄) on hatching of Heterodera avenae. Two concentrations of each fertilizer were tested in lab for which 10 cysts and 5 ml of each concentration were taken in 5 cm diameter Petri plates. Observations were recorded at weekly intervals up to six weeks. Urea, DAP, SSP and MOP inhibited hatching and ZnSO₄ increased it. After six weeks, hatching was least (5.45%) in higher dose of urea and greatest (46.9%) in higher dose of ZnSO₄. In pot experiment, two doses of urea and single dose of SSP, MOP, and ZnSO₄ were applied in H. avenae-infested soil and WH-1105 wheat was sown. Observations on nematodes in roots, soil and remaining cyst contents were recorded 40 days after sowing. Among all the fertilizers, least nematodes in soil and roots were found at higher dose of urea and greatest number in ZnSO₄.     

Genetic variation in natural populations of the cereal cyst nematode (Heterodera avenae Woll.) submitted to resistant and susceptible cultivars of cereals

The purpose of the present work was to study the genetic characteristics of cereal cyst nematode (Heterodera avenae) populations re-established after the long-term use of resistant oat cultivars in field conditions. Population features were analyzed through fitness components and variation in enzymatic polymorphism (esterase and malate dehydrogenase loci). The longest (6 year) use of the same resistance genes (oat cv Panema) at high frequency (Rotation IB) led to the selection of a resistance-breaking pathotype and to a decrease in viability which suggested either a founder effect or a lower reproductive potential for the new pathotype. Analysis of esterase allelic frequencies led to the conclusions that: (1) the genetic constitution of this pathotype was different from the reference population maintained on the susceptible host (oat cv Peniarth), and (2) that the esterase locus may develop a disequilibrium linkage with loci involved in virulence. Random mating was recorded at the whole-field level but not always at the single-plant level, suggesting that cultivation practices such as annual ploughing could play a major role in homogenizing subpopulations developed in the vicinity of a plant. These phenomena showed that the long-term use of highly effective resistance could provok marked genetic modifications in populations. These risks should be taken into account when deciding strategies for optimal use of resistance genes in nematode management programs.

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The effect of inbreeding on potato cyst-nematode, Heterodera rostochiensis

Two populations, Duddingston and Feltwell, of potato cyst-nematode were inbred for seven and six generations respectively. The Duddingston population maintained a high fecundity in some lines for three generations, but in subsequent generations the fecundity as measured by cyst production declined in spite of selection for high fecundity. By selecting for fecundity, some lines of the Feltwell population still produced many cysts after five generations of inbreeding. In the Duddingston population it was possible to increase the number of cysts produced by an inbred line of poor fecundity by using paired cysts plus selection for fecundity. A similar change may have been obtained for unprolific lines of the Feltwell population. Selection for production of few cysts quickly produced lines with poor fecundity. One cause of poor fecundity was the small number of larvae in cysts used for inoculations. Yellow eggs were found in large numbers in some cysts but these were not a cause of loss of fecundity following repeated inbreeding.     

Observations on the invasion and development of the pea cyst-nematode Heterodera goettingiana in winter field beans (Vicia faba)

A plot experiment to investigate the invasion, development and number of generations of Heterodera goettingiana on winter-sown field beans showed that, although second-stage juveniles invaded in October, white females did not appear until mid-May and only one generation occurred. The results are discussed in the context of temperature requirements for the development of this nematode.     

The effect of formalin on potato cyst-nematode, Heterodera rostochiensis

Applying 5 ml of a 38% aqueous solution of formaldehyde to 1500 ml of sandy loam prevented potato cyst-nematode, Heterodera rostochiensis, from increasing on the roots of Arran Banner potatoes planted in the soil and increased the weight of tubers produced. A comparable dosage applied to plots of sandy loam and sandy clay in winter did not prevent the nematode from multiplying on Pentland Crown potatoes but increased tuber yields. Smaller amounts of formalin had less effect on tuber yields and drenching dilute formalin solutions onto the soil was no more effective than incorporating undiluted formalin into the soil by rotavation 15 cm deep.     

Hatching agents for the potato cyst-nematode, Heterodera rostochiensis Woll

Of 444 compounds tested for their ability to stimulate eggs of Heterodera rostochiensis to hatch, forty-five did so to varying extents. Many compounds that hatched H. schachtii Schm. eggs did not hatch H. rostochiensis eggs. The most effective compounds with H. rostochiensis were picrolonic acid, anhydro-tetronic acid and vanadates. Tests with analogues of picrolonic and anhydro-tetronic acids suggested some structures that may be concerned in hatching H. rostochiensis and H. schachtii. The simple analogues hatched the eggs of H. schachtii but not those of H. rostochiensis. Suitably placed polarisable atoms seemed to be an essential feature. Picrolonic acid contains pairs of polarizable atoms, about 4 and 6·7 Å apart, that may respectively be associated with the hatching of H. schachtii and H. rostochiensis. Other features of the more active hatching agents were a cis and co-planar arrangement of the two polarizable atoms with respect to the rest of the molecule. The function of hatching agents is discussed.     

The chemical composition of the cyst wall of the potato cyst-nematode, Heterodera rostochiensis

1. Cyst walls of the potato cyst-nematode (Heterodera rostochiensis Woll.) were isolated by sieving a suspension of crushed cysts. About 12mg. of dried cyst walls was obtained from 1000 cysts. 2. The cyst walls contained mainly protein (72%, calculated from nitrogen content). On acid hydrolysis about 77% of the cyst wall went into solution. Of 19 amino acids present, proline, glycine, and alanine were the most abundant, and made up about 50% by weight of the total amino acids. The amino acid composition suggested that collagen-like proteins predominated in the cyst wall and larval cuticle. 3. A small amount of glucosamine (1.5%) was present in the hydrolysates, but chitin was not detected in the cyst walls. 4. Other components of the cyst walls were lipid (2%), carbohydrate (0.5%) and a small amount of inorganic matter (ash, 5%). Polyphenols (2% by wt. of the cyst walls) occurred in the acid hydrolysates. The dark pigments of the cyst wall were not indole-containing melanins.     

The composition of the cyst wall of the beet cyst-nematode Heterodera schachtii

1. Cyst walls of the beet cyst-nematode (Heterodera schachtii Schmidt) were obtained by sieving a suspension of crushed cysts; about 15mg of dried cyst walls was obtained from 1000 cysts. 2. The cyst walls contained 68% protein calculated from nitrogen content. Glutamic acid, glycine, proline and hydroxyproline made up about 54% by weight of the amino acids obtained on acid hydrolysis. 3. Minor constituents of the cyst wall were hexosamine (3.3%), lipid (6%), carbohydrate (2%) and phenols (2%). The hexosamine was identified as galactosamine. 4. The cyst walls contained inorganic material (ash 17%), most of which was extractable with EDTA, but not with water. Major inorganic components were calcium and phosphorus (1.7% and 1.5% respectively, by weight). Carbon dioxide (about 1% by weight) was liberated from the cyst walls on acidification. 5. The cyst walls of H. schachtii and the potato cyst-nematode (Heterodera rostochiensis) contained different amounts of the same amino acids. They also differed in their inorganic content and in the nature of the hexosamine present.     

Breeding for resistance to the white potato cyst-nematode, Heterodera pallida

Progenies bred from material derived from the wild potato, Solanum vernei and from the cultivated potato, S. tuberosum ssp. andigena, clone CPC 2775, were compared for their resistance to Heterodera pallida, pathotype E. The influence of additional resistance derived from the wild species, S. multidissectum, was also investigated. Both S. vernei and CPC 2775 gave progenies with variable levels of resistance and there was often no clear segregation into resistant and susceptible categories. Incorporation of gene H₂ derived from S. multidissectum increased resistance to pathotype E of H. pallida for resistant material bred from both S. vernei and clone CPC 2775. The results indicate that adequate resistance to all British populations of potato cyst-nematodes can best be obtained by combining the factors for resistance from the two Andigena clones, CPC 1673 (gene H₁) and CPC 2775 (gene H₃), and from S. multidissectum (gene H₂).     

Observations on the introduction of Verticillium chlamydosporium and other parasitic fungi into soil for control of the cereal cyst-nematode Heterodera avenue

Isolates of Verticillium chlamydosporium and a sterile fungus added to soil on ground oat grain reduced the numbers of Heterodera avenae on wheat by between 26 and 80%. Nematode populations in uninoculated soil increased from 15 eggs/g soil before planting to 218 eggs/g after harvest. V. chlamydosporium was isolated from oat grain that had been air-dried and milled before introduction into soil. Applications of the fungi on attapulgite clay or as suspensions of mycelium and spores in water had no effect on nematode multiplication. The effect of the fungi on numbers of H. avenae eggs was similar in autoclaved and non-sterilised soil. V. chlamydosporium added on attapulgite clay to a calcareous sand and a calcareous silty loam could be re-isolated after at least 6 months. Some isolates colonised the roots of wheat without causing lesions or affecting the dry weights of shoots or roots. These results indicate that V. chlamydosporium may be useful for the biological control of cyst nematode pests.