Analysis of the transcriptional mechanisms of yellowfin tuna (Thunnus albacares) juveniles in response to acute cold stress in brain and muscle tissues.

Splenic tissue injury and physiological response mechanisms in juvenile yellowfin tuna (Thunnus albacares) under acute cold stress.

To understand the impacts of salinity stress on the antioxidation of yellowfin tuna Thunnus albacares, 72 fishes (646.52 ± 66.32 g) were randomly divided into two treatments (32‰ and 29‰) and sampled at four time points (0 h, 12 h, 24 h, and 48 h). The salinity of the control group (32‰) was based on natural filtered seawater and the salinity of the stress group (29‰) was reduced by adding tap water with 24 h aeration to the natural filtered seawater. The superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) from liver, gill, and muscle tissues were used as the antioxidant indexes in this study. The results showed that the changes of SOD and GSH-Px in the gills were first not significantly different from the control group (p > 0.05) and finally significantly higher than the control group (SOD: 50.57%, GSH-Px: 195.95%, p < 0.05). SOD activity in fish liver was not significantly changed from 0 h to 48 h (p > 0.05), and was not significantly different between the stress group and control group (p > 0.05). With the increase in stress time, GSH-Px and MDA activities in the liver of juvenile yellowfin tuna increased first (GSH-Px: 113.42%, MDA: 137.45%) and then reduced (GSH-Px: −62.37%, MDA: −16.90%) to levels similar to the control group. The SOD activity in the white and red muscle of juvenile yellowfin tuna first decreased (white muscle: −27.51%, red muscle: −15.52%) and then increased (white muscle: 7.30%, red muscle: 3.70%) to the level of the control group. The activities of GSH-Px and MDA in white and red muscle increased first (white muscle GSH-Px: 81.96%, red muscle GSH-Px: 233.08%, white muscle MDA: 26.89%, red muscle MDA: 64.68%) and then decreased (white muscle GSH-Px: −48.03%, red muscle GSH-Px: −28.94%, white muscle MDA: −15.93%, red muscle MDA: −28.67%) to the level observed in the control group. The results from the present study indicate that low salinity may lead to changes in the antioxidant function of yellowfin tuna juveniles. In contrast, yellowfin tuna juveniles have strong adaptability to the salinity of 29‰. However, excessive stress may consume the body’s reserves and reduce the body’s resistance.

We studied the mechanical properties of deep red aerobic muscle of yellowfin tuna (Thunnus albacares), using both in vivo and in vitro methods. In fish swimming in a water tunnel at 1-3 L s(-1) (where L is fork length), muscle length changes were recorded by sonomicrometry, and activation timing was quantified by electromyography. In some fish a tendon buckle was also implanted on the caudal tendon to measure instantaneous muscle forces transmitted to the tail. Between measurement sites at 0.45 to 0.65 L, the wave of muscle shortening progressed along the body at a relatively high velocity of 1.7 L per tail beat period, and a significant phase shift (31+/-4 degrees ) occurred between muscle shortening and local midline curvature, both suggesting red muscle power is directed posteriorly, rather than causing local body bending, which is a hallmark of thunniform swimming. Muscle activation at 0.53 L was initiated at about 50 degrees of the tail beat period and ceased at about 160 degrees , where 90 degrees is peak muscle length and 180 degrees is minimum length. Strain amplitude in the deep red fibres at 0.5 L was +/-5.4%, double that predicted from midline curvature analysis. Work and power production were measured in isolated bundles of red fibres from 0.5 L by the work loop technique. Power was maximal at 3-4 Hz and fell to less than 50% of maximum after 6 Hz. Based on the timing of activation, muscle strain, tail beat frequencies and forces in the caudal tendon while swimming, we conclude that yellowfin tuna, like skipjack, use their red muscles under conditions that produce near-maximal power output while swimming. Interestingly, the red muscles of yellowfin tuna are slower than those of skipjack, which corresponds with the slower tail beat frequencies and cruising speeds in yellowfin.

The scaling of red muscle with body weight and the distribution of red muscle within the body were compared in seven scombrid fish species to determine relationships between red muscle function and the maintenance of endothermy by tunas. In ectothermic Sarda chiliensis and Scomber japonicus, red muscle occurs along the body edge, is concentrated posteriorly, and the total amount of this tissue is proportional to body weight raised to a power significantly greater than 1.0. In five endothermic tunas, Auxis thazard, Euthynnus lineatus, Katsuwonus pelamis, Thunnus albacares, and T. alalunga, red muscle scaling coefficients are 1.0 or less, and red muscle is positioned deep and anterior in the body. The power needed to overcome drag increases with fish body size (weight and length) and velocity and is reflected in the red muscle scaling relationships of both Sarda and Scomber. By contrast, decreasing relative amounts of red muscle in larger tunas suggest these fishes increase propulsion efficiency as they grow. This may be a result of either or both greater muscle efficiency and reduced division of labor between red and white muscle to which both endothermy and thermoregulation could contribute.

<i>Circulation:</i> Clinical Summaries

![Figure][1] Arend Bonen Department of Human Health and Nutritional Sciences, University of Guelph, Guelph, Ontario, Canada abonen{at}uoguelph.ca ![Figure][1] Adrian Chabowski Department of Physiology, Medical University of Bialystok, Bialystok, Poland ![

To learn about muscle function in two species of tuna (yellowfin Thunnus albacares and skipjack Katsuwonus pelamis), a series of electromyogram (EMG) electrodes was implanted down the length of the body in the internal red (aerobic) muscle. Additionally, a buckle force transducer was fitted around the deep caudal tendons on the same side of the peduncle as the electrodes. Recordings of muscle activity and caudal tendon forces were made while the fish swam over a range of steady, sustainable cruising speeds in a large water tunnel treadmill. In both species, the onset of red muscle activation proceeds sequentially in a rostro-caudal direction, while the offset (or deactivation) is nearly simultaneous at all sites, so that EMG burst duration decreases towards the tail. Muscle duty cycle at each location remains a constant proportion of the tailbeat period (T), independent of swimming speed, and peak force is registered in the tail tendons just as all ipsilateral muscle deactivates. Mean duty cycles in skipjack are longer than those in yellowfin. In yellowfin red muscle, there is complete segregation of contralateral activity, while in skipjack there is slight overlap. In both species, all internal red muscle on one side is active simultaneously for part of each cycle, lasting 0.18T in yellowfin and 0.11T in skipjack. (Across the distance encompassing the majority of the red muscle mass, 0.35-0.65L, where L is fork length, the duration is 0.25T in both species.) When red muscle activation patterns were compared across a variety of fish species, it became apparent that the EMG patterns grade in a progression that parallels the kinematic spectrum of swimming modes from anguilliform to thunniform. The tuna EMG pattern, underlying the thunniform swimming mode, culminates this progression, exhibiting an activation pattern at the extreme opposite end of the spectrum from the anguilliform mode.

The musculature of yellowfin tuna (Thunnus albacares) exhibits distinct functional specialization, with slow-twitch oxidative red muscle and fast-twitch glycolytic white muscle demonstrating marked disparities in energy metabolic characteristics. To elucidate the molecular mechanisms underlying these functional divergences, this study implemented an integrated approach incorporating ultrastructural analysis via transmission electron microscopy (TEM), transcriptomic profiling, and enzymatic activity assays of key metabolic regulators. TEM imaging revealed that red muscle fibers contain larger mitochondria and prominent lipid droplets compared to white muscle fibers. Our transcriptome analysis identified 3,162 genes with significant expression differences-1,515 were up-regulated, and 1,647 were down-regulated. Functional enrichment analysis demonstrated significant association of red muscle DEGs with oxidative phosphorylation, tricarboxylic acid cycle, and fatty acid β-oxidation, while white muscle preferentially enriched glycolysis/gluconeogenesis pathways. Enzymatic validation revealed red muscle exhibited higher citrate synthase activity (2.3-fold) and elevated β-hydroxyacyl-CoA dehydrogenase levels (1.8-fold), whereas white muscle showed greater hexokinase activity (4.7-fold) and increased lactate dehydrogenase activity (3.2-fold). These findings provide novel insights into the physiological adaptations underlying the distinctive swimming strategies of scombroid fishes, revealing evolutionary optimization of muscle metabolic pathways corresponding to their sustained cruising capacity and burst swimming performance.

Rice (Oryza sativa L.) cultivars show impairment of growth in response to environmental stresses such as cold at the early seedling stage. Locally adapted weedy rice is able to survive under adverse environmental conditions, and can emerge in fields from greater soil depth. Cold-tolerant weedy rice can be a good genetic source for developing cold-tolerant, weed-competitive rice cultivars. An in-depth analysis is presented here of diverse indica and japonica rice genotypes, mostly weedy rice, for cold stress response to provide an understanding of different stress adaptive mechanisms towards improvement of the rice crop performance in the field. We have tested a collection of weedy rice genotypes to: 1) classify the subspecies (ssp.) grouping (japonica or indica) of 21 accessions; 2) evaluate their sensitivity to cold stress; and 3) analyze the expression of stress-responsive genes under cold stress and a combination of cold and depth stress. Seeds were germinated at 25°C at 1.5- and 10-cm sowing depth for 10d. Seedlings were then exposed to cold stress at 10°C for 6, 24 and 96h, and the expression of cold-, anoxia-, and submergence-inducible genes was analyzed. Control plants were seeded at 1.5cm depth and kept at 25°C. The analysis revealed that cold stress signaling in indica genotypes is more complex than that of japonica as it operates via both the CBF-dependent and CBF-independent pathways, implicated through induction of transcription factors including OsNAC2, OsMYB46 and OsF-BOX28. When plants were exposed to cold + sowing depth stress, a complex signaling network was induced that involved cross talk between stresses mediated by CBF-dependent and CBF-independent pathways to circumvent the detrimental effects of stresses. The experiments revealed the importance of the CBF regulon for tolerance to both stresses in japonica and indica ssp. The mechanisms for cold tolerance differed among weedy indica genotypes and also between weedy indica and cultivated japonica ssp. as indicated by the up/downregulation of various stress-responsive pathways identified from gene expression analysis. The cold-stress response is described in relation to the stress signaling pathways, showing complex adaptive mechanisms in different genotypes.

Cold and drought stresses severely limit crop production and can occur simultaneously. Although some transcription factors and hormones have been characterized in plants subjected each stress, the role of metabolites, especially volatiles, in response to cold and drought stress exposure is rarely studied due to lack of suitable models. Here, we established a model for studying the role of volatiles in tea (Camellia sinensis) plants experiencing cold and drought stresses simultaneously. Using this model, we showed that volatiles induced by cold stress promote drought tolerance in tea plants by mediating reactive oxygen species and stomatal conductance. Needle trap microextraction combined with GC-MS identified the volatiles involved in the crosstalk and showed that cold-induced (Z)-3-hexenol improved the drought tolerance of tea plants. In addition, silencing C. sinensis alcohol dehydrogenase 2 (CsADH2) led to reduced (Z)-3-hexenol production and significantly reduced drought tolerance in response to simultaneous cold and drought stress. Transcriptome and metabolite analyses, together with plant hormone comparison and abscisic acid (ABA) biosynthesis pathway inhibition experiments, further confirmed the roles of ABA in (Z)-3-hexenol–induced drought tolerance of tea plants. (Z)-3-Hexenol application and gene silencing results supported the hypothesis that (Z)-3-hexenol plays a role in the integration of cold and drought tolerance by stimulating the dual-function glucosyltransferase UGT85A53, thereby altering ABA homeostasis in tea plants. Overall, we present a model for studying the roles of metabolites in plants under multiple stresses and reveal the roles of volatiles in integrating cold and drought stresses in plants.

Proximate composition was determined in different body parts (skin, white muscle, red muscle, head muscle and belly flap of five species of tuna; Katsuvonus pelamis (skipjack, balaya), Thunnus Albacares (yellow fin tuna, kellawalla), Auxis rochei (Bullet tuna, ragoduwa), Auxis thazard (frigate tuna, alagoduwa) and Euthvnnus affinis (kawakawa, attawalla) obtained from the Negambo fish landing site. Fatty acid profiles were also analyzed in the akin, red and white muscle of the five species. No significant differences between the tuna species were observed with respect to protein, total fat. and moisture contents. The ash content in Frigate tuna and Kawakawa were significantly higher than the other species. The muscle tissue in all the species was rich in protein (20-25%) and low in fat (

Physiological responses to cold and starvation stresses in the liver of yellow drum (Nibea albiflora) revealed by LC-MS metabolomics.

Overwinter mortality in yellow drum (Nibea albiflora): Insights from growth and immune responses to cold and starvation stress

Plants enhance their cold stress tolerance by cold acclimation, a process which results in vast reprogramming of transcriptome, proteome and metabolome. Evidence suggests nitric oxide (NO) production during cold stress which regulates genes (especially the C-repeat binding factor (CBF) cold stress signalling pathway), diverse proteins including transcription factors (TFs) and phosphosphingolipids. About 59% (redox), 50% (defence/stress) and 30% (signalling) cold responsive proteins are modulated by NO-based post translational modifications (PTMs) namely S-nitrosylation, tyrosine nitration and S-glutathionylation, suggesting a cross-talk between NO and cold. Analysis of cold stress responsive deep proteome in apoplast, mitochondria, chloroplast and nucleus suggested continuation of this cross-talk in sub-cellular systems. Modulation of cold responsive proteins by these PTMs right from cytoskeletal elements in plasma membrane to TFs in nucleus suggests a novel regulation of cold stress signalling. NO-mediated altered protein transport in nucleus seems an important stress regulatory mechanism. This review addresses the NO and cold stress signalling cross-talk to present the overview of this novel regulatory mechanism.

Low-temperature stress is an increasing problem for the cultivation of tea (Camellia sinensis), with adverse effects on plant growth and development and subsequent negative impacts on the tea industry. Methyl jasmonate (MeJA), as a plant inducer, can improve the cold-stress tolerance in tea plants. R2R3-MYB transcription factors (TFs) are considered potentially important regulators in the resistance to cold stress in plants. However, the molecular mechanisms, by which MYB TFs via the jasmonic acid pathway respond to cold stress in the tea plant, remain unknown. In this study, physiological and biochemical assays showed that exogenous MeJA application could effectively promote ROS scavenging in the tea plant under cold stress, maintaining the stability of the cell membrane. Sixteen R2R3-MYB TFs genes were identified from the tea plant genome database. Quantitative RT-PCR analysis showed that three CsMYB genes were strongly induced under a combination of MeJA and cold-stress treatment. Subcellular localization assays suggest CsMYB45, CsMYB46, and CsMYB105 localized in the nucleus. Exogenous MeJA treatment enhanced the overexpression of CsMYB45, CsMYB46, and CsMYB105 in E. coli and improved the growth and survival rates of recombinant cells compared to an empty vector under cold stress. Yeast two-hybrid and bimolecular fluorescence complementation experiments confirmed that CsMYB46 and CsMYB105 interacted with CsJAZ3, CsJAZ10, and CsJAZ11 in the nucleus. Taken together, these results highlight that CsMYB45, CsMYB46, and CsMYB105 are not only key components in the cold-stress signal response pathway but also may serve as points of confluence for cold stress and JA signaling pathways. Furthermore, our findings provide new insight into how MYB TFs influence cold tolerance via the jasmonic acid pathway in tea and provide candidate genes for future functional studies and breeding.