Plant Physiol.
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Plant Physiology 65:309-313 (1980)
© 1980 American Society of Plant Biologists

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Transcription in Isolated Wheat Nuclei

II. CHARACTERIZATION OF RNA SYNTHESIZED IN VITRO1

Dawn Sywassink Luthe2 and Ralph S. Quatrano3

Department of Botany and Plant Pathology, Oregon State University, Corvallis, Oregon 97331

Nuclei free of RNase activity were isolated from 3-hour-imbibed wheat (var. Yamhill) embryos by centrifugation through a discontinuous gradient of Percoll. The maximum rate of RNA synthesis observed in these nuclei was approximately 5 picomoles [3H]UTP per milligram DNA per minute. Two monovalent cation optima were found when measured in the presence of 15 millimolar MgCl2 or 2 millimolar MnCl2; 15 and 75 millimolar (NH4)2SO4. At the high monovalent cation optimum, the rate of RNA synthesis was linear for the first 10 to 15 minutes of incubation and still increasing after 3 hours. RNA synthesized in vitro (30-minute pulse followed by a 30-minute chase) showed distinct 18 and 26S RNA peaks comprising 13 and 17% of the total radioactivity, respectively. The over-all pattern of RNA synthesized in vitro was similar to the in vivo pattern. Approximately 40 to 50% of the RNA synthesized was inhibited by {alpha}-amanitin at 4 micrograms per milliliter. The newly synthesized 6 to 10S RNA appeared to be selectively inhibited by {alpha}-amanitin.

2 Present address: Department of Biochemistry, P.O. Drawer BB, Mississippi State University, Mississippi State, Mississippi 39762.

3 To whom reprint requests should be addressed.

1 This research was supported by the Science and Education Administration of the United States Department of Agriculture under Grant 5901-0410-8-0196-0 from the Competitive Research Grants Office, and by the United States Public Health Service under Grant GM 19247.






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ASPB Publications PLANT PHYSIOLOGY® THE PLANT CELL
Copyright © 1980 by the American Society of Plant Biologists