Variability in Complementarity for Chloroplastic and Cytoplasmic Ribosomal Ribonucleic Acid among Plant Nuclear Deoxyribonucleic Acids

Matsuda, Kazutoshi; Siegel, Albert; Lightfoot, Donald

doi:10.1104/pp.46.1.6

Cited by 21 publications

(8 citation statements)

References 32 publications

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“…These results are strikingly different from those reported in other systems. For example, the number of cytoplasmic rRNA genes in the nuclear DNA (nDNA) varies considerably among the different angiosperm species (4,5). Similarly, distantly related bacterial species have been shown to contain rRNA genes which show only partial sequence homology (6).…”

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confidence: 99%

Conservation of 70S Ribosomal RNA Genes in the Chloroplast DNAs of Higher Plants

Thomas

Tewari

1974

Proc. Natl. Acad. Sci. U.S.A.

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Chloroplast DNAs of higher plants have been found to contain two chloroplast ribosomal RNA genes. The base sequences of these chloroplast ribosomal RNA genes in chloroplast DNAs have been studied by molecular hybridization. The results have shown that these genes have undergone little divergence in the evolution of either monocotyledonous or dicotyledonous plants.Chloroplast DNA (ctDNA) has been shown to contain genes for the 70S ribosomal RNA of chloroplasts (1,2). Detailed hybridization studies using pea ctDNA and ct-rRNA isolated from 70S ribosomes from pea leaves have shown that there are two ct-rRNA genes in the ctDNA (3). In this communication, we have further studied the ct-rRNA genes in ctDNAs from bean, lettuce, spinach, corn, and oats. The experiments have been carried out by hybridizing the ctDNAs from the above plants with 32P-labeled ct-rRNA from pea chloroplasts. The amount of hybridization in each case accounted for approximately two ct-rRNA gene equivalents in these DNAs. Competition hybridization and thermal stability experiments involving homologous and heterologous ct-rRNA * DNA combinations have shown that within the limits of this technique the base sequences of these genes are very similar in all the higher plants studied. These results are strikingly different from those reported in other systems. For example, the number of cytoplasmic rRNA genes in the nuclear DNA (nDNA) varies considerably among the different angiosperm species (4, 5). Similarly, distantly related bacterial species have been shown to contain rRNA genes which show only partial sequence homology (6). MATERIALS AND METHODSIsolation of ctDNAs and ct-rRNAs. ctDNAs were obtained from different plants in large quantities by the method described for pea leaves (7). This method consisted of making a cell free homogenate, centrifuging at 100 X g for 10 min to remove nuclei, and collecting the crude chloroplast fraction by centrifuging the supernatant solution for 15 min at 1020 X g. The lysed chloroplasts were centrifuged at 17,000 X g for 30 min and the top threefourths of the supernatant solution above the loosely packed pellet was collected. Aliquots containing 60-80 A2eo units were then loaded on 10-30% linear sucrose gradients. After 4 hr of centrifugation at 27,000 rpm in a SW-27 rotor, the gradients were fractionated for 70S ribosomes, and the RNA from the pooled fractions of ribosomes was extracted by adding 30 ml of buffer containing 0.05 M Tris * HCI-25 mM MgCl2-25 mM KCI-2% sodium dodecyl sulfate-2% Triton X-100 (pH 7.5) and 30 ml of water-saturated phenol. After three such extractions, two volumes of 95% ethanol were added to the final aqueous phase and the precipitated RNA was suspended in buffer containing 2 mM Mg(OAc)2-0.1 mM ethylenediaminetetraacetate (EDTA)-0.05 M Tris-HCI (pH 8.0). The same procedure was used for the isolation of ct-rRNA from other plants.Hybridization. ctDNA was denatured by alkali, neutralized, and fixed on Schleicher & Schuell B-6 membrane filters as reported (8). The hybridization co...

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confidence: 99%

Conservation of 70S Ribosomal RNA Genes in the Chloroplast DNAs of Higher Plants

Thomas

Tewari

1974

Proc. Natl. Acad. Sci. U.S.A.

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“…It should be pointed out that although data have been published which indicate that the only functional genes for the production of 23S and 16S rRNA in Chlamydomonas are in the chloroplasts (35), data have not been forthcoming showing this to be the case in higher plant cells. In fact, recent evidence indicates that nuclear DNA isolated from several different higher plant species contains sequences complementary to both chloroplastic and cytoplasmic rRNA (28). Therefore, the possibility for the existence of functional 23S and 16S rRNA genes in the nucleus of the higher plant cell cannot, at this time, be ruled out.…”

Section: Discussionmentioning

confidence: 99%

“…Although many studies concerning nucleic acid synthesis during germination have been reported (5,16,27,28,40), little consideration has been given to the cellular configuration of the samples studied (i.e., are the cells G1 [2C] or G2 [4C], or are they a mixed population?). Work which has taken this into account has been of a cytochemical nature (3,4,6,15,37) and few if any biochemical determinations have been made.…”

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confidence: 99%

Nucleic Acid Metabolism in Germinating Onion

Melera

1971

Plant Physiol.

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Nucleic acid synthesis in the GI cell population of the 1-millimeter apex of the Allium cepa embryo was studied during the initial 73 hours of germination. Quantitative data indicate that the total amount of RNA per cell began to increase after 18 hours of germination while the initial DNA per cell increase did not occur until some 20 hours later. Polyacrylamide gel electrophoresis patterns of 3H-uridine-labeled total nucleic acid samples indicated that synthesis of all detectable RNA fractions present in the pre-emergent 1-millimeter apex (i.e., cytoplasmic and "chloroplast-like" RNA) (3,4,6,15,37) and few if any biochemical determinations have been made.The Allium root tip offers a unique opportunity to study a population of higher plant cells which, while in the early stages of germination, are in an extended G1 phase (3).This report presents the results of experiments designed to map the pattern of nucleic acid synthesis in the 1 mm apex of the A llium root tip during the initial 73 hr of germination.The apical 1 mm of the A llium embryo contains several tissues including the leaf primordium, cotyledon, hypocotyl, promeristem, provascular tissue, and root cap (11). These tissues are associated with the apical 1 mm of the axis throughout germination until the emergent axis is approximately 5 mm in length, by which time the leaf primordium and cotyle- Philadelphia, Pa.) were sown on moist filter paper in covered glass Petri dishes and placed in a dark incubator at 24 C. At the appropriate time seeds were removed and taken to a 24 C darkroom where they were harvested under dim green light. Care was taken to keep the seeds in the dark at all times prior to harvesting. All samples were sown to be harvested between the hours of 8 and 10 AM.Seeds were sown in the manner described. At 36 hr the lengths of all emergent roots were estimated to the nearest 0.1 mm. All those roots between 0.8 mm and 1.2 mm in lengths were transferred to new dishes and measured at subsequent 24-hr intervals. The mean length and standard deviation for each root was recorded and the data used to plot a standard 1-mm growth curve.The pre-emergent intervals analyzed were defined by the length of time the seeds were exposed to germinative conditions and included 4-, 12-, 18-, and 24-hr seed. Postemergent intervals included 1-, 2-, 5-, and 10-mm roots, all of which were harvested at the optimal time designated by the growth curve. In all cases the apical 1 mm of the root was used as sample. After being subjected to the bacterial decontamination step outlined below, the pre-emergent samples were harvested by gently squashing the seeds with a spatula (thereby forcing the embryo from the seed at the micropylar end), and then excising the apical 1 mm of the root with a scalpel. The postemergent samples were treated with the decontamination procedure, and the apical 1 mm of the root was excised directly.Extraction of the Tips for TNA3 and Subsequent Prepara-

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“…Indeed, in some plants, sequences complementary to ribosomal RNA appear to be concentrated in the satellite band (4,10). These have been estimated to account for perhaps 15%o of the satellite in muskmelon (8), and to contain cistrons for both cytoplasmic and chloroplastic rRNAs in pumpkin (11). Other rapidly reannealing and repetitive satellite DNA sequences whose functions were not determined have also been described (7,8).…”

Section: Discussionmentioning

confidence: 99%

“…In this regard, several higher plants, including cucumber, have previously been shown to contain particularly large amounts (up to 44%) of satellite DNAs in neutral CsCl (4)(5)(6)(7)(8). These large plant satellites-in analogy with the satellite DNAs of animal cells (9)-have variously been reported to be rapidly renaturing (4,7,8,10), associated with the genes for rRNA (4,11) and of nuclear origin (7,10,12). The relationship between cytoplasmic organelle DNA and the large plant satellites, however, was not seriously investigated.…”

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Satellite-Rich DNA in Cucumber: Hormonal Enhancement of Synthesis and Subcellular Identification

Kadouri¹,

Atsmon²,

Edelman³

1975

Proc. Natl. Acad. Sci. U.S.A.

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Cucumber hypocotyl DNA in neutral CsCl distributes into a mainband comprising 59% of the total, and two large satellite bands which contribute 41%to the DNA pattern. Organelle enrichment studies show that the densities of mitochondrial and chloroplast DNA coincide with those of the satellite bands. At least 12-19% of total cucumber DNA is associated with the cytoplasmic organelles. These In prior studies, gibberellin-induced elongation of the cucumber hypocotyl was correlated with an increased incorporation of [3H]thymidine into DNA (1). Autoradiographic and fractionation experiments (2) further suggested that much of this synthesis might be extranuclear, reflecting a hormoneenhanced multiplication of chloroplasts in the elongating zone (3). This possibility has now been investigated at the level of different DNA density peaks by CsCl gradient centrifugation. In this regard, several higher plants, including cucumber, have previously been shown to contain particularly large amounts (up to 44%) of satellite DNAs in neutral CsCl (4-8). These large plant satellites-in analogy with the satellite DNAs of animal cells (9)-have variously been reported to be rapidly renaturing (4,7,8,10), associated with the genes for rRNA (4, 11) and of nuclear origin (7,10,12). The relationship between cytoplasmic organelle DNA and the large plant satellites, however, was not seriously investigated.In the present report we demonstrate that a considerable fraction of the large satellite DNAs in cucumber are associated with chloroplasts and mitochondria. We further show that in elongating hypocotyl tissue the rates of synthesis of organelle-rich satellite DNA and mainband DNA respond in vivo to hormone treatments by increasing markedly. Alternatively (method 2), hypocotyl sections were processed as described by Tautvydas (17) except that steeping was carried out at 0-2°in 4% gum arabic, 0.4 M sucrose, 6 mM Mg acetate, 0.3 mM dithiothreitol, and 5 mM morpholinoethane sulfonate, pH 6.4. Resuspension buffer was without gum arabic and contained 0.15% 2-ethyl-1-hexanol. Method 3 was identical to method 2 except that 0.1% Triton X-100 was included during the steeping procedure. For the isolation of chloroplasts the filtered homogenate was centrifuged at 1000 X g, and the pellet was dispersed in modified Honda medium supplemented with 40 mM Mg+2 and incubated with DNase (70 ug/ml) at 0°for 15-20 min. After addition of 0.1 M EDTA, discontinuous sucrose gradient centrifugation was carried out (2). For the isolation of mitochondria filtered homogenate was centrifuged at 5000 X g for 5 min and the supernatant was recentrifuged at 23,500 X g for 20 min.The 23,500 X g pellet was treated with DNase as described for chloroplasts and then rinsed in Tris-saline-EDTA.

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Variability in Complementarity for Chloroplastic and Cytoplasmic Ribosomal Ribonucleic Acid among Plant Nuclear Deoxyribonucleic Acids

Cited by 21 publications

References 32 publications

Conservation of 70S Ribosomal RNA Genes in the Chloroplast DNAs of Higher Plants

Conservation of 70S Ribosomal RNA Genes in the Chloroplast DNAs of Higher Plants

Nucleic Acid Metabolism in Germinating Onion

Satellite-Rich DNA in Cucumber: Hormonal Enhancement of Synthesis and Subcellular Identification

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