Regulation of ADPglucose pyrophosphorylase in cucumber plants infected with the cucumber mosaic virus

ADPglucose pyrophosphorylase level and mechanisms regulating its activity were studied in cucumber plants infected with the cucumber mosaic virus at the stage of chronic infection. Studies carried out with partially purified preparations of the enzyme have shown that there was no substantial difference in the regulatory influence of the ratio 3-PGA/P₁, or in the number of binding sites of the effectors on the enzyme, but that the virus infection reduced the level of the enzyme in the tissues to 74% of the control and the 3-PGA/P₁ ratio to one half which resulted in a further decrease in ADPglucose pyrophosphorylase activity. In crude homogenate prepared from diseased plants, activity of the enzyme was reduced to 42% of the healthy control. The level of UDPglucose pyrophosphorylase was three times higher in cucumber leaf tissues than the level of ADPglucose pyrophosphorylase which was inhibited by both 3-PGA and P₁. Inhibitory effects of both these effectors were cumulated. The enzyme isolated from healthy plants was inhibited by inorganic phosphate more strongly than the enzyme isolated from diseased plants. UDPglucose pyrophosphorylase activity was increased in crude homogenate of diseased plants to 127% of the healthy control when the level of the enzyme was the same in the tissues of both healthy and diseased plants which was presumably connected with the enhanced rate of sucrose catabolism.     

Comparative analysis of expressed sequence tags in resistant and susceptible ecotypes of Arabidopsis thaliana infected with cucumber mosaic virus

Arabidopsis thaliana ecotype Columbia (Col-0) is susceptible to the yellow strain of cucumber mosaic virus [CMV(Y)], whereas ecotype C24 is resistant to CMV(Y). Comprehensive analyses of approximately 9,000 expressed sequence tags in ecotypes Col-0 and C24 infected with CMV(Y) suggested that the gene expression patterns in the two ecotypes differed. At 6, 12, 24 and 48 h after CMV(Y) inoculation, the expression of 6, 30, 85 and 788 genes, respectively, had changed in C24, as opposed to 20, 80, 53 and 150 genes in CMV(Y)-infected Col-0. At 12, 24 and 48 h after CMV(Y) inoculation, the abundance of 3, 10 and 55 mRNAs was altered in both ecotypes. However, at 6 h after CMV(Y) inoculation, no genes were co-induced or co-suppressed in both ecotypes. This differential pattern of gene expression between the two ecotypes at an early stage of CMV(Y) infection indicated that the cellular response for resistance may differ from that resulting in susceptibility at the level detectable by the macroarray. According to the expression pattern at various stages of infection, the expression of many genes could be grouped into clusters using cluster analysis. About 100 genes that encode proteins involved in chloroplast function were categorized into clusters 1 and 4, which had a differentially lower expression in CMV(Y)-inoculated C24. The expression of various genes encoding proteins in the endomembrane system belonged to clusters 2 and 4, which were induced in CMV(Y)-inoculated C24 and Col-0 leaves. Characterization of CMV(Y)-altered gene expression in the two ecotypes will contribute to a better understanding of the molecular basis of compatible and incompatible interactions between virus and host plants.     

Evolutionary relationship of alfalfa mosaic virus with cucumber mosaic virus and brome mosaic virus

The amino acid sequences of the non-structural protein (molecular weight 35,000; 3a protein) from three plant viruses — cucumber mosaic, brome mosaic and alfalfa mosaic have been systematically compared using the partial genomic sequences for these three viruses already available. The 3a protein of cucumber mosaic virus has an amino acid sequence homology of 33.7% with the corresponding protein of brome mosaic virus. A similar protein from alfalfa mosaic virus has a homology of 18.2% and 14.2% with the protein from brome mosaic virus and cucumber mosaic virus, respectively. These results suggest that the three plant viruses are evolutionarily related, although, the evolutionary distance between alfalfa mosaic virus and cucumber mosaic virus or brome mosaic virus is much larger than the corresponding distance between the latter two viruses.     

Interaction of cucumber mosaic virus and potato virus y with tobacco mosaic virus

A study was performed on the interaction of cucumber mosaic virus (CMV) of potato virus Y (PVY) with tobacco mosaic virus (TMV). Interference was evaluated using tobacco plantsNicotiana tabacum cv. Java responding to CMV and PVY with a systemic infection and to TMV with local necrotic lesions. The decrease in TMV — induced lesion number gave evidence of a decrease in susceptibility caused by the previous infection with CMV or PVY, the decrease of lesion enlargement demonstrated a decreased TMV reproduction in the plants previously infected with CMV or PVY. The interference concerned was incomplete, as evaluated from reproduction of the challenging TMV and from the decrease in susceptibility of the host to TMV brought about by the first infection with CMV or PVY.     

Plant growth-promoting rhizobacteria systemically protect Arabidopsis thaliana against Cucumber mosaic virus by a salicylic acid and NPR1-independent and jasmonic acid-dependent signaling pathway

Arabidopsis thaliana ecotype Columbia plants (Col-0) treated with plant growth-promoting rhizobacteria (PGPR) Serattia marcescens strain 90-166 and Bacillus pumilus strain SE34 had significantly reduced symptom severity by Cucumber mosaic virus (CMV). In some cases, CMV accumulation was also significantly reduced in systemically infected leaves. The signal transduction pathway(s) associated with induced resistance against CMV by strain 90-166 was determined using mutant strains and transgenic and mutant Arabidopsis lines. NahG plants treated with strains 90-166 and SE34 had reduced symptom severity indicating that the resistance did not require salicylic acid (SA). Strain 90-166 naturally produces SA under iron-limited conditions. Col-0 and NahG plants treated with the SA-deficient mutant, 90-166-1441, had significantly reduced CMV symptom severity with reduced virus accumulation in Col-0 plants. Another PGPR mutant, 90-166-2882, caused reduced disease severity in Col-0 and NahG plants. In a time course study, strain 90-166 reduced virus accumulation at 7 but not at 14 and 21 days post-inoculation (dpi) on the non-inoculated leaves of Col-0 plants. NahG and npr1-1 plants treated with strain 90-166 had reduced amounts of virus at 7 and 14 dpi but not at 21 dpi. In contrast, no decrease in CMV accumulation occurred in strain 90-166-treated fad3-2 fad7-2 fad8 plants. These data indicate that the protection of Arabidopsis against CMV by strain 90-166 follows a signaling pathway for virus protection that is independent of SA and NPR1, but dependent on jasmonic acid.     

Effects of salicylic and jasmonic acids on the components of pro/antioxidant system in potato plants infected with late blight

The effects of salicylic acid (SA) and jasmonic acid (JA) on plant defense responses were studied with aseptic potato (Solanum tuberosum L.) plantlets infected with Phytophthora infestans (Mont.) de Bary. Plant treatment with 10⁻⁶ M SA or 10⁻⁷ M JA induced plant resistance; the mixture of these acids was most efficient. After treatment with these compounds, phenolic compounds were accumulated and peroxidase was activated in the sites of pathogen localization, and this might be the reason of resistance enhancement. In addition, more H₂O₂ was accumulated in infected plants treated with JA or its mixture with SA but not in plants treated with SA alone. It might occur because of observed inhibition of catalase and activation of isoperoxidase with the isoelectric point (pI) of ∼9.3, which manifests an affinity for the pathogen cell wall. The data obtained allow us to recommend the application of these compounds for potato plant protection against late blight.     

Interactive effects of jasmonic acid, salicylic acid, and gibberellin on induction of trichomes in Arabidopsis

Leaf trichomes protect plants from attack by insect herbivores and are often induced following damage. Hormonal regulation of this plant induction response has not been previously studied. In a series of experiments, we addressed the effects of artificial damage, jasmonic acid, salicylic acid, and gibberellin on induction of trichomes in Arabidopsis. Artificial damage and jasmonic acid caused significant increases in trichome production of leaves. The jar1-1 mutant exhibited normal trichome induction following treatment with jasmonic acid, suggesting that adenylation of jasmonic acid is not necessary. Salicylic acid had a negative effect on trichome production and consistently reduced the effect of jasmonic acid, suggesting negative cross-talk between the jasmonate and salicylate-dependent defense pathways. Interestingly, the effect of salicylic acid persisted in the nim1-1 mutant, suggesting that the Npr1/Nim1 gene is not downstream of salicylic acid in the negative regulation of trichome production. Last, we found that gibberellin and jasmonic acid had a synergistic effect on the induction of trichomes, suggesting important interactions between these two compounds.     

Metabolic alterations in cotyledons of Cucurbita pepo infected by cucumber mosaic virus

Changes in the capacities of enzymes in various metabolic pathwayshave been measured during infection of cotyledons of Cucurbitapepo L. with cucumber mosaic virus (CMV). Starch accumulationand low sucrose content, which are characteristic of the earlystages of infection, are reversed in the later stages of infection.The decline in starch correlated with a reduced capacity forstarch synthesis (ADP-glucose pyrophosphorylase) and a risein the capacity for starch degradation (total starch hydrolase,starch phosphorylase). ¹⁴CO₂ feeding experiments, conductedat saturating CO₂ concentration, show that the newly-assimilatedcarbon was lost at a lower rate from infected cotyledons andless was incorporated into structural carbohydrates, phosphorylatedintermediates plus organic acids, more into soluble sugars,amino acids and proteins. At a later stage of infection therewere dramatic increases in respiratory capacity and a substantialalteration of carbohydrate metabolism. The infection had a largestimulatory effect on the capacity for oxidative pentose-phosphatepathway (glucose-6-phosphate dehydrogenase, 6-phospho-gluconatedehydrogenase), glycolysis (ATP- and pyrophosphate-dependentphosphofructokinases), tri-carboxylic acid cycle (isocitratedehydrogenase, fumarate hydratase), anaplerotic reactions (NAD-dependentmalic enzyme, phosphoenolpyruvate car-boxylase) and oxidativeelectron transport (cytochrome c oxidase). While there wereno overall changes in photosynthetic rate (measured in saturatingCO₂), infection either reduced (Rubisco and glycerate kinase)or did not affect (chloroplastic fructose bis-phosphatase andhydroxypyruvate kinase) the capacities of the photosyntheticcarbon reduction pathway or the photosynthetic carbon oxidationpathway. Key words: Plant-virus interaction, sucrose, starch, enzymes, ¹⁴CO₂ incorporation, O₂ flux     

Specific enrichment of miRNAs in Arabidopsis thaliana infected with Tobacco mosaic virus.

RNA silencing is a broadly conserved machinery and is involved in many biological events. Small RNAs are key molecules in RNA silencing pathway that guide sequence-specific gene regulations and chromatin modifications. The silencing machinery works as an anti-viral defense in virus-infected plants. It is generally accepted that virus-specific small interfering (si) RNAs bind to the viral genome and trigger its cleavage. Previously, we have cloned and obtained sequences of small RNAs from Arabidopsis thaliana infected or uninfected with crucifer Tobacco mosaic virus. MicroRNAs (miRNAs) accumulated to a higher percentage of total small RNAs in the virus-infected plants. This was partly because the viral replication protein binds to the miRNA/miRNA* duplexes. In the present study, we mapped the sequences of small RNAs other than virus-derived siRNAs to the Arabidopsis genome and assigned each small RNA. It was demonstrated that only miRNAs increased as a result of viral infection. Furthermore, some newly identified miRNAs and miRNA candidates were found from the virus-infected plants despite a limited number of examined sequences. We propose that it is advantageous to use virus-infected plants as a source for cloning and identifying new miRNAs.     

Response of tomato plants infected with cucumber mosaic virus to foliar sprays of gibberellic acid

The effect of spraying tomato plants infected with cucumbermosaic virus (CMV) with gibberellic acid (GA) 1 and 5 ppm hasbeen studied in a glasshouse. Measurements showed that the differencein size between GA-treated healthy and infected plants eitherdisappeared or became insignificant and this was true at bothhigh and low planes of nutrition. Virus content was not reducedby GA. Virusinduced checks to leaf production and stem elongationtended to disappear in unsprayed plants but leaf area and shootweight did not show natural recovery. The stimulating effectof GA was primarily on leaf expansion and secondarily on shootweight.     

Resistance of tomato infected with cucumber mosaic virus satellite RNA to potato spindle tuber viroid

Tomato (Lycopersicon esculentum cvs Rutgers and Lichun) plants were firstly pre-inoculated either with a cucumber mosaic virus (CMV) isolate containing satellite RNA (CMV-S52) or with a CMV isolate without satellite RNA, and then challenged 14 days later with a severe strain of potato spindle tuber viroid (PSTVd). Also, tomato plants transformed with CMV satellite cDNA and non-transgenic control plants were directly inoculated with PSTVd. Protection effects were assessed by the observation of symptoms and by assay of PSTVd accumulation in tomato plants using return polyacrylamide gel electrophoresis and silver staining. The results indicated that the satellite-transgenic plants and plants pre-inoculated with CMV-S52 showed much milder symptoms of PSTVd infection than the respective control plants. The concentration of PSTVd RNA in the satellite-transgenic plants and CMV-S52 pre-inoculated plants was reduced to about 0.02–0.03 of the controls. PSTVd infection did not increase the amount of satellite ds-RNA in plants. It is concluded that the plant resistance to PSTVd is induced by the presence of satellite RNA rather than the CMV infection. It is suggested that as there is considerable sequence similarity between satellite RNA and PSTVd, base pairings may be a cause of reduction of both symptoms and the accumulation of PSTVd.     

Resistance of tomato infected with cucumber mosaic virus satellite RNA to potato spindle tuber viroid

Tomato (Lycopersicon esculentum cvs Rutgers and Lichun) plants were firstly pre-inoculated either with a cucumber mosaic virus (CMV) isolate containing satellite RNA (CMV-S52) or with a CMV isolate without satellite RNA, and then challenged 14 days later with a severe strain of potato spindle tuber viroid (PSTVd). Also, tomato plants transformed with CMV satellite cDNA and non-transgenic control plants were directly inoculated with PSTVd. Protection effects were assessed by the observation of symptoms and by assay of PSTVd accumulation in tomato plants using return polyacrylamide gel electrophoresis and silver staining. The results indicated that the satellite-transgenic plants and plants pre-inoculated with CMV-S52 showed much milder symptoms of PSTVd infection than the respective control plants. The concentration of PSTVd RNA in the satellite-transgenic plants and CMV-S52 pre-inoculated plants was reduced to about 0.02-0.03 of the controls. PSTVd infection did not increase the amount of satellite ds-RNA in plants. It is concluded that the plant resistance to PSTVd is induced by the presence of satellite RNA rather than the CMV infection. It is suggested that as there is considerable sequence similarity between satellite RNA and PSTVd, base pairings may be a cause of reduction of both symptoms and the accumulation of PSTVd.     

Isolates of cucumber mosaic virus from spontaneously infected plants ofChelidonium majus andImpatiens parviflora

Cucumber mosaic virus (CMV) isolates obtained from spontaneously infected (1)Chelidonium majus L. and (2)Impatiens parviflora DC. plants growing in the same part of a scree forest in central Bohemia showed differences in ten test plants, but not in further 26 plant species tested and in TIP. Slight quantitative differences between both isolates were obtained in serological tests. Properties of both isolates did not suit precisely any proposed CMV strain classification.     

外源水杨酸和茉莉酸诱导巨峰葡萄抗根瘤蚜

【目的】水杨酸和茉莉酸在植物诱导防御虫害反应中发挥着重要作用。本研究旨在探讨水杨酸和茉莉酸诱导葡萄对根瘤蚜的抗性。【方法】以盆栽巨峰为试材,在接种葡萄根瘤蚜Daktulosphaira vitifoliae的同时喷施水杨酸和茉莉酸,测定和评估了对根瘤蚜生长发育及产卵量的影响,以及对葡萄根系抗氧化相关酶［过氧化物酶(POD)和过氧化氢酶(CAT）］活性、丙二醛（MDA）含量、新梢生长量及光合速率的影响。【结果】水杨酸和茉莉酸诱导处理显著降低了根瘤蚜卵量及下代1,2龄若蚜总数,接种35 d后根瘤蚜的产卵量分别减少了41.35％和50.00％,1龄和2龄若蚜总数分别减少了42.31％和50.00％;根瘤蚜侵染后根系中POD和CAT活性均呈先升高后降低的趋势,且水杨酸和茉莉酸诱导处理在各测定时期均高于仅接种根瘤蚜处理;水杨酸和茉莉酸处理的根系中MDA含量在各测定时期均低于仅接种根瘤蚜处理;水杨酸和茉莉酸处理降低了根瘤蚜侵染对植株地上部生长及光合的抑制。接种处理后第30天,仅接种根瘤蚜处理的植株地上部生长量减少了48.11%,光合速率降低了58.77％,而水杨酸和茉莉酸处理后的新梢生长量分别减少了31.57％和25.71％,光合速率分别降低了32.89％和24.67％。【结论】叶片喷洒茉莉酸和水杨酸能够降低根瘤蚜种群密度,并提高葡萄根系活性氧清除能力和防御酶活性,缓解树势衰退。     

Mild mosaic of spiraea caused by cucumber mosaic virus

A disease of spiraea(Spiraea xvanhouttei) manifested in leaves by very mild, mostly hardly perceptible mosaic, was found to be caused by cucumber mosaic virus (CMV) infection. The proof was given on the basis of responce of differential plants after virus transmission, by immunosorbent electron microscopy and ELISA.     

Comparison of the nucleotide sequences of cucumber mosaic virus and brome mosaic virus

Cucumber mosaic virus (CMV) and brome mosaic virus (BMV) are isometric plant viruses. Although biologically distinct, they share many common chemical properties. An analysis of the partial genomic RNA sequence available for these two viruses reveals that they are evolutionarily related. Different segments of the genome exhibit different evolutionary rates. The coat proteins, which serve as carriers of genetic material, possess little or no homology. In contrast, the 3a proteins show over 35% homology. The non-coding regions of the genome also exhibit extensive but variable homology suggesting the functional importance of the nucleic acid.     

The effect of 2,4-dichlorophenoxyacetic acid on the metabolic utilization of free carbohydrates in cucumber mosaic virus infected cucumber plants

The application of 2,4-dichlorophenoxyacetic acid at a concentration of 1.0 × 10⁻³ M toCucumis sativus brings about a decrease in the activity of carbohydrate catabolizing enzymes in the leaves of experimental plants. On the contrary, in CMV infected plants the activity of sucrase, glucokinase, fructokinase, phosphoglucoisomerase and glucose-6-phosphate dehydrogenase are enhanced at the same time. The application of 2,4-D to virus infected plants promotes this effect further, so that the activities of the enzymes investigated are twice as high as those in the control.