Nucleic Acid Metabolism During Early Development of Pollinated and Auxin-induced Parthenocarpic Watermelon Fruits

Following pollination or induction of parthenocarpy by NAA¹treatment watermelon ovaries synthesize large quantities ofRNA. Hormone-treated ovaries increase their rate of RNA synthesisover untreated controls by 6 h after treatment whereas pollinatedovaries do so by 24 h. The lag period with pollinated ovarieshas been related to the growth of pollen tubes toward the ovulesand it is suggested that auxin secretion by the growing pollentubes may be the stimulus for increased RNA synthesis. Evidenceis presented that the cells in the flesh of developing ovariesbecome polyploid as they enlarge.     

Effect of Selenium on Lipid and Amino Acid Metabolism in Yeast Cells

Biological Trace Element Research - This article discusses the effect of selenium in aqueous solutions on aspects of lipid and amino acid metabolism in the cell biomass of Saccharomyces cerevisiae...     

Reduction of Nucleic Acid Content in Candida Yeast Cells by Bovine Pancreatic Ribonuclease A Treatment

Yeast as a source of protein for human consumption is limited by its relatively high nucleic acid content. In this study, we developed an enzymatic method of decreasing the nucleic acid content. Candida utilis cells, heat-shocked at 80 C for 30 sec, were treated with bovine pancreatic ribonuclease A. Maximum leakage of nucleic acid was observed when the incubation temperature was between 55 and 65 C, the pH of the system from 6.75 to 8.0, and the enzyme-to-cell ratio 1:10,000 on a weight-by-weight basis. Other factors, such as yeast strain, age of cells, and method of propagation, did not influence the susceptibility of the yeast cells to the action of ribonuclease. Buffers and monovalent cations had no inhibiting effects. Magnesium and calcium ions at concentrations greater than 0.001 m showed marked inhibition on the rate of nucleic acid leakage. This enzymatic method reduced the nucleic acid content of yeast cells from 7.5 to 9.0% to 1.5 to 2.0% with no significant concomitant loss of protein.     

Comparison of Auxin-induced and Acid-induced Elongation in Soybean Hypocotyl

Acid-induced growth was compared to auxin-induced growth. After a transient pH 4-induced increase in the elongation rate was completed, auxin could still induce an enhanced rate of elongation in soybean (Glycine max) hypocotyl segments. This auxin response occurred both when the medium was changed to pH 6 before auxin addition, and when the auxin was added directly to the pH 4 medium. This postacid response to auxin was persistent, and quite unlike a postacid response to acid, which was again shortlived. One mm N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (pH 7) inhibited the first response to auxin (the first response to auxin being similar to the acid response), but not the second response. This did not appear to be simply a hydrogen ion neutralizing effect, however, since a 50-fold increase in buffer concentration at pH 6 did not inhibit the first response. Decrease in the pH of the external medium, previously shown to occur with excised soybean hypocotyl segments, was not affected by auxin. Furthermore, this pH drop, during which the cells appear to be adjusting their external pH to about 5.4, did not result in an increased rate of elongation. Addition of auxin after the equilibrium pH had been attained did not alter the pH, but it did increase the rate of elongation, eliciting a normal auxin response. It was concluded that hydrogen ions do not mediate in long term auxin-induced elongation in soybean hypocotyl.     

Protein and Nucleic Acid Metabolism in Germinating Peas

Protease, peptidase, and ribonuclease activities were demonstratedin germinating pea cotyledons and axis tissues. These activitiesindicate that the enzymatic machinery for the hydrolysis ofprotein and nucleic acid reserves are present in the germinatingcotyledon. The fate of hydrolytic products was determined byinjecting leucine-¹⁴C or adenine-8-¹⁴C into the cotyledons.At most, 20 per cent of the leucine-¹⁴C and 10 per cent of theadenine-8-¹⁴C administered were transported from the cotyledonto axis tissues. Both compounds were extensively metabolizedand the labelling patterns suggest that different metabolicpathways are in operation in the two organs. The amounts ofadenine incorporated into nucleic acids and of leucine incorporatedinto protein in the cotyledons suggest that synthesis and turnoverwere occurring at a rapid rate. The adenine transported fromthe cotyledon was not readily available for nucleic acid synthesisin the axis whereas transported leucine was readily incorporatedinto axis protein.     

Circadian Rhythms of Nucleic Acid Metabolism in Neurospora crassa

Wild-type, band, and fluffy strains of Neurospora crassa exhibit circadian rhythms of ribonucleic acid and deoxyribonucleic acid content in the growth-front hyphae of cultures grown on a solid medium. There is also a rhythm of (3)H-uridine incorporation into the nucleic acids of the band strain. Maximum incorporation precedes the peaks of nucleic acid content which occur during conidiation. As cultures age, ribonucleic acid content decreases rapidly and deoxyribonucleic acid content decreases gradually in standing, shake, and bubble cultures. A reduction of ribonuclease activity with age is also noted in standing and shake cultures. The nucleic acid content, nuclease activity, and changes associated with age vary with the culture conditions.     

Correction to: Effect of Selenium on Lipid and Amino Acid Metabolism in Yeast Cells

Biological Trace Element Research - The authors forgot to include the following information in Materials and Methods.     

Nucleic Acid Metabolism in Germinating Onion: I. Changes in Root Tip Nucleic Acid during Germination

Nucleic acid synthesis in the G₁ 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) began at approximately the same time (18 hours). Synthesis of the various cytoplasmic RNA fractions continued throughout the germination period. Data indicating synthesis of the “chloroplast-like” RNA were obtained only for the initial 36 hours of germination. Specific radioactivity of ³H-uridine-labeled total nucleic acid increased during the first 41.5 hours of germination but then decreased while the accumulation of RNA per cell continued to increase throughout the 73-hour period. In addition, a method is described which reduced the bacterial contamination of Allium seed to a level not detectable by incorporation of radioactive precursors into bacterial ribosomal RNA.     

Growth-limiting Proteins in Relation to Auxin-induced Elongation in Lupin Hypocotyls

The role of protein synthesis in auxin-induced cell elongation in lupin hypocotyl segments was studied using cycloheximide. Cycloheximide inhibited protein synthesis by 9 minutes. Experiments adding cycloheximide at various times before and after indolyl-3-acetic acid are reported. Estimates of the relative amounts of growth-limiting protein(s), and a first order rate constant for the apparent turnover of the growth-limiting protein(s) were made. It was shown that there is a sizeable growth promotion by auxin after protein synthesis has essentially ceased. It is concluded that the initial phases of auxin action do not require protein synthesis but that its action depends on the existing pool of growth-limiting proteins which is rapidly depleted, and protein synthesis is then required for continued elongation.     

Nucleic Acid Metabolism During Cytokinin Induced Cellular Differentiation

Edstrom's microphoresis technique has been employed to determine the quantitative alterations in nucleic acid content and base composition of individual cells associated with the initiation of bud primordia in Funaria hygrometrica. The filamentous protonema of this moss initiates bud cells which through repeated divisions form the leafy gametophores. The cytokinin, 6-furfurylaminopurine (kinetin), was used to induce the differentiation of bud cells from protonematal cells. The total RNA content of kinetin-induced bud cells (22.0 μμg/cell) was nearly 15 times that of protonematal cells (1.6 μμg/cell). The same dramatic increase in total RNA was apparent in bud cells which developed spontaneously in older cultures. As would be predicted, the adenine (A) to guanine (G) ratio for DNA from bud and protonematal cells was identical (0.7). The A:G ratio for RNA from bud cells (1.0) was much lower than that from protonematal cells (1.7). Thus, kinetin-induced differentiation in this system involves a dramatic increase in total RNA, the base composition of which approaches that of DNA. The base composition (A:G ratio) of DNA remains constant.     

基于PC/Linux的核酸序列电子延伸系统的构建及其应用

新基因全长cDNA序列的获得常常是分子生物学工作者面临的难题。人类基因组计划及其相关计划的实施导致了大量表达序列标签(EST)的产生。利用一定的生物信息学算法,这些EST序列往往可用来对新基因片段进行延伸。采用Linux操作系统,利用Blast软件和Phrap软件以及EST数据库在微机上构建了EST序列的电子延伸系统,并对来自于人胎肝的11386条EST序列和511条插入片段全长cDNA序列进行了电子延伸,结果显示8373条EST序列和389条插入片段全长cDNA序列得到了程度不等的延伸,部分结果通过RACE实验得到证实。该套系统可高效地、规模化进行EST序列的延伸,可为通过实验获得新基因全长cDNA序列提供重要线索。 Abstract:Normally it is difficult to obtain full-length cDNA sequence of novel genes.More and more expressed sequence tags(ESTs) have been obtained since the start-up of human genome project.Powerful system is badly needed for data mining on these EST sequences.Based on a personal computer coupled with Linux operating system and EST database,the Blast software and Phrap software were used to construct a platform for in silico elongation of ESTs in our lab.The performance was tested using 11386 EST sequences and 511 partial-length cDNA sequences.Results demonstrated that 8373 EST and 389 cDNA sequence were elongated using this system.Thus the platform seems to be a fast way for full-length cDNA sequence cloning of new genes.     

Requirement of RNA for the Auxin-induced Elongation of Oat Coleoptile

Using etiolated oat coleoptile segments the following results were obtained. Actinomycin D pretreatment for one hour produced about 50 per cent inhibition of RNA synthesis (labeled uracil incorporation), but the elongation caused by IAA was not inhibited in the following 5 hours at least. Actinomycin D pretreatment for three hours produced about 75 per cent inhibition of RNA synthesis and almost complete inhibition of subsequent IAA-induced elongation, which is accompanied by the inhibition of IAA-induced increase in cell wall extensibility. The inhibiting effect of actinomycin D seemed to be reduced when IAA was given within a certain period.     

Nucleic Acid Metabolism in Chlamydomonas moewusii Undergoing Daily Cell Doubling

Nucleic acid metabolism was studied in synchronized cultures(12 h light, 12 h dark) of Chlamydomonas moewusii Gerloff inwhich cell number approximately doubled each 24 h. Analysisfor DNA showed a small but significant rise between 5 to 7 hand a major increase from 11 to 16 h after first exposure tolight. Cellular RNA levels increased from 0·5 to 1·1pg within 1 h after exposure to light, dropped after 2 h toa plateau value of about 0·9 pg, declined from the 10thto the 17th hour, and subsequently increased a second time duringthe dark period. Investigations using gel electrophoresis showedthat RNAs of chloroplastic ribosomes were only a minor componentin extracts obtained in the first 8 h of light. It is suggestedthat the time of initiation of synthesis and degradation aswell as the nature and amounts of nucleic acids produced inChlamydomonas are regulated by conditions of growth.     

Membrane Rafts Are Involved in Intracellular Miconazole Accumulation in Yeast Cells

Azoles inhibit ergosterol biosynthesis, resulting in ergosterol depletion and accumulation of toxic 14α-methylated sterols in membranes of susceptible yeast. We demonstrated previously that miconazole induces actin cytoskeleton stabilization in Saccharomyces cerevisiae prior to induction of reactive oxygen species, pointing to an ancillary mode of action. Using a genome-wide agar-based screening, we demonstrate in this study that S. cerevisiae mutants affected in sphingolipid and ergosterol biosynthesis, namely ipt1, sur1, skn1, and erg3 deletion mutants, are miconazole-resistant, suggesting an involvement of membrane rafts in its mode of action. This is supported by the antagonizing effect of membrane raft-disturbing compounds on miconazole antifungal activity as well as on miconazole-induced actin cytoskeleton stabilization and reactive oxygen species accumulation. These antagonizing effects point to a primary role for membrane rafts in miconazole antifungal activity. We further show that this primary role of membrane rafts in miconazole action consists of mediating intracellular accumulation of miconazole in yeast cells.     

Reevaluation of the Effect of Calcium Ions on Auxin-induced Elongation

The mechanism by which calcium ions inhibit cell elongation has been reinvestigated. Growth-inhibiting levels of calcium, when applied to isolated walls (in vitro treatment) do not decrease cell wall extensibility as measured by the Instron technique. Thus, the hypothesis that calcium inhibits growth by forming wall-stiffening calcium bridges must be abandoned. Treatment of living auxin-treated sections with calcium (in vivo treatment) does cause a decrease in the subsequently measured wall extensibility, but this decline appears to be simply a consequence of the growth inhibition rather than its cause. Growth-inhibiting levels of calcium do not appreciably reduce the rate of auxin-enhanced H(+) excretion. Pretreatment with calcium does not reduce the capacity of walls to undergo acid-activated wall loosening in the absence of calcium. High concentrations of CaCl(2) (0.02 m) cause an initial elastic shrinkage of Avena sections comparable to that caused by the same osmolarity of mannitol, but the subsequent growth inhibition is too great to be explained by an osmotic inhibition. Calcium ions do inhibit H(+)-induced extension of frozen-thawed sections under tension. The growth-inhibitory effects of calcium, then, may be ascribed to a direct inhibition exerted by calcium ions on the H(+)-induced wall-loosening process.     

玉米中植酸代谢研究的策略

作物尤其是玉米的种子中积累了丰富的植酸。早先的研究侧重于降低种子中植酸的含量,但是随着人们对植酸认识的深入,发现植酸对于动、植物而言具有不可替代的生物功能。对于人和动物而言,植酸有抗营养作用,但也是重要的健康因子;对于植物而言,植酸及其代谢中间体的生物学功能却缺乏明确的研究。若要明确把握植酸的育种方向,就必须对植酸在植物中的合成过程有明确的认识。但自植酸被发现至今,人们对于其在高等植物中的合成过程仍然知之甚少,对其生物学功能更是缺乏全面的了解。本文综述了植酸代谢研究的现状,分析并总结了植酸的代谢通路,指出了植酸代谢研究的突破点,结合植酸代谢的研究特点和进展,比较了基因同源克隆、关联分析等4种最具潜力的研究策略。     

Simple Process for the Reduction in the Nucleic Acid Content in Yeast

A simple one-step process for the nucleic acid reduction in Rhodotorula glutinis is described. The process consists of submitting the yeast cells to a heat treatment in an acidic (pH 2) spent medium. The optimal temperature for pH 2 medium is 90 C and the final nucleic acid content in treated yeasts was 1.2%. Heat treatment at acidic pH is preferred to that at alkaline pH because it offers a better protection for amino acids and crude protein, while being more efficient in lowering the nucleic acid level. The new process is economic and rapid and could be easily used for industrial application.     

Effects of Phosfon-S on Nucleic Acid Metabolism in Pisum sativum Alaska

Phosfon-S, a substance which inhibits stem elongation, alters nucleic acid metabolism in Pisum sativum Alaska. Methylated albumin kieselguhr (MAK) columns were used to fractionate ³²P-labeled nucleic acids. Phosfon-S treatment of the plants resulted in a decrease in soluble RNA and an increase in ribosomal RNA. Specific activities of the various nucleic acid fractions were lower as a result of treatment. The nucleic acids from treated tissues were more resistant to RNase degradation, and endogenous RNase activity was lower in treated tissues. When RNase treated nucleic acids were fractionated on MAK columns, the DNA-RNA fractions from treated plants had a higher specific activity than that of the control, which was not true before nuclease treatment. Spectrophotometric examination of this fraction revealed a difference in absorption spectra, possibly indicating a Phosfon-S nucleic acid complex. It is suggested that these alterations in nucleic acid metabolism could in turn alter a wide variety of metabolic processes, resulting in retarded growth.