Further studies on double-stranded RNA species from raspberry plants infected with aphid-borne viruses

Analysis by electrophoresis in polyacrylamide gels, followed by silver staining, of dsRNA extracted from many samples of raspberry leaves infected with raspberry leaf mottle virus (RLMV) and/or raspberry leaf spot virus (RLSV) failed to detect reliably any significant quantities of dsRNA species in excess of 1·0 × 10⁶mol. wt. This contrasts with results reported from Canada where three dsRNA species of estimated mol. wt 2·6 × 10⁶1·6 × 10⁶and 1·1 × 10⁶were consistently associated with infection with RLSV but none were associated with RLMV. However, in Scotland, four dsRNA species of estimated mol. wt 2·4 × 10⁶1·6 × 10⁶0·7 × 10⁶and 0·3 × 10⁶were detected in raspberry infected with apple mosaic ilarvirus. These results suggest that the dsRNA species reported from Canada are not those of RLSV but are probably those of a second virus, possibly an ilarvirus, which occurs together with RLSV and/or induces similar symptoms. A few samples from plants infected with RLMV and RLSV contained very small amounts of two dsRNA species of estimated mol. wt 4·7 × 10⁶and 4·5 × 10⁶. It is not known whether these species are those of RLMV and RLSV.     

RNA沉默与植物病毒

植物中RNA沉默（RNAsilencing)亦称为转录后基因沉默（PTGS）或共抑制,是植物抵抗外来核酸（转座子、转基因或病毒）入侵,并保护自身基因组完整性的一种防御机制。RNA沉默是近十年来发现的植物界中普遍存在的现象,已成为植物分子生物学领域的一个新的研究方向。对RNA沉默特点和机制的研究表明,植物病毒与（转基因）植物内发生的RNA沉默有着密切的联系,作者从病毒对RNA沉默的诱导、抑制、防御等方面,简述了RNA沉默与病毒的关系。并对病毒载体所诱导的RNA沉默在植物发育和基因组功能分析等方面的应用价值进行了讨论。     

Epigenetic modification of plants with systemic RNA viruses

Knowledge of gene function is critical to the development of new plant traits for improved agricultural and industrial applications. Viral expression vectors offer a rapid and proven method to provide epigenetic expression of foreign sequences throughout infected plants. Expression of these sequences from viral vectors can lead to gain- or loss-of-function phenotypes, allowing gene function to be determined by phenotypic or biochemical effects in the infected plant. Tobacco mosaic virus and barley stripe mosaic virus expression vectors have been developed to express foreign gene sequences in dicotyledonous and monocotyledonous hosts, respectively. Large-scale application of both viral vector systems for gene function discovery in Nicotiana and barley hosts resulted in high infection rates and produced distinctive visual phenotypes in approximately 5% of transfected plants. Novel genes expressing potential herbicide target proteins in addition to genes promoting stem elongation, leaf development and apical dominance were identified in the large-scale screening. This report illustrates the adaptability of viral vectors for gene function discovery in higher plants.     

Biosynthesis and metabolism of ascorbic acid in plants

The biosynthesis of L‐ascorbic acid in plants differs from that encountered in ascorbic acid‐synthesizing animals. Enzymic details are sparse, but in vivo studies with tracers clearly establish the stereochemical detail of both processes. Examples of each process are found in separate classes of algae. Plants utilize L‐ascorbic acid as the carbon source for the biosynthesis of two important plant acids, oxalic acid and L‐tartaric acid. Here, cleavage of L‐ascorbic acid between carbons 2 and 3 releases the 2 and 4 carbon intermediates. A second L‐tartaric acid‐synthesizing process peculiar to vitaceous plants, i.e., grape, cleaves ascorbic acid between carbons 4 and 5. The physiological significance of these metabolic interconversions is discussed. Other metabolic processes such as the oxidation/reduction properties of L‐ascorbic acid are also considered.     

Expanding diversity and ecological roles of RNA viruses

While the diversity of global environmental RNA viruses has remained largely unexplored, recent advances have reported on the discovery of over 10⁶ RNA viral contigs from both terrestrial and marine ecosystems that will help us to better understand the diversity, evolution, ecological roles, and transmission of RNA viruses.     

The presence of glutathione and glutathione reductase in chloroplasts: A proposed role in ascorbic acid metabolism

Both glutathione and an NADPH-dependent glutathione reductase are present in spinach (Spinacia oleracea L.) chloroplasts. It is proposed that glutathione functions to stabilise enzymes of the Calvin cycle, and it may also act to keep ascorbic acid in chloroplasts in the reduced form.Abbreviations GSH tripeptide glutathione - GSH reduced form of glutathione - GSSG oxidised form of glutathione     

Localization of ascorbic acid, ascorbic acid oxidase, and glutathione in roots of Cucurbita maxima L

To understand the function of ascorbic acid (ASC) in root development, the distribution of ASC, ASC oxidase, and glutathione (GSH) were investigated in cells and tissues of the root apex of Cucubita maxima. ASC was regularly distributed in the cytosol of almost all root cells, with the exception of quiescent centre (QC) cells. ASC also occurred at the surface of the nuclear membrane and correspondingly in the nucleoli. No ASC could be observed in vacuoles. ASC oxidase was detected by immunolocalization mainly in cell walls and vacuoles. This enzyme was particularly abundant in the QC and in differentiating vascular tissues and was absent in lateral root primordia. Administration of the ASC precursor L-galactono-gamma-lactone markedly increased ASC content in all root cells, including the QC. Root treatment with the ASC oxidized product, dehydroascorbic acid (DHA), also increased ASC content, but caused ASC accumulation only in peripheral tissues, where DHA was apparently reduced at the expense of GSH. The different pattern of distribution of ASC in different tissues and cell compartments reflects its possible role in cell metabolism and root morphogenesis.     

Hemin-mediated Hemolysis in Erythrocytes： Effects of Ascorbic Acid and Glutathione

In the present work,we investigated the effect of ascorbic acid and glutathione on hemolysisinduced by hemin in erythrocytes.Ascorbic acid not only enhanced hemolysis,but also induced formationof thiobarbituric acid-reactive substances in the presence of hemin.It has been shown that glutathioneinhibits hemin-induced hemolysis by mediating hemin degradation.Erythrocytes depleted of glutathionebecame very sensitive to oxidative stress induced by hemin and ascorbic acid.H_2O_2 was involved in hemin-mediated hemolysis in the presence of ascorbic acid.However,a combination of glutathione and ascorbicacid was more effective in inhibiting hemolysis induced by hemin than glutathione alone.Extracellular andintracellular ascorbic acid exhibited a similar effect on hemin-induced hemolysis or inhibition of hemin-induced hemolysis by glutathione.The current study indicates that ascorbic acid might function as anantioxidant or prooxidant in hemin-mediated hemolysis,depending on whether glutathione is available.     

Cytochrome oxidase and ascorbic acid oxidase activities in cereal plants

Cytochrome oxidase and ascorbic acid oxidase activities were investigated in rye, wheat, barley and oat plants. The variations in the activity of both enzymes was followed in the course of the initial 28 days of growth, as well as at the phase of milk ripeness, namely in the cytoplasmic and mitochondrial cell fractions of roots, leaves and spikes. Both enzymes were active in all measurements. Cytochrome oxidase mostly exhibited a higher activity than ascorbio acid oxidase. The activity of the former enzyme was substantially higher in the mitochondrial fraction of leaves, roots and spikes of the four experimental plants in comparison with the cytoplasmic fraction. On the contrary, the ascorbic acid oxidase activity varied in both cell fractions according to the plant species, organ and growth phase. The variations in the activity of both enzymes exhibited on the whole a course similar to that of the respiration rate. During the first 14 to 21 days of growth the enzyme activities increased up to the maximum. This was thon followed at first by a rapid, later on by a slow decrease. The course of variations in the enzyme activities was, with certain exceptions, alike in all the four plant species investigated.     

Kinetics of ozone reaction with uric acid, ascorbic acid, and glutathione at physiologically relevant conditions

This study quantified the reaction kinetics of O3 with three low molecular weight antioxidants-uric acid (UA), ascorbic acid (AH2), and glutathione (GSH)-found in respiratory mucous. Using a semi-batch reactor in which a 500 ml/min flow of air containing 1-5 parts per million of O3 contacted 3 ml of well-stirred physiological saline solution containing 100-200 microM antioxidant, we found that: (1) mass transfer resistances in the gas and liquid phases were successfully eliminated by the reactor design; (2) the reaction of O3 with UA, AH2 and GSH had stoichiometries of 1:1, 1:1, and 1:2.5, respectively; (3) the reactivity between O3 and antioxidants was in the order UA approximately AH2>GSH. Simulating the measured amounts of O3 absorbed and antioxidant consumed with a mathematical model, reaction rate constants of O(3) with UA, AH2, and GSH were found to be 5.83 x 10(4) M(-1) s(-1), 5.5 x 10(4) M(-1) s(-1), and 57.5 M(-0.75) s(-1), respectively.