坛紫菜自由丝状体响应琼胶寡糖的激发

为了研究琼胶寡糖对坛紫菜自由丝状体藻丝生长和壳孢子囊枝形成的激发的影响,实验采用琼胶寡糖激发坛紫菜自由丝状体,以液相氧电极检测坛紫菜丝状体净光合放氧速率的变化;以对羟基苯乙酸(POHPAA)化学发光法检测坛紫菜丝状体的H_2O_2释放量;利用LC-MS检测红藻糖苷含量变化;利用实时定量PCR技术检测坛紫菜丝状体H_2O_2产生相关基因(Phrboh、Ph SOD)和红藻糖苷合成相关基因(Phnho1、Phgpdh、Phtps)的表达情况;并利用显微镜观察法检测壳孢子囊枝数量的变化。结果发现,琼胶寡糖能够激发坛紫菜自由丝状体的系列响应,表现在净光合放氧速率以及光合同化产物红藻糖苷的含量和生物合成出现增加;H_2O_2释放量持续增加,与产生H_2O_2相关的2个酶基因Phrboh和Ph SOD的表达增强。此外,琼胶寡糖也能够促进在坛紫菜自由丝状体的发育,在培养第30天时,琼胶寡糖处理组的壳孢子囊枝形成率达到59%,显著高于对照组46%。综上所述,琼胶寡糖能够增加坛紫菜自由丝状体的光合速率和光合同化产物,并通过形成H_2O_2的酶的表达来诱导活性氧的释放;琼胶寡糖还能促进坛紫菜自由丝状体的繁殖发育。     

琼胶寡糖诱导坛紫菜活性氧爆发

研究了琼胶寡糖诱导坛紫菜氧爆发的响应及其活性氧产生的位点。用琼胶寡糖诱导坛紫菜,以微呼吸测量系统检测坛紫菜氧消耗的变化;以对羟基苯乙酸(POHPAA)化学发光法检测坛紫菜的H2O2释放量;利用DCFH-DA染色观察活性氧在细胞上的定位;并利用实时定量PCR检测坛紫菜NADPH氧化酶基因Phrboh的差异表达;通过2-AMAC标记琼胶寡糖,观察其在坛紫菜的结合部位。结果显示,100μg/mL琼胶寡糖处理坛紫菜,出现两次呼吸爆发,分别在4min和10min左右,强度分别是基础呼吸的4倍和14.1倍;5min内出现H2O2的积累,15min达到峰值,比对照组高出近10倍。10μmol/LDPI可部分抑制H2O2的产生。3min内Phrboh的表达已上调,并持续近10min。DCFH-DA染色显示,活性氧主要产生在膜系统上。荧光标记琼胶寡糖的定位发现,坛紫菜细胞膜上存在琼胶寡糖的结合位点。综上所述,琼胶寡糖能识别细胞膜上的结合位点,并诱导坛紫菜膜系统上的H2O2爆发,H2O2的产生与NADPH氧化酶相关。     

坛紫菜养殖技术之二坛紫菜优质纯系苗种培育技术研究

坛紫菜在自然分类系统中属于红藻门、原红藻纲、红毛菜目、红毛菜科、紫菜属,在其生活史中具有2个明显的发育阶段,即叶状体阶段和丝状体阶段.叶状体是由丝状体放散出来的壳孢子萌发形成的,即日常食用的紫菜;丝状体是由叶状体放散出来的果孢子萌发形成的,多数在贝壳中生长.坛紫菜的育苗就是指叶状体成熟后形成的果孢子,通过采果孢子培育贝壳丝状体,丝状体在秋季成熟后放散壳孢子的过程.南日木平养殖场自2003年1月开始承担"坛紫菜优质纯系苗种培育技术研究"项目,现将技术总结介绍如下:     

坛紫菜优质纯系苗种培育技术研究

坛紫菜在自然分类系统中属于红藻门、原红藻纲、红毛菜目、红毛菜科、紫菜属,在其生活史中具有2个明显的发育阶段,即叶状体阶段和丝状体阶段。叶状体是由丝状体放散出来的壳孢子萌发形成的,即日常食用的紫菜;丝状体是由叶状体放散出来的果孢子萌发形成的,     

浅谈坛紫菜丝状体培育过程

<正>坛紫菜是我国特有的暖温带种类,也是我国南方养殖的主要经济红藻。坛紫菜的叶状体和丝状体是其生活史中的两个重要阶段。紫菜栽培生产中,苗种供应是生产的关键环节,而获得健康优良的丝状体则尤为重要。坛紫菜丝状体有两种培养方式,即自由丝状体和贝壳丝     

紫菜人工栽培技术

1人工栽培紫菜的生长过程紫菜的人工栽培分丝状体培育和叶状体养成2个阶段。丝状体培育阶段：春末取成熟的叶状体经阴干刺激后,放入盛有海水的容器内放散果孢子,当放散量达到要求时,捞出叶状体。把孢子水按一定比例喷洒到已放有贝壳的培育池内，果孢子即附着到贝壳上，萌发钻人贝壳内生长成丝状体。秋季水温下降时，贝壳内丝状体成熟并放散壳孢子，将维尼仑网帘放入培养池内采苗，     

二氧化碳和阳光紫外辐射对坛紫菜丝状体光合生理特性的影响

为了探索大型海藻生活史丝状体阶段对于海洋酸化与紫外辐射的响应,实验选取经济海藻坛紫菜的自由丝状体作为实验材料。实验设置两个CO2浓度,正常CO2浓度(390 ppmv)和高CO2浓度(1 000 ppmv);3种辐射处理,PAR处理(仅接受可见光)、PA处理(滤除UV-B)、PAB处理(全波长辐射)。研究结果表明,高CO2显著提高了坛紫菜自由丝状体的生长速率,但高CO2处理下坛紫菜自由丝状体中藻红蛋白、藻蓝蛋白、叶绿素a、类胡萝卜素及紫外吸收物质UVACs分别降低了7.3%、9.3%、19.8%、16.5%和18.7%。高CO2处理的坛紫菜自由丝状体最大光化学效率Fv/Fm,光能利用效率(α)及最大相对电子传递速率(rETRmax)都显著高于正常CO2处理。太阳模拟器下处理的坛紫菜自由丝状体,PAR与PA处理下的抑制率,正常CO2与高CO2处理间无显著差异,然而在PAB处理下,高CO2处理的抑制率要高于正常CO2处理,这可能与其体内紫外吸收物质含量下降有关。PAR处理下的抑制率,无论是在正常CO2还是在高CO2处理下,都显著低于PA及PAB处理,而PA与PAB之间无显著差异。在未来海洋持续酸化的背景下,UV辐射的增加将会影响到坛紫菜自由丝状体的光合生理及生长。     

紫菜属海藻色素突变的研究

使用化学诱变剂MNNG(N-甲基-N’-硝基-N-亚硝基胍)对条斑紫菜、坛紫菜、少精紫菜和华北半叶紫菜的叶状体、丝状体和壳孢子进行诱变,研究与分析了紫菜属海藻色素突变的发生与表达。结果显示,在合适的诱变剂量范围内,紫菜各生长阶段藻体均发生较高的突变率,但突变的表达特征不同。叶状体以细胞为单位发生点状突变,突变细胞的生长形成了各种突变斑块的无序镶嵌,显示单倍体细胞的致变结果。经诱变的丝状体虽表现较低的突变率,然而诱变引发的突变却主要在后代叶状体中表达,显示为二倍体细胞的突变表达规律。壳孢子突变随萌发个体的细胞分裂被立即表达,显示细胞倍性发生变化的遗传特征。本文还讨论了色素突变表达规律显示的减数分裂过程及其对藻体形态建成的影响。     

条斑紫菜南移福建养殖的研究

条斑紫菜系我国长江以北的养殖品种,1986年和1987年南移福建连江进行了丝状体育苗和叶状体养殖,均获成功。叶状体产量折合亩产干品67.9公斤。条斑紫菜南移存在的主要问题是:丝状体育苗时间长,温度高;叶状体则是生长期短,产量低。实行坛紫菜和条斑紫菜轮养,可提高紫菜的产量和经济效益。     

条斑紫菜自由丝状体扩增的意义及影响因子

紫菜的生活史由两个形态截然不同的阶段组成,即宏观的叶状体阶段和微观的丝状体阶段.条斑紫菜(Porphyra yezoensis Ueda)的丝状体是其生活史中的一个重要阶段.通常果孢子是粘附在含钙的贝壳等基质上萌发,并钻入壳内生长、发育成贝壳丝状体(conchocelis),如果不钻壳,在培养液中悬浮生长,这种丝状体称为自由丝状体(free-living conchocelis)[1,2].     

饲料中添加不同诱食剂对南美白对虾生长和饲料消化的影响

两种等氮但不同鱼粉水平 (4 0 %和 35 % )的基础饲料中分别添加 0 2 %、 0 5 %甜菜碱、 0 2 %甜菜碱 +0 1%牛磺酸或一种市售添加剂配置成 8种实验饲料。饲料试验在循环系统的玻璃钢桶 (2 6 0L)中进行。结果显示饲料中添加不同诱食剂 ,有利于对虾的摄食和生长 ,甜菜碱诱食效果最佳 ,牛磺酸和甜菜碱的协同诱食效果并不明显。南美白对虾对无机盐、粗蛋白质有较高的表观消化率 ,但是粗脂肪的表观消化率较低只有 6 0 %左右。本研究表明甜菜碱是一种较好的诱食剂 ,其适宜添加量为 0 2 %。诱食剂在两种等氮等能但不同鱼粉水平 (4 0 %和 35 % )的饲料中作用并不明显 ,进一步研究诱食剂在鱼粉含量低、低蛋白饲料中的作用 ,研究南美白对虾脂类营养及调控机理十分必要     

团头鲂性腺发育不同时期组织学观察及Kiss2／Kiss2r基因表达分析

Kiss基因（Kisspeptin）及其受体Kissr基因（Kiss receptor）组成的系统参与下丘脑-垂体-性腺轴功能的调节,在脊椎动物青春期启动的过程中发挥重要作用。该研究通过组织切片技术观察了团头鲂（Megalobrama amblycephala）卵巢和精巢的不同分期,进一步采用荧光定量PCR技术分析比较了团头鲂Kiss2和Kiss2r基因在雌、雄性腺不同时期对应的脑、垂体、性腺组织中的表达量。组织切片观察明确鉴别出团头鲂卵巢和精巢发育Ⅰ期至Ⅵ期的性腺组织结构,基因荧光定量PCR分析结果表明Kiss2/Kiss2r系统主要是通过增加团头鲂卵巢发育Ⅱ期时脑和垂体的表达量以及Ⅲ期精巢的表达量来发挥生殖启动作用。     

三角帆蚌不同蚌龄外套膜细胞的增殖能力及其矿化功能

研究了淡水珍珠贝——三角帆蚌(Hyriopsis cumingii)在不同蚌龄下外套膜组织的细胞增殖及生物矿化相关因子活性。选择5组不同蚌龄三角帆蚌(0.5龄、1龄、2龄、3龄、4龄),通过流式细胞技术分析了外套膜细胞的增殖指数(proliferation index,PrI)和胞内Ca~(2+)浓度,并应用实时荧光定量PCR检测了与生物矿化相关的碳酸酐酶(carbonic anhydrase, CA)、碱性磷酸酶(alkaline phosphatase, ALP)基因的表达并测定其酶活性。结果表明, 2龄蚌的外套膜细胞PrI及胞内Ca~(2+)浓度显著高于其他蚌龄(P0.05);生物矿化相关基因在不同蚌龄的外套膜组织中表达量不同, 1龄三角帆蚌CA基因表达量和CA酶活性最高(P0.05), ALP基因表达量和ALP酶活在0.5龄三角帆蚌中显著高于其他蚌龄(P0.05)。本研究旨为深入探讨三角帆蚌外套膜细胞增殖能力及人工育珠过程中供体蚌的选择奠定基础。     

异源铜盐对仿刺参幼参急性毒性及组织形态学影响

采用急性毒性试验方法研究异源铜盐对仿刺参（Apostichopus japonicus）幼参的毒性影响,通过24h、48h、72h和96h的致死数量统计分析硫酸铜（CuSO4）/氯化铜（CuCl2）对幼参的半致死质量浓度（LC50）和安全质量浓度（SC）;利用石蜡切片显微定性观察不同铜盐对刺参幼参后肠的毒性情况。结果表明,CuSO4来源的Cu^2＋对幼参24h、48h和72h的LC50均大于CuCl2来源的Cu^2＋对幼参24h、48h和72h的LC50,而CuCl2对幼参96h LC50大于CuSO4对幼参96hLC50;异源铜盐在暴露前48h内对刺参的毒性影响极其显著,随着暴露时间的延长其对刺参的毒性影响不显著;2种铜盐（以铜离子质量浓度为0.06 mg·L^-1计）对幼参的消化道均有不同程度的腐蚀作用,导致结缔层与上皮层分离,CuCl2对刺参肠道的毒性（类似腐蚀作用）大于CuSO4的毒性;不同的阴离子对Cu^2＋的毒性有拮抗作用。     

汞暴露对草鱼器官组织中碱性磷酸酶活性的影响

测定暴露于不同汞离子质量浓度(0 mg/L、0.05 mg/L、0.10 mg/L、0.15 mg/L、0.20 mg/L、0.25 mg/L)下草鱼(Ctenopharyngodon idella)血清、鳃、肝胰脏、脾、肾脏和肌肉等组织器官中碱性磷酸酶(alkaline phosphatase,AKP)活性。研究目的在于评价汞离子对草鱼生理代谢的影响。暴露周期为21 d。结果显示,与对照组相比,血清AKP活性在0.05mg/L、0.10 mg/L和0.15 mg/L汞暴露组无显著性变化(P>0.05),而在0.20 mg/L、0.25 mg/L组显著下降(P<0.01);鳃AKP活性在0.10 mg/L和0.15 mg/L汞暴露组显著上升(P<0.05或P<0.01),而0.05 mg/L、0.20 mg/L和0.25mg/L组无显著变化(P>0.05);肝胰脏和脾脏AKP活性在所有实验组均显著下降(P<0.01);肾脏AKP活性在0.05mg/L组无显著变化,其他实验组均显著升高(P<0.01);肌肉AKP活性在所有实验组均无显著性变化(P>0.05)。本研究说明草鱼暴露于不同汞离子浓度下,具有不同生理功能的器官组织的AKP活性变化存在较大差异     

The pigmenting effect of different carotenoids on fancy carp (Cyprinus carpio)

The optimal concentration of a panel of individual and combined carotenoid sources on skin pigmentation in fancy carp was investigated by nine experimental diets that were formulated and supplemented with astaxanthin at 25 mg kg^?1, lutein at 25 and 50 mg kg^?1, β‐carotene at 25, 50 and 75 mg kg^?1, and lutein combined with β‐carotene at 25 : 25 and 50 : 50 mg kg^?1, while a diet without supplemented carotenoid served as a control. The results showed that serum TC of fish fed diets containing supplemented with lutein plus β‐carotene at 25 : 25; 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet were higher than the other treatments (P ≤ 0.05). Serum TC of the respective treatments was 6.2 ± 2.0, 7.8 ± 3.3 and 7.3 ± 1.9 μg mL^?1 serum, respectively. Fish fed diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet had serum astaxanthin concentrations similar to fish fed the diet with astaxanthin alone at 25 mg kg^?1. Serum astaxanthin concentrations was 0.7 ± 0.01, 0.9 ± 0.01, 0.4 ± 0.02 and 1.7 ± 0.18 μg mL^?1 serum, respectively. The chromaticity of fish body skin of red and white position was assessed by colourimetry using the CIE L*a*b (CIELAB) system. Pigmentation response of skin redness of fancy carp fed with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were higher than other treatments (P ≤ 0.05) but they were similar to fish fed with 25 mg kg^?1 astaxanthin diet. The redness (a* values) of fish fed diets with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were 28.3 ± 0.53, 29.9 ± 1.38, 28.8 ± 3.95 and 28.5 ± 2.49, respectively. After 3 weeks of feeding the experimental diets, the fish fed on a diet without carotenoid supplement for one week demonstrated that the same three groups still retained their redness and had an overall tendency to improve skin colouring. Finally, concentrations 50 mg kg^?1 of lutein, or the combination of lutein and β‐carotene at 25 : 25 mg kg^?1 showed the highest efficiency for improving skin pigmentation and redness of skin.     

罗非鱼不同养殖系统对产出效果及异味物质含量的影响

对鱼菜共生系统、底排污系统、底排污+鱼菜共生3种养殖系统的水质变化和罗非鱼产出效果进行了分析,并利用微波蒸馏-吹扫捕集-气相色谱质谱联用技术检测了各系统水体中和罗非鱼肌肉中异味物质的含量变化。经过12周的养殖实验,对终重、生物量、特定生长率、饵料系数、增重率等指标进行测量比较,结果发现,罗非鱼的产出效果从优到差依次为:鱼菜共生组、底排污+鱼菜共生组、底排污组、对照组;底排污+鱼菜共生系统水体中溶解氧在各时期均最高,其亚硝氮、氨氮含量在各时期均最低;底排污或鱼菜共生系统均能有效降低养殖水体中异味物质2-MIB和GSM含量;对照组罗非鱼肌肉中异味物质含量最高,2-MIB和GSM分别达到(0.67±0.02)μg/kg和(0.870±0.018)μg/kg,而底排污+鱼菜共生组罗非鱼肌肉中异味物质含量在各时期最低,2-MIB和GSM分别为(0.31±0.02)μg/kg和(0.53±0.042)μg/kg。结果表明,在罗非鱼精养池塘中,可通过底排污和鱼菜共生结合,降低水体中和罗非鱼体内异味物质的累积,提高其经济价值。     

Sperm motility and fertilization performance of Nodipecten nodosus (L., 1758) exposed at two different cryoprotectants

Cryopreservation is a valuable tool for aquaculture as it provides continuous seed production, regardless of the spawning season of the brood stock. The selection of a suitable cryoprotectant with low toxicity and high water solubility is important to avoid membrane injuries and intracellular ice crystallization. This study has been aimed at the assessment of the toxic effects of two usually applied cryoprotectants, 1‐2 propylene glycol (PG) and methanol (MetOH), on spermatozoa of the of lion‐paw scallop Nodipecten nodosus, by evaluating the sperm motility and the development of D larvae after fertilization procedure. Sperm was exposed at room temperature (22°C) for 10, 20 and 30 min to different concentration ranges of two cryoprotectants. Regarding the sperm motility, PG5%, PG7%, MetOH4% and MetOH6% did not show differences compared to control (semen incubated in seawater) (P < 0.05). The development of D larvae was not affected by the exposition to PG5%, MetOH 4% and MetOH 6%. These results indicate the potential use of both cryoprotectants for cryopreservation procedures.     

Induction of mRNAs in response to acclimation of trout cells to different osmolalities

The direct effect of osmolality on growth and mRNA population were investigated in the rainbow trout cell line (RTG-2). These cells can grow in the media of osmolalities ranging from 200 to 600 mosmol kg-1. With two-dimensional electrophoresis, the in vitro translation of poly(A⁺) RNA isolated from these cells showed osmoresponsive changes in the population of translatable mRNAs. Using differential mRNA display polymerase chain reaction, however, we identified inducible cDNA products in hyper-osmotic and hypo-osmotic media as third component of complement, and as homologues of known genes: an atypical protein kinase regulated by the thyrotropin-dependent mitogenic pathway, nucleolin and CHD3. The remaining cDNAs have no significant homology in GenBank. Northern blots demonstrate that their mRNA levels were induced in hyper-osmotic and hypo-osmotic media, but not by other stresses. The expressed proteins of these mRNAs may be involved directly or indirectly in the adaptation of RTG-2 cells to different osmolalities probably through the osmotic signal transduction and adjustment in cellular metabolism to osmotic stress.     

Effect of dietary calcium on growth and survival of silver catfish fingerlings, Rhamdia quelen (Heptapteridae), exposed to different water pH

The purpose of this study was to verify the effect of dietary Ca²⁺ on the growth and survival of silver catfish fingerlings (Rhamdia quelen) exposed to different water pH (5.5, 7.5 and 9.0). Silver catfish fingerlings were randomly placed in a thermoregulated water re‐use system with twelve 250 L‐tanks, two 1000 L‐biofilters and a 2000 L‐reservoir with a medium flow of 3.84 L min^?1 tank. Stocking density was 0.16 fingerlings L^?1. To prepare the treatment diets, the control diet (0.8 g kg^?1 Ca²⁺) was supplemented with CaCO₃ to yield experimental diets with 6.4, 9.5 and 23.9 g kg^?1 Ca²⁺. There were three replicates/treatments. Survival was more than 93.9% in all treatments. Exposure of silver catfish fingerlings to alkaline or acid water reduced growth, and this effect was not ameliorated by dietary Ca²⁺ supplementation. Moreover, when fingerlings were maintained in water with pH 7.5, the best dietary Ca²⁺ range for silver catfish fingerling growth was 0.8–6.4 g kg^?1.