长心卡帕藻原生质体分离、培养研究

In this study, protoplasts were successfully isolated from Kappaphycus alvarezii using snail enzymes, abalone enzymes and cellulase. The optimum enzymic ratio was fixed to be 20% of abalone enzyme, 12% of cellulase and the osmotic stabilizer was 2.0 mol/L glucose. The optimum enzymic hydrolysis conditions were found to be dark enzymolysis at 30°C continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×104 mL-1, 65.20×104 g-1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20°C, light intensity of 1 500–2 000 lx and photoperiod of 12 h/d, two developmental pathways were investigated:(1) callus-like cell mass and regenerated plantlet occurred on protoplast;(2) young shoots and calluslike cell mass occurred in tissue blocks after enzymolysis.     

厦门大嶝岛潮间带底栖微藻叶绿素荧光对温度和光照变化的响应

采用PAM叶绿素荧光技术,于2013年5月在厦门大嶝岛潮间带,对不同底质类型（泥质、泥砂质和砂质）底栖微藻的光合作用特征进行了初步研究.结果显示：底栖微藻的光合系统II最大光量子效率（Fv/Fm）呈泥质（0.33）〉泥砂质（0.28）〉砂质（0.01）;底栖微藻叶绿素a含量亦呈上述变化趋势.不同温度（15-35℃）和不同光照透射率（0-100%）的短期（0.5-2.0 h）培养实验表明,底栖微藻光合作用的最适温度（27℃左右）接近于环境温度,体现了其对生长环境的适应;过高的温度（35℃及以上）会抑制光合作用活性,进而限制其生长;此外,光照实验表明底栖微藻可通过自身的运动和变化来适应光照强度的变化,不同底质类型可能存在不同的光适应策略.     

侧孢短芽孢杆菌溶藻活性代谢产物部分特性的研究

研究了不同因素对侧孢短芽孢杆菌（Brevibaci Uuslaterosporus）无菌滤液溶藻效果的影响,并对无菌滤液进行了紫外光谱扫描分析,探讨侧孢短芽孢杆菌溶藻活性代谢产物的物质特性、探究物质的组成成分．结果显示：无菌滤液分别经不同温度、pH值、蛋白酶和紫外照射时间处理后,均仍具有极为显著的溶藻能力（P〈0．01）,溶藻实验7d后颤藻（Oscillatoria sp．）叶绿素a质量浓度的平均下降率分别为：58．65％、63．88％、61．50％和62．61％,藻体干重的平均下降率分别为：39．77％、39．30％、36．49％和37．02％,表明溶藻活性代谢物质具有耐酸、耐碱、耐高温、抗蛋白酶水解、抗紫外辐射的特性;同时,分别在4、28℃环境中贮存28d的无菌滤液,始终保持着极高的溶藻活性,溶藻实验7d后颤藻叶绿素a质量浓度的平均下降率分别为：50．84％和60．39％,藻体干重的平均下降率分别为：37．82％和37．54％,说明该物质的结构极其稳定,在环境中可长久存在;另外,加热与未加热处理的无菌滤液的紫外光谱最大吸收峰都在290～300nm之间．分析表明,溶藻活性代谢产物极大程度上属于具有极强稳定性的肽类物质．     

硅浓度对筒柱藻B169生长、叶绿素荧光参数、总脂含量及脂肪酸组成的影响

本文以底栖硅藻筒柱藻(Cylindrotheca sp.)为实验材料,研究了一次性培养过程中,不同硅浓度即:0mg/L(Si0)、8.35mg/L(Si1)、33.4mg/L(Si2)、133.6mg/L(Si3)、534.4mg/L(Si4)和1068.8mg/L(Si5)对藻细胞密度、叶绿素荧光参数、叶绿素含量、总脂含量及脂肪酸组成的影响。结果表明:第5~9d,筒柱藻的细胞密度随硅浓度的增加而增加,硅浓度较高的Si3、Si4和Si5处理组的细胞密度显著高于硅浓度较低的Si0、Si1和Si2处理组的细胞密度。第1~9d,Si5处理组的荧光参数F_v/F_m(PSII的最大光能转换效率)和F_v/F_o(PSII的潜在活性)均显著低于其它处理组。第3~6d,Si3处理组的上述荧光参数均显著高于其它处理组。第1~7d,Si4和Si5处理组的荧光参数φPSII(PSII的实际光能转化效率)和rETR(相对电子传递效率)均显著低于其它处理组,其中Si5处理组的上述荧光参数最低,其次是Si4处理组。筒柱藻的叶绿素含量随着硅浓度的增加而增加,到Si3处理组叶绿素含量达最高值,之后叶绿素含量下降,Si3处理组的叶绿素含量最高,Si0处理组的叶绿素含量最低。筒柱藻的总脂含量随硅浓度的增加而降低,Si0处理组的总脂含量最高,Si5处理组的总脂含量最低。筒柱藻的主要脂肪酸是1 4:0(5.1 5~8.7l%)、16:0(22.40~42.27%)、1 6:1n-7(3l.40~33.12%)、16:3n-4(1.89~8.45%)、20:4n-6(5.95~11.39%)和20:5n-3(EPA)(3.47~8.07%)。饱和脂肪酸含量随着硅浓度的增加而降低,多不饱和脂肪酸含量随着硅浓度的增加而增加,硅浓度对单不饱和脂肪酸的影响差异不显著。经济价值较大的20:4n-6和20:5n-3含量均在Si3和Si4处理组达到最高值。     

微丝解聚剂及微管阻断剂对藓羽藻细胞重建过程的影响

某些种类的海洋多核绿藻的原生质被挤出后,其细胞器在人工海水中可立即发生聚集,随后其细胞组分发生重建,形成新的细胞膜与细胞壁,这些重建的细胞在最佳培养条件下,最多有40%能够发育成成熟藻体。细胞骨架系统是细胞内有序性的维系者,也是细胞及细胞器运动的动力源,这些过程都涉及细胞骨架的解聚与重聚合的动态过程。本文利用微丝解聚剂细胞松弛素B及微管阻断剂秋水仙素对藓羽藻(Bryopsis hypnoides)细胞重建过程的影响进行了研究。结果显示,细胞器聚集及聚集体变圆的动力不来自于细胞骨架系统,而原始微丝体系的解聚是细胞器高效聚集的基本条件;细胞松弛素B及秋水仙素都显著抑制细胞膜的形成及细胞内组分有序性重建等后继发育过程,表明细胞骨架系统在这些过程中扮演着重要角色。     

营养盐输入方式对3种微藻生长及种间竞争的影响

在不同的营养盐输入方式下(一次性输入、每2d输入、每5d输入)对杜氏盐藻(Dunaliella salina)、三角褐指藻(Phaeodactylum tricornutum)和米氏凯伦藻(Karenia mikimotoi)进行单种培养和混合培养,研究不同的营养盐输入方式对这3种海洋微藻生长及种间竞争的影响。结果表明:单种培养时,盐藻在一次性输入方式下的最终细胞密度和平均比生长速率均显著大于其它2种输入方式;三角褐指藻和米氏凯伦藻在每5d输入方式下的最终细胞密度显著大于其它2种输入方式。混合培养时,盐藻在一次性输入方式下的最终细胞密度和平均比生长速率均显著大于其它2种输入方式,最终细胞密度占总细胞密度的47.3%;三角褐指藻在一次性输入方式下的最终细胞密度和平均比生长速率均显著小于每2d输入和每5d输入方式,后2个处理组的最终细胞密度分别占总细胞密度的90.3%和79.9%;在3种输入方式下米氏凯伦藻的最终细胞密度及占总细胞密度的百分比、平均比生长速率均显著小于其它2种微藻。研究表明,无论单种培养还是混合培养,营养盐一次性输入方式均能促进盐藻的生长,而脉冲输入方式能促进三角褐指藻的生长。脉冲输入方式下混合培养时,三角褐指藻在种群中占优势。     

Effect of Fe3+ on the growth and lipid content of Isochrysis galbana

Inducing lipid accumulation in microalgae cells without suppressing cell growth is vital to the economical production of biodiesel from microalgae. In two experiments, we demonstrate that the cell concentration and lipid content of marine microalgae Isochrysis galbana depend upon the iron concentration in the growth media. In Experiment I, adding chelated FeCl 3 to the medium at the late exponential growth phase prolonged this phase and increased the lipid content in I. galbana cells. The fi nal cell density and lipid content of I. galbana supplemented with chelated FeCl 3 was approximately 2 and 1.65 times higher than that of non-supplemented cultures, respectively. In Experiment II, I. galbana cells in the late exponential phase were collected and re-inoculated into new media containing Fe 3+ at various concentrations. The fi nal cell concentration and lipid content were maximized at the highest iron concentration(38% biomass by dry weight at 1.2×10-5 mol/L Fe 3+). In this study, intracellular neutral lipid storage was evaluated by fl uorescent spectrophotometry using fl uorochrome Nile red, and the measurement conditions were optimized.     

Characterization of calcium deposition induced by Synechocystis sp. PCC6803 in BG11 culture medium

Calcium carbonate （CaCO3） crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803 （inoculated BG11）. In this study, the features of calcium carbonate deposition were investigated. Inoculated BGll in different calcium ion concentrations was used for the experimental group, while the BGll culture medium was used for the control group. The surface morphologies of the calcium carbonate deposits in the experimental and control groups were determined by scanning and transmission electron microscopy. The deposits were analyzed by electronic probe micro-analysis, Fourier transform infrared spectrum, X-ray diffraction, thermal gravimetric analysis and differential scanning calorimetry. The results show that the surfaces of the crystals in the experimental group were hexahedral in a scaly pattern. The particle sizes were micrometer-sized and larger than those in the control group. The deposits of the control group contained calcium （Ca）, carbon （C）, oxygen （O）, phosphorus （P）, iron （Fe）, copper （Cu）, zinc （Zn）, and other elements. The deposits in the experimental group contained Ca, C, and O only. The deposits of both groups contained calcite. The thermal decomposition temperature of the deposits in the control group was lower than those in the experimental group. It showed that the CaCO3 deposits of the experimental group had higher thermal stability than those of the control group. This may be due to the secondary metabolites produced by the algae cells, which affect the carbonate crystal structure and result in a close-packed structure. The algae cells that remained after thermal weight loss were heavier in higher calcium concentrations in BGll culture media. There may be more calcium- containing crystals inside and outside of these cells. These results shall be beneficial for understanding the formation mechanism of carbonate minerals.     

Nutrient-controlled growth of Skeletonema costatum: an applied model

To model Skeletonema costatum blooms and their relationship with environmental parameters in situ, a S. costatum-specifi c zero-dimensional box model based on the mechanistic model Eco3M was established using physiological features. The parameters were calibrated using experimental counterparts, and simulations were compared with published laboratory fi ndings. The resulting normalized objective function(NOF) values are less than 1.0(and in most cases less than 0.58) and the values for the slope γ(between 0.656 7–1.127 4) and R 2(between 0.806 8–0.971) are close to 1.0 for most of the sub-fi gures. This indicates good agreement between simulated and measured data and suggests that the model reproduces the general characteristics of S. costatum growth and use of nutrients under different N- or P-limiting conditions. The model is appropriate for further applications and can be used to test more scenarios using other nutrients.     

Seasonal Changes in Phytoplankton Biomass and Dominant Species in the Changjiang River Estuary and Adjacent Seas：General Trends Based on Field Survey Data 1959-2009

The characteristics of seasonal variation in phytoplankton biomass and dominant species in the Changjiang River Estuary and adjacent seas were discussed based on field investigation data from 1959 to 2009. The field data from 1981 to 2004 showed that the Chlorophyll-a concentration in surface seawater was between 0.4 and 8.5 ktg dm-3. The seasonal changes generally presented a bimodal trend, with the biomass peaks occurring in May and August, and Chlorophyll-a concentration was the lowest in winter. Seasonal biomass changes were mainly controlled by temperature and nutrient levels. From the end of autumn to the next early spring, phytoplankton biomass was mainly influenced by temperature, and in other seasons, nutrient level （including the nutrient supply from the terrestrial runoffs） was the major influence factor. Field investigation data from 1959 to 2009 demonstrated that dia- toms were the main phytoplankton in this area, and Skeletonerna costatum, Pseudo-nitzschia pungens, Coscinodiscus oculus-iridis, Thalassinoema nitzschioides, Paralia sulcata, Chaetoceros lorenzianus, Chaetoceros curvisetus, and Prorocentrum donghaiense Lu were common dominant species. The seasonal variations in major dominant phytoplankton species presented the following trends： 1） Skeletonema （mainly S. costatum） was dominant throughout the year; and 2） seasonal succession trends were Coscinodiscus （spring） →Chaetoceros （summer and autumn） → Coscinodiscus （winter）. The annual dominance of S. costatum was attributed to its environmental eurytopicity and long standing time in surface waters. The seasonal succession of Coscinodiscus and Chaetoceros was associated with the seasonal variation in water stability and nutrient level in this area. On the other hand, long-term field data also indicated obvious interannual variation of phytoplankton biomass and community structure in the Changjiang River Estuary and adjacent seas： average annual phytoplankton biomass and dinoflagellate proportion both presented inc