Cross-talk between high light stress and plant defence to the two-spotted spider mite in Arabidopsis thaliana.

Little is known about how plants deal with arthropod herbivores under the fluctuating light intensity and spectra which occur in natural environments. Moreover, the role of simultaneous stress such as excess light (EL) in the regulation of plant responses to herbivores is poorly characterized. In the current study, we focused on a mite-herbivore, specifically, the two-spotted spider mite (TSSM), which is one of the major agricultural pests worldwide. Our results showed that TSSM-induced leaf damage (visualized by trypan blue staining) and oviposition rate (measured as daily female fecundity) decreased after EL pre-treatment in wild-type Arabidopsis plants, but the observed responses were not wavelength specific. Thus, we established that EL pre-treatment reduced Arabidopsis susceptibility to TSSM infestation. Due to the fact that a portion of EL energy is dissipated by plants as heat in the mechanism known as non-photochemical quenching (NPQ) of chlorophyll fluorescence, we tested an Arabidopsis npq4-1 mutant impaired in NPQ. We showed that npq4-1 plants are significantly less susceptible to TSSM feeding activity, and this result was not dependent on light pre-treatment. Therefore, our findings strongly support the role of light in plant defence against TSSM, pointing to a key role for a photo-protective mechanism such as NPQ in this regulation. We hypothesize that plants impaired in NPQ are constantly primed to mite attack, as this seems to be a universal evolutionarily conserved mechanism for herbivores.

A novel superoxide dismutase with a high isoelectric point in higher plants. expression, regulation, and protein localization.

Several isoforms of superoxide dismutase (SOD) with a high isoelectric point (pI) have been identified by isoelectric focusing chromatography in protein extracts from Scots pine (Pinus sylvestris) needles. One of these isoforms, a CuZn-SOD with a pI of about 10 and thus denoted hipI-SOD, has been isolated and purified to apparent homogeneity. A cDNA encoding the hipI-SOD protein was cloned and sequenced. Northern hybridization of mRNA isolated from different organs and tissues showed that hipI-SOD has a markedly different pattern of expression compared with chloroplastic and cytosolic SOD. Furthermore, the transcript levels of hipI-SOD and cytosolic SOD were found to respond differently to mechanical wounding, treatment with oxidized glutathione, paraquat, and ozone. Immunogold electron microscopy localized the hipI-SOD in the plasma membrane of sieve cells and the Golgi apparatus of albuminous cells. Moreover, high protein density was also detected in extracellular spaces such as secondary cell wall thickenings of the xylem and sclerenchyma and in intercellular spaces of parenchyma cells.

Are diverse signalling pathways integrated in the regulation of arabidopsis antioxidant defence gene expression in response to excess excitation energy?

When low-light-grown Arabidopsis rosettes are partially exposed to excess light (EL), the unexposed leaves become acclimated to excess excitation energy (EEE) and consequent photo-oxidative stress. This phenomenon, termed systemic acquired acclimation (SAA), is associated with redox changes in the proximity of photosystem II, changes in foliar H2O2 content and induction of antioxidant defences. The induction of extra-plastidial antioxidant systems is important in the protection of the chloroplast under EL conditions. A larger range of transcripts encoding different antioxidant defence enzymes may be induced in the systemically acclimated leaves and these include those encoded by the glutathione peroxidase (GPX2) and glutathione-S-transferase (GST) genes, which are also highly induced in the hypersensitive response and associated systemic acquired resistance (SAR) in incompatible plant-pathogen interactions. Furthermore, the expression of the SAR-inducible pathogenesis-related protein gene, PR2, is enhanced in SAA leaves. Wounded leaf tissue also shows enhanced systemic induction of a cytosolic ascorbate peroxidase gene (APX2) under EL conditions. These and other considerations, suggest H2O2 and other reactive oxygen species (ROS) could be the common factor in signalling pathways for diverse environmental stresses. These effects may be mediated by changes in the level and redox state of the cellular glutathione pool. Mutants with constitutive expression of a normally EL-inducible APX2 gene have much reduced levels of foliar glutathione. The expression of APX1 and APX3, encoding cytosolic and peroxisome-associated isoforms, respectively, are also under phytochrome-A-mediated control. The expression of these genes is tightly linked to the greening of plastids in etiolated seedlings. These data suggest that part of the developmental processes that bring about the acclimation of leaves to high light includes the configuration of antioxidant defences. Therefore, the linkage between immediate responses of leaves to EL, acclimation of chloroplasts to EEE and the subsequent changes to leaf form and function in high light could be mediated by the activity of foliar antioxidant defences and changes in the concentration of ROS.

Antagonistic effects of hydrogen peroxide and glutathione on acclimation to excess excitation energy in Arabidopsis.

The redox status of the quinone B (Q(B)) and plastoquinone (PQ) pools plays a key role in the cellular and systemic signalling processes that control acclimatory responses in plants. In this study, we demonstrate the effects of hydrogen peroxide and glutathione on acclimatory responses controlled by redox events in the proximity of the Q(B)-PQ pools. Our results suggest that the chloroplast is a sink for H2O2 and that, paradoxically, high concentrations of H2O2 in the chloroplast protect the photosynthetic apparatus and the plant cell from photoinhibition and photooxidative damage. Excess glutathione, however, caused an effect antagonistic to that observed for high H2O2. An explanation of this apparent paradox and a hypothetical redox-signalling model are suggested.

Systemic signaling and acclimation in response to excess excitation energy in Arabidopsis.

Land plants are sessile and have developed sophisticated mechanisms that allow for both immediate and acclimatory responses to changing environments. Partial exposure of low light-adapted Arabidopsis plants to excess light results in a systemic acclimation to excess excitation energy and consequent photooxidative stress in unexposed leaves. Thus, plants possess a mechanism to communicate excess excitation energy systemically, allowing them to mount a defense against further episodes of such stress. Systemic redox changes in the proximity of photosystem II, hydrogen peroxide, and the induction of antioxidant defenses are key determinants of this mechanism of systemic acquired acclimation.

The chlB gene encoding a subunit of light-independent protochlorophyllide reductase is edited in chloroplasts of conifers.

ChlB is one of three chloroplast genes shown so far to be required for light-independent chlorophyll synthesis. It occurs in some algae, lower plants, and gymnosperms, but not in angiosperms. We have demonstrated, for the first time in conifer chloroplasts, the presence of two internal C to U editing sites in this transcript. In the chlB transcript of Pinus sylvestris, the editing of the second position in a CCG codon leads to an amino-acid substitution from proline to leucine. Editing of a nearby CGG codon, resulting in an arginine to tryptophan substitution, has also been observed. The nucleotide sequence of this region has been compared with other species of gymnosperms. Out of seven species analysed, editing at both sites has only been detected in spruce, while in Larix only the editing which results in the Arg to Trp substitution was found. In other cases, both leucine and tryptophan are encoded by cpDNA, suggesting that conservation of these amino acids, through encoding by DNA or by editing of the RNA, is critical for the protein function. Transcripts are partially edited at the CGG codon and the relative abundance of cDNA molecules with the edited C is species-specific. The possible involvement of RNA editing in the regulation of gene expression in different organs of pine seedlings is discussed.

Photosynthetic electron transport regulates the expression of cytosolic ascorbate peroxidase genes in Arabidopsis during excess light stress.

Exposure of Arabidopsis plants that were maintained under low light (200 mumol of photons m-2 sec-1) to excess light (2000 mumol of photons m-2 sec-1) for 1 hr caused reversible photoinhibition of photosynthesis. Measurements of photosynthetic parameters and the use of electron transport inhibitors indicated that a novel signal transduction pathway was initiated at plastoquinone and regulated, at least in part, by the redox status of the plastoquinone pool. This signal, which preceded the photooxidative burst of hydrogen peroxide (H2O2) associated with photoinhibition of photosynthesis, resulted in a rapid increase (within 15 min) in mRNA levels of two cytosolic ascorbate peroxidase genes (APX1 and APX2). Treatment of leaves with exogenous reduced glutathione abolished this signal, suggesting that glutathione or the redox status of the glutathione pool has a regulatory impact on this signaling pathway. During recovery from photooxidative stress, transcripts for cytosolic glutathione reductase (GOR2) increased, emphasizing the role of glutathione in this stress.

The genes encoding subunit 3 of NADH dehydrogenase and ribosomal protein S12 are co-transcribed and edited in Pinus sylvestris (L.) mitochondria.

The nucleotide sequence of the region encoding NADH dehydrogenase subunit 3 and ribosomal protein S12 from Pinus sylvestris (L.) mitochondrial DNA (mtDNA) has been determined. A sequence comparison of this region with six individual cDNA clones prepared by RT-PCR revealed 35 C-to-T differences, showing the occurrence of RNA editing. All but one of these alterations in mRNA sequence change codon identities to specify amino acid better conserved in evolution. Most of these modifications take place within the nad3 gene changing 20% of the amino-acid sequence, which is much more than in angiosperms. Of six cDNA clones investigated, four clones of nad3 were differentially edited, but the editing of the rps12 sequences was identical. As in angiosperms, the two genes are separated by a short sequence of 52 bp, which is not edited. Two transcripts of about 0.9 kb and 1.2 kb, each encoding both proteins, have been detected by Northern hybridisation. The hybridisation of nad3 and rps12 probes with pine mtDNA digested with different restriction enzymes indicates that both genes are present in a single copy in pine mtDNA. The analysis of PCR amplification products with gene-specific primers shows a conserved order of these genes in a wide range of gymnosperms.

[Dexamethasone test in the assessment of depression grade in patients with low back pain]. / Zastosowanie testu deksametazonowego dla oceny stopnia depresji u chorych z bólami krzyza.

The results of the dexamethasone test were compared with the depression depth and pain intensity in patients with low back pain caused by degenerative changes and/or disc prolapse. In the light of these studies no correlation was demonstrated between the positive results of the dexamethasone test and depression symptoms and the intensity and duration of pain.

Organization of the 18S, 5S, 4S rRNA genes and the tRNA-like repeat in the mitochondrial genomes of three lupin species.

Southern blots of mitochondrial (mt) DNAs of three Lupinus species cleaved with three restriction enzymes were probed with Lupinus luteus mtDNA fragments containing 18S, 5S rRNA genes or a tRNA-like repeat. Comparison of the number of hybridizing bands and their intensity suggested that the mt 18S and 5S rRNA genes occur mostly in one copy in the genomes of three lupin species. The exception concerned the Lupinus angustifolius 5S rRNA gene showing two hybridizing bands of unequal intensity. The results of hybridization of the lupin mitochondrial genomes with a probe specific for the Lupinus luteus tRNA-like repeat pointed to the presence of such a repeat in other parts of the genomes besides the vicinity of the 18S rRNA gene. Northern hybridization analysis showed the presence of 18S, 5S and tRNA-like repeat transcripts similar in size in all lupin species.

[Analgesic effect of imipramine in the treatment of ischium pain and circadian rhythm of cortisol release into the serum]. / Efekt przeciwbólowy imipraminy w leczeniu rwy kulszowej a okolodobowy rytm wydzielania kortyzolu w surowicy.

In the paper the results are presented of treatment of ischialgia with Imipramine, taking into account the circadian rhythm of cortisol release into the serum. The obtained results encourage to the use of tricyclic antidepressants in the treatment refractory cases of ischialgia. No significant differences were observed in the circadian rhythm of cortisol release into the serum before and after the treatment with Imipramine.

Molecular Responses to Photooxidative Stress in Pinus sylvestris (L.) (II. Differential Expression of CuZn-Superoxide Dismutases and Glutathione Reductase.

The influence of photooxidative stress on genes expressing superoxide dismutase (Sod) and glutathione reductase (Gor) was analyzed in needles of top and side shoots of 3-year-old Pinus sylvestris (L.) seedlings. The study was carried out in the field during spring recovery. From mid-April the top shoots of seedlings protruded above the snow and thus were exposed to sunlight, whereas the side shoots were covered with snow until May 4. Needles were sampled from top and side shoots on five different occasions. At the beginning of May the mRNA levels for cytosolic CuZn-Sod were significantly higher in top-shoot needles than in side-shoot needles. Similar results were obtained for chloroplastic CuZn-Sod mRNA. After May 6 we could not detect any significant differences between top- and side-shoot needles for either CuZn-Sod mRNA level. Transcript accumulation for the chloroplastic CuZn-Sod was up to 4-fold higher than for cytosolic CuZn-Sod in both types of shoots. On June 1 minimum transcript levels were observed for both CuZn-SOD isoforms. Protein activity analysis for CuZn-SOD isozymes did not reveal any significant differences between top- and side-shoot needles during the whole period of measurements. The mRNA level for chloroplastic Gor was similar in both types of shoots. However, the total GR activity was significantly higher in top-shoot needles than in side-shoot needles at the beginning of May. The analysis of mRNA accumulation for chloroplastic CuZn-Sod and Gor indicates that transcript levels were at least 5- to 20-fold higher for CuZn-Sod than for chloroplastic Gor. The differential expressions of Sod and Gor genes are discussed in relation to regulation of the enzymic scavenging system during photooxidative stress conditions.

[Assessment of dexamethasone effectiveness in the treatment of ischalgia]. / Ocena skutecznosci leczenia rwy kulszowej deksametazonem.

The authors studied 43 patients admitted to hospital with acute episodes of ischalgia or recurrences caused by intervertebral disc disease. The patients were randomly selected into two groups treated with dexamethasone in high but rapidly decreasing doses, or tramadol. A comparison of the results in both groups showed that dexamethasone was effective and safe when given by this method to patients with ischalgia.

Lupin chloroplast and mitochondrial tRNA populations and their genes.

Transfer RNAs isolated from lupin chloroplasts and mitochondria were compared by two-dimensional gel electrophoresis. Twenty chloroplast and 24 mitochondrial tRNA species were identified. The saturation hybridization between lupin chloroplast DNA and 125I-labelled lupin chloroplast tRNAs pointed to the presence of about 34 tRNA genes in lupin chloroplast DNA. The number of mitochondrial tRNA genes estimated by the same method was about 30 genes. EcoRI restriction digest of lupin mitochondrial DNA probed with 32P-labelled lupin mitochondrial tRNAs revealed only a small number of positive restriction fragments. Some of these mitochondrial restriction fragments hybridized with 32P-labelled chloroplast tRNA.

The effect of atmospheric pollution on α-glycerophosphate dehydrogenase activity in the satin moth (Leucoma salicis (L.)).

The purpose of this study was to investigate how various combinations of atmospheric pollutants may affect α-GPDH activity in the satin moth. Enzyme activity was measured in crude larva and pupa as well as in hoogenates from the fat body and thoracic muscles of adults. The insects were divided into five groups and treated with dust (containing heavy metals) and gaseous pollutants (SO2, NOx) in different concentrations. Specific activity of α-GPDH was the highest in thoracic muscles of adult satin moth and was significantly higher in males, regardless of their origin. Low concentration of SO2 caused stimulation of enzyme activity but simultaneous action of SO2, NOx and heavy metal containing dust with accompanied acid precipitation caused inhibition of α-GPDH activity, probably as the result of sulphite bounds with pyridine and flavin nucleotides.