Microarray Analysis of Late Response to Boron Toxicity in Barley (Hordeum vulgare L.) Leaves

DNA microarrays, being high-density and high-throughput, allow quantitative analyses of thousands of genes and their expression patterns in parallel. In this study, Barley1 GereChip was used to investigate transcriptome changes associated with boron (B) toxicity in a sensitive barley cultivar (Hordeum vulgare L. cv. Hamidye). Eight-day-old aseptically grown seedlings were subjected to 5 or 10 mM boric acid (B(OH)(3)) treatments for 5 days and expression profiles were determined with DNA microarrays using total RNA from leaf tissues. Among the 22,840 transcripts - each represented with a probe set on the GeneChip - 19,424 probe sets showed intensity values greater than 20(th) percentile in at least one of the hybridizations. Compared to control (10 mu M B(OH)(3)), 5 mM B(OH)(3) treatment resulted in differential expression of 168 genes at least by twofold. Moreover, 10 mM B(OH)(3) treatment resulted in at least twofold induction or reduction in expression of 312 transcripts. Among these genes, 37 and 61 exhibited significantly (P <0.05) altered levels of expression under 5 and 10 mM B(OH)(3) treatments, respectively. Differentially expressed genes were characterized using expression-based clustering and HarvEST:Barley. Investigations of expression profiles revealed that B toxicity results in global changes in the barley transcriptome and networks of signaling or molecular responses. A noticeable feature of response to 8 was that it is highly interconnected with responses to various environmental stresses. Additionally, induction of jasmonic acid related genes was found to be an important late response to B toxicity. Determination of responsive genes will shed light on successive studies aiming to elucidate molecular mechanism of B toxicity or tolerance. To the best of our knowledge, this is the first report on global expression analysis of barley seedlings under B toxicity

Anatomy of a microearthquake sequence on an active normal fault

The analysis of similar earthquakes, such as events in a seismic sequence, is an effective tool with which to monitor and study source processes and to understand the mechanical and dynamic states of active fault systems. We are observing seismicity that is primarily concentrated in very limited regions along the 1980 Irpinia earthquake fault zone in Southern Italy, which is a complex system characterised by extensional stress regime. These zones of weakness produce repeated earthquakes and swarm-like microearthquake sequences, which are concentrated in a few specific zones of the fault system. In this study, we focused on a sequence that occurred along the main fault segment of the 1980 Irpinia earthquake to understand its characteristics and its relation to the loading-unloading mechanisms of the fault system

Superoxide dismutase activity of hexaploid and tetraploid wheat cultivars subjected to heat and chilling stress

Effect of heat (37degreesC, 24 hours) and chilling (4degreesC, 24 hours) stress on SOD activity was investigated in shoots and roots of hexaploid and tetraploid wheat cultivars. In order to analyse the changes of SOD isozymes against heat and chilling stress, shoot and root extracts were subjected to native PAGE. Extracts of both tissues revealed the presence of Mn and Cu/Zn isozymes of SOD. Cu/ZnSOD activity comprised 90% of total SOD activity in both tissues. Heat stress caused a significant increase in SOD activity both in root and shoot tissues of the hexaploid cultivars whereas a decrease was observed in the tetraploid cultivar. Under cold stress no significant change in SOD activity was observed in shoot tissues of all cultivars. On the contrary, SOD activities in the roots of the hexaploid cultivars were significantly enhanced while a decrease was observed in the roots of tetraploid cultivar. Our data suggest that enhanced SOD activity might be one of the reasons of relatively greater stress tolerance of bread wheat cultiars over durum cultivars under heat and chilling stress conditions

Superoxide dismutase activity in salt stressed wheat seedlings

We investigated the effect of salt stress on enzymatic activity of superoxide dismutase (SOD) isozymes in shoot and root tissues of salt tolerant and sensitive wheat (Triticum aestivum L. and Triticum durum Defs.) cultivars. Ten day old seedlings were subjected to 0.7 M NaCl stress for 3 and 5 days. Seedlings treated in the same manner without salt stress served as controls. Activity of SOD isozymes in root and shoot extracts was determined by activity staining of native polyacrylamide gels. In both shoot and root extracts of examined cultivars two isozymes of SOD, namely MnSOD and Cu/ZnSOD were identified. Cu/ZnSOD activity comprised 90 % of total SOD activity in both root and shoot tissues. Salt stress caused 1-1.5 fold increase in MnSOD activity of shoots in tolerant cultivars when compared with non-stressed controls. Under stress conditions, compared to controls all cultivars exhibited reduced MnSOD activity in root tissues. Cu/ZnSOD activity, on the other hand, was remarkably enhanced (3-4 fold) in root extracts of the tolerant cultivars, whereas it was reduced in the sensitive ones

Antioxidant responses of shoots and roots of lentil to NaCl-salinity stress

The effect of salt stress (100 mM and 200 mM NaCl) on antioxidant responses in shoots and roots of 14-day-old lentil (Lens culinaris M.) seedlings was investigated. Salt stress caused a significant decrease in length, wet-dry weight and an increase in proline content of both shoot and root tissues. In leaf tissues, high salinity treatment resulted in a 4.4 fold increase in H2O2 content which was accompanied by a significant level of lipid peroxidation and an increase in electrolyte leakage. Root tissues were less affected with respect to these parameters. Leaf tissue extracts exhibited four activity bands, of which two were identified as Cu/Zn-SOD and others as Fe-SOD and Mn-SOD. Fe-SOD activity was missing in root extracts. In both tissues Cu/Zn-SOD activity comprised 70-75% of total SOD activity. Salt stress did not cause a significant increase in total SOD activity of leaf tissues but a significant enhancement (88%) was observed in roots mainly due to an enhancement in Cu/ZnSOD isoforms. Compared to leaf tissues a significantly higher constitutive ascorbate peroxidase (APX) and glutathion reductase (GR) activity was observed in root tissues. Upon salt stress no significant change in the activity of APX, catalase (CAT) and GR was observed in root tissues but a higher APX activity was present when compared to leaf tissues. On the other hand, in leaf tissues, with the exception of CAT, salt stress caused significant enhancement in the activity of other antioxidant enzymes. These results suggested that, root tissues of lentil are protected better from NaCl stress induced oxidative damage due to enhanced total SOD activity together with a higher level of APX activity under salinity stress. To our knowledge this is the first report describing antioxidant enzyme activities in lentil

Microarray Analysis of Late Response to Boron Toxicity in Barley (Hordeum vulgare L.) Leaves

DNA microarrays, being high-density and high-throughput, allow quantitative analyses of thousands of genes and their expression patterns in parallel. In this study, Barley1 GereChip was used to investigate transcriptome changes associated with boron (B) toxicity in a sensitive barley cultivar (Hordeum vulgare L. cv. Hamidye). Eight-day-old aseptically grown seedlings were subjected to 5 or 10 mM boric acid (B(OH)(3)) treatments for 5 days and expression profiles were determined with DNA microarrays using total RNA from leaf tissues. Among the 22,840 transcripts - each represented with a probe set on the GeneChip - 19,424 probe sets showed intensity values greater than 20(th) percentile in at least one of the hybridizations. Compared to control (10 mu M B(OH)(3)), 5 mM B(OH)(3) treatment resulted in differential expression of 168 genes at least by twofold. Moreover, 10 mM B(OH)(3) treatment resulted in at least twofold induction or reduction in expression of 312 transcripts. Among these genes, 37 and 61 exhibited significantly (P <0.05) altered levels of expression under 5 and 10 mM B(OH)(3) treatments, respectively. Differentially expressed genes were characterized using expression-based clustering and HarvEST:Barley. Investigations of expression profiles revealed that B toxicity results in global changes in the barley transcriptome and networks of signaling or molecular responses. A noticeable feature of response to 8 was that it is highly interconnected with responses to various environmental stresses. Additionally, induction of jasmonic acid related genes was found to be an important late response to B toxicity. Determination of responsive genes will shed light on successive studies aiming to elucidate molecular mechanism of B toxicity or tolerance. To the best of our knowledge, this is the first report on global expression analysis of barley seedlings under B toxicity

Evaluation of photosynthetic performance of wheat cultivars exposed to boron toxicity by the JIP fluorescence test

kayihan, ceyhun/0000-0003-1684-4147; Oz, Mehmet Tufan/0000-0002-0042-2671WOS: 000345394500010The changes in growth and photosynthetic performance of two wheat (Triticum aestivum L.) cultivars (Bolal-2973 and Atay-85) differing in their sensitivity to boron (B) toxicity were investigated under toxic B conditions. Eight-day old seedlings were exposed to highly toxic B concentrations (5, 7.5, and 10 mM H3BO3) for 5 and 9 days. Fast chlorophyll a fluorescence kinetics was determined and analysed using JIP test. Growth parameters, tissue B contents, and membrane damage were measured at two stress durations. The photochemical performance of PSII was hindered more in the sensitive cultivar (Atay-85) than that of the tolerant one (Bolal-2973) under B toxicity. The increase in the B concentration and stress duration caused membrane leakage in both cultivars. However, higher membrane damage was observed in Atay-85 compared to Bolal-2973. Additionally, significant reduction of growth parameters was observed in both cultivars at toxic B concentrations. The accumulation of B was higher in shoots than in roots of both cultivars. Nevertheless, Atay-85 translocated more B from roots to leaves compared to Bolal-2973. The advantages of certain JIP test parameters were demonstrated for evaluation of PSII activity in plants exposed to B stress. Evaluation of photosynthetic performance by JIP test as well as assessment of growth and tissue B content might be used to determine the effects of B toxicity in wheat. The results indicated lesser sensitivity to B toxicity in Bolal-2973 compared to Atay-85.National Boron Research Institute, Turkey [BOREN-2009-C0217]This work was supported by the National Boron Research Institute, Turkey (Project BOREN-2009-C0217)