Octadecanoid-derived alteration of gene expression and the "oxylipin signature" in stressed barley leaves. Implications for different signaling pathways.

Stress-induced gene expression in barley (Hordeum vulgare cv Salome) leaves has been correlated with temporally changing levels of octadecanoids and jasmonates, quantified by means of gas chromatography/mass spectrometry-single ion monitoring. Application of sorbitol-induced stress led to a low and transient rise of jasmonic acid (JA), its precursor 12-oxophytodienoic acid (OPDA), and the methyl esters JAME and OPDAME, respectively, followed by a large increase in their levels. JA and JAME peaked between 12 and 16 h, about 4 h before OPDA and OPDAME.

The jasmonate-induced 60 kDa protein of barley exhibits N-glycosidase activity in vivo.

Upon jasmonate treatment barley leaf segments express a putative ribosome-inactivating protein (JIP60). The influence of this protein on translation in planta has been analysed by using barley plants and tobacco plants transformed with a barley cDNA encoding JIP60. In both plant systems JIP60 exhibited N-glycosidase activity in vivo.

Characterization of a methyljasmonate-inducible lipoxygenase from barley (Hordeum vulgare cv. Salome) leaves.

We found three methyl jasmonate-induced lipoxygenases with molecular masses of 92 kDa, 98 kDa, and 100 kDa (LOX-92, -98 and -100) [Feussner, I., Hause, B., Vörös, K.

Amino acid conjugates of jasmonic acid induce jasmonate-responsive gene expression in barley (Hordeum vulgare L.) leaves.

Leaves of barley (Hordeum vulgare L. cv. Salome) treated with jasmonic acid (JA), its methyl ester (JM), or its amino acid conjugates exhibit up-regulation of specific genes and down-regulation of house-keeping genes.

Purification and Characterization of Allene Oxide Cyclase from Dry Corn Seeds.

Allene oxide cyclase (AOC; EC 5.3.99.

Temporal pattern of jasmonate-induced alterations in gene expression of barley leaves.

Leaf tissues of barley (Hordeum vulgare L. cv. Salome) respond to methyl jasmonate (JaMe) treatment with a characteristic pattern of gene expression.

Genomic sequence and mapping of a methyljasmonate-induced O-methyltransferase from barley (Hordeum vulgare L.).

We have isolated a genomic clone corresponding to a caffeic acid O-methyltransferase (COMT) from barley (Hordeum vulgare L.) using a cDNA for a previously described jasmonate-regulated mRNA showing homology to COMT. Primer extension was used to characterize the 5' end of the mRNA while the 3' end, intron/exon structure and other features of the sequence were deduced by comparison to the cDNA sequence and/or conserved motifs.

Expression of the ribosome-inactivating protein JIP60 from barely in transgenic tobacco leads to an abnormal phenotype and alterations on the level of translation.

In this paper we report the in-planta activity of the ribosome-inactivating protein JIP60, a 60-kDa jasmonate-induced protein from barley (Hordeum vulgare L.), in transgenic tobacco (Nicotiana tabacum L.) plants.

Developmental and tissue-specific expression of JIP-23, a jasmonate-inducible protein of barley.

Developmental expression of a 23 kDa jasmonate-induced protein (JIP-23) of barley leaves (Hordeum vulgare cv. Salome) was studied by measuring the time-dependent accumulation of transcript and protein during germination. Tissue-specific expression of JIP-23 was analyzed immunocytochemically and by in situ hybridizations, respectively.

Jasmonate signalling can be uncoupled from abscisic acid signalling in barley: identification of jasmonate-regulated transcripts which are not induced by abscisic acid.

Jasmonate and abscisic acid induce several identical mRNAs and proteins in barley. In order to study whether both hormones act through the same signalling pathway, we identified four transcripts induced by jasmonic acid methylester (JM) in leaf segments of barley (Hordeum vulgare L. cv.

Cytosolic and plastid forms of 5-enolpyruvylshikimate-3-phosphate synthase in Euglena gracilis are differentially expressed during light-induced chloroplast development.

The enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase (EC 2.5.1.

JIP60, a methyl jasmonate-induced ribosome-inactivating protein involved in plant stress reactions.

Plant tissues treated with the naturally occurring cyclopentanone compound methyl jasmonate or exposed to stress causing in planta jasmonate accumulation express distinctive proteins and, concomitantly, reduce the synthesis of most preexisting proteins. One of the recently identified jasmonate-induced proteins, designated JIP60, in barley is a ribosome-inactivating protein that cleaves polysomes of both animal and plant origin into their ribosomal subunits. By attacking foreign and self ribosomes, respectively, JIP60 appears to be both a defense protein and a potent regulator of protein synthesis in stressed plant tissues.

Overproduction by gene amplification of the multifunctional arom protein confers glyphosate tolerance to a plastid-free mutant of Euglena gracilis.

Cells of the plastid-free mutant line of Euglena gracilis var. bacillaris, W10BSmL, can be adapted to glyphosate [N-(phosphonomethyl)glycine] by gradually increasing the concentration of the herbicide in the culture medium. The molecular basis of glyphosate tolerance is the selective ca.

Methyl jasmonate-regulated translation of nuclear-encoded chloroplast proteins in barley (Hordeum vulgare L. cv. salome).

The naturally occurring plant growth regulator (-)-jasmonic acid methyl ester (JaMe) induces the formation of novel abundant proteins in excised barley leaf segments. Concomitantly, this substance depresses the translation of most preexisting ("control") leaf mRNAs, including those for nuclear-encoded chloroplast proteins such as the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (SSU, rbcS gene product) and several light harvesting chlorophyll protein complex apoproteins (LHCPs, cab gene products). The changes in protein synthesis observed for SSU and LHCPs did not correspond to equivalent alterations in the rbcS and cab transcript levels.

A methyl jasmonate-induced shift in the length of the 5' untranslated region impairs translation of the plastid rbcL transcript in barley.

The plant growth substance (-)-jasmonic acid methyl ester (methyl jasmonate, JaMe) affects plastid gene expression at the protein and mRNA levels when applied exogenously to detached leaf segments of Hordeum vulgare L. cv. Salome.

The identification of leaf thionin as one of the main jasmonate-induced proteins of barley (Hordeum vulgare).

Jasmonic acid (JA) and its methyl ester (JA-Me) are able to introduce the accumulation of several specific polypeptides in cut leaf segments of barley. Two of the most prominent JA-induced proteins of M(r) 15,000 and 23,000 have been characterized by isolating and sequencing complete cDNA sequences. While the sequence of the M(r) 23,000 polypeptide shows no similarity to published sequences, the sequence of the M(r) 15,000 polypeptide corresponds to the higher-molecular-weight precursor of a leaf thionin previously characterized.

N-(phosphonomethyl)glycine (glyphosate) tolerance in Euglena gracilis acquired by either overproduced or resistant 5-enolpyruvylshikimate-3-phosphate synthase.

Photoautotrophic cells of Euglena gracilis can be adapted to N-(phosphonomethyl)glycine (glyphosate) by cultivation in media with progressively higher concentrations of the herbicide. Two different mechanisms of tolerance to the herbicide were observed. One is characterized by the overproduction and 40-fold accumulation of the target enzyme.

Translational regulation of plastid gene expression in Euglena gracilis.

Translation of plastid messenger RNAs depends on aminoacyl-tRNAs formed by charging plastid-encoded tRNAs with cognate amino acids. The enzymes involved, chloroplast aminoacyl-tRNA synthetases, are encoded in the nucleus. Both the tRNAs and the aminoacyl-tRNA synthetases are stimulated in synthesis if dark-grown cells are exposed to light.

In-vitro transport of chloroplast proteins in a homologousEuglena system with particular reference to plastid leucyl-tRNA synthetase.

In-vitro translations of total or polyadenylated RNA from chemoorganotrophic and photoautotrophicEuglena gracilis showed no substantial differences in the polypeptide patterns of the two cell types. By contrast, the corresponding patterns of in-vivo labelling indicated that posttranscriptional control of abundant cellular proteins occurred in illuminated cells. This type of control was confirmed forEuglena chloroplast proteins of cytoplasmic origin.

Jasmonate-induced alteration of gene expression in barley leaf segments analyzed by in-vivo and in-vitro protein synthesis.

Jasmonic-acid methylester promotes barley leaf senescence without changing the average synthesizing capacity for bulk leaf proteins in the treated tissues. This protein balance is the result of a massive formation of jasmonate-induced proteins (JIPs), which cannot be detected in controls (water-treated leaf segments). Jasmonate-induced proteins synthesized in vivo are virtually identical to the respective polypeptides translated in a wheat-germ system if programmed with the RNA of jasmonate-treated leaf segments.

Synthesis of diadenosine 5',5'''-P1,P4-tetraphosphate by organellar and cytoplasmic phenylalanyl-tRNA synthetases of Euglena gracilis.

Purified phenylalanyl-tRNA synthetases present in chloroplasts, mitochondria and cytoplasm of green and bleached Euglena gracilis strains, respectively, are able to synthesize diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A). Ap4A synthesis is strictly dependent on zinc ions. This is the first evidence that chloroplasts should be able to synthesize Ap4A.

Polypeptides of DNA-dependent RNA polymerase of spinach chloroplasts: characterization by antibody-linked polymerase assay and determination of sites of synthesis.

Using solid-phase ;Sandwich' immunoassays we studied DNA-dependent RNA polymerase of spinach chloroplasts with regard to (i) polypeptide composition of the multimeric enzyme; (ii) immunological cross-reaction with Escherichia coli RNA polymerase; (iii) sites of synthesis of polymerase polypeptides. Our main results are as follows. (i) All polypeptides of isolated chloroplast RNA polymerase (150, 145, 110, 102, 80, 75 and 38 kd) are labeled by an antibody-linked polymerase assay (ALPA), i.

Gene expression in cytokinin-and light-mediated plastogenesis of Cucurbita cotyledons: ribulose-1,5-bisphosphate carboxylase/oxygenase.

The effects of a cytokinin (N(6)-benzyladenine, BA) and light on plastogenesis have been studied in detached Cucurbita cotyledons using the key enzyme of photosynthetic CO2 fixation, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCase), as an example of a coordinated program of plastid and nucleo-cytoplasmic gene expression. Treatment of etiolated cotyledons with either BA in darkness or in light or light alone results in a marked and correlated stimulation of enzyme activity, quantity and biosynthesis (in-vivo [(14)C]leucine incorporation into immunoprecipitated enzyme protein), indicating an increase of de-novo synthesis under the influence of the two effectors. Cell-free translation of non-polyadenylated (poly(A)(-))RNA in an Escherichia coli system and total RNA in a wheat-germ system likewise demonstrate a light and hormone-dependent increase in the amounts of translatable mRNAs for the large (LS) and small subunits (SS) of RuBPCase (among other polypeptides).

Peptide hydrolase activities in seedlings and hormone-treated cotyledons of pumpkin (Cucurbita pepo).

Enzymes hydrolyzing Gly-Ala-, Met-Met- and Pro-4-phenylazo-phenylamides, and N-benzoyl-L-arginine-4-nitroanilide have been identified in germinating seeds and cotyledons of pumpkin (Cucurbita pepo). The enzyme activities per cotyledon increase markedly during the germination process, but the proportion of enhancement depends on the type of enzyme species. The increase in enzyme activities is due to de novo synthesis as shown by cycloheximide treatment and is influenced by phytohormones (cytokinins and abscissic acid).