By J. A. Thompson (auth.), I. Potrykus, C. T. Harms, A. Hinnen, R. Hütter, P. J. King, R. D. Shillito (eds.)

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Plasmid of Agrobacterium tumefaciens,or that the DNA maydis of opposIng matIng types (kindly supplied bYcopy of the dsRNA may be Involved In pathogenesis. Day, PBI, CamTo determine the role of the VLP's, determination of br~dge) were grown to early stationasy phase (approx. maydis isolate is being 10 cells/ml) in shake cultures at 28 C in minimal correlated with the presence or absence of the dsRNA's liquid culture medium (2). Stationary phase cultures in each isolate. If any correlation is found then the of opposing mating types were mixed in equal concenpracticability of incorporating the relevant dsRNA's trations with one drop of Tween 80 and 1 ml of the directly into lea mays protoplasts to induce divisions combined isolates were pipetted into the leaf whorl of 10 day old lea mays (var.

Nnitol (cpw 13M) for 1 hour. Leaves were peeled and the leaf segments were placed in 30 ml of CPW 13 M for 1 hour. In both cases epw 13 }.! 5% Macerozyme in CPW 13 M. After overnight incubation (about 14 hours) the protoplasts were isolated and counted following the usual methods. 5 mg/1) and BAP (1 mg/1). The protoplasts were then incUbated at 25°c with an illumination of about 1000 lux (16 hours photoperiod). Of all the cell densities tried, 5 X 104 cells/ml showed best growth. ~ The yield of protoplasts in mesopnyll and hypocotyl segments were 5 X 105 and 7 X 105/ml respectively.

69,854-58. n-propanol, n-butanol), before embedding in glycol 5) Billet,E. (1974), The host-paraSite physiology of methacrylate (3). l/um sections were cut on a Reichert Ustilago maydis, PhD thesis, Oxford University. Ultra Microtome. H. Bot. maydis were grown to statio403-410. nary phase. The mycelium was then broken by shaking the suspension with 0,45 mm glass beads (lg/ml) on an orbital shaker at 400 rpm for 15' in the presence of 10 mM Dipyrocarbonate. Shillito for (4). The dsRNA's were electrophorised in a 1% agarose discussions and 6t"h nrlvice.

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