The buoyant densities of bean yellow mosaic virus (BYMV) B25, pea mosaic virus (PMV) E198, lettuce mosaic virus (LMV), and potato virus yN (pVyN) were 1.318, 1.321, 1.330, and 1.326 g/ml, respectively. Their S values were 143, 140, 143, and 145 S. The particle morphology of BYMV B25, PMV E198, and LMV could reversibly be changed by magnesium ions. PVY N particles were broken in the presence of magnesium ions. The molecular weight of the coat protein subunit of the four viruses was 34,000 daltons. In many preparations also a 28,000 daltons component was present. This must be considered to be a breakdown product, derived from the 34,000 daltons component by proteolytic activity.
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.
Stunt viroid can be detected in chrysanthemums with the polyacrylamide gelelectrophoresis (PAGE) method developed by Morris and Smith (1977) for potato spindle tuber viroid. The time of sample preparation can even be shortened considerably. The reliability of the short and the complete PAGE method proved to be similar to that of the biological 'Mistletoe' test in a parallel experiment. Combined samples can be tested in the complete PAGE method easily permitting the detection of one diseased chrysanthemum top in a total of ten.Although potato spindle tuber viroid is not known to occur in the Netherlands we searched for methods to detect possible infections. Artificial infections of tomato and potato plants and o f sprouts of potato tubers could readily be detected by Morris and Smith's method. Using this method it was possible to demonstrate infections by severe and weak isolates even when not yet producing symptoms. In tomato plants the viroid could be detected four to eight days before symptoms appeared.
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