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Hormonal control of tobacco protoplast nucleic acid metabolism during in vitro culture

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Abstract

Tobacco mesophyll protoplasts cultivated in vitro do not synthesize a measurable quantity of chloroplastic ribosomal RNA, but actively synthesize cytoplasmic ribosomal RNA, polyadenylated RNA, and proteins. These syntheses are essentially independent of the presence of hormones in the culture medium and are thus related to the ageing phenomenon induced by isolation from the plant and in-vitro culture. At all stages of culture and in all culture media, protoplasts incorporate low levels of thymidine into their DNA. However, the incorporation of considerable quantities of thymidine, indicative of the S phase, only takes place after 25–30 h and requires the presence of auxin and cytokinin.

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Abbreviations

6-BA:

6-benzyladenine

2,4-D:

2,4 dichlorophenoxyacetic acid

DPC:

diethylpyrocarbonate

OD:

optical density; oligo-dT cellulose-oligothymidylic acid-cellulose

poly A+ RNA:

polyadenylated RNA

poly A- RNA:

non-polyadenylated RNA

tRNA:

ribosomal RNA

SDS:

sodium dodecyl sulphate

TCA:

trichloroacetic acid

Tris buffer:

Tris (hydroxymethyl)aminomethane

tRNA:

transfer RNA

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Cooke, R., Meyer, Y. Hormonal control of tobacco protoplast nucleic acid metabolism during in vitro culture. Planta 152, 1–7 (1981). https://doi.org/10.1007/BF00384976

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  • DOI: https://doi.org/10.1007/BF00384976

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