Oxygenation of European seabass (Dicentrarchus labrax L.) in cages through aeration and effect on lipid metabolism.

Two groups of the same batch of European seabass were used in an industrial-scale trial in sea cages in Vorios Evoikos, Greece. For about one month, one of the two cages was oxygenated by use of compressed air injected in seawater through an AirX frame (Oxyvision A/S, Norway) at 3.5 m depth, while oxygen concentration and temperature were monitored every 30 min. The liver, gut, and pyloric ceca samples were taken from fish of both groups for measurement of gene expression of phospholipase A2 (PLA2) and hormone-sensitive lipase (HSL), and for histological analysis at the middle and end of the experiment. Real-time qPCR was performed with housekeeping genes ACTb, L17, and EF1a. The expression of PLA2 increased in pyloric caeca samples from the oxygenated cage, suggesting that aeration improved the absorption rate of dietary phospholipids (p < 0.05). Expression of HSL increased significantly in liver samples from the control cage, in comparison with the aerated cage (p < 0.05). Histological examination of sea bass samples revealed an increased fat accumulation in the hepatocytes of fish in the oxygenated cage. The results of the present study showed increased lipolysis induced by low DO levels in farmed sea bass in cages.

Genetic and Epigenetic Responses of Autochthonous Grapevine Cultivars from the 'Epirus' Region of Greece upon Consecutive Drought Stress.

Within the framework of preserving and valorizing the rich grapevine germplasm of the Epirus region of Greece, indigenous grapevine (Vitis vinifera L.) cultivars were characterized and assessed for their resilience to abiotic stresses in the context of climate change. The cultivars 'Debina' and 'Dichali' displayed significant differences in their response to drought stress as judged by morpho-physiological analysis, indicating higher drought tolerance for Dichali. Hence, they were selected for further study aiming to identify genetic and epigenetic mechanisms possibly regulating drought adaptability. Specifically, self-rooted and heterografted on 'Richter 110' rootstock plants were subjected to two phases of drought with a recovery period in between. Gene expression analysis was performed for two stress-related miRNAs and their target genes: (a) miRNA159 and putative targets, VvMYB101, VvGATA-26-like, VvTOPLESS-4-like and (b) miRNA156 and putative target gene VvCONSTANS-5. Overall, grafted plants exhibited a higher drought tolerance than self-rooted plants, suggesting beneficial rootstock-scion interactions. Comparative analysis revealed differential gene expression under repetitive drought stresses between the two cultivars as well as between the self-rooted and grafted plants. 'Dichali' exhibited an up-regulation of most of the genes examined, which may be associated with increased tolerance. Nevertheless, the profound down-regulation of VvTOPLESS-4-like (a transcriptional co-repressor of transcription factors) upon drought and the concomitant up-regulation of miRNA159 highlights the importance of this 'miRNA-target' module in drought responsiveness. DNA methylation profiling using MSAP analysis revealed differential methylation patterns between the two genotypes in response to drought. Further investigations of gene expression and DNA methylation will contribute to our understanding of the epigenetic mechanisms underlying grapevine tolerance to drought stress.

Temperature-Biased miRNA Expression Patterns during European Sea Bass (Dicentrarchus labrax) Development.

Environmental effects and, particularly, temperature changes have been demonstrated to influence the activity, function, and well-being of teleosts. Temperature may change seasonally in the wild, and in captivity under aquaculture operations. Moreover, climate change is expected to shift temperature profiles worldwide. MicroRNAs (miRNA) are important temperature-sensitive gene-expression regulators acting at the post-transcriptional level. They are known to be key regulators in development, reproduction, and immune responses. Therefore, early larval development of the European sea bass (Dicentrarchus labrax), one of the most extensively cultured species in Mediterranean aquaculture, was investigated at early rearing temperatures, i.e., 15, 17.5, and 20 °C, in regard to the impact of temperatures on miRNAs through sncRNA high-throughput sequencing but also at the phenotypic level in terms of growth, sex, vision, and skeletal deformities. Expression profiling revealed stage- and temperature-specific miRNA expression targeting genes with roles in reproduction and immune response mainly at the flexion and all-fins stages. Similar stage- and temperature-specific results were also observed concerning the number of rod cells and lower jaw elongation. The present work presents for the first time highly promising results on the influence of early rearing temperature at the post-transcriptional level during European sea bass development, with a putative impact on reproduction and immune response, as well as regarding teleost vision and larval development.

Enriched HeK4me3 marks at Pm-0 resistance-related genes prime courgette against Podosphaera xanthii.

Powdery mildew (PM) disease, caused by the obligate biotrophic fungal pathogen Podosphaera xanthii, is the most reported and destructive disease on cultivated Cucurbita species all over the world. Recently, the appearance of highly aggressive P. xanthii isolates has led to PM outbreaks even in resistant crops, making disease management a very difficult task. To challenge this, breeders rely on genetic characteristics for PM control. Analysis of commercially available intermediate resistance courgette (Cucurbita pepo L. var. cylindrica) varieties using cytological, molecular, and biochemical approaches showed that the plants were under a primed state and induced systemic acquired resistance (SAR) responses, exhibiting enhanced callose production, upregulation of salicylic acid (SA) defense signaling pathway genes, and accumulation of SA and defense metabolites. Additionally, the intermediate resistant varieties showed an altered epigenetic landscape in histone marks that affect transcriptional activation. We demonstrated that courgette plants had enriched H3K4me3 marks on SA-BINDING PROTEIN 2 and YODA (YDA) genes of the Pm-0 interval introgression, a genomic region that confers resistant to Cucurbits against P. xanthii. The open chromatin of SA-BINDING PROTEIN 2 and YDA genes was consistent with genes' differential expression, induced SA pathway, altered stomata characteristics, and activated SAR responses. These findings demonstrate that the altered epigenetic landscape of the intermediate resistant varieties modulates the activation of SA-BINDING PROTEIN 2 and YDA genes leading to induced gene transcription that primes courgette plants.

Boron toxicity in higher plants: an update.

MAIN CONCLUSION: In this review, emphasis is given to the most recent updates about morpho-anatomical, physiological, biochemical and molecular responses adopted by plants to cope with B excess. Boron (B) is a unique micronutrient for plants given that the range of B concentration from its essentiality to toxicity is extremely narrow, and also because it occurs as an uncharged molecule (boric acid) which can pass lipid bilayers without any degree of controls, as occurs for other ionic nutrients. Boron frequently exceeds the plant's requirement in arid and semiarid environments due to poor drainage, and in agricultural soils close to coastal areas due to the intrusion of B-rich seawater in fresh aquifer or because of dispersion of seawater aerosol. Global releases of elemental B through weathering, volcanic and geothermal processes are also relevant in enriching B concentration in some areas. Considerable progress has been made in understanding how plants react to B toxicity and relevant efforts have been made to investigate: (I) B uptake and in planta partitioning, (II) physiological, biochemical, and molecular changes induced by B excess, with particular emphasis to the effects on the photosynthetic process, the B-triggered oxidative stress and responses of the antioxidant apparatus to B toxicity, and finally (III) mechanisms of B tolerance. Recent findings addressing the effects of B toxicity are reviewed here, intending to clarify the effect of B excess and to propose new perspectives aimed at driving future researches on the topic.

Changes in sugar metabolism associated to stem bark thickening partially assist young tissues of Eriobotrya japonica seedlings under boron stress.

Boron (B) toxicity frequently affects plant performances and productivity, especially in arid and semi-arid environments. In this experiment, loquat seedlings were subjected to 25 µM (control) or 400 µM B (B excess) to test the hypothesis that (i) B alters sugar/polyol metabolism in polyol-producing tree species as loquat and (ii) changes of leaf and stem anatomy assist young tissues against toxic effect of B. Gas exchange was monitored from the beginning of the experiment (FBE) till one week after the first visible symptoms of B toxicity appeared in the upper part of the stems (147 d FBE). At 147 FBE, plant biometric parameters and pattern of B accumulation, leaf and stem anatomy, chlorophyll a fluorescence kinetics as well as biochemical measurements were assessed in top (asymptomatic) leaves and upper stem bark. Boron accumulated principally (in the row) in top leaves > top bark > top wood in B-stressed plants, but no changes in allocation pattern were found between controls and B-stressed plants. Excess B promoted the increase in the spongy layer of top leaves and caused the development of cork and numerous collenchyma cells with increased cell wall thickness. This mechanism, which has never been described before, can be considered an attempt to store excessive B in tissues where B ions are less harmful. The accumulation of sorbitol (B-complexing polyol) in top leaves and stem bark can be considered as a further attempt to detoxify B excess. However, B toxicity drastically affects the photosynthetic rate of top leaves, mainly due to non-stomatal limitations, i.e., reduction of ambient CO2 use efficiency and of photosystem II (PSII) efficiency, modification of the partitioning excess energy dissipation in PSII, thus leading to an increased level of lipid peroxidation. Our results suggest that changes in sugar metabolism associated with leaf and stem bark thickening partially assist (but not totally preserve) young tissues of loquat plants under B stress.

Mn-induced changes in leaf structure and chloroplast ultrastructure of Citrus volkameriana (L.) plants.

Seedlings of Citrus volkameriana (L.) were grown hydroponically for 43 days in order to study the effect of Mn concentration (0, 2, 14, 98 and 686 microM) in the nutrient solution on leaf anatomy and mesophyll chloroplast ultrastructure. Increasing Mn concentration stimulated leaf lamina thickness. The size of mesophyll chloroplasts decreased and increased under 0 and 686 microM Mn, respectively, compared to the intermediate Mn concentrations, similar with regard to the number of chloroplasts per mesophyll cell area. Thylakoid membranes of plants grown under 0 microM Mn were somewhat swelled, while those in other Mn treatments did not present any visible malformation. The relative volume of starch grains per chloroplast was significantly smaller under 0-98 microM Mn (12.8-16.0%) than in the treatment with 686 microM Mn (67.6%). Further, under 686 microM Mn, dark deposits were found in vacuoles. The existence of a cell adaptation mechanism to excessive Mn availability (686 microM Mn) by increasing the size of chloroplasts as well as their number per cellular area, is discussed.