Expression and functional characterization of glutamine synthetase from giant freshwater prawn (Macrobrachium rosenbergii) under osmotic stress

The freshwater prawn, Macrobrachium rosenbergii naturally lives in the freshwater, though it migrates to the brackish water environment during spawning that claimed to be resistant on a broad range of saline fluxes. However, little is known about the osmoregulatory patterns and the effect of an enzyme glutamine synthetase (GS) in M. rosenbergii under stress. Here, we described the identification and functional characterization of GS from M. rosenbergii (Mr‐GS) at molecular and protein levels. The identified Mr‐GS was comprised of 361 amino acids that phylogenetically shared the highest identity with other crustaceans and predicted to contain Gln‐synt_C and Gln‐synt_N domains at the respective terminal regions. Tissue distribution analysis in M. rosenbergii revealed that the Mr‐GS was highly expressed in muscle, and commonly existed in other examined tissues in the following order gills > heart > stomach > brain > haemolymph. Whereas, the mRNA of Mr‐GS was significantly up‐regulated in the muscle and gill tissues following challenges with either hyper (0 → 13‰), or hypo (13 → 0‰) osmotic stress at 3, 6 and 12 hr. Furthermore, the level of Glutamine concentration was positively correlated with the GS mRNA and protein expression patterns in hyper‐osmotic stress, whereas in hypo‐osmotic stress a slight decrease in the gills and maintained a level in the muscle tissues at 3, 6 and 12 hr post‐treatments. Our findings suggest that Mr‐GS potentially exhibited the osmoregulation responses in the gill and muscle tissues of M. rosenbergii throughout the time of osmotic stress, which will benefit for future study on osmoregulation.     

Rapid salinity changes affect the survival and physiology of a penaeid prawn: Implications of flood events on recruitment to the fishery

Estuaries act as nurseries for many penaeid prawns, but these habitats are highly susceptible to salinity decline through flooding. The rate of salinity decline and duration of exposure to non‐optimal salinity may affect survival and subsequent recruitment of prawns to the fishery. This study aimed to determine the effect of salinity fluctuations observed in local estuaries during flood events using a novel dilution approach. Mortality of juvenile Melicertus plebejus (Hess) was assessed after 24 hr exposure to 24 rates of salinity decline ranging from 0.01% to 20% per hr. After the salinity decline, prawns were held at the final salinities for five days before again assessing mortality as well as aerobic metabolic rate and prawn water content. Salinity decline from 36 to ~0.8 led to 50% mortality, but continued exposure to low salinity for five days increased mortality at this salinity to 99% and shifted the 50% mortality point to salinity ~5. Aerobic metabolic rate and water content data suggested the cause of mortality due to exposure to salinities < 5 was osmoregulatory failure. Rapid salinity declines over 24 hr and sustained low salinity due to flooding could compromise the survival of juvenile prawns, potentially reducing recruitment to the fishery.     

Effects of salinity on growth characteristics and osmoregulation of juvenile cobia,Rachycentron canadum (Linnaeus 1766), reared in potassium‐amended inland saline groundwater

The suitability of inland saline groundwater as a medium to culture juvenile cobia, Rachycentron canadum, was assessed. In the first experiment, juvenile cobia stocked in raw (unamended) saline groundwater at salinities of 5, 10, and 15 g/L exhibited complete mortality after 108, 176, and 195 hr, respectively. The second experiment evaluated the rearing of juvenile cobia (mean weight ~9.23 ± 0.12 g) in potassium (K⁺)‐amended saline groundwater (100% K⁺ fortified) and reconstituted seawater at salinities of 5, 10, and 15 g/L to assess growth and osmoregulation in distinct culture media. Following 60 days of culture, all fish survived the experimental period. Final mean bodyweight of cobia reared in K⁺‐amended saline groundwater (103.2–115.8 g) and seawater (111.2–113.8 g) of different salinities did not vary significantly (p > .05). No differences (p > .05) were observed in specific growth rate, weight gain (%), and feed conversion ratio between treatment groups. Serum osmolality increased with salinity and was significantly higher (p < .05) for fish in K⁺‐amended saline groundwater (353–361 mOsmol/Kg) than in reconstituted seawater (319–332 mOsmol/Kg), although differences were not observed between salinities by water type. Cobia stocked in saline groundwater of different salinities were osmoregulating normally, and the higher values observed may be because of variations in ionic composition and other interfering ions in saline groundwater. Trial results suggest that juvenile cobia can achieve optimal growth in K⁺‐amended saline groundwater of low and intermediate salinities.     

盐度对珍珠龙胆石斑鱼幼鱼渗透调节与耗氧率的影响

为研究盐度对珍珠龙胆石斑鱼（）渗透调节与耗氧率的影响，设计实验一，将（162.5±12.1）g珍珠龙胆石斑鱼置于不同盐度（6、12、18、24、30）下养殖10 d，测定血清渗透压及Na⁺、Cl^-、K⁺离子浓度。实验结果表明，随盐度的升高血清渗透压及Na⁺、Cl^-、K⁺离子浓度也随着升高，各组[Na⁺]：[Cl^-]比值无显著差异（>0.05）；经回归分析得到血清等渗点渗透压为365.95 mOsm/kg，所对应盐度为12.75。实验二，将（26.4±2.7）g幼鱼置于不同盐度（6、12、18、24、30）下养殖30 d，测定在开始暴露后0 h、3 h、24 h、72 h鳃Na⁺/K⁺-ATPase活性及表达和第30天耗氧率，结果表明鳃Na⁺/K⁺-ATPase活性随盐度的增大呈“U”形变化；鳃Na⁺/K⁺-ATPase α1基因表达量波动较大，在72 h后随盐度增大先降低后增加，变化趋势与酶活性一致；第30天耗氧率随盐度的增加先降低后增加又降低。综上所述，珍珠龙胆石斑鱼幼鱼10 d内能够完全适应6~30盐度急性变化，耗氧率除了受离子渗透调节的影响，还可能与其生活史阶段有关。     

花鲈cox6a基因与相关miRNA的鉴定及其在盐度适应中的表达调控分析

细胞色素c氧化酶（COX）是线粒体电子传递链的末端酶，COX6A是细胞色素c氧化酶的核编码亚基之一。为丰富花鲈（基因及相关miRNAs在花鲈渗透调节机制中的潜在作用，本研究开展了花鲈cox6a基因的miRNAs，并对其与）。系统进化树分析进一步确认了两个花鲈在垂体、脑、肝脏和肾脏中的表达量较高，基因的miRNA：miR-30e-5p、miR-223、miR-200a-3p和miR-155-5p，但没有发现可能靶定基因以及miR-30e-5p、miR-223、miR-200a-3p都呈现显著差异表达，而miR-155-5p仅在适应高盐环境时出现了显著的差异表达。这表明，基因和4个miRNAs在花鲈的渗透调节过程中发挥潜在作用。其中miR-30e-5p、miR-223和miR-200a-3p的表达趋势与cox6a2基因的表达调控。本研究为探讨花鲈适应不同盐度环境的渗透调节机制提供了基础数据。     

Fluctuating salinity effect on Sphoeroides annulatus (Jenyns 1842) physiological responses

Euryhaline fish, such as the Bullseye puffer Sphoeroides annulatus (Jenyns 1842), experience sudden salinity changes in their natural environment, which is more common than the exception, so they must adapt to survive and cope with extreme salt conditions. Therefore, Bullseye puffer juveniles were exposed to short‐term stress (39 hr) by fluctuating salinity conditions (41, 35, 29, 23, 17, 11, 5, 11, 17, 23, 29, 35, 41 psu) with a 3‐hr interval between each point at 26 ± 1ºC in a respirometer chamber and acclimation reservoirs. Responses to oxygen consumption rate (OCR: 23–35 mg O₂ h^–1 kg^–1), ammonium excretion rate (AER: 1–1.85 mg NH4⁺ h^?1 kg^?1), oxygen‐nitrogen atomic ratio (O:N 17–30), osmoregulatory pattern (blood osmotic pressure from 342.4 to 332.8 mmol/kg) and changes in expression levels of Na⁺/K⁺‐ATPase in the gills (higher values at higher salinities) were measured. Although some signs of stress were detected below the iso‐osmotic point (11.4 psu), the puffer fish is a strong euryhaline fish that survives under these conditions. Nonetheless, it could recover when salinity returned to the initial acclimation point because Sphoeroides annulatus is able to live in a wide range of environments with wide natural salinity fluctuations; thus, a common practice in aquaculture has been to expose fish to low salinity for several reasons discussed in this study. This capacity reveals its high plasticity to saline adaptation from 41 to 5 psu an up from 5 to 41 psu, all in less than 2 days.     

Foliage‐applied sodium nitroprusside and hydrogen peroxide improves resistance against terminal drought in bread wheat

Terminal drought is threatening the wheat productivity worldwide, which is consumed as a staple food by millions across the globe. This study was conducted to examine the influence of foliage‐applied stress signalling molecules hydrogen peroxide (H₂O₂; 50, 100, 150 μm ) and nitric oxide donor sodium nitroprusside (SNP; 50, 100, 150 μm ) on resistance against terminal drought in two bread wheat cultivars Mairaj‐2008 and BARS‐2009. These stress signalling molecules were applied at anthesis stage (BBCH 61); drought was then imposed by maintaining pots at 35% water holding capacity. Terminal drought caused significant reduction in grain yield of both tested bread wheat cultivars; however, foliage application of both stress signalling molecules at either concentration improved the performance of both bread wheat cultivars. Maximum improvement in 100‐grain weight (12.2%), grains per spike (19.7%), water‐use efficiency (WUE; 19.8%), chlorophyll content index (10.7%), total soluble phenolics (21.6%) and free leaf proline (34.3%), and highest reduction in leaf malondialdehyde contents (20.4%) was recorded when H₂O₂ was foliage‐applied at 100 μm . Foliage application of SNP enhanced the grains per spike, 100‐grain weight and grain yield by 14.9%, 11.3% and 20.1%, respectively, than control. The foliage‐applied stress signalling molecules improved the accumulation of soluble phenolics, proline and glycine betaine with simultaneous reduction in malondialdehyde contents, which enabled wheat plants to sustain the biological membranes under stress resulting in better stay green (high chlorophyll contents) under drought. This helped improving the grain number, grain weight, grain yield, WUE and transpiration efficiency. In crux, foliage‐applied H₂O₂ and SNP, at pre‐optimized rate, may be opted to lessen the drought‐induced yield losses in bread wheat in climate change conditions.     

Physiological responses of pink abalone Haliotis corrugata (Gray, 1828) exposed to different combinations of temperature and salinity

Physiological responses of pink abalone Haliotis corrugata were determined under different temperature and salinity conditions. Oxygen consumption rate was not affected by temperature and salinity. Ammonium excretion of pink abalone was inversely related to salinity. The O:N ratio indicated that abalone maintained in lower salinities had an interval of 4.9–7.7, which is indicative of a protein‐dominated metabolism, whereas the O:N in 35‰ was 28.8–35.5 for both temperatures, suggesting that carbohydrates were used as energy substrate. Haemolymph osmolality of abalone exposed to 20 and 24 °C was slightly hyperiso‐osmoconformic in salinity ranges of 20–35‰. The results of this study suggested that for optimized culture, pink abalone should be cultivated at 24 °C at a salinity of 35‰.     

硝普钠(SNP)浸种对渗透胁迫下不同紫花苜蓿种子萌发生理特性的影响

以紫花苜蓿阿尔岗金(Medicago sativa‘Algonquin’)、金皇后(M. sativa‘Golden empress’)和三得利(M.sativa‘Sanditi’)为材料,探讨外源一氧化氮(NO)对PEG胁迫下紫花苜蓿种子萌发期丙二醛和渗透调节物质含量及抗氧化酶活性的影响。结果表明:(1) SNP浸种能有效缓解PEG胁迫下紫花苜蓿萌发期MDA含量的积累,在第6 d SNP+PEG与PEG处理相比,阿尔岗金、金皇后和三得利种子MDA含量分别降低了23. 11%,21. 81%和30. 38%。(2)随PEG胁迫时间延长,3个苜蓿品种渗透调节物质含量呈先增后降趋势。SNP+PEG与PEG处理相比,阿尔岗金中脯氨酸、可溶性糖和可溶性蛋白含量分别在处理第4 d,第2 d和第4 d达到最大值,分别提高了13.71%,18.57%和9.23%;金皇后各含量均在第2 d有最大值,分别提高了20.96%,16.84%和5.48%;三得利各含量分别在第4 d,第2 d和第2 d达到最大值,分别提高了17.87%,17.52%和13.40%。(3) 3个品种紫花苜蓿SOD、POD、CAT、APX活性表现为先升后降趋势,且均在第4 d活性最大。与PEG处理相比,SNP+PEG处理下三得利SOD、POD活性增加率是阿尔岗金的2.76倍和1.21倍,是金皇后的4.10倍和1.21倍。渗透调节物质含量增加和抗氧化酶活性增强是紫花苜蓿适应干旱的生理基础,外源NO可提高紫花苜蓿对干旱的防御,3个品种紫花苜蓿受外源NO调节强弱顺序为:三得利金皇后阿尔岗金。     

油菜素内酯对干旱胁迫下羊草渗透调节及抗氧化酶的影响研究

受全球气候变化影响,我国牧区干旱现象日益严重,极大限制了草原生产力的提高。本文采用盆栽人工控水的方法,研究不同浓度油菜素内酯(BR)处理对羊草生长和抗旱特性的影响。结果表明干旱显著抑制植株的生长,而BR能有效减缓干旱胁迫对羊草造成的伤害。经过不同浓度的BR处理后,与干旱胁迫相比,株高、叶面积、干重、含水量、叶绿素a、叶绿素b、类胡萝卜素、根系活力、脯氨酸、可溶性蛋白、可溶性糖和5种抗氧化酶活性均随浓度增加而升高,但高浓度下又有降低的趋势,而丙二醛(MDA)和叶片电导率则相反,其中以BR浓度为0.1 mg/L时的抗旱效果最好。研究发现适当浓度的BR提高羊草株高、叶面积和光合色素含量,促进干物质积累和根系活力,与其降低膜脂过氧化产物MDA与质膜透性,提高脯氨酸、可溶性蛋白、可溶性糖等渗透调节物质含量,增强超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)等抗氧化酶活性密切相关。