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Large Scale Hatching, Disinfestation, and Storage of Heterodera schachtii Larvae. E. D. Whitney, Research Plant Pathologist, Crops Research Division, ARS, USDA, Salinas, California 93901; D. L. Doney, Geneticist, Crops Research Division, ARS, USDA, Salinas, California 93901. Phytopathology 60:1191-1194. Accepted for publication 11 March 1970. DOI: 10.1094/Phyto-60-1191.

Methods of hatching, disinfesting, and storing large numbers of second-stage Heterodera schachtii larvae without loss of infectivity are described. For greenhouse studies, larvae are hatched by incubating screened cysts for 3-5 days in 4 mm zinc chloride, 10 ppm ethoxyethyl mercury chloride, 0.01% dioctyl sodium sulphosuccinate, 1 mg/ml streptomycin sulfate, and 1,000 units/ml penicillin G potassium. The larvae are further treated in the same solution for 72 hr to reduce contamination. For pure culture studies, the larvae are hatched by incubating recently matured hand-picked cysts as described for greenhouse studies. The larvae are further disinfested, however, by placing them in the above solution plus neomycin sulfate (1 mg/ml) for 7 days. Surface disinfestation of the larvae was complete after storage at 24 C for 30 days as indicated on a wide range of media. Infectivity of the larvae was determined by staining and counting the number of larvae infecting 3-week-old sugarbeet seedlings grown in sand culture.