细胞质膜在罗非鱼和叉尾斗鱼低温驯化过程中的功能（英文）

研究通过一系列科学的指标,比如死亡温度、累计温度处理时间和存活率曲线,检测并比较了吉富罗非鱼和叉尾斗鱼的低温耐受能力,然后系统地检测了在两种鱼中,低温对一系列与细胞膜相关的生理指标的影响,包括细胞膜流动性、内吞作用和膜蛋白Na~+,K~+-ATPase的活性。实验结果显示叉尾斗鱼比吉富罗非鱼有着明显优良的耐寒力。两种鱼内与细胞膜相关的生理指标都对温度敏感,但是具体的敏感程度却在耐寒种(叉尾斗鱼)和不耐寒种(吉富罗非鱼)之间存在显著差异。在具体每个物种内部,与细胞膜相关的一系列生理功能对温度的敏感程度是一致的。所有这些种间差异性和种内一致性都使得细胞膜流动性在物种耐寒过程中的重要性凸显出来。结果揭示细胞质膜,尤其是质膜流动性可能在这两种鱼对低温适应过程中扮演重要的角色。     

斗鱼属鱼类亲缘关系的Cyt b基因序列和RAPD分析

采用mtDNA Cyt b基因序列分析和RAPD两种分子标记技术,研究了中国分布的叉尾斗鱼(Macropodus opercularis)、圆尾斗鱼(M.chinensis)和香港斗鱼(M.hongkongensis)以及越南的红鳍斗鱼(M.erythropterus)4种鱼类之间的亲缘关系.获得4种斗鱼14条Cyt b基因全序列(1 155 bp),结合GenBank中搜索到的近缘物种同源序列进行分析.从133条随机引物中筛选到36条引物,在优化的反应条件下,10个群体96个个体共扩增出清晰稳定的条带749条,构建矩阵进行分析和聚类.基于Cyt b全序列以邻接法和最小进化法构建的系统树及RAPD数据UPGMA聚类分析的结果都显示,香港斗鱼和红鳍斗鱼先聚为一分支,再与叉尾斗鱼聚类,圆尾斗鱼处于外缘.本研究结果反映了圆尾斗鱼与其他斗鱼的亲缘关系较远,种间遗传距离为0.184 5～0.225 3(Cyt b)和0.653 6～0.746 5(RAPD),两者为同一单系群中两个独立演化的自然类群;香港斗鱼与叉尾斗鱼间遗传分化明显,Cyt b碱基差异为11.00%,RAFD遗传距离达0.577 7,支持其为独立物种的观点,且香港斗鱼群体间遗传差异较大,Cyt b碱基差异为3.12%,RAPD遗传距离0.060 1;叉尾斗鱼群体间Nei's基因多样度和Shannon信息指数分别为0.058 2和0.086 9,而各群体内的数值分别为0.016 1～0.031 7和0.023 5～0.046 7,表明遗传差异主要来自群体间,并按分布流域分别聚类.     

越南黑叉尾斗鱼展示行为的偏侧化及其对镜面影像的响应

动物行为的偏侧化现象广泛存在于各个类群,特别是具有仪式化展示和打斗行为的鱼类。越南黑叉尾斗鱼(Macropodus spechti)是一种雄性个体间具有模式化展尾行为小型淡水鱼类。本文主要研究越南黑叉尾斗鱼展尾行为或眼睛使用的偏侧化现象,以及镜面影像对其行为的影响。研究发现,越南黑叉尾斗鱼中在个体(94.6%,n=35)和群体(左侧化指数:LⅠ=0.25±0.19)水平均具有明显的左侧化倾向;镜面影像会降低斗鱼的展示强度和左侧化倾向(LⅠ=0.10±0.23),但是,对水面呼吸的频次没有显著影响。这表明,越南黑叉尾斗鱼的左侧化倾向是对"头-尾"相对展示的一种适应,而镜面影像中"头-尾"展示姿势的变化对越南黑叉尾斗鱼的展示行为具有明显影响,但这种影响在反应强度(换气次数)上并没有差异,由此认为越南黑叉尾斗鱼对镜面影像缺乏自我识别能力。同时,由于偏侧化现象的存在,使得使用镜面影像无法获得和真实个体一致的行为响应结果。本研究对未来行为学研究中如何使用镜面影像研究动物行为具有重要的指导意义。     

尼罗罗非鱼TCP-1-beta和TCP-1-eta的分子特征及其低温诱导表达

为了研究尼罗罗非鱼耐寒性状的分子基础并为耐寒品种选育提供参考,研究从尼罗罗非鱼中克隆了HSP60家族TCP-1-beta和TCP-1-eta基因并对其在低温诱导下的表达特征进行了分析。尼罗罗非鱼TCP-1-beta cDNA长度为1755 bp,包括1605 bp的完整开放阅读框,编码534个氨基酸;尼罗罗非鱼TCP-1-eta cDNA长度1651 bp,包括1638 bp的完整开放阅读框,编码545个氨基酸。与其他物种同源基因的蛋白序列比对结果显示,TCP-1-beta和TCP-1-eta蛋白在物种间同源性很高,且都具有保守的ATP结合结构域等,预示其在物种间功能的保守性。实时荧光定量PCR结果表明:TCP-1-beta和TCP-1-eta在各组织中呈遍在表达,但在肌肉中表达量最高;诱导温度从22℃降至12℃,不同低温诱导48h后TCP-1-beta和TCP-1-eta均呈上调表达,在18℃时表达开始上调,随着低温胁迫程度加强,表达上调幅度增大,至12℃时表达量达到最高,TCP-1-beta和TCP-1-eta上调幅度分别达到常温的12.2倍和10.7倍。这些结果预示在尼罗罗非鱼中,TCP-1-beta和TCP-1-eta是潜在的耐寒相关基因。     

叉尾斗鱼的胚胎和幼鱼发育的研究

叉尾斗鱼Macropodus opercularis(Linnaeus)分类上属于鲈形目Per-ciformes攀鲈科Anabantidae,是一种广泛分布于我国南方及东南亚各国的内陆淡水小鱼。有关其分类位置、形态、习性和分布等问题,Myers,G.S.(1932),Sterba,G.(1963)等先后作了颇详细的记述;Tomita,G.et al.(1936,1938)曾对早期阶段的叉尾斗鱼对光的色素效应以及早期的行为发育做了研究和观察。有关叉尾斗鱼的胚胎和幼鱼发育尚未见系统地报道;在1979—1981年,作者对叉尾斗鱼的胚胎和幼鱼发育进行多次的观察和研究。     

多不饱和脂肪酸对微生物低温适应性的影响

地球生物圈75%以上的环境温度常年低于5℃,在这种低温环境中栖息着多种适应低温的微生物。在长期进化过程中低温微生物从细胞到分子水平形成一套独特的低温环境适应机制,而通过增加细胞膜膜脂中多不饱和脂肪酸含量来维持低温条件下最佳的细胞膜流动性是其中的一种。从多不饱和脂肪酸对微生物低温生长、细胞膜流动性细胞膜蛋白的组成和表达水平的影响来探讨多不饱和脂肪酸与微生物低温适应性的关系,总结多不饱和脂肪酸低温合成调节机制的研究进展,为相关的基础和应用开发研究提供参考。     

温度、盐度和pH对尼罗罗非鱼性别分化的影响

采用Box-Behnken设计及响应曲面法研究了温度(20～36℃)、盐度(0 ～16)和pH(5.5 ～8.5)3个主要环境因子对吉富品系尼罗罗非鱼性别分化的影响.结果表明:温度的一次和二次效应分别对吉富罗非鱼性别分化的影响显著.盐度和pH的一次和二次效应分别对吉富罗非鱼性别分化的影响不显著,3个环境因子中任意2个因子之间的互作效应都不显著.采用响应曲面法分析发现,随着温度的升高,吉富罗非鱼的雄性率呈逐渐上升的趋势;温度为36℃、盐度为8、pH为8.5时,吉富罗非鱼最大雄性率可达80％.建立雄性率与温度、盐度和pH三者之间关系的模型方程,并剔除相关不显著的因子后,得到雄性率与温度之间的模型方程,可用于预测吉富罗非鱼雄性率的变化.     

温度、盐度和pH对尼罗罗非鱼性别分化的影响

采用Box Behnken设计及响应曲面法研究了温度(20～36 ℃)、盐度(0～16)和pH(5.5～8.5)3个主要环境因子对吉富品系尼罗罗非鱼性别分化的影响.结果表明： 温度的一次和二次效应分别对吉富罗非鱼性别分化的影响显著.盐度和pH的一次和二次效应分别对吉富罗非鱼性别分化的影响不显著,3个环境因子中任意2个因子之间的互作效应都不显著.采用响应曲面法分析发现,随着温度的升高,吉富罗非鱼的雄性率呈逐渐上升的趋势;温度为36 ℃、盐度为8、pH为8.5时,吉富罗非鱼最大雄性率可达80%.建立雄性率与温度、盐度和pH三者之间关系的模型方程,并剔除相关不显著的因子后,得到雄性率与温度之间的模型方程,可用于预测吉富罗非鱼雄性率的变化.     

香港斗鱼海南种群的分子鉴定和亲缘关系分析

香港斗鱼（Macropodus hongkongensis）是一种斗鱼科（Belontiidae）斗鱼属（Macropodus）的淡水鱼,记录于香港、广东等地,分布稀少。本研究通过线粒体COI、Cytb及16S rRNA基因对海南万泉河水系发现的疑似香港斗鱼进行分子鉴定和亲缘关系分析。基于3种基因的比对鉴定显示,该斗鱼和香港斗鱼广东鹤山种群的亲缘关系最近,海南万泉河水系发现的斗鱼为香港斗鱼海南种群。本文中几种斗鱼的种间遗传距离在0.136～0.238,大于香港斗鱼海南种群种内遗传距离（0.000～0.006(COI)）的10倍,与10倍理论一致,满足物种的有效鉴定。香港斗鱼不同地理种群之间遗传差异明显,海南种群和广东鹤山种群之间的种内遗传距离为0.036。NJ进化树也显示,香港斗鱼海南种群和香港斗鱼广东种群聚为一支,接着与红鳍斗鱼（M.erythropteru）聚为一支,再与叉尾斗鱼（M. opercularis）聚为一支,最后与圆尾斗鱼（M. chinensis）聚为一支。本文为海南岛香港斗鱼种群的首次研究报道,有关结果将为我国香港斗鱼的演化和保护研究提供重要参考。     

不同光照条件下叉尾斗鱼仔鱼摄食节律

在自然光照、持续光照和持续黑暗3种光照条件下,研究了孵化后第5、8和12日龄的叉尾斗鱼仔鱼的摄食节律。结果表明:自然光照下,3个日龄的叉尾斗鱼仔鱼在午后12:00—16:00表现出明显的摄食高峰,而持续光照组与持续黑暗组未表现出明显的摄食峰谷。持续光照下,叉尾斗鱼仔鱼的昼夜摄食活动均很活跃,全日各时段平均摄食量显著增加,尤其是夜晚20:00—翌日4:00各时段平均摄食量极显著高于同一时段自然光照组。而在持续黑暗下,与自然光照相比全日各时段平均摄食量明显减少,尤其是白天8:00—16:00各时段的平均摄食量极显著降低。叉尾斗鱼仔鱼属于典型的白天摄食类型,其摄食节律与光照条件密切相关。     

Cold acclimation strategy is highly variable among the sunfishes (Centrarchidae)

We tested the hypothesis that the physiological strategy for acclimating to low body temperature is similar among closely related fish. Largemouth bass (Micropterus salmoides), green sunfish (Lepomis cyanellus), bluegill sunfish (Lepomis macrochirus), black crappie (Pomonix nigromaculatus), and white crappie (Pomonix annularis), all members of the family Centrarchidae, were acclimated to 5 degrees and 25 degrees C. Morphometric variables (total mass, total length, organ masses) and enzyme activities (hexokinase; lactate dehydrogenase; and cytochrome oxidase in heart, liver, and muscle) were measured in 5 degrees C- and 25 degrees C-acclimated fish at 5 degrees and 25 degrees C assay temperatures. Each species displayed a distinct physiological response to cold acclimation that differed among tissues. These data suggest that the response to cold acclimation is highly variable within families. Our findings are consistent with other studies suggesting that acclimation responses are labile and may evolve independently even among closely related species.     

The role of abscisic acid and low temperature in chickpea (Cicer arietinum) cold tolerance. II. Effects on plasma membrane structure and function

The frost hardiness of many plants such as chickpea can be increased by exposure to low non-freezing temperatures and/or the application of abscisic acid (ABA), a process known as frost acclimation. Experiments were conducted to study the response over a 14 d period of enriched plasma membrane fractions isolated from chickpea plants exposed to low temperature and sprayed with exogenous ABA. Measurement of the temperatures inducing 50% foliar cell death (LT50), and subsequent statistical analysis suggest that, like many plants, exposure to low temperatures (5/-2 degrees C; day/night) induces a significant level (P <0.05) of frost acclimation in chickpea when compared with control plants (20/7 degrees C; day/night). Spraying plants with exogenous ABA also increased frost tolerance (P <0.05), but was not as effective as low temperature-induced frost acclimation. Both pre-exposure to low temperatures and pre-treatment with ABA increased the levels of fatty acid desaturation in the plasma membrane (measured as the double bond index, DBI). Exposure of chickpea plants to low temperatures increased the DBI by 15% at day 4 and 19% at day 14 when compared with untreated control plants. Application of ABA alone did not increase the DBI by more than 6% at any time; the effects of both treatments applied together was more than additive, inducing a DBI increase of 27% at day 14 when compared with controls. There was a good correlation (P <0.05) between the DBI and LT50, suggesting that the presence of more unsaturated lipid in the plasma membrane may prevent cell lysis at low temperatures. Both pre-exposure to low, non-freezing temperatures and pre-treatment with ABA induced measurable changes in membrane fluidity, but these changes did not correlate with changes in LT50, suggesting that physical properties of the plasma membrane other than fluidity are involved in frost acclimation in chickpea.     

Environmental regulation and physiological basis of freezing tolerance in woody plants

Cold acclimation of plants is a complex process involving a number of biochemical and physiological changes. The ability to cold acclimate is under genetic control. The development of freezing tolerance in woody plants is generally triggered by non-freezing low temperatures but can also be induced by mild drought or exogenous abscisic acid, as well as by short photoperiod. In nature, the extreme freezing tolerance of woody plants is achieved during sequential stages of cold acclimation the first of which is initiated by short photoperiods and non-freezing low temperatures, and the second by freezing temperatures. Although recent breakthroughs have increased our knowledge on the physiological molecular basis of freezing tolerance in herbaceous species, which acclimate primarily in response to non-freezing low temperatures, very little is known about cold acclimation of woody plants. This article attempts to review our current understanding of the physiological aspects that underline cold acclimation in woody plants.     

温度驯化对尖头鳉热耐受特征的影响

为了研究不同驯化温度对尖头鰂(Rhynchocypris oxycephalus)热耐受特征的影响, 本研究设置4组水温(14℃、19℃、24℃和29℃), 对尖头鰂驯化两周, 采用临界温度法观察尖头鰂的耐受温度。结果显示: 尖头鰂的热耐受性受到温度驯化的影响, 表现为高温驯化可以升高最大临界温度(CT_max), 4个驯化组的平均CT_max分别为32.29℃、33.23℃、33.40℃和35.71℃; 低温驯化可以降低最小临界温度(CT_min), 平均CT_min分别为0.00、0.10℃、2.10℃和5.27℃; 在适中的温度(19℃)驯化条件下具有最高的温度耐受范围(33.13℃)。在高温条件下的温度驯化具有较高的驯化反应率, 最大值出现在24—29℃内(0.46); 低温驯化反应率最大值出现在29—24℃内, 为0.63。尖头鰂在本研究的驯化区间(14—29℃)内的热耐受区域面积为478.98℃2, 与温水性鱼类的温度耐受性相当, 说明尖头鰂具有较强的温度适应能力。     

Temperature sensing and cold acclimation

The fundamental question in cold acclimation is how do plants perceive the low but nonfreezing temperatures that activate cold acclimation responses. New findings in the past year suggest that changes in membrane fluidity, cytoskeleton rearrangement, and calcium influxes are among the earliest events taking place in plants upon exposure to low nonfreezing temperatures. In the cyanobacterium Synechocystis PCC6803, temperature change is detected by at least two separate sensors. One of these measures membrane fluidity using a classical two-component system involving histidine kinases and a response regulator in a His-to-Asp phosphorelay. Although these Synechocystis results may not be directly relevant to cold acclimation, they can guide our thinking as we search for biological thermometers in higher plants.     

Changes in leaf ultrastructure and carbohydrates inArabidopsis thaliana L. (Heyn) cv. Columbia during rapid cold acclimation

Summary We studied cell ultrastructure and carbohydrate levels in the leaf tissue ofArabidopsis thaliana L. (Heyn) cv. Columbia during rapid cold acclimation. Freezing tolerance of the leaves from 26 day old plants was determined after 48 h and 10 days at 4°C. Acclimation treatment of 48 h decreased the lethal freezing temperature from –5.7°C to –9.4°C. Freezing tolerance was not altered further by acclimation at 4 °C for 10 days. Ultrastructural changes in the parenchyma cells were evident after 6 to 24 h of cold acclimation. The plasma membrane showed signs of extensive turnover. Evidence of membrane invaginations and sequestering of membrane material was observed. In addition, numerous microvesicles, paramural bodies, and fragments of endoplasmic reticulum were noticed in the vicinity of plasma membrane. Modifications in the structure of cell membranes were evident after 5 days of exposure to low temperature. Small, darkly stained globules were seen on the plasma membrane, tonoplast, chloroplast envelope membrane, mitochondrion outer membrane, dictyosome cisternae membrane, and microvesicle membrane. As far as we are aware, this type of membrane modification has not been described previously in plant cells exposed to low temperature. We propose to call these structures membraglobuli. Acclimation treatment also increased the concentrations of soluble sugars and starch. These observations suggest that cold acclimation inA. thaliana induces changes in both plasma membrane properties and carbohydrate composition.     

秦岭细鳞鲑代谢及低氧耐受能力对温度驯化的响应

为考察秦岭细鳞鲑（Brachymystax lenok tsinlingensis）代谢及低氧耐受能力对温度驯化的响应,将实验鱼于6℃、12℃和18℃下驯化4周后,采用密闭式呼吸代谢测定仪对其静止代谢率（Resting metabolic rate,RMR）和临界氧压（Critical oxygen press,P_crit）等参数进行测定。结果发现,RMR随驯化温度升高而升高,在6-12℃、12-18℃温度内,RMR的Q₁₀值分别为2.59和2.77,表明该物种对温度的敏感性较高;P_crit值随驯化温度升高而升高且与RMR显著正相关,在驯化温度范围内（6-18℃）,P_crit提升了76.2%。研究结果提示:秦岭细鳞鲑应对低氧环境的生理可塑性较差,在高温下RMR增加可能是导致其低氧耐受能力降低的内在机制之一。     

植物抗寒及其基因表达研究进展

植物经过逐渐降低的温度从而提高抗寒能力 ,这个过程被人们称为低温驯化。植物低温驯化过程是一个复杂的生理、生化和能量代谢变化过程 ,这些变化主要包括膜系统的稳定性、可溶性蛋白的积累和小分子渗透物质 ,比如脯氨酸、糖等 ,这些变化中的一些是植物抗寒必需的 ,而另外一些变化不是必需的。主要对冷害和低温生理生化变化、低温诱导表达基因的功能和作用、低温驯化的调节机制及其信号转导方面进行了综述。通过差别筛选 c DNA文库的方法已经鉴定了许多低温诱导表达、进而提高植物抗寒能力的基因 ,其中有脱水素、COR基因和 CBF1转录因子等。低温信号的感受、转导和调节表达是低温驯化的关键环节 ,低温信号的转导过程与干旱胁迫之间具有一定的交叉 ,这为利用 ABA等来提高植物抗寒能力成为可能 ,相信不久的将来人们可以通过提高植物抗寒能力从而增加经济产量成为现实。     

TEMPERATURE ACCLIMATION AND THE NERVOUS SYSTEM

1. The conduction velocity of the compound action potential of peripheral nerves shows compensatory acclimation to temperature in a fish, a snail, a crab, and probably also in the frog. The heat and cold tolerances of peripheral conduction are probably both increased by cold acclimation in the frog. 2. The properties of compound action potentials are not suitable for temperature acclimation studies, since different neuronal populations in the same nerve have been found to exhibit different temperature characteristics. 3. Single but septate giant nerve fibres of earthworms show compensatory temperature acclimation of the conduction properties, the form of the action potential and of the axonal cable properties, especially below 13–19 °C. 4. The fatty acids and the plasmalogen aldehydes of the phospholipids of the goldfish brain are more unsaturated at lower acclimation temperatures. 5. The Na⁺-K⁺ ATPase activity of the earthworm nerve cord shows compensatory acclimation at low temperatures. 6. The spontaneous activity of the central nervous system of insects is altered in a compensatory manner by temperature acclimation. In fish, the cold tolerance of simple and complex reflexes and of conditioning is adaptively altered by temperature acclimation. The role of the central nervous system, especially of the thermoregulatory centre, in the temperature acclimation of homeotherms is established. 7. There are adaptive isoenzymes of acetylcholinesterase in the brain of the rainbow trout. These isoenzymes differ from each other in respect of the temperature dependence of their enzyme-substrate affinity. The synthesis of acetylcholine receptor molecules may also be affected by temperature acclimation. 8. The metabolism of putative synaptic neurotransmitters (5-hydroxytryptamine, adrenaline, noradrenaline) is altered in the frog and mouse brains during the early phases of temperature acclimation. These changes may initiate the physiological processes connected with temperature acclimation. 9. The neuromuscular transmission in the frog shows after acclimation to cold, increased resistance to it and some indications of temperature compensation. 10. Changes in neurosecretion seem to be involved in temperature acclimation both in poikilotherms and homeotherms. The fast axonal transport of proteins shows compensatory acclimation to cold in the frog.