The response of individual adult males of the potato cyst nematodes, Globodera rostochiensis and G. pallida, to sex pheromones from adult females was investigated using electrophysiological techniques. Each male nematode was pierced with an electrode close to the cephalic region and then exposed to pheromones from virgin females. Cellular responses in the form of action potentials were recorded as spike activity. The spike frequency produced by G. rostochiensis and G. pallida males increased significantly after the application of their homospecific pheromone. The spike frequency produced by G. pallida males also increased significantly after the application of G. rostochiensis female sex pheromone. In contrast, males of G. rostochiensis showed no significant response to G. pallida female sex pheromone. The electrophysiological results support and considerably extend information from agar plate behavioural bioassays.

At field rates (ca 0·1–10 μg active ingredient/ml), non-fumigant nematicides such as oxamyl do not kill plant-parasitic nematodes directly but act by impairing neuromuscular activity. The present study has shown that neutral lipid levels in 2nd-stage (infective) juveniles of Globodera rostochiensis, which had been incubated with the oximecarbamate nematicide oxamyl (1 μg a.i./ml) for between 8 and 35 days, were significantly (P < 0·05) greater when compared with controls maintained in water alone. The difference between treatments was found to be due principally to changes in the lipid content in the posterior half of the body which appears to act as the major long-term lipid store. When nematodes were allowed to recover for 72 h following incubation for 35 days with oxamyl, their infectivity was not significantly (P > 0·05) different from that of controls. This work is discussed in relation to the action of oxamyl and similar nematicides and to the use of controlled-release formulations as a means of increasing their efficacy.

Cysteine proteinases from the fruit and latex of plants, such as papaya, pineapple and fig, have previously been shown to have substantial anthelmintic efficacy, in vitro and in vivo, against a range of animal parasitic nematodes. In this paper, we describe the in vitro effects of these plant extracts against 2 sedentary plant parasitic nematodes of the genera Meloidogyne and Globodera. All the plant extracts examined caused digestion of the cuticle and decreased the activity of the tested nematodes. The specific inhibitor of cysteine proteinases, E-64, blocked this activity completely, indicating that it was essentially mediated by cysteine proteinases. In vitro, plant cysteine proteinases are active against second-stage juveniles of M. incognita and M. javanica, and some cysteine proteinases also affect the second-stage juveniles of Globodera rostochiensis. It is not known yet whether these plant extracts will interfere with, or prevent invasion of, host plants.

It has been demonstrated that the surface lipophilicity of the plant-parasitic nematode Globodera rostochiensis decreases when infective larvae are exposed to the phytohormones indole-3-acetic acid (auxin) or kinetin (cytokinin). In the present study, it was shown that inhibition of phospholipase C (PLC) or phosphatidylinositol 3 kinase (PI3-kinase) reversed the effect of phytohormones on surface lipophilicity. The signalling pathway(s) involved in surface modification were investigated using ‘caged’ signalling molecules and stimulators or inhibitors of different signalling enzymes. Photolysis of the ‘caged’ signalling molecules, NPE-caged Ins 1,4,5-P3, NITR-5/AM or caged-cAMP to liberate IP3, Ca2+ or cAMP respectively, decreased the surface lipophilicity. Activation of adenylate cyclase also decreased the surface lipophilicity. In contrast, inhibition of PI3-kinase using Wortmannin, LY-294002 or Quercetin, and inhibition of PLC using U-73122 all increased the surface lipophilicity. Two possible signalling pathways involved in phytohormone-induced surface modification are proposed.

The direct effects of phytohormones (auxin and kinetin) and root diffusates on the surface lipophilicity of the plant parasitic nematodes Globodera rostochiensis and Meloidogyne incognita were investigated. The fluorescent lipid probe AF18 (5-N-(octodecanoyl) aminofluorescein) was used to detect surface changes. Root diffusates increased AF18 uptake by G. rostochiensis while it had no effect on M. incognita. Kinetin and auxin decreased AF18 uptake in G. rostochiensis, while they had the opposite effect on M. incognita. Auxin/kinetin ratio was also found to be important in triggering the surface changes, especially at high concentrations. Whether plant nematodes have auxin and/or kinetin binding proteins is discussed as well as the mechanism behind the surface lipophilicity changes due to root diffusates and phytohormones.

Secretions were induced from second (invasive) stage juveniles (J2s) of the potato cyst nematode Globodera rostochiensis by exposing them to 5-methoxy-N,N-dimethyl tryptamine oxalate (DMT). Secretions were collected from J2s in sufficient quantity to allow direct analysis. Gel electrophoresis followed by monochromatic silver staining demonstrated the presence of at least 10 proteins. The presence of several enzymes, including superoxide dismutase and proteases, was demonstrated using Western blots and activity assays. Antisera raised against the secretions recognized bands on Western blots consistent in molecular mass with those identified on silver stained gels. The antisera recognized structures implicated in the production of secretions including the subventral gland cells and surface of J2s.

The total lipid content of the dry weight of whole cysts and 2nd-stage juveniles (J2) of Globodera rostochiensis was 17·1% in dry cysts, 20·9% in cysts soaked in distilled water (DW), 20·3% in cysts that had been in potato root diffusate (PRD) for 7 days, 7·3% in cysts that had been in PRD for 28 days and 29·2% for hatched J2. The fatty acid composition of the total lipid did not differ between dry cysts, cysts in DW and cysts in PRD for 7 days. However, major differences in the fatty acid composition of all lipid classes were found between rehydrated cysts in PRD and freshly hatched J2. After hatching, the degree of saturation and the percentage of monounsaturated fatty acids decreased and the percentage of polyunsaturated fatty acids increased considerably, especially in the free fatty acid fraction, where C20[ratio ]1 showed an 8-fold decrease and C20[ratio ]4 a 33-fold increase. There was a difference in the fatty acid composition of cysts in PRD for 7 days and cysts in PRD for 28 days (after most of the J2 had hatched); with increased time in PRD the percentage of polyunsaturated fatty acids increased and the percentage of monounsaturated fatty acids decreased in all lipid classes. Differences in the fatty acid profiles between cysts in PRD for 28 days and hatched J2 were found mainly in the free fatty acid and the non-acidic phospholipid fractions. The free fatty acid fraction of the cysts was dominated by monounsaturated fatty acids (52%), whereas the same fraction of hatched J2 contained large amounts of polyunsaturated fatty acids (60%). These polyunsaturated fatty acids, especially C20[ratio ]4, might play an important part in nematode–plant interactions during infection.

Electrophoresis of secretions collected from Globodera pallida revealed a smeared region between 25 and 50 kDa, and a single band of <20 kDa. The secretions were used to raise an antiserum (LW1). Immunoblotting of parasite homogenates with LW1 differentiated G. pallida from its sibling species G. rostochiensis and revealed differences between different populations of G. pallida and G. rostochiensis. Indirect immunofluorescence studies with LW1 indicated that at least some of the secretions were surface localized and that antibody binding to the nematode surface was periodate sensitive. Periodate sensitivity indicated that these differences could be due to glycosylation differences. Glycosylation differences were also detected by blotting nematode homogenates with the lectin wheat germ agglutinin (WGA). WGA was also able to differentiate between G. rostochiensis which gave 2 bands at 130 kDa and 110 kDa, and G. pallida which produced 2 bands present at 120 kDa and 110 kDa. Further localization studies using immunoelectron microscopy demonstrated that antibody binding could be seen to secretions found in the pump chamber of the metacorpal bulb at the base of the stylet. From further specimens it could be observed that the contents of the subventral glands were heavily labelled, indicating that the material seen in the metacorpal bulb had originated from the subventral glands.

Electrophysiological techniques and behavioural assays were used to determine the response of males of Globodera rostochiensis to 4 fractions of the homospecific sex pheromone from virgin females, separated using reverse-phase high-pressure liquid chromatography. Sex pheromone activity was confined to 2 of the fractions. The sex pheromone of G. rostochiensis appears to be composed of several weakly basic, polar compounds.

The potato cyst nematode (PCN) Globodera rostochiensis, like other parasitic nematodes, needs to synchronize its life-cycle with that of its host. This synchrony is achieved by the invasive-stage juvenile remaining dormant within its egg until stimulated to hatch by the presence of root diffusates of its host. Root diffusates may induce changes in gene expression in PCN, some of which may be important in the transition to a parasitic mode of existence. We have used a range of techniques including differential display to examine gene expression during stimulation and hatching of PCN. We find that few changes in gene expression appear to be induced directly by root diffusates. Instead, changes in gene expression seem to occur during or immediately after the hatching process. These results are discussed in the context of the host–parasite relationship.