Diversity of RNA Components in Green Plant Tissues

Loening, Ulrich E.; Ingle, John

doi:10.1038/215363a0

Cited by 300 publications

(99 citation statements)

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“…The filters were air-dried, and the radioactivity was determined after suspending filters in an appropriate scintillator. The sedimentation coefficients of rapidly labeled RNA fractions were determined as described by Martin and Ames (30), using the coefficients of light and heavy ribosomal RNA as 18 and 25 S (27).…”

Section: Methodsmentioning

confidence: 99%

Characterization of Rapidly Labeled Ribonucleic Acid from Dwarf Peas

Johri

Varner

1970

Plant Physiol.

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The ribonucleic acid synthesized by excised shoots of dwarf pea (Pisum sativum L. cv. Progress No. 9) during short labeling periods has been characterized. Thirty per cent of the total 32Pi incorporated in 1 hour is found in the ribosomal fraction. This labeled RNA was polydisperse (6-18 Svedberg units) and after chromatography on a methylated albumin-kieselguhr column about 80% of the radioactivity appeared in two peaks. One of these appeared on the shoulder of heavy ribosomal RNA ("mRNA") while the other was tenaciously bound to the column (TB-RNA).In the presence of high NaCl concentration, about half of the polydisperse RNA interacted with ribosomal RNA and eluted as "mRNA" while the remainder eluted as TB-RNA.This interaction in the presence of salt seems to result in the alteration of secondary structure because the "mRNA" fraction had a high sedimentation coefficient (45-50 Svedberg units). The polydisperse RNA approaches DNA in low cytidylate and guanylate content. After short periods of labeling TB-RNA showed higher adenylate content than "mRNA." The radioactivity from the "mRNA" peak can be chased, and these counts may represent a class of shortlived messenger RNA molecules with an average half-life of 10 to 15 minutes. The other component, TB-RNA, could not be chased and accumulated radioactivity during the chase period.In bacteria and viruses the information for the sequence of amino acids in a polypeptide chain is encoded into messenger RNA. Presence of a similar fraction has been suggested in some higher plants (5,6,21,26,39). All of our knowledge about messenger RNA from higher plants is presumptive and based on analogy to microorganisms; in no case has this fraction been purified or characterized, nor has its rate of synthesis or its stability been studied.We chose a dwarf pea as the experimental material because we have been investigating the RNA synthesized by nuclei isolated from shoots of this plant (19), and the knowledge obtained from the system in vivo may be applicable to the results obtained with isolated nuclei. MATERIALS AND METHODS Uptake and Incorporation of Labeled Precursors. Dwarf pea seeds (Pisum sativum L., cv. Progress No. 9) were surfacesterilized for 7 to 10 min in 1% sodium hypochlorite, implanted in 500-ml Erlenmeyer flasks containing moist, sterile vermiculite, and allowed to germinate at 23 C in continuous light. At the end of 5 or 6 days, shoots 0.8 to 1 cm long were excised and 10 or 15 shoots were incubated in 25-ml Erlenmeyer flasks under aseptic conditions with labeled uridine or 32Pj (carrier-free H3u2PO4, neutralized) in 2 ml of water containing 100 ,ug of chloramphenicol. Flasks were transferred to a shaker, and, after incubation for various times, the shoots were washed thoroughly in ice-cold 0.05 M KH2PO4 and deionized water and were then extracted for nucleic acids.In experiments designed to study the relationship between uptake and incorporation, the labeled shoots were homogenized in ethanol, 80% (v/v,) and the nucleic acids were determined (33). The ...

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Section: Methodsmentioning

confidence: 99%

Characterization of Rapidly Labeled Ribonucleic Acid from Dwarf Peas

Johri

Varner

1970

Plant Physiol.

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“…Tobacco mosaic virus (TMV) and cowpea mosaic virus (CPMV) were purified by methods similar to those of Boedtker & Simmons (1958) and van Kammen & De Jager (1978), respectively. Tobacco leaf RNA was isolated from leaves of uninoculated plants by the method of Loening & Ingle (1967).…”

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confidence: 99%

VPg-mediated Aggregation of Potyviral RNA

Luciano¹,

Murphy²,

Rhoads

et al. 1991

Journal of General Virology

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“…Indeed, this material brings on several difficulties, like the cellular heterogeneity, the non uniform labelling by radioactive precursors and the bacterial contamination encountered with naturally growing plants. Moreover, the presence in higher plants of several kinds of organelles adds to the above difficulties; particularly chloroplasts have been shown to contain specific 23 S and 16 S rRNA molecules [9,10]. This paper reports for the first time, as far as we know, the identification and characterization of mt rRNA extracted from plant cells, asceptically cultured in liquid nutrient medium.…”

Section: Introductionmentioning

confidence: 99%