The hatching factor of the potato-root eelworm

1. The hatching factor of the potato-root eelworm was concentrated from potato-root leachings by adsorption on charcoal. The crude material, extracted from charcoal with acetone, was purified by partition between ethyl acetate and m-potassium dihydrogen phosphate, and ethyl acetate and a solution of potassium metabisulphite (disulphite). Neutral material was removed by extraction from phosphate buffer at pH7.8. The yield of purified material, after chromatography on silica, DEAE-cellulose and Perlon columns, was 18mug./l. of potato-root leachings, and 0.04% by weight of the acetone-soluble material extracted from charcoal; 20% of the activity of the original potato-root leachings was recovered. 2. The purified material was a colourless gum. On paper chromatograms it gave a single weakly fluorescing spot, which reacted with 2,4-dinitrophenylhydrazine and aniline-xylose reagents. It was confirmed that the hatching factor is an acid, and it is inactivated at values above pH9. Other properties were a ready oxidation by permanganate and reaction with certain carbonyl reagents. Colour tests for alpha- and beta-unsaturated gamma-lactones were negative. The hatching factor showed no light-absorption maxima at wavelengths above 210mmu; absorption bands in the infrared region (670-4000cm.(-1)) are given. Our purified material, although a gum, gave a similar hatch to a ;crystalline' product at 640 times its concentration.     

The inhibition of hatching of potato root eelworm (Heterodera rostochiensis Woll.) in partially sterilized soil

Experiments on the hatching of Heterodera rostochiensis have shown that the addition of ammonium carbonate to potato root water markedly inhibits hatching when the concentration of ammonia introduced is approximately 100 p.p.m. The strong acid salts of ammonia in equivalent amounts have no such inhibitory effect.
These observations are linked with experiments on the effect of partial sterilization of soil on the hatching of H. rostochiensis , and it is demonstrated that delay of hatching in such soils is only effective so long as the ammonia concentration within the soil is maintained at a sufficiently high level.     

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.     

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.     

The resistance of potato varieties to Heterodera schachtii Schmidt, the potato-root eelworm

The potato varieties Epicure and Doon Star permitted the production of fewer and smaller cysts of Heterodera schachtii on their roots than the varieties Golden Wonder and Majestic, when all were grown under standard conditions. Under the stimulus of potato-root excretion the number of larvae which emerged from Epicure and Doon Star cysts was less than that from Majestic or Golden Wonder cysts of the same volume. Accepting Lapage's (1938) criteria of resistance to nematode infestation in animals and applying them to plants, it is concluded that this deleterious effect of the variety on the eelworm is probably of the nature of a degree of resistance to eelworm attack specific to each variety and is physiological and not anatomical in action. Of the varieties tested, Epicure is judged the most resistant, closely followed by Doon Star. Majestic is fairly susceptible but not so much so as Golden Wonder on which the eelworm forms large cysts that are extremely productive of larvae.     

Assessment of the damage caused to potatoes by potato cyst eelworm, Heterodera rostochiensis Woll

Thirty field trials were carried out from 1960 to 1967, mainly on peat and silt soils in the eastern counties of England, to study the relationship between potato yield and numbers of H. rostochiensis. The population range of most interest to advisers (0–60 full cysts/100 g (0–150 eggs/g)) was encountered by chance fairly frequently. The relationship between yield and initial eelworm densities over this range could be expressed equally well untransformed or transformed (using square root transformation for cyst counts and log. transformation for egg counts). Sigmoid curves could not often be fitted to the graphs of yield against log. egg count because densities large or small enough were not often encountered and small numbers cannot be estimated with sufficient accuracy. Assuming straight-line regressions of yield on untransformed eelworm densities certain generalizations can be made. The slopes vary somewhat, but on average losses are 1·2 tons/acre per ten full cysts/100 g and 0·85 tons/acre per twenty eggs/g. Linear regressions tend to be parallel over a large range of potential yields which means that yield loss is independent of potential yield. In five trials there was some evidence that the yield had reached a minimum varying between 5 and 10 tons/acre when eelworm populations were very dense. In two trials, estimates of maximum and minimum yield were obtained. Both gave the difference (maximum loss) as 7 tons/acre and the ratio of minimum to maximum as 0·50 and 0·44 respectively. Yield losses were often great without obvious symptoms appearing in the haulm.     

Partial purification of hatching activity for Globodera rostochiensis from potato root diffusate

The potential of several chromatographic methods for isolating hatching factors for potato cyst nematodes from potato root diffusate was investigated using a bioassay based on emergence of juveniles from cysts. Gel filtration provided an overall estimate of molecular weight of 437 Da for the hatching activity and ion exchange chromatography indicated that at least 60% of the recovered activity was anionic in nature. Material less polar than the hatching activity could be removed by passing potato root diffusate through a reversed-phase Sep-Pak C₁₈ cartridge and the elutant showed 83.3 ± 4.4% (mean of 32 cysts) of the initial hatching activity. High performance liquid chromatography (HPLC) with a reversed phase, C₁₈ column and gradient elution (0–80% CH₃OH in water) confirmed that much of the hatching activity was polar and that it was not retained by this method of separation. A weak anion exchange resin achieved slight retention of much of the hatching activity and an ion pairing reagent lowered the polarity sufficiently to allow some retention in subsequent reversed phase HPLC on a C_IS column. Both ion exchange and ion pairing HPLC suggested that hatching activity was not chromatographed as a single compound and indicated that fractions able to influence the nucleolus of the nucleus within the dorsal pharyngeal gland cell did not always show hatching activity.     

Resolution of natural hatching factors for golden potato cyst nematode, Globodera rostochiensis

The hatching responses of Globodera rostochiensis (golden potato cyst nematode) to purified and partially-purified preparations of natural (including the potato glycoalkaloids solanine and α-chaconine) and artificial hatching factors (HFs) were bimodal. At least 10 HFs, mostly anionic, were resolved from potato root leachate by a combination of gel permeation and ion-exchange chromatography. Whereas potato roots were the principal source of HFs, haulm leachate also contained such chemicals. Root leachate from aseptically-grown potato plants lacked several HFs which were present in conventionally-produced leachate.     

The behaviour of populations of potato cyst eelworm Heterodera rostochiensis Woll. Towards resistant potato varieties derived from Solarium tuberosum ssp. andigena Juz & Buk

Many populations of Heterodera rostochiensis mostly from the main potato growing areas of England and Wales, were tested with eelworm-resistant potato varieties (andigena hybrids). Except in the Eastern Region, where resistant varieties were planted alongside commercial varieties in infested fields, tests were done in pots inoculated with a standard number of cysts extracted from a single soil sample from each field. The tests showed that England and Wales could be divided into areas according to the percentage of cysts(< 10%, 11–70%,> 70%) produced on the resistant compared with a commercial non-resistant variety. On the peat soils of the East Anglian fens and in Bedfordshire, Essex and Kent (area A) most populations produce few cysts on resistant varieties. In much of the East Midlands and Yorkshire (area D), populations produce many cysts on resistant varieties which behave much as ordinary susceptible potatoes. Populations in the rest of England and Wales (areas B, C) vary and in many districts every field must be tested separately. If the pathotypes occurring in Britain had separate origins in South America, the present distribution in England and Wales is probably due to chance introductions dispersed with seed and following the patterns of trade in seed potatoes. As all commercial varieties, before the release of eelworm-resistant ones, were about equally susceptible to the different pathotypes, the varieties themselves probably played little part in determining the present distribution of pathotypes.     

The interrelationships of the causal fungus of brown root rot of tomatoes and potato root eelworm, Heterodera rostochiensis Woll.

In a study of the association between the causal fungus of brown root rot of tomatoes and Heterodera rostochiensis Woll., it was demonstrated that the nematode did not increase the susceptibility of the roots to invasion by the fungus; however, the fungus decreased the hatch of the potato root eleworm, the invasion of the host plant by the nematode, and number of new cysts subsequently produced.