SOME OBSERVATIONS ON THE HATCHING RESPONSES OF THE CABBAGE-ROOT EELWORM, HETERODERA CRUCIFERAE FRANKLIN

Samples of root diffusate from sprouts ( Brassica oleracea var. gemmifera ), swedes ( B. napus var. napobrassica ), rape kale ( B. napus var. arvmsis ), and white mustard ( Sinupis alba ), all stimulated larval emergence from cysts of Heterodera cruciferae. When mustard-root diffusate was added to leachings from the other species tested there was no apparent effect on larval emergence. In half dilution with tap water, mustard leachings were comparable in activity with similar dilutions of leachings from the other species under observation. Leachings from soil were less active than those from the four species of plants tested, but showed significant activity in comparison with tap water.
The form of the hatching curve was studied using diffusates from both sprouts and mustard roots, and in each case when cumulative hatch was plotted against time on a logarithmic scale, the sigmoid curve obtained closely fitted the theoretical sigmoid calculated from a probit analysis of the data.
Estimates were obtained of the log activity values of leachings from the four species of plants tested and from soil, following an examination of the dilution curves for mustard and sprouts root diffusates. These curves were closely parallel with one another, and it was therefore considered justifiable to derive log activity values for these root diffusates, although the cysts used in the hatching tests and the root-diffusate samples were each obtained from a single source. For this reason some caution is perhaps called for in the interpretation of the results presented in this paper.     

Field studies of potato-root eelworm, Heterodera rostochiensis Wollenweber, 1938-40

Field studies of potato crops under different soil and fertility conditions and exposed to attack by diverse concentrations of potato-root eelworm led to the following conclusions:
In the north-western agricultural advisory province potato-root eelworm is not a major cause of failure or partial failure of potato crops on land where the viable cyst concentration is low (under 10/10 c.c. of soil). On such land increased yields can be obtained by improving conditions for plant growth, widening the rotation and increasing the fertility of the soil.
The production of satisfactory crops on land with a low eelworm population is likely to be accompanied by considerable increases in the numbers of root eelworms in the soil.
Crops free from signs of 'potato sickness' or 'potato-root eelworm disease' can be grown on land heavily infested with root eelworms (viable cyst concentration exceeding 25/10 c.c.), if no other adverse condition, such as unsuitable weather, lack of available food or incidence of fungus disease, affects the growth of the crop, but yields from heavily infested land are not likely to be more than moderate.
The treatments used against eelworms in the trials described here did not give results commensurate with their cost.
The practical difficulties of incorporating lethal substances with soil in the field are considerable, and this may have contributed to the disappointing results obtained.
The study of the influence of host variety on cyst production suggests that varietal differences in the host plants may influence the numbers of cysts formed on the crop, but the evidence is not conclusive.     

PROVISIONAL LISTS OF HOST PLANTS OF SOME ROOT EELWORMS (HETERODERA SPP.)

In host-range tests carried out at Cambridge, 1951-3, beet eelworm was found to parasitize nearly all Cruciferae tested, while other hosts were found amongst the Amaranthaceae, Caryophyllaceae, Chenopodiaceae, Labiatae, Phytolaccaceae, Polygonaceae, Scrophulariaceae and Tropaeolaceae. Cabbage eelworm parasitized nearly all Cruciferae tested, and a few Labiatae. Hosts of Galeopsis and clover eelworms were found amongst the Caryophyllaceae, Labiatae, Leguminosae, Polygonaceae and Scrophulariaceae. Potato eelworm cysts were found only on Lycopersicum and Solanum ; carrot eelworm only on Daucus ; and hop eelworm only on Urtica (Urticaceae) and Cannabis and Humulus (Cannabinaceae). Pea eelworm appeared to parasitize only the genera Lathyrus, Lens, Pisum and Vicia (Leguminosae), while hosts of oat eelworm appeared to be distributed more or less at random within the Gramineae.     

MOVEMENT OF EELWORMS

Tracks were plotted of about 300 individual eelworms comprising six species among water droplets on a glass surface. Measurements of the tracks indicated that the product of length and activity of an eelworm divided by its speed was a constant. This supports the hypothesis that the speed of an eelworm among water droplets is a function of its length and activity. This principle can only be applied to movement in soil where the length of the eelworm is less than about three times the particle diameter. Under such conditions the eelworms move in thin films or water droplets over particles. Among smaller-sized particles the speed of the eelworms is influenced by particle size. With increasing eelworm length there is an increase in soil particle size for maximum mobility.     

ON THE STEM AND BULB EELWORM (ANGUILLULINA DIPSACI KUHN) WITH SPECIAL REFERENCE TO ITS OCCURRENCE ON WEEDS OF ARABLE LAND

The importance of wild hosts of Anguillulina dipsaci Kühn as carriers of infection needs no emphasis. The occurrence of the eelworm in the weeds, cleavers ( Galium aparine ) and duckweed ( Stellaria media ) taken in crops of oats and field beans which were themselves suffering from stem eelworm disease, was recorded by Johnson (1936) and Johnson & Thompson (1937); Walton (1937) confirmed the occurrence of infested cleavers plants in eelworm-infested oats. Walton also stated that experiments indicated that seed oats failed to carry eelworm infection, and from circumstantial evidence similar to that recorded by Johnson (1936) stated that "infestations of oat crops which occur, even after the absence of the crop for more than six years, are in all probability attributable to weeds rather than to infested seed". The present writer failed to obtain living eelworms from oat seed from infested crops in Yorkshire. This paper summarizes the results of field observations and experiments accumulated mainly during the past three years.     

MOVEMENT OF EELWORMS.: IV. THE INFLUENCE OF WATER PERCOLATION

A study was made of the relationship between rate of flow of water through sands of different particle size and the downward velocity of eelworms of various lengths and activities. The relationship between rate of flow of water and velocity of eelworms is not linear. For rates of flow less than about 500 cm./hr. the velocity of the eelworms relative to the velocity of water is greater than for flow rates greater than about 500 cm./hr. For rates of flow greater than 500 cm./hr. an equation is derived which relates velocity of eelworms, rate of water flow, length of eelworms and pore diameter in the sand. The following facts were established: the velocity of the eelworms increases as pore diameter increases; the velocity approaches zero when the eelworms' length exceeds four times the pore diameter; the relationship between velocity of eelworm and velocity of water is independent of the eelworms' own activity; the equation of eelworm movement is valid for both continuous and discontinuous saturated flow. With flow rates less than about 500 cm./hr., the velocity of the eelworms increases as the eelworms' activity increases and eelworms appear to pass through smaller pores than at high flow rates. Dead or inactive eelworms do not progress very far in sand even at high flow rates and observations suggest that slight flexing movements of the eelworm body is essential for passage.     

MOVEMENT OF EELWORMS

The optimum crumb sizes for movement of potato-root eelworm larvae in a sandy loam, a heavy clay and a peat soil were 150–250 and 250–400 μ. Mobility was very similar in clay and sandy loam, in both of which there was an optimum suction for movement. In the peat, however, mobility increased with suction and no optimum suction was established. Larvae may be able to move in peat at high suctions because friction between the larvae and the peat crumbs is less than between clay or sand crumbs. Larvae moved to the wet end of a moisture gradient in sand, the number increasing with the steepness of the gradient. The rate of spread of larvae in sand 150–250 μ diameter varied between 2 and 3 cm. a day, depending on suction. As pore size increases, any upward movement in a moisture gradient is opposed by falling under gravity. Larvae do not respond to a moisture gradient or fall under gravity in sand where the width of the pore approximates to the diameter of the larva. The presence of host roots also counteracted the response to a moisture gradient; the degree of orientation to the roots increased with the time the roots were in the sand. Direct observation on larvae, newly emerged from cysts, in the presence of host plant roots, suggests that larvae orientate themselves at a distance from the root and do not reach the root by random movement. Many of the movements of eelworms are explicable by considering the relationship between pore size, eelworm diameter and water distribution, and a diagram relates movement and various soil factors.     

PÓT TESTS OF NEMATICIDES AGAINST POTATO-ROOT EELWORM

Details are given of nematicidal tests, directed against the potato-root eelworm, in 81. glazed pots of infested soil. The soil was sampled a few weeks after injection to secure data on the kill achieved, by counting the eelworm larvae hatching in root diffusate. A potato tuber was then planted in each pot and yielded information on phytotoxic or soil-amendment effects. A second soil sample at the end of the season showed the degree to which the eelworm population had recovered after treatment. With only duplicate pots, and single batches of 100 cysts from each pot, clear-cut results were achieved with D–D mixture and ethylene dibromide. In the present pilot test, the data were handled by ordinary analysis of variance methods, by which the lowest significant percentage kill was 73%, but the methods could be adapted for purposes of probit analysis.     

SOME OBSERVATIONS ON THE CEREAL-ROOT EELWORM POPULATION OF FIELD PLOTS OF CEREALS WITH DIFFERENT SOWING TIMES AND FERTILIZER TREATMENTS

In the autumn of 1953 an experiment was begun to follow changes in the cereal-root eelworm population of small plots on a field in Shropshire. The plots were cropped with either oats, wheat, barley or rye, sown in the autumn and spring, and some plots had fertilizer. Each plot received the same treatment for 3 years; in the fourth year an indicator crop of spring oats was grown on all the plots.
Under rye and autumn-sown wheat the eelworm population fell to a level which permitted a good oat crop in 1957. Autumn-sown wheat, barley and rye generally produced lower eelworm populations than their spring-sown counterparts, but autumn-sown oats proved to be the most efficient host. The order of host efficiency was oats (best), barley, wheat, rye. The eelworm populations were generally higher on plots receiving fertilizer treatment. In this experiment all oat plots, and spring-sown barley plus fertilizer, produced populations which severely damaged the 1957 oat crop.     

Genetic-physiological compatibility and karyotypes of stem nematode]

The paper presents literary and the authors own data. The potato stem eelworm Ditylenchus destructor has a set of chromosomes 2n=44-48 and does not cross with the close species D. dipsaci from onion, garlic, parsley and parsnip. Most stem eelworms of the collective species D. dipsaci have n=12 and cross with each other producing fertile progeny. Some forms of stem eelworms, e.g. those of broad beans, plantain, dandelion and falcaria are polyploid. Intraspecies and intrapopulation polymorphism according to the number of chromosomes was recorded. Some races do not cross, some cross but unilaterally, some races produce sterile, non-viable or little viable hybrids. The species D. dipsaci should be regarded not only as a complex of species but as a group of forms at different stages of intraspecies differentiation.     

Soil teachings and the hatching of larvae of the potato-root eelworm, Heterodera rostochiensis Wollenweber

Leachings were taken from fallow soil brought in from an outside plot at monthly intervals throughout a year, and tested for their ability to stimulate hatching of the potato-root eelworm. Some of the leachings showed stimulatory action, the results indicating a seasonal pattern of activity with a peak in August. As noted previously, this suggests that soil organisms produce substances able to stimulate hatching and that these are probably responsible for the so-called ‘spontaneous’ hatching.     

Comparative Responses of Globodera rostochiensis and G. pallida to Hatching Chemicals

Globodera rostochiensis and G. pallida responded similarly to hatch stimulation by potato root leachate, but proportionally more second-stage juveniles (J2s) of G. rostochiensis hatched than of G. pallida in response to picrolonic acid, sodium thiocyanate, alpha-solanine, and alpha-chaconine. Fractionation of the potato root leachate identified hatching factors with species-selective (active toward both species but stimulating greater hatch of one species than the other), -specific (active toward only one species), and -neutral (equally active toward both species) activities. In a comparison of two populations of each of the two potato cyst nematode (PCN) species, however, greater similarity in response to the individual hatching factors was observed among populations of different species produced under the same conditions than among different populations of the same PCN species. Smaller numbers of species-specific and species-selective hatching factor stimulants and hatching inhibitors than of hatching factors were resolved. In a study to determine whether the different hatching responses of the two species to the same root leachate were associated with different ratios of species-selective and species-specific hatching factors, G. rostochiensis pathotype Ro1 exhibited greater hatch than did G. pallida pathotype Pa2/3 in response to leachate from older plants (more than 38 days old), while G. pallida exhibited greater hatch in response to leachate from younger plants (less than 38 days old); the response of G. pallida pathotype Pal with respect to plant age was intermediate between the other two populations. Combined molecular exclusion-ion exchange chromatography of the root leachates from plants of different ages revealed an increase in the proportion of G. rostochiensis-specific and -selective hatching factors as the plants aged.     

STUDIES IN THE RELATIONSHIP OF EELWORMS AND BACTERIA TO CERTAIN PLANT DISEASES

Two types of organism are known to be present in strawberry plants suffering from cauliflower disease, namely, eelworms, Aphelenchoides ritzema-bosi and/or Aphelenchoides fragariae , and a bacterium, Corynebacterium fascians. Neither type, inoculated separately, has consistently reproduced the disease.
An experimental investigation of a possible obligate etiological relationship between the eelworm and the bacterium is described, in which Aphelenchoides ritzema-bosi and several strains of Corynebacterium fascians were inoculated together and separately.
No abnormalities appeared in the absence of the eelworm. Typical cauliflower symptoms were reproduced only by a combination of the eelworm and strains of C. fascians originally isolated from cauliflower strawberries. Small alaminate leaves and enations appeared in all treatments where the eelworm was introduced, including those where no bacteria were co-inoculated. Subsequent isolations, however, demonstrated the presence of contaminant strains of C. fascians in the latter plants. The severity of the enation symptom was related to the presence of a particular strain of C. fascians , but the alaminate leaves showed no such correlation. Evidence on the source of the contaminant strains found in the experimental plants is discussed.
In parallel field investigations it was established that eelworm-infested strawberries showing no cauliflower symptom frequently bore alaminate leaves, with which C. fascians was consistently associated. It is suggested therefore that cauliflower is simply the least frequent of a range of symptoms of a bacterial disease endemic in strawberry plants infested with Aphelenchoides spp.     

Detection of hatching inhibitors and hatching factor stimulants for golden potato cyst nematode, Globodera rostochiensis, in potato root leachate

In a comparison of four potato varieties, in-soil hatch of the golden potato cyst nematode (Globodera rostochiensis) was positively correlated to in vitro hatch in response to potato root leachate (PRL). The in-soil hatch of cysts of G. rostochiensis to two of the four varieties was significantly less than that of the control (cysts in gravel without potato plants) in the first 2 wk after plant emergence, suggesting the production of hatching inhibitors (HIs) by young potato plants. The hatching factor: hatching inhibitor ratio of PRL was positively associated with the net hatching activity of the PRL. Four zones of HI activity were resolved following gel permeation chromatography of PRL on Sephadex G-10. Hatch-inactive chemicals, which stimulated the activity of hatching factors (HFs) in PRL (hatching factor stimulants, HSs), were also isolated from PRL, hatch levels induced by individual HFs responding differently to the same HS preparation. The complex interactions between individual HFs and other hatching chemicals in PRL was illustrated when addition of the hatch-active potato glycoalkaloid α-solanine caused both inhibition and stimulation of PRL-induced hatch, depending on the α-solanine concentration.     

Plant–parasite coevolution: A weak signature of local adaptation between Peruvian Globodera pallida populations and wild potatoes

Plant–parasite coevolution has generated much interest and studies to understand and manage diseases in agriculture. Such a reciprocal evolutionary process could lead to a pattern of local adaptation between plants and parasites. Based on the phylogeography of each partner, the present study tested the hypothesis of local adaptation between the potato cyst nematode Globodera pallida and wild potatoes in Peru. The measured fitness trait was the hatching of cysts which is induced by host root exudates. Using a cross‐hatching assay between 13 populations of G. pallida and root exudates from 12 wild potatoes, our results did not show a strong pattern of local adaptation of the parasite but the sympatric combinations induced better hatching of cysts than allopatric combinations, and there was a negative relationship between the hatching percentage and the geographical distance between nematode populations and wild potatoes. Moreover, a strong effect of the geographic origin of root exudates was found, with root exudates from south of Peru inducing better hatching than root exudates from north of Peru. These results could be useful to develop new biocontrol products or potato cultivars to limit damages caused by G. pallida.     

Changes in the second stage juveniles of Globodeva rostochiensis prior to hatching in response to potato root diffusate

cAMP levels in eggs of G. rostochiensis and the diameter of the nucleolus of the nucleus within the dorsal pharyngeal gland cell of the second stage juvenile have been measured as indicators of the response of the nematode to the hatching stimulus in potato root diffusate. The nucleolus increased from 2.72 ± 0.103 μm for unhatched individuals to 3.28 ± 0.14 μm and 3.88 ± 0.15 μm after soaking eggs in potato root diffusate for 3 and 4 days respectively. Juveniles expressed from unstimulated eggs in water to potato root diffusate for 4–5 days showed a similar increase in size of the nucleolus to 3.94 ±0.15 μm but those released into water for this time had smaller nucleoli of 3.20 ± 0.98 μm. The change in diameter of the nucleolus is probably related to the accumulation of secretions in this gland cell before hatching. Preliminary results with dibutyryl analogues of CAMP and cGMP showed some inhibition of hatch in 10% potato root diffusate. Theophylline had a similar effect but NaF was dissimilar in that the effect of this inhibitor was not reversible. A standard radioimmunoassay showed that significant changes in cAMP levels occurred in the unhatched juveniles within cysts after treatment with potato root diffusate for 2.5 or 8 h compared with values for cysts kept in water. This change occurs before other known responses of the juveniles to potato root diffusate and it defines the period of interest for future work on the initial action of hatching factor.     

Observations on the biology of the potato-root eelworm, Heterodera schachtii Schmidt

Results are presented of experiments on the effect of low temperature and of soil type upon the potato-root eelworm, Heterodera schachtii.
In some experiments exposure of cysts, to low temperatures had a definite lethal effect, but in others the cysts were unharmed. The reason for this variation was not apparent. Exposure of the larvae of die nematode to even a relatively slight degree of frost resulted in the death of the larvae, provided mat die surrounding medium was completely frozen. In this respect the larvae appear to differ markedly from those of the beet strain of H. schachtii. That the larvae had actually been killed was shown by the fact that they produced no cysts on the roots of potato plants and also by their almost immediate absorption of iodine.
It was confirmed that H. schachtii cysts produced on the roots of potato plants grown in a heavy medium are smaller in size and fewer in number than those formed on the roots of plants grown in a light medium. The heavy medium used was pure clay, the light one pure sand, while equal numbers of eelworm larvae served in each case as inoculum. The difference in cyst size was significant, and fewer larvae emerged from the cysts formed in clay than from those produced in sand.     

THE PERMEABILITY TO THE HATCHING FACTOR OF THE CYST WALL OF THE POTATO-ROOT EELWORM, HETERODERA ROSTOCHIENSIS WOLLENWEBER

Experiments are described in which a particle of concentrated hatching factor is separated by cyst wall from eelworm eggs. Comparison of the hatching of these and control eggs showed that the cyst wall is permeable to the hatching stimulant. The experiments also demonstrated that the cyst wall is permeable to dissolved air.     

MOVEMENT OF EELWORMS: V. OBSERVATIONS ON APHELENCHOIDES RITZEMA-BOSI (SCHWARTZ, 1912) STEINER, 1932 ON FLORISTS' CHRYSANTHEMUMS

Adults of Aphelenchoides ritzema-bosi tend to migrate up the stems of chrysanthemum plants in stationary water films possibly by a negatively geotropic response. A current of water down the stem opposes such an upward movement. Greatest mobility occurred in thick films of water in places with a high concentration of epidermal hairs as at the top of the stem and on the undersurface of leaves. Ciné films of movement in thick and thin films showed that there were fundamental differences in the type of locomotion in these two environments. Invasion of leaves via stomata was observed and the method of movement is described. The presence of A. ritzema-bosi in leaves appears to render the epidermis permeable to water. During dry weather there is little movement inside the leaf, but after rainfall activity increases as water enters the leaf. Spread of eelworm infestation in the leaf occurs in the mesophyll and across veins although initially these act as barriers. Emergence occurs via the stomata, chiefly on the undersurface. When the leaf is wet, about 50% of the eelworms emerge in the first hour. During wet weather many eelworms were recovered from the surfaces of leaves and it is suggested that eelworms spread mostly under these conditions.