A comparison was made of methods for viroid detection. Molecular hybridization using cDNA is a very sensitive method that can handle large quantities of samples at the same time but it has the disadvantage that only small amounts of the sample can be applied to the nitrocellulose filter. The method therefore can only detect viroid in plants when its concentration is 1&20 ng g-' of leaves, using 32P as a marker system. Bi-directional electrophoresis can detect viroid in plants when its concentration is 10 ng g-' of leaves, because it uses larger samples. It does not need hazardous chemicals like 32P and formamide, and the reading of the results of the test is less liable to failures because it is based on two criteria (position and intensity of RNA band). The Dutch Plant Protection Service and the Dutch General Inspection Service for Ornamentals therefore use a modified bi-directional electrophoresis method to detect potato spindle tuber viroid and chrysanthemum stunt viroid, respectively.

Results are reported for several international collaborative experiments which examined methods of assessing degrees of partial resistance in potato and virulence in potato cyst nematode (PCN, Globodera pallida). It was demonstrated that absolute rates of multiplication can be extremely variable on both susceptible and partially resistant clones, even when the same population and test procedures are employed. It was therefore concluded that, on clones with quantitatively inherited resistance (i.e. partially resistant), absolute rates of multiplication cannot be used to separate pathotypes. Expression of these rates as percentages of those on the susceptible controls reduced the absolute differences between tests, but the values obtained were still too variable for statutory use. However, whatever the environment or nematode population used, it was observed that the resistance of the test clones and virulence of the nematode populations were generally ranked in a similar order. The main exceptions to this were: (1) a Petri‐dish test where cv. Vantage was less resistant than in canisters or pots and (2) a pot test where cv. Darwina tended to be more interactive with environmental factors than the other test clones. It was also observed that with some populations of PCN in pots the susceptible cv. Bintje was a less good host than cv. Désirée. On the basis of these results it is suggested that certain partially resistant clones should be used as internal references in statutory, recommended list and breeders' assessment tests against which the resistance of the test clones are compared. For international comparability it is necessary that the different centres conducting such tests use the same reference clones and nematode populations, and similar test methods.

A standard test method for detecting viroids was designed, to be applied on imported plant material, for which a zero-tolerance exists in the Netherlands towards potato spindle tuber viroid (PSTV).Partial purification of nucleic acids after homogenizing leaf material with a Polytron homogenizer, followed by increasing the viroid concentration by inoculation of an intermediate tomato host, and complete purification of the small nucleic acids from the tops of these plants, followed by polyacrylamide gelectrophoretic analysis, proved successful. With this procedure, now used as a standard method, more samples could be handled than with other methods tested.Desalting by Sephadex filtration proved to be superior to dialysis. An attempt to develop a serological test for PSTV failed. Albinism, induced in PSTV-infected tomato plants by certain environmental conditions, was not of diagnostic value.Additional keywords: potato spindle tuber viroid, chrysanthemum stunt viroid, Sephadexdesalting, serology, viroid-induced albinism.

In The Netherlands germplasm and plants collected in the wild for breeding and scientific purposes may only be imported with a special licence. For this material specific post‐entry import requirements are prescribed. A glasshouse with special quarantine facilities has been built on the premises of the Plant Protection Service at Wageningen. In this paper the special facilities needed to carry out post‐entry quarantine are described.

The polyacrylamide gel electrophoresis (PAGE) test of Morris & Smith (1977) was evaluated for detection of potato spindle tuber viroid (PSTV) in breeding material. Number, density and mobility of nucleic acid bands in the electropherograms were influenced by genotype and growing temperature. So direct testing of genotypes was not reliable. After an intermediate viroid multiplication in tomato host plants at about 30oC and high irradiance, PSTV was reliably detectable with PAGE in inocula of potato samples of diverse origin. A 4‐week incubation period proved to be suitable for inocula with low and high concentrations of a mild strain of PSTV (m‐PSTV) as well as a severe strain of PSTV (s‐PSTV). If incidence of PSTV is expected to be low, testing can be speeded up by bulking samples. With the combined tomato‐intermediate/ PAGE assay, one m‐PSTV or one s‐PSTV infected leaf disk in 200 healthy ones was consistently detectable. Occasionally gels with a nucleic acid band of about the same relative mobility as the viroid band were found. Evidence that these bands were not caused by viroid is presented. A procedure to resolve such questionable test results is described. Infectivity of s‐PSTV was higher than that of m‐PSTV. Concentration of viroids in the inoculum influenced appearance of mild or severe symptoms and the rate of symptom production.