Isolierung und Charakterisierung schnell markierter,hochmolekularer RNS aus frei suspendierten Calluszellen der Petersilie (Petroselinum sativum)

Summary By culturing of callus tissue originating from root explants of Petroselinum sativum in a synthetic liquid medium under aeration, freely suspended single cells and small clusters consisting of mostly five cells were obtained. The rapidly dividing cells did not exhibit any morphogenesis. Their nucleic acid metabolism was investigated by pulse experiments with ³²P-orthophosphate. Rapidly labelled RNA was prominently found associated with high molecular RNA. During the fractionation of the total nucleic acids on MAK columns it was eluted after the ribosomal RNA components. Its base ratio, however, differed from the latter in that the AMP content was higher than the GMP content. Sucrose gradient centrifugation and polyacrylamide gel electrophoresis resulted in the separation of the ribosomal RNA from the rapidly labelled RNA, thus proving the higher molecular weight of the latter. Based upon the migration in the gel a sedimentation coefficient of approximately 32S was calculated. The possible function of the heavy rapidly labelled RNA component as precursor of ribosomal RNA is discussed.     

Growth and Dormancy Cycles in Citrus Bud Cultures and Their Hormonal Control

An in vitro bud culture method was devised in order to better understand the control mechanism of Citrus bud development. This technique offers a new approach to the study of hormonal control of growth, dormancy and flowering cycles in perennial plants. Buds were excised from orchard trees throughout the year, cultured on defined media for prolonged periods, and their vegetative growth responses to various growth hormones were determined. The buds proceeded with their vegetative development in vitro and achieved sprouting on a basal medium. The various growth regulators affected both the time required for sprouting (TRS) and the type of growth. In summer buds, IAA delayed sprouting, while GA enhanced it and caused shoot elongation. Cytokinins specifically induced the formation of numerous adventitious buds, whereas ABA completely inhibited sprouting; this inhibition, however, was reversible. A marked decrease in total protein and in the rate of its synthesis was evident during the first 20 days of sprouting induction and early bud growth. The annual growth rhythm was determined in spring buds sampled and cultured throughout the year, and an innate dormancy of citrus buds was revealed. Both the dormancy and the sprouting periods of buds in vitro corresponded to the natural periods occurring under field conditions. The effect of exogenous IAA, GA and cytokinins on the TRS varied at different periods along the season, suggesting the concept of “critical levels” in the endogenous balance of hormones.     

Occurrence of Indole-3-Acetic Acid in Buds of Pinus silvestris

The major ether-soluble, growth-stimulating substance detected by the Avena coleoptile straight-growth test in extract from sprouting buds of Scots pine (Pinus silvestris L.) was identified as indole-3-acetic acid by Rf values in 5 solvent systems and by its elution volume in ethanol on a Sephadex LH-20 column. When the substance was applied to the growth solution of wheat roots in a special test the growth in length of the roots was at first inhibited, but growth was recovered after about 6 hours in the same manner as when small quantities of IAA were applied. The extracts also contained large amounts of growth inhibitors which interfered with the auxin response if they were not removed.     

Phytochrome Controlled RNA Changes in Terminal Buds of Dark Grown Peas (Pisum sativum cv. Alaska)

The terminal buds of intact dark grown Alaska pea plants (Pisum sativum) respond to low intensity red light by an enhanced rate of expansion of their leaflets. Twenty four hours after irradiation, red light was found to have increased the ribosomal fraction of the RNA content of the terminal buds. The increase in total RNA liter did not occur if the red light was immediately followed by irradiance with low intensity far red light.     

Low temperature stress-induced biochemical changes affect stubble bud sprouting in sugarcane (<Emphasis Type="Italic">Saccharum</Emphasis> spp. hybrid)

A laboratory experiment was conducted to study the effect of low temperature stress on stubble bud sprouting and associated biochemical changes in sugarcane (Saccharum spp. hybrid). At 25°C, stubble bud sprouting was about 80%, whereas at 15 and 6°C, it was 56% and 23%, respectively. In stubble buds, the levels of reducing sugars and acid invertase were low, while IAA, total phenols and proline contents were high at low temperatures, as compared to normal temperature (25°C). Similarly, the specific activities of antioxidant enzymes, viz., catalase and peroxidase in stubble buds were higher at low temperatures than at normal temperature. The results indicate that poor sprouting of stubble buds at low temperatures appears to be due to a reduced availability of reducing sugars concomitant with a lower activity of acid invertase. An increased level of IAA together with toxicity build-up in situ due to an accumulation of total phenols may be responsible for the maintenance of dormancy in stubble buds at low temperatures. On the other hand, higher activities of catalase and peroxidase enzymes may protect stubble buds from an oxidative damage, while proline accumulates to act as an osmoprotectant under low temperature stress.     

In vitro Protein Synthesis by Polyribosomes of Peach Flower Buds at Different Physiological Stages

The in vitro phenylalanine incorporation by polyribosomes of peach flower buds (Prunus persica Stokes) during dormancy, dormancy break and flowering was investigated. Protein synthesis was measured using as catalyst either calf liver soluble factors or the ribosomal supernatant from the peach flower buds in the presence or the absence of the synthetic mRNA, polyuridylic acid. In the presence of polyuridylic acid, the activity of protein synthesis of dormant ribosomes is the same as that of ribosomes during dormancy break and flowering. The absence of synthetic messenger did not cause a change in activity. The ribosomal supernatant of dormant buds, but not of flowering buds, reduces the phenylalanine incorporation by polyribosomes from buds harvested at dormancy break.     

STUDIES ON THE LABELLING PATTERNS OF RNA FROM CEREBRAL CORTEX NUCLEI

RNA labelling patterns in nuclei from rat cerebral cortex were investigated subsequent to intracerebral injection of [³H]uridine. Although there was a rapid uptake of label into the ‘heavy’ regions when nuclear RNA was analysed in density gradients, it was not possible to show conclusive evidence for 455 ribosomal precursor RNA. Methyl-ation of 18S and 28S nuclear RNA became evident only after 2 hr and did not appear to involve the intermediacy of RNA species of higher molecular weight.     

Changes in the Protein Composition of the Shoot Apical Bud of Sinapis alba in Transition to Flowering

Vegetative plants of Sinapis alba L. grown in short days were induced to flower by expsoure to one or continuous long days. In both inductive conditions, the first flowers were initiated about 60 h after the start of the treatment. Soluble protein extracts were prepared from apical buds and just-expanded leaves of both vegetative and induced plants. Rabbit antisera were prepared using extracts from vegetative and reproductive buds. Immunodiffusion tests were performed. Analysis of the precipitin bands indicated that: (1) one antigenic protein was present in the vegetative buds and disappeared from the buds of induced plants between 96 and 240 h after the start of the inductive treatment; (2) the concentration of a another antigenic protein increased in buds of induced plants 30 h after the start of the inductive treatment; (3) the concentration of a third antigenic proteín increased in buds of induced plants at 96 h.     

Isolation of Messenger Ribonucleic Acid for Myosin Heavy Chain

A chicken embryonic polysome fraction that contains 50–60 monoribosomes and synthesizes the heavy chains of myosin is separated from other polysomes of smaller sizes by centrifugation through two cycles of discontinuous and continuous sucrose gradients. The unique properties of the polyadenylic acid segment present at the 3′-end of eukaryotic messenger RNA (mRNA) were used to purify the mRNA for myosin heavy chain from the phenol-extracted total RNA obtained from this polysome fraction. The total RNA was filtered thro ugh millipore filters resulting in partition of the riboscmal RNA (rRNA) and mRNA species. This millipore-bound RNA fraction, which consists of the mRNA and some ribosomal RNAs, was eluted from the filters with sodium dodecyl sulfate (SDS). Subsequent chromatography of this fraction on a cellulose column gave two well-separated peaks: an unadsorbed peak of ribosomal RNAs which was eluted with buffers of high ionic strength and an adsorbed peak of mRNA which was eluted only with a buffer of low ionic strength. Polyacrylamide gel electrophoresis of the mRNA peak fraction showed a single band with no detectable amounts of other RNAs, the mRNA migrating slower than 28S rRNA. The product of in vitro translation of the purified mRNA using a homologous cell-free system was identified as the myosin heavy chain by the following criteria: coprecipitation with carrier myosin at low ionic strength; elution properties on DEAE-cellulose column; and comigration with the heavy chain in polyacrylamide gel electrophoresis. In order to demonstrate the fidelity of translation of the mRNA, ¹⁴C-labeled products of the in vitro translation were copurified with unlabeled myosin heavy chains added as a carrier. The mixture of polypeptides was then cleaved with CNBr and the resulting peptides were separated by molecular sieving. The correlation between the radioactivity and the UV absorbance in the separated peptides indicates that total synthesis of the myosin heavy chain was achieved.     

The chromosomal localization of a heavy satellite DNA in the testis of Plethodon c. cinereus

When DNA from blood or liver of Plethodon c. cinereus is centrifuged to equilibrium in cesium chloride it separates out into 2 components. The smaller or satellite component is relatively rich in G + C and is therefore heavy, and it amounts to about 2% of the total DNA. The heavy satellite does not include the ribosomal cistrons, and it is unrelated to the nucleolar organizer. When squash preparations of cells from the testis of P. c. cinereus are incubated in synthetic E³RNA complementary to the satellite DNA, the RNA anneals specifically to the centromeric heterochromatin of spermatogonia, spermatocytes, and spermatids, and to the centromeric regions of all discernible chromosomes. RNA/DNA hybrids were located by autoradiography. H³RNA complementary to the major component of the DNA anneals to all nuclei and to all parts of the chromosomes. H³RNA complementary to nucleolar DNA from Xenopus laevis anneals specifically to the chromatin associated with nucleoli in nuclei at various stages of the meiotic divisions. The nature of the centromeric heterochromatin and its role in the meiotic divisions are discussed.     

Bark structure and mode of canopy regeneration in trees of Melia azedarach L.

Summary The bark texture of Melia azedarach L. changes from smooth to furrowed as trees age. In trees that were cut down, those with smooth bark sprouted below the cut from suppressed buds; trees with thick, furrowed bark sprouted at the edge of the cut surface from adventitious buds. The trees that had thin, furrowed bark sprouted mainly at the edge of the cut from adventitious buds, but sometimes also from suppressed buds in cracks. The relationship between sprouting pattern and tree architecture are discussed.     

Identification of a ∼30S size non-ribosomal Saccharomyces cerevisiae RNA that is rapidly labeled on its 3′ end by ATP or UTP

Cell-free extracts prepared from S. cerevisiae cells were incubated in the presence of [-³²P]-labeled ATP, CTP, GTP or UTP. An RNA larger than ribosomal 25S RNA with an apparent size of approximately 30S was prominently labeled on its 3 end in the presence of ATP or UTP but not with CTP or GTP. This labeled RNA was not hybrid-selected by cloned yeast ribosomal DNA; in addition, this 30S RNA was not cleaved by RNase H in the presence of complementary deoxyribooligonucleotides to rRNA. These two lines of evidence show that this 30S RNA is not structurally related to ribosomal RNA gene repeat. The cell-free extracts prepared from yeast cells containing temperature-sensitive poly(A) polymerase adenylated this novel yeast RNA at restrictive temperature with efficiency similar to extracts prepared from wild-type yeast cells. These data show that the enzyme responsible for adenylation of this 30S RNA is distinct from mRNA poly(A) polymerase. While the human SRP RNA 3 adenylating enzyme in the HeLa cell extract adenylated human SRP or Alu RNAs, the yeast adenylating enzyme did not adenylate the human SRP or Alu RNAs in vitro; these data indicate species specificity for this adenylating enzyme.     

Adaptive significance of sprouting ofEuptelea polyandra,a deciduous tree growing on steep slopes with shallow soil

We investigated growth characteristics ofEuptelea polyandra Sieb. et Zucc. (Eupteleaceae), a Japanese endemic deciduous tree species growing on unstable ground such as that of very steep slopes with thin soil.Euptelea polyandra began to sprout at the juvenile stage and had a multiple-stemmed tree form. There was a positive correlation between diameter of the maximumsized stem within a stool (DMS) and the number of stems within the stool. Many stools had suffered from disturbances as shown by the fact that uprooting scars were found on 31.4% and 42.4%, respectively, of the stools of the two populations studied. Sprouting played a significance role in repairing damaged stems and stools, and at least 15.5% and 18.2% of the stools of the two populations, respectively, had apparently avoided death by sprouting. Sprouted stems gradually inclined with the increase in their relative sizes within each stool, and this seemed to facilitate the establishment of younger sprouted stems. The small younger sprouted stems had their own roots. There were dormant buds on stems which originated from axillary buds, and secondary dormant buds occurred by branching. The total number of dormant buds in a stool increased with DMS. It is concluded thatE. polyandra accumulates dormant buds for sprouting in order to respond to disturbances quickly.     

Nascent ribosomal and messenger RNA in DNA-membrane-complexes of Escherichia coli

Summary From Escherichia coli, DNA-membrane-complexes have been isolated which contain about 40% of the ribosomes, about 95% of the DNA and nearly all of the nascent RNA. The kinetic data on pulse labeled RNA show an average time of turnover of about 60 sec both for nascent messenger- and nascent ribosomal RNA. A proportion of the polysomes with nascent messenger RNA as well as most of the nascent ribosomal RNA is found in association with membranes, as has been shown by subfractionations of the DNA-membrane-complex involving treatment with DNase and desoxycholate. In this early transient stage, ribosomal precursor RNA already acquires some ribosomal proteins, as has been shown by arginine pulse label. Data on partial release of DNA from the DNA-membrane-complex by treatment with extremely low doses of DNase indicate that messenger RNA synthesis occurs in clusters on the DNA.The results support models in which, at any given time, RNA synthesis proceeds mainly in sections of the DNA close to the membrane. Thereby the DNA is linked to the membrane via nascent RNA contained in ribosomal precursors as well as via nascent messenger RNA on membrane-bound polysomes.     

Effect of Reciprocal Bud-Graft Transfers Between Old Home and Bartlett Pears and Centrifugatiou on Trans-location of Endogenous Growth Substances in Hardwood Cuttings

An investigation was made of the effect of bud-graft exchanges between two pear varieties—the easily-rooted Old Home and the hard-to-root Bartlett—on the root-promoting activity of extracts obtained from basal segments of hardwood cuttings of each variety. The effect of a centrifugal force obtained from 2000 rpm for 30 minutes on the root-promoting activity of diffusates and extracts from bases of centrifuged cuttings of these two varieties was also studied. Bud exchange treatments showed no appreciable influence on the root-promoting or inhibiting activity of basal extracts unless the transbudded shoots (used for making cuttings) were girdled one month prior to the sampling date; in such cases an influence did appear. The centrifugation treatment increased the number of buds sprouting on cuttings. It also increased the root-promoting activity of extracts from the basal parts of cuttings and of their growth-active components (separated by paper chromalography) when bioassayed by the mung bean rooting test. Apparently some inhibitory materials from the buds were removed through the bases of the cuttings by centrifuging.     

Osservazioni Anatomo-Fisiologiche Sulla Kleinia Articulata Haworth

Abstract

Ultrastructural modifications of plastids in leaflets of Larix decidua and Picea excelsa during sprouting of buds.—Ultrastructural modifications of plastids in leaflets of Larix decidua and Picea excelsa during sprouting of buds kept in different light conditions were observed.

While in quiescent buds of both species typical plastids with magnograna are present, fully expanded leaflets kept in the light have plastids with an organized lamellar apparatus.

When the buds are kept in darkness the cells of the fully expanded, etiolated leaflets have hardly differentiated plastids with prolamellar bodies partially modified into short tubules and vesicles.

Plastids of Picea and Larix buds, in their development, behave almost identically both in darkness and in the light.

The differences previously observed in dark grown seedlings of the two species are not to be found in buds.     

The bud and root sprouting capacity of Alternanthera philoxeroides after over-wintering on sediments of a drained canal

Alternanthera philoxeroides (Mart.) Griseb. is one of many aggressive invasive plants that can grow in diverse habitats. Aquatic A. philoxeroides forms dense floating mats over the water surface. However, when water levels decrease during winter, some mats become stranded on exposed sediments and are thus exposed to air. Do the stems of these mats possess the capacity to develop new shoots during the next growing season? In this study, we examined the sprouting of sediment-stranded over-wintering mats of A. philoxeroides. Stems of the over-wintering mats were divided into three types (dry, withered, and fresh stems) depending on moisture content and were immersed in water for 4 weeks to observe the sprouting of axillary buds and roots. The results showed that withered stems yielded much more biomass than dry or fresh stems. Stem moisture content significantly affected the sprouting rate and the length growth rate of buds and roots. Dry stems lacked reproductive capacity. The sprouting rate and length growth rate of the buds and roots were higher in fresh stems than in withered stems. Furthermore, the mean values of the bud sprouting rate and the bud length growth rate were highest during the first week, i.e., most of buds sprouted within 1 week or less. Our results suggest that more than 70% (on a dry weight basis) of the stems in stranded mats possessed rapid sprouting capacity even after over-wintering on the sediment for more than 2 months. This strategy may be an adaptation to the fluctuations inherent in many aquatic habitats, and it possibly explains why A. philoxeroides can flourish even after a dry winter. Handling editor: S. M. Thomaz     

Control of Deoxyribonucleic Acid and Ribonucleic Acid Synthesis in Pyrimidine-Limited Escherichia coli

The effects of pyrimidine limitation on chromosome replication and the control of ribosomal and transfer ribonucleic acid syntheses were investigated. Chromosome replication was studied by autoradiography of (3)H-thymine pulse-labeled cells. Pyrimidine limitation did not affect the fraction of cells incorporating radioactive thymine during a short pulse, indicating that when growth is limited by the supply of pyrimidine, the time required for chromosome duplication increases in proportion to the time required for cell duplication. Control of ribosomal RNA and transfer RNA syntheses was examined by chromatographing cell extracts on methylated albumin kieselguhr columns. When growth was controlled by carbon-nitrogen limitation, the ratio of tRNA to total RNA remained roughly constant at growth rates above 0.5 doublings per hour. During pyrimidine limitation, however, the control of rRNA synthesis was apparently dissociated from the control of tRNA synthesis: the ratio of tRNA to total RNA increased as the growth rate decreased.     

Assessment of Regeneration Potential in the Clonal Macrophyte Miscanthus sacchariflorus (Poaceae) after Burial Disturbance Based on Bud Bank Size and Sprouting Capacity

The demography of the bud bank and its sprouting capacity are important for understanding the population dynamics of clonal plants and their potential responses to disturbances. To this end, we investigated the size and composition of the bud bank of Miscanthus sacchariflorus (Maxim.) Hack. immediately after flooding (November), in winter (January), in spring (March), and before flooding (May) in the wetlands of Dongting Lake. We then examined the sprouting capacity of axillary buds after sediment burial at 0, 5, 10, 15, and 20 cm. Total bud density of M. sacchariflorus ranged from 2524 buds m^-2 in November to 4293 buds m^-2 in March. Rhizome segments with inactive axillary buds, which represented the majority of the bud population (88.7% in November, 93.3% in May), did not sprout during the 140 days of the experiment (n = 250). The sprouting ratio was the highest for active axillary buds buried at 0 cm (64%) and decreased when buried at 10–20 cm (34%–40%). Due to the large number of active axillary buds in the bud bank (211–277 buds m^-2 from November to the following March), M. sacchariflorus could completely replace its aboveground shoot population, except in May (142 buds m^-2). Increasing burial depth delayed bud emergence and reduced the growth period of shoots; however, burial depth did not affect the resulting plant height and only reduced the accumulated biomass at 20 cm. Therefore, the belowground bud bank and its strong sprouting capacity are important factors in the maintenance of local populations and colonization of new habitats for M. sacchariflorus after burial disturbances. The present methodology, which combined measurements of bud bank demography and sprouting capacity, may reflect the regeneration potential of clonal plants after burial disturbances.     

Adventitious sprouting enables the invasive annual herb Euphorbia geniculata to regenerate after severe injury

Euphorbia geniculata, an annual weed of arable land native to America and invasive in subtropical and tropical regions, is able to regenerate from seeds and is also able to produce adventitious buds on the hypocotyl. Whether sprouting from adventitious buds represents a mechanism for surviving severe injury, and whether this ability is crucial for species invasion is, however, not known. The significance of such sprouting was investigated with a field survey and a pot experiment. Among 897 plants in 25 field populations surveyed in Indonesia, only a few exhibited marks of injury and sprouting from adventitious buds. When seeds were collected from 12 of the populations and used in a pot experiment, however, the seedlings were able to survive severe injury (removal of all tissue above the hypocotyl) by sprouting from adventitious buds on the hypocotyl and were able to set seed, although they produced less vegetative and generative (flowers and fruits) biomass than control plants. Growth but not fitness of plants in the pot experiment was population specific but neither growth characteristic correlated with disturbance level assessed in the field. Although the pot experiment indicates that E. geniculata can cope with severe injury by adventitious sprouting from the hypocotyl, the survey data are inconsistent with the hypothesis that such adventitious sprouting is important for the plant??s invasion in tropical regions.