氮、磷营养盐对东海原甲藻生长和硝酸还原酶活性的影响

在实验室培养条件下,研究了不同氮、磷浓度及氮磷比结构对东海原甲藻生长和藻细胞硝酸还原酶活性(NRA)的影响.结果表明,各培养组藻细胞在接种后d 2即进入指数生长期,但受氮、磷浓度及结构的影响,各培养组的比生长率和藻密度存在一定的差异.同东海原甲藻的生长类似,藻细胞的NRA也受氮、磷浓度及结构的影响.研究进一步发现,东海原甲藻硝酸还原酶活性的最大值(NRAmax)都出现在指数生长期,与最大比生长率出现时间基本一致,且当N/P=16时,酶活力有最大值,表明东海原甲藻的硝酸还原酶活性存在一定阈值.此外,藻的生长速率与营养盐的同化速率并不一致,存存一定的滞后效应.     

东海原甲藻Prorocentrum donghaiense Lu生物学研究进展

东海原甲藻(Prorocentrum donghaiense Lu)是中国沿海频繁发生的大规模赤潮的原因种之一。文章对东海原甲藻的命名、生理生态和分子生物学等方面的研究成果作了综述。东海原甲藻与在日本、韩国等亚洲海域分布的被报道为Prorocentrum dentatum的原甲藻以及来自美国CCMP的具齿原甲藻(Prorocentrum dentatum)为同一种。东海原甲藻是一种喜长光照的赤潮藻类,其生长的最适温度为22℃,最适盐度为25~31。东海原甲藻能利用铵盐、硝酸盐和亚硝酸盐,最适生长的w(N)/w(P)比范围在8~20之间。作者构建了东海原甲藻cDNA文库并进行了EST分析,发现了22个与东海原甲藻生长发育、物质转换和能量代谢相关的基因标签。认为从基因表达及调控角度研究赤潮形成与消亡机制并阐明它与环境因子的关系是必要的。     

鼠李糖脂对东海典型甲藻赤潮生物生长抑制作用的影响

探讨了铜绿假单胞菌产鼠李糖脂类生物表面活性剂对东海典型甲藻赤潮生物锥状斯式藻(Scrippsiella trochoidea)、塔玛亚历山大藻(Alexandrium tamarense)、海洋原甲藻(Prococentrum marinum)和微小原甲藻(P.minium)生长的抑制作用,并对其不同抑制作用的机理进行了初步探讨.结果表明,鼠李糖脂在一定浓度范围内对几种甲藻的生长表现出明显的抑制作用,对几种甲藻生长抑制作用的强弱顺序为:锥状斯式藻>微小原甲藻>塔玛亚历山大藻>海洋原甲藻.对不同属的甲藻,这种抑制作用可能与藻细胞本身表面的附属物、内部物质组成及个体大小有关;而对于同一属的甲藻则与各甲藻的生物膜脂肪酸组成尤其是其多不饱和脂肪酸的含量有关,即甲藻的多不饱和脂肪酸含量越低,鼠李糖脂对藻细胞生长的抑制作用越明显,反之亦然.图1表2参18     

海洋底栖甲藻——利玛原甲藻（Prorocentrum lima）产毒特征的研究（Studies on Toxin Production of Prorocentrum lima）

为了解海洋底栖甲藻——利玛原甲藻(Prorocentrum lima)的产毒特征,阐明腹泻性贝毒(Diarrhetic shellfishpoisoning,DSP)复杂的生物合成机制,采用酶联免疫试剂盒,对利玛原甲藻不同生长期、不同营养盐条件下毒素生成情况进行了研究.结果显示,不同生长期单个利玛原甲藻细胞中大田软海绵酸(Okadaic acid,OA)的含量明显不同,平台期时OA含量最高,与对数中期存在显著差异(p<0.05).低N、P条件下藻的生长状况较差,所能到达的藻密度明显低于对照组(f/2培养基);藻细胞中OA的总含量也低于对照组,但单个藻细胞中OA的含量明显高于对照组(p<0.05).以上结果提示,利玛原甲藻DSP毒素的产生可能与藻所处的环境和藻的生理状态等有关.     

丙烯酰胺对水环境中典型微藻的毒性效应

作为可疑致癌物,丙烯酰胺(acrylamide,AM)是目前各国政府和广大民众普遍关注的重要污染物.为探究AM对水体生物的毒性效应及可能存在的生态风险,本文以海洋微藻东海原甲藻(Prorocentrum donghaiense)和淡水微藻莱茵衣藻(Chlamydomonas reinhardtii)为研究对象,采用室内培养法,测定了不同暴露浓度的AM对2种微藻生长、形态和生理状态的影响.结果显示,AM对2种藻类生长均有显著的抑制作用(P0.05),96 h半抑制质量浓度(EC_(50))分别为22.79 mg·L~(-1)和161.8 mg·L~(-1);最高无抑制浓度(NOEC)分别为1.04 mg·L~(-1)和9.84 mg·L~(-1).不同微藻对AM胁迫的响应存在较大的差异性,与莱茵衣藻相比,东海原甲藻对AM更敏感.扫描电镜(SEM)和透射电镜(TEM)结果显示,当受到AM影响时,2种藻细胞严重变形,表现出塌陷、质壁分离、空泡数量增多和叶绿体片层结构少量断裂等现象;叶绿素含量和F_v/F_m(PSⅡ最大光化学量子产量)测试表明AM可以通过破坏微藻的光合系统而抑制光合作用.AM对东海原甲藻属于中毒性物质,对莱茵衣藻属于低毒性物质.     

2014年春季东海大面积甲藻赤潮的生态特征

为准确掌握东海近岸海域赤潮发生情况,深入了解东海海域赤潮发生时和未发生时的生态环境条件,根据浙江省舟山海洋生态环境监测站对东海近岸海域常年的监测结果,选定从杭州湾外侧向南到温州南麂列岛的东海近岸海域为赤潮高发区,开展生态环境研究.2014 年4 月和5 月利用专业海洋调查船,按《海洋监测规范》对东海赤潮高发海域进行了二次综合海洋生态环境调查,结果发现调查海域氮、磷营养盐含量普遍较高,4 月研究海域无机氮均值为0.406 mg·L^-1,活性磷酸盐均值0.013 mg·L^-1;5 月无机氮均值0.244 mg·L^-1,活性磷酸盐均值0.004 mg·L^-1.大多数样品无机氮超一类海水水质标准,尤其是4 月91.7%的样品超-类海水标准,5 月也有35.7%样品超-类海水标准.硝酸盐氮是无机氮的主要存在形式.2014 年5 月调查期间发生了大面积的赤潮,赤潮原因种是东海原甲藻（Prorocentrum donghaiense）,细胞密度平均在107/L 以上.分析赤潮发生前后的水环境变化,5 月调查海域海水温度有较大上升,各海域水温均超过了20 ℃,海域平均温度从4 月的15.9 ℃上升到5 月的21.9 ℃.通过本次研究发现在营养盐浓度普遍较高的调查海域,水温上升是引发赤潮的关键因素.使用专业统计软件SPSS17 发现赤潮藻类的细胞密度与水温、化学需氧量和溶解氧存在着极显著的正相关（P〈0.01）,与活性磷酸盐、硝酸盐氮和无机氮存在着显著负相关（P〈0.05）.另外本次调查还发现,大面积东海原甲藻赤潮暴发后,活性磷酸盐被大量消耗.综合目前有关东海原甲藻赤潮期间对活性磷酸盐的吸收动力学研究,发现今后应进一步加强这方面的调查研究.     

4种典型有害赤潮原因种的溶血特性研究

对4种典型赤潮原因种的细胞和培养水中的溶血特性进行了研究.藻种分别是:赤潮异弯藻(Heterosigma akashiwo)的两个品系,微小亚历山大藻(Alexandrium minutum),东海原甲藻(Prorocentrum donghaiense)和微小卡罗藻(Karlodinum micrum).对各藻类的去藻上清液和超声破碎细胞提取液进行了溶血性检测,发现:几种赤潮种对鲫鱼红血细胞的溶血特性具有种间差异,同藻种的细胞与其培养水中的溶血强度存在差异.具体表现为:赤潮异弯藻具有最强的溶血性,去藻上清液的溶血性(60￣100%)远大于超声破碎细胞提取液的溶血性(30￣60%);微小卡罗藻超声破碎细胞提取液的溶血性(70%)大于去藻上清液的溶血性(30%),且是各藻超声破碎细胞提取液溶血性中最强的;东海原甲藻和微小亚历山大藻溶血性较弱,微小亚历山大藻去藻上清液的溶血性(30%)略大于超声破碎细胞提取液的溶血性(10%),东海原甲藻两部分的溶血性差异不显著.赤潮种类及其细胞内外的溶血特性在一定程度上显示了赤潮原因种在赤潮致害过程中的特征.     

投喂不同密度卤虫无节幼体对灰海马幼体生长和存活的影响

为比较投喂不同密度卤虫对灰海马幼体生长和存活的影响,共设3个卤虫无节幼体密度组[5(A)、10(B)和20(C)个/mL],用初始密度为2尾/L的灰海马幼体进行为期50 d的养殖研究.结果显示,总体而言,3个饵料密度对灰海马幼体的存活、生长有显著影响(P<0.05).在前10 d,3组海马间的成活率和生长速度差异均不显著(P>0.05);d20～50,B、C两组海马的成活率均显著高于A组(P<0.05),而B、C两组之间无显著差异(P>0.05).d 20时,B、C两组的生长速度显著快于A组(P<0.05);d 30往后,A、C两组的生长速度开始加快,随后3组海马生长速度间的差距进一步缩小,直至A组的体质量和体高超过B组和C组(P<0.05),而B、C两组间的差异显著性消失(P>0.05).随着投放卤虫密度的增加,各时间点水体的pH和溶氧含量呈显著下降的趋势(P<0.05);C组氨态氮含量显著高于A组和B组(P<0.05);除第24 h外,3组间亚硝态氮的含量差异不显著.综合结果表明,卤虫的最适投喂密度应随着灰海马的生长进行适时调整.从初孵至d 10,饵料密度以10个/mL最佳;从d 10起,将饵料密度上调至20个/mL;d 30后当海马密度降至0.5～1尾/L时,卤虫密度可降至10个/mL左右.图2表1参21     

浮游植物细胞程序化死亡研究进展

过去,动物摄食和沉降被认为是浮游植物死亡的原因。后来人们发现病毒感染以及逆境条件下的主动死亡(或程序化死亡),也是引起浮游植物死亡的重要原因。文章介绍了浮游植物存在细胞程序化死亡的实验证据和半胱氨酰天冬氨酸特异蛋白酶(caspase)在浮游植物细胞程序化死亡中可能起的作用及浮游植物基因组中的caspase直系同源物。程序化死亡是细胞自主死亡的过程,具有坏死的形态和生物化学过程。浮游植物具有一套后生动物caspase的直系同源物蛋白。作者构建了东海原甲藻cDNA文库,其中有两组EST分别与两个细胞程序化死亡的关键蛋白同源,它们分别是半胱氨酰天冬氨酸特异蛋白酶和分裂细胞核抗体。因此作者推断东海原甲藻可能存在细胞程序化死亡过程。尽管已从细胞生物学特征角度,获得了真核微藻程序化死亡的证据,并从基因组序列中预测出和大规模表达序列标签中鉴定出细胞程序化死亡相关蛋白的编码基因,但离全面认识单细胞真核浮游藻程序化死亡的分子机制以及其生态学意义相差甚远。     

Effects of photoperiod on photosynthetic characteristics and quality of Tetragonia tetragonoides (Pall.) Kuntze in a plant factory北大核心CSCD

为探究光周期对番杏[Tetragonia tetragonoides(Pall.)Kuntze]生长及品质的影响,利用植物工厂中光源高度可控的优势对番杏进行不同光照时间(8、10、12、14、16 h)处理,测定分析番杏的生长、光合特性及营养品质.结果表明:(1)番杏各农艺性状随光照时间延长而增长,在光照时间14 h与16 h处理之间未呈显著性差异(P<0.05);(2)适宜光周期有利于光合色素积累,在16 h处理下叶绿素a、总叶绿素和类胡萝卜素含量最大,叶绿素b含量则在14 h光照时最大;净光合速率(P_(n))、蒸腾速率(T_(r))和气孔导度(G_(s))随光照时间延长上升,其中净光合速率在14 h与16 h光照之间变化不显著,胞间CO_(2)(C_(i))则呈先降后升的趋势;光周期变化对最大光化学效率F_(v)/F_(m)与光化学淬灭系数q_(P)未造成显著影响,非光化学淬灭系数NPQ随光照时间延长呈先降后升的趋势,14-16 h光照处理之间变化不显著,电子传递效率ETR则随光周期延长而降低;(3)不同光周期对番杏各营养成分含量的影响有所差异,可溶性蛋白变化差异较小,硝酸盐随光照时间延长而降低,14 h光照下可溶性糖含量最大.本研究表明14-16 h光照处理有利于番杏生长和营养物质积累.(图5表2参37)     

Can photoperiod manipulation affect gonad development of a boreo-arctic echinoid (Strongylocentrotus droebachiensis) following exposure in the wild after the autumnal equinox?

A laboratory experiment was conducted in the winter of 2003–2004 to assess the effect of varying photoperiod regime on consumption rate, assimilation rate, absorption efficiency, and gonad development of the green sea urchin, Strongylocentrotus droebachiensis. Adult individuals were collected from the wild after they had been exposed to the ambient autumn photoperiod cue (which is the extraneous trigger thought to elicit gametogenesis in this species) and placed at ambient temperature for 12 weeks under five different photoperiod regimes: (1) 24 h light:0 h dark=“0D”, (2) 16 h light:8 h dark=“8D”, (3) 8 h light:16 h dark=“16D”, (4) 0 h light:24 h dark=“24D”, and (5) ambient photoperiod (range: 10.50–15.25 h dark). Urchins in these five treatments were fed ad libitum with bull kelp, Nereocystis luetkeana. A sixth treatment consisted of starved individuals held under 0D conditions. Various gonad factors including gonad index, percent gonad water, gonad colour (CIE lightness or L*, CIE hue or a*, and CIE chroma or b*), percent area occupation of the gonad by various cell types (nutritive phagocytes, spermatozoa, and secondary oocytes/ova), and stage of development were assessed at the beginning of the experiment and at weeks 4, 8, and 12 of the study. Consumption and assimilation rates were assessed at weeks 4 and 12 and absorption efficiency at week 12 of the experiment. Urchins were predominantly in the growing and premature stages at the beginning of the experiment, but by week 4 at least 20% of individuals in all treatments receiving food were classified as mature. Spawning occurred during all these treatments between weeks 4 and 8, as evidenced by significant decreases in spermatozoa and secondary oocytes/ova and a significant decrease in percent gonad water, but was not accompanied by major declines in gonad indices. Greater than 90% of individuals in all five of the fed treatments were in the recovering and growing stages at the end of the experiment. The 16D treatment had by far the greatest percentage of urchins in the growing stage. In contrast, individuals that were starved were predominantly in the mature stage at weeks 4, 8, and 12, with only ~30% reaching the spawning stage by the end of the experiment. Photoperiod significantly affected gonad indices at the termination of the experiment with gonad index being the highest in the 16D treatment; this was significantly greater than in the 8D and ambient treatments. Photoperiod did not significantly affect gonad percent water, gonad lightness, or gonad hue. Gonad chroma was significantly affected by photoperiod, urchins held under ambient conditions having significantly lower b* readings than individuals in any other treatment. Photoperiod had little or no affect on consumption rate, assimilation rate, or absorption efficiency. Thus, differences among treatments in regards to gonad index, gonad chroma, and stage of development cannot be attributed to variations in feeding, absorption, or assimilation. The results of this experiment indicate that once gametogenesis is initiated, photoperiod manipulation cannot prevent ultimate spawning. However, photoperiod regime can affect the rate at which urchins move through the various stages of the gametogenic cycle. Urchins placed on short days under artificial lighting (16D) moved through the spawning stage into recovering and growing stages the fastest. This photoperiod regime also produced the highest gonad index at the end of the experiment. Since the commercial urchin market prefers large gonads in the growing and premature stages (i.e. before the mature stage is reached and gonads start leaking sperm and eggs), short day-lengths under artificial lighting (16D) appear to be the best photoperiod conditions for optimizing marketability.     

Morphological plasticity of submerged macrophyte Potamogeton wrightii Morong under different photoperiods and nutrient conditions

The morphological plasticity of the submerged macrophyte Potamogeton wrightii under different nutrient conditions and photoperiods was measured in a laboratory controlled experiment for 70 days in Japan. Six treatments were used in this experiment (3 × 2 factorial design with three replications) which consisted of three photoperiods and two nutrient conditions. Both photoperiod and nutrient condition had a pronounced effect on shoot and leaf morphology in P. wrightii. New shoot recruitment, and the length of main and new shoots gradually decreased with shortening photoperiod under both nutrient treatments. Plants under an 8 h photoperiod and high nutrient levels generated significantly more dead leaves (7.42 leaf·shoot^?1) and decomposed shoots (1.3 shoots·pot^?1) than plants under other treatments. Under short photoperiods (12 and 8 h) plants failed to produce flowering spikes in both nutrient conditions. In high nutrient conditions, P. wrightii produced shorter shoots, fewer leaves with shorter and narrower laminas, and smaller petioles compared with plants in the low nutrient condition. This may be adaptive under high nutrient conditions because it lowers foliar uptake and, thus, nutrient toxicity.     

Cell cycle and toxin production in the benthic dinoflagellate Prorocentrum lima

Profiles of diarrhetic shellfish poisoning (DSP) toxins produced throughout the growth cycle and the cell cycle of the toxigenic marine dinoflagellate Prorocentrum lima were studied in triplicate unialgal batch cultures. Cells were pre-conditioned at 18 ± 1 °C, under a photon flux density (PFD) of 90 ± 5 μmol m⁻² s⁻¹ on a 14 h light:10 h dark photoperiod. In exponential growth phase, cultures were synchronized in darkness for 17 d. After dark synchronization, cultures were transferred back to the original photoperiod regime. Cells were harvested for DSP toxin analysis by LC-MS (liquid chromatography with mass spectrometry), and double-stranded (nuclear) DNA was quantified by flow cytometry. The cell populations became asynchronous within approximately 3 d after transition from darkness to the 14 h light:10 h dark photoperiod. This may be due to the prolonged division cycle (5 to 7 d) that is not tightly phased by the photoperiod. Unlike other planktonic Prorocentrum spp., cytokinesis in P. lima occurred early in the dark and ceased by “midnight”. Cellular levels of the four principal DSP toxins, okadaic acid (OA), OA C8-diol-ester (OA-D8), dinophysistoxin-1 (DTX1) and dinophysistoxin-4 (DTX4), ranged from 0.37 to 6.6, 0.02 to 1.5, 0.04 to 2.6, and 1.8 to 7.8 fmol cell⁻¹, respectively. No toxin production was evident during the extended period of dark synchronization nor during the initial period when NH₄ was consumed as the major nitrogen source. Soon after the cells were returned to the 14 h light:10 h dark cycle and they began to take up NO₃, cellular levels of all four toxins gradually increased. This increase in DSP toxins usually occurred in the light, marked by a rise in DTX4 levels that preceded an increase in the cellular concentration of OA and DTX1 (delayed by 3 to 6 h). Thus, DTX4 synthesis is initiated in the G1 phase of the cell cycle and persists into S phase (“morning” of the photoperiod), whereas OA and DTX1 production occurs later during S and G2 phases (“afternoon”). No toxin production was measured during cytokinesis, which happened early in the dark. The evidence indicates that toxin synthesis is restricted to the light period and is coupled to cell cycle events. Received: 3 September 1998 / Accepted: 30 March 1999     

Annual and spatial variation in recruitment of labroid fishes among seven reefs in the Capricorn/Bunker Group,Great Barrier Reef

Recruitment of labroid fishes to a lagoonal habitat within each of 7 reefs in the Capricorn/Bunker Group on the Great Barrier Reef was recorded for 2 yr (April 1982–April 1983). The density of recruitment of a species on one reef was not a good indicator of its density on other reefs. There were significant differences in the levels of recruitment between years. For several species, the ranking of reefs according to density of recruitment changed between years and the ranking of the two years changed among reefs. Recruitment varied differently for each species. Southern reefs were ranked more highly than northern reefs overall, but no North-South gradient was apparent. Populations of labroid fishes in the Capricorn/Bunker Group experience unpredictable variations in recruitment.     

Observations on the diel migration of marine dinoflagellates off the Baja California coast

The vertical distribution of Gonyaulax polyedra, Ceratium furca, Gymnodinium sp., Ceratium dens, Gonyaulax digitale, Prorocentrum micans, Polykrikos kofoidii, and Peridinium depressum was followed during two 36 h time-series stations, off the Baja California coast. Water samples were taken at 6 h intervals, at 7 light penetration depths, for phytoplankton identification and for chemical and biological analysis. The results indicate that the dinoflagellates were able to perform vertical migration against the physical water movements, such as upwelling internal waves and wind mixing, that existed in the area during this study. Attempts to correlate the vertical dstribution of the dinoflagellates with the seawater density and nitrate distribution lead to the conclusion that even a weak density gradient acted as a barrier to the downward migration, and that nitrate assimilation in darkness was not required for their growth. Differences in the vertical distribution at 12.00 hrs of each species supports the hypothesis that migration is of a phototactic nature. A comparison of this study with previous ones on vertical migration in other areas suggests that a species should not be defined in general as a positive or negative phototactic organism, but that the direction of the migration is related to the light intensity at the sea surface.Contribution No. 77-004 of the Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, Matne 04575, USA.