条斑紫菜色彩突变体的诱导_分离和特性分析

条斑紫菜叶状体的原生质体经化学诱变剂－Ｎ－甲基－Ｎ’－硝基－Ｎ－亚硝基胍（ＭＮＮＧ）处理后,在它们的再生叶状体中,出现了少量色彩发生变异的点状异体。它们的色彩变异细胞呈点状无规则地与野生细胞或其它变异细胞镶嵌在一起。这类变异体的单孢子萌发成单色变异（或野生色）叶状体以及与母体相似的点状变异体。利用单色变异叶状体分离出绿色（ｙｅｌ）,浅桔黄色（ｏｃｈ）以及深桔红色（ｂｕｓ）等三个突变株。突变株的叶状体活体吸收光谱以及藻红蛋白和藻蓝蛋白的含量与野生型相比均存在着明显的差异。经突变体支生型杂交实验证明,突变体ｙｅｌ和ｏｃｈ均含一个与色彩变化有关并且符合孟德尔遗传规律的隐性变异基因。相关变异基因的着丝粒距离,在突变体ｙｅｌ和ｃｏｈ中分别为１１．５和１５．８图距。     

Identification and characterization of a green-type mutant of Porphyra tenera Kjellman var. tamatsuensis Miura (Bangiales, Rhodophyta)

We identified two green-colored conchocelis strains of cultivated Porphyra, ‘Oba-green’ and HGT-6 (F₁ strain of ‘Oba-green’), based on PCR-RFLP and sequence analyses of the nuclear ITS region and the plastid RuBisCO spacer region. These molecular analyses confirmed that the two strains were green-type mutants of the endangered species Porphyra tenera, although their blade color was not green. Under the same culture conditions, blade length increase was significantly higher in HGT-6 than in a wild strain HGT-1 of P. tenera, and the blade shape of HGT-6 was extremely elongate compared with that of HGT-1. From the growth characteristics and ITS-1 sequence data, HGT-6 was confirmed to be a green-type mutant of P. tenera var. tamatsuensis, a vigorously growing cultivar. Although photosynthetic pigment contents were lower in HGT-6 than in HGT-1, total content of four major free amino acids was higher and the blade thickness was almost the same in HGT-6 and HGT-1. These results suggest that the green-type mutant HGT-6 has potential as breeding material for further development of Porphyra breeding.     

条斑紫菜自由丝状体的形态结构观察

以光镜和电镜观察条斑紫菜（Porphyra yezoensis）自由丝状体生长过程中，从营养藻丝、壳孢子囊枝到成熟的壳孢子囊枝细胞形态结构的变化，结果表明，光镜下自由丝状体在生长过程中细胞外部形态、色素体位置和颜色均发生变化；电镜观察显示了3个不同时期细胞的超微结构，主要包括细胞壁、细胞核、色素体、类囊体、内质网、蛋白核、线粒体、液泡、红藻淀粉和质体小球等。在自由丝状体的生长过程中细胞超微结构的主要变化为细胞壁增厚并产生脊突；色素体数量减少、分布位置从连续排列在细胞两侧到分散排列在细胞边缘，并且在类囊体膜之间出现空隙；红藻淀粉量增多并充满细胞间质。对这些变化与自由丝状体生长环境水温、光照强度和光照时间发生变化之间的关系进行探讨，综合分析认为，条斑紫菜自由丝状体在生长过程中，生长环境经由夏季高温、光照时间长、光照强度高到秋季温度下降、光照时间缩短、光照强度下降的变化，丝状体细胞发生细胞壁增厚、疏松，色素体数量减少，内质网面积增加，液泡、线粒体数量增多，红藻淀粉量大大增加等结构的变化。     

条斑紫菜叶状体细胞的发育与分化

条斑紫菜壳孢子经室内10～80d培养后长成的叶状体分别被酶解成单个细胞，后者在液体培养基中发育成正常叶状体、畸形叶状体、细胞团、性细胞囊等10种不同类型的再生体，它们的形态与结构、细胞排列与大小，放散单孢子难易程度及最终发育结果等均不同。来自日龄不同的种藻的细胞，其再生体类型的数目和各类再生体的百分数不同。当种藻日龄为10～30d时，在它们的单离细胞再生体中出现了极少的细胞团，正常叶状体和具假根畸形叶状体的百分数随着种藻日龄的增加，急剧减少，但不具假根．的畸形叶状体百分数却随着种藻日龄的增加而增加。当种藻日龄大于40d以后，它们的单离细胞就几乎不能直接再生成正常叶状体。随着种藻日龄的增加，细胞再生体中不具假根的畸形叶状体百分数也显著减少，而细胞团的百分数急剧上升，精子囊和果胞子囊的数量也逐渐上升。来自同一藻体不同部位的细胞，在它们的再生体中，随着细胞所处藻体部位的上移，正常叶状体和畸形叶状体的百分数急剧减少，而细胞团百分数却不断增加。上述结果说明，在离体条件下，条斑紫菜叶状体的单离细胞发育成不同类型的再生体，是由于它们在种藻里处于不同分化阶段的缘故；同时，初步得出在条斑紫菜叶状体的发育过程中，从壳孢子细胞分化成性原细胞的过程可分为8个不同的分化阶段。     

Comparison of horse mackerel and tilapia surimi gel based on rheological and <Superscript>1</Superscript>H NMR relaxation properties

ABSTRACT:   Horse mackerel and tilapia surimi were subjected to six different heat and pressure treatments in order to compare gelation characteristics of easy- and difficult-setting gels, that is, temperature dependence, by observing rheological properties and microscopic molecular mobility. The stress–relaxation and proton spin–spin relaxation time (¹H T ₂) of water were measured for all treated gels. Horse mackerel gel demonstrated higher elasticity, large distribution of the stress–relaxation process, and smaller water ¹H T ₂ than tilapia in both heat and pressure treatments. The water ¹H T ₂ was steeply increased in the pressure treatment at around 294 MPa for both fishes. In contrast, the ¹H T ₂ rarely changed in the heat treatment in spite of the considerable change in rheological properties. From the experimental results, it is considered that the gelation of horse mackerel (easy setting) surimi is induced by highly unfolding and re-aggregation of protein, which contributes to the formation of a strong network structure compared with tilapia in both heat and pressure treatments, and that pressure treatments hardly improve the gel strength of tilapia (difficult setting) surimi. The water ¹H T ₂ measurement was used effectively in order to study gelation characteristics of easy- and difficult-setting fish through observing its molecular dynamics.     

Identification of currently cultivated Porphyra species by PCR-RFLP analysis

ABSTRACT:   Porphyra yezoensis and P. tenera are the representative species of the marine crop Porphyra (nori) in Japan. Since the two species are extremely similar to each other in morphology, nori breeders have tentatively classified many strains of cultivated Porphyra into the two species without strict species identification. In order to facilitate rapid and reliable identification of the many strains of Porphyra currently cultivated in various Japanese regions, 24 conchocelis strains were examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis using the plastid RuBisCo spacer and the nuclear internal transcribed spacer regions. From the results, all the strains were identified as P. yezoensis f.  narawaensis , and P. tenera was not detected. Hence, it was confirmed that most of the Porphyra strains currently cultivated in Japan are P. yezoensis f.  narawaensis , and that intensive selective breeding has led to a reduced genetic diversity in the stock used for nori cultivation.     

5个紫菜属物种丝状体的微卫星初步遗传分析

文章利用68对微卫星引物对坛紫菜（Porphyra haitanensis）、半叶紫菜（P.katadai）、条斑紫菜（P.yezoensis）、少精紫菜（P.oligospermatangia）和皱紫菜（P.crispata）5个紫菜属物种的丝状体进行初步的遗传分析,共筛选出20对高多态性适用引物,在10份紫菜丝状体样品扩增位点的多态性比例为95.24%,每对引物可扩增的等位基因位点数为1.22～4.05,平均为2.37;各样品之间的遗传多样性指数为0.254 6～0.899 4;不同物种的遗传距离为0.106 0～1.406 6,同一物种的丝状体材料优先聚类为一支,表明筛选出的微卫星引物具有良好的遗传分辨能力。     

条斑紫菜优良品系(LC-14)的筛选与特性分析

从条斑紫菜(Pyropia yezoensis)绿色突变体和红色突变体种内杂交产生的F_1叶状体中,分离出新品系LC-14。该品系的F_1叶状体在生长速度和品质等方面均显著优于条斑紫菜野生型栽培品系(WT)。日龄70 d的叶状体平均长度和湿重,LC-14品系分别为106.53 cm和3.07 g,分别是WT品系的4.39和10.27倍。培养至第55天,LC-14品系的3种主要光合色素(叶绿素a、藻红蛋白和藻蓝蛋白)含量分别为9.85 mg/g、52.09 mg/g和23.26 mg/g,分别比WT品系提高了65%、65%和49%,而其藻体的平均厚度(20.89μm)反而降低了32%。此外,LC-14品系的壳孢子放散总量为761.32×10~4 ind/壳,是WT品系的1.28倍。上述结果表明,LC-14品系具有生长快、品质优、壳孢子放散量大的优点,藻体颜色与野生色(棕褐色)相近,是条斑紫菜的优良品系,有望在生产中应用。     

In vitro hydroxylation of vitamin D3 and 25-hydroxy vitamin D3 in tissues of Atlantic salmon Salmo salar, Atlantic mackerel Scomber scombrus, Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua

The in vitro metabolism of ¹⁴CD₃ and ³H25OHD₃ was investigated in different tissues from Atlantic salmon Salmo salar , Atlantic mackerel Scomber scombrus , Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua . The tissues analysed were liver, kidney, head kidney, gills, spleen and intestine. The metabolites were extracted in methanol–chloroform and separated by normal-phase high-pressure liquid chromatography (HPLC) followed by scintillation counting. Identification of the metabolites was by comigration with standards on normal and reversed-phase HPLC systems and by protein-binding assays. All tissues from all species analysed produced hydroxylated derivatives identified as 25OHD₃, 24,25(OH)₂D₃ and 1,25(OH)₂D₃. In addition, some unidentified derivatives were recorded, one probably being 25,26(OH)₂D₃. Organs producing great amounts of one metabolite also produced considerable amounts of the other possible derivatives, suggesting a lower degree of specificity in fish organs than in human organs. The predominating metabolite was 24,25(OH)₂D₃ in all organs from salmon and mackerel during incubation with ¹⁴CD₃ and within most organs from all species during ³H25OHD₃ incubation. The latter observation probably results from the need for decreasing rather than increasing the calcium absorption in these species, which live at least some periods of life in a marine environment.     

高温胁迫下条斑紫菜叶状体HSP70基因的表达谱分析

采用实时荧光定量RT-PCR技术检测了条斑紫菜叶状体在25℃高温胁迫下PyHSP70-1基因的相对表达量变化。定量分析显示PyHSP70-1基因表达受到高温胁迫的显著诱导。在持续高温胁迫下,PyHSP70-1表达量在8 h时达到最大,此后一直维持在较高水平,且与对照差异显著。在短暂高温胁迫结束后PyHSP70-1表达量最大,随着恢复时间的增加,其mRNA量逐渐减少,在恢复的24 h时间内,mRNA量仍然显著高于对照。结果表明PyHSP70-1是诱导型HSP,受热激诱导表达,在热激胁迫中维持细胞蛋白与膜的稳定,对细胞具有重要的保护作用。     

生态因子对条斑紫菜自由丝状体生长的影响

报道了生态因子对条斑紫菜自由丝状体生长的影响,结果表明,条斑紫菜自由丝状体生长的最适宜盐度范围为２０￣３０,低于１０的盐度不能生长,最适宜温度范围为１５℃￣２５℃,高限为３０℃,适宜光辐射照度１５μＥｍ％－２Ｓ＾－１￣６０μＥｍ＾２Ｓ＾－１,添加的适宜Ｎ浓度２０ｇ／ｍ＾３￣６０ｇ／ｍ＾３,Ｐ为１．５ｇ／ｍ＾３￣５ｇ／ｍ＾３。     

A pharmacokinetic study of flumequine in sea bass, Dicentrarchus labrax (L.), after a single intravascular injection

The pharmacokinetic properties of flumequine following a single intravascular injection (10 mg kg^–1 fish) were studied in sea bass, Dicentrarchus labrax (L.), 120 g held at 18 °C. The absorption half life ( t _1/2α) and the elimination half life ( t _1/2β) of the drug were calculated to be 1.05 and 10.71 h, respectively. Tissue penetration of flumequine seemed to be moderate because both the apparent volume of distribution of the drug at steady-state ( V _d(ss)) and the extensive apparent volume of the central compartment ( V _c) were found to be small (1.51 and 0.626 L kg^–1). The mean residence time ( MRT ) was short (09.73 h) and the total clearance (CL_T) of the drug was rapid (0.156 L kg^–1 h^–1).     

Biochemical composition of the wild and cultured seahorses, Hippocampus kuda Bleeker and Hippocampus trimaculatus Leach

The biochemical composition of wild and cultured seahorses Hippocampus kuda Bleeker and Hippocampus trimaculatus Leach was analysed in this study. The crude protein contents (% dry weight) of wild seahorses were high: 72.7±2.5% in H. kuda and 78.5±4.2% in H. trimaculatus . The crude lipid contents (% dry weight) of wild and cultured seahorses (1.1±0.1 vs. 1.4±0.2 for H. kuda and 1.2±0.1 vs. 1.2±0.2 for H. trimaculatus ) were low and did not differ significantly ( F _3, 12=1.461, P =0.12). The essential amino acids and flavour-enhancing amino acids in wild seahorses were higher than those in cultured seahorses, although the essential amino acid index in wild and cultured H. kuda and H. trimaculatus were generally low. The total amino acid content decreased with growth in cultured H. trimaculatus ( F _3, 12=14.927, P <0.05). The poly-unsaturated fatty acid (PUFA) contents in wild and cultured H. kuda and H. trimaculatus were 21.69%, 37.72%, 21.39% and 33.89% respectively. The fatty acid content in seahorses first decreased and then increased with growth both in wild and in cultured H. trimaculatus . The PUFA/saturated fatty acid ratio was 1.31 in cultured H. kuda and 0.87 in cultured H. trimaculatus . The trace metal concentrations in the wild seahorses were relatively low. The results presented in this study provide biochemical information necessary for understanding the medicinal value of H. kuda and H. trimaculatus and for elucidating the potential dietary requirements for their culture.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Haematological responses in the European eel, Anguilla anguilla L., to sublethal infestation by Anguillicola crassus in a thermal effluent of the Swedish Baltic

Abstract. Haematological variations were studied in a European eel population, Anguilla anguilla L., naturally infested by the sanguivorous nematode Anguillicola crassus. This occurred off the Swedish Baltic coast in an area receiving heated cooling water from a nuclear power station. The erythrocyte count, haematocrit, leucocrit, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentrations, the various cell types in peripheral blood, serum iron concentration, the amount of total serum protein, and scrum protein fractions α ₁ , β₁ , α ₂, β ₂ and γ were all measured. Each variable was correlated to: (1) the intensity of infestation, i.e. number of parasites per host; and (2) a parasitization index, i.e. the weight of parasites per somatic weight of the host. Most variables showed no or only minor reactions to the infestation. However, a marked increase in the γ -fraction of scrum proteins, reduced lymphocyte numbers and increased granulocyte numbers are indicative of a humoral and cellular immune response.     

Induced Meiotic Gynogenesis of Paddlefish Polyodon spathula

Viable, diploid gynogenetic (gynogenotes) paddlefish Polyodon spathula were produced by activating eggs with ultraviolet-irradiated shovelnose sturgeon Scaphirhynchus platorynchus spermatozoa and heat shocking. Without irradiation treatment, sturgeon spermatozoa appeared to activate the eggs (up to gastrulation), but did not result in any viable hybrids. Experiment 1 determined that heat-shock treatment of 35 C for a 2-min duration within the interval of 2–22 min post-activation resulted in highest yield of gynogenotes (12–19%) from eggs incubated at 18 C. Experiment 2 applied the heat shock treatment at 35 C from 14.0 to 28.0 min in 2-min intervals after activation at 18 C for a larger scale of gynogenetic production. Both experiments showed that the best yields of gynogenotes were obtained when the heat shock treatment occurred at 16, 18, and 20 min after activation. When these times were expressed in terms of τ₀. units (duration of one mitotic cycle of synchronous cell division related to water temperatures), optimal activations were 0.26, 0.29, and 0.32τ₀ (τ₀@ 18 C = 63.5 min). Experiment 3 tested the utility of τ₀. at two different pre-shock incubation water temperatures of 18 C and 16 C, and determined that there was no significant interaction in percentage of viable gynogenotes between two different incubation temperatures and the mitotic intervals (0.21, 0.26, 0.31, 0.36, 0.41τ₀) tested. Best survival of gynogenotes occurred when eggs held at either pre-shock incubation water temperatures were shocked at 0.26τ₀ All gynogenotes examined were histologically confirmed to have ovarian tissue and were determined to have similar oocyte development to that of normal female (control) paddlefish.     

Nephrocalcinosis in rainbow trout Salmo gairdneri Richardson; the effect of exposure to elevated CO2 concentrations

Abstract. Rainbow trout exposed to environmental free C0₂ concentrations ranging from 12–60 mg/1 for a minimum of 275 days developed nephrocalcinosis. Gross, histopathological and chemical examinations demonstrated that prevalence and severity of nephrocalcinosis increased with increasing CO₂ concentrations. The condition was characterized by calcification and granuloma formation in kidney and stomach. Diet, but not water, PO₄ -P concentration appeared to modify the prevalence and severity at each CO₂ concentration tested.     

Kinetics of Nitrogen Excretion by Cultured Flounder Paralichthys olivaceus

This study was undertaken to quantify the rate of nitrogen excretion by the cultured Japanese flounder as a function of body weight and water temperature. Measurements of the did rate of nitrogen excretion for the cultured flounder were made at mean water temperature of about 16, 19, and 23 C. The excretion rate of nilmgen was estimated by adding the nitrogen excreted in the forms of ammonia and urea nitrogens. The diurnal pattern in the excretion rate of nitrogen was governed mainly by the feeding time. The proportions of excreted ammonia to total nitrogen were 77.9% at 16 C, 84.5% at 19 C, and 83.2% at 23 C. The average was 81.9%. The relationship between the body weight and excretion rate of nitrogen ( E_N , mg N/fish per h) is expressed as E_N =α,.W', where α, = nitrogen excretion rate at temperature 0 . The parameter α, is dependent on water temperature, such that α, =α₂₀ Q₀₁^(0–200) where α₂₀= nitrogen excretion rate at 20 C, and Q₀₁, = temperature coefficient. The values of the kinetics parameters for the cultured flounder were found to be γ = 0.57, α₂₀= 0.060, and Q₀₁= 1.12.     

Growth and mortality of the catfish, Hemisynodontis membranaceus (Geoffroy St. Hilaire), in the northern arm of Lake Volta, Ghana

Estimates of growth and mortality of the catfish, Hemisynodontis membranaceus (Geoffroy St. Hilaire), in Lake Volta were obtained from length composition data compiled in 1995 and 1996. The von Bertalanffy growth function (VBGF) estimates were: L _∞=44.5 cm standard length; K= 0.62 year⁻¹; and t ₀=−0.23 years. Natural mortality rate, M , was 1.20 year⁻¹. Total mortality rate, Z , was computed as 4.39 year⁻¹ and the exploitation ratio ( E = F / Z ) was 0.72. Although the fish is estimated to have longevity of about 5 years, those exploited are normally less than 2 years of age, which is indicative of growth over-fishing. In order to arrest over-exploitation of the species, there is a need to establish 'lake reserves'. In addition, the fisheries management should be devolved from the state to the local level to compel fishermen to take greater responsibility for the sustainability and conservation of the fisheries.     

二氧化锗对坛紫菜自由丝状体生长发育的影响

本文研究了二氧化锗(GeO_2)对我国特有的坛紫菜自由丝状体的生长发育,及对一些附生性硅藻类和绿藻的影响,论证了二氧化锗在本实验浓度范围内,对坛紫菜自由丝状体无叨显毒理影响,而能抑制一些附生性硅藻和绿藻的生长;并探讨了二氧化锗对硅藻、绿藻的毒性机理。由于在培养坛紫菜自由丝状体时很容易附生硅藻 因而研究结果将为坛紫莱自由丝状体及其他藻种的培养和保存提供依据。