Optimization of conditions for tetraspore release and assessment of photosynthetic activities for different generation branches of Gracilaria lemaneiformis Bory

Gracilaria lemaneiformis Bory is an economically important alga that is primarily used for agar production. Although tetraspores are ideal seeds for the cultivation of G. lemaneiformis, the most popular culture method is currently based on vegetative fragments, which is labor-intensive and time-consuming. In this study, we optimized the conditions for tetraspore release and evaluated the photosynthetic activities of different colonies formed from the branches of G. lemaneiformis using a PAM (pulse-amplitude-modulated) measuring system. The results showed that variations in temperature and salinityhad significant effects on tetraspore yield. However, variations in the photon flux density (from 15 μmol m⁻² s⁻¹ to 480 μmol m⁻² s⁻¹) had no apparent effect on tetraspore yield. Moreover, the PAM-parameters Y(I), Y(II), ETR(I), ETR(II) and F _v/F _m of colonies formed from different branches showed the same trend: parameter values of first generation branches>second generation branches>third generation branches. These results suggest that the photosynthetic activities of different colonies of branches changed with the same trend. Furthermore, photosynthesis in G. lemaneiformis was found to be involved in vegetative reproduction and tetraspore formation. Finally, the first generation branches grew slowly, but accumulated organic compounds to form large numbers of tetraspores. Taken together, these results showed that the first generation branches are ideal materials for the release of tetraspores.     

Involvement of cyclic electron flow in irradiance stress responding and its potential regulation of the mechanisms in <Emphasis Type="Italic">Pyropia yezoensis</Emphasis>

Pyropia yezoensis, belongs to the genus of Porphyra before 2011, inhabit on intertidal zone rocks where irradiation changes dramatically, implying that the seaweed has gained certain mechanisms to survive a harsh environment. Based on the photosynthetic parameters with or without the inhibitors determined by a Dual-PAM-100 apparatus, we investigated the photosynthetic performance and the changes in electron flow that occurred during the algae were stressed with different light intensities previously. When the irradiation saturation was approaching, the CEF around PS I became crucial since the addition of inhibitors usually led to an increase in non-photochemical quenching. The inhibitor experiments showed that there were at least three different CEF pathways in Py. yezoensis and these pathways compensated each other. In addition to maintaining a proper ratio of ATP/NAD(P)H to support efficient photosynthesis, the potential roles of CEF might also include the regulation of different photoprotective mechanisms in Py. yezoensis. Under the regulation of CEF, chlororespiration is thought to transport electrons from the reduced plastoquinone (PQ) pool to oxygen in order to mitigate the reduction in the electron transfer chain. When irradiation was up to the high-grade stress conditions, the relative value of CEF began to decrease, which implied that the NADP⁺ pool or PQ⁺ pool was very small and that the electrons were transferred from reduced PS I to oxygen. The scavenging enzymes might be activated and the water-water cycle probably became an effective means of removing the active oxygen produced by the irradiation stressed Py. yezoensis. We believe that the different mechanisms could make up the photoprotective network to allow Py. yezoensis for survival in a highly variable light stress habitat, which may enlighten scientists in future studies on irradiance stress in other algae species.     

Combined effects of light intensity and NH^＋4-enrichment on growth,pigmentation, and photosynthetic performance of Ulva prolifera （Chlorophyta）

The aim of this study was to investigate the effects of light intensity and enhanced nitrogen supply on the growth and photosynthesis of the green-tide macroalga, Ulva prolifera. Thalli of U. prolifera were grown in natural or NH 4 +-enriched seawater under two different light intensities for 7 days, and then the growth rate, pigmentation, and photosynthetic performance of the thalli were evaluated. The results show that the relative growth rate(RGR) was markedly higher under the high light level than under the low light level. Enrichment with NH 4 + enhanced the RGR under high light intensity, but did not affect RGR under low light intensity. In low light conditions, NH 4 +-enrichment resulted in a marked decrease in the maximal photosynthetic rate( P m) and the maximum carbon fixation rate( V max), but it did not affect the half saturation constant for carbon( K 0.5) or the ratio of V max to K 0.5, which reflects the carbon acquisition efficiency. In high light conditions, P m, K 0.5, and the dark respiration rate( R d) increased under NH 4 + enrichment, but V max and the V max / K 0.5 ratio decreased. Regardless of the light intensity, NH 4 +-enrichment did not affect the apparent photosynthetic efficiency( α), which refl ects the ability of the alga to use light energy at low light levels. Under both low and high light intensities, the chlorophyll a(Chl a), chlorophyll b(Chl b), and carotenoids(Car) contents in thalli were higher in NH 4 +-enriched than in natural seawater, except that there was a decrease in the Chl b content of thalli in NH 4 +-enriched seawater under low light intensity. Therefore, NH 4 + enrichment improved the growth and photosynthetic performance of U. prolifera under high light intensity, but not under low light intensity. We discuss the possible mechanisms underlying these physiological responses.     

Identification of two Skeletonema costatum-like diatoms by fluorescence in situ hybridization

A harmful algae bloom (HAB) is a dense aggregation of algae in a marine or aquatic environment that can result in significant environmental problems. To forecast the occurrence of HAB, development of a rapid and precise detection method is urgently required. In this study, two Skeletonema costatum-like diatoms (SK-1 and SK-2), were identified morphologically under a light microscope, and detected using fluorescent in situ hybridization (FISH). Strain SK-1 was isolated from a frequently HAB affected area of the East China Sea, and strain SK-2 from an aquatic farm in Qingdao, China. Fluorescent DNA probes were designed that were complementary to the ITS sequence (including 5.8S rDNA) of strain SK-1. After hybridization, strong green fluorescence was observed in cells of strain SK-1 under an epifluorescence microscope; however, no such fluorescence was observed with strain SK-2, which indicates that probes hybridized only the DNA of the target strain, SK-1, in species-specific manner, and that the two strains do not belong to a same species. This finding was confirmed by ITS sequence analysis. The FISH technique used in this study was sensitive, simple, and rapid, and is a promising tool for detecting target HAB species in natural environments.     

Characterization of a new lectin involved in the protoplast regeneration of <Emphasis Type="Italic">Bryopsis hypnoides</Emphasis>

A group of coenocytic marine algae differs from higher plants,whose totipotency depends on an intact cell(or protoplast).Instead,this alga is able to aggregate its extruded protoplasm in sea water and generate new mature individuals.It is thought that lectins play a key role in the aggregation process.We purified a lectin associated with the aggregation of cell organelles in Bryopsis hypnoides.The lectin was ca.27 kDa with a pI between pH 5 and pH 6.The absence of carbohydrate suggested that the lectin was ...     

条斑紫菜类囊体膜蛋白质组双向电泳研究方法

以蓝绿温和胶电泳为工具,首次在国内用于条斑紫菜(Porphyra yezoensis)类囊体膜色素蛋白质复合物的研究。结果显示:(1)利用非离子型去污剂十二烷基麦芽糖苷(DM)增溶条斑紫菜类囊体膜,DM/Chla(w/w)15︰1产生的效果较好,在BN-PAGE胶中可以分离到较多较清晰的条带。(2)选取蔗糖密度层50%条带制备得到的类囊体膜样品进行第一向BN-PAGE和第二向SDS-Urea-PAGE电泳实验。第一向电泳分离出4个蛋白复合物,对第二向电泳胶上的15个蛋白点切取做质谱鉴定,检测到PSⅡ47ku,PSⅡ44ku,cytochrome f,PSⅡD2,PSⅡD1等蛋白。本实验证实了温和胶电泳与SDS电泳结合对于条斑紫菜这种原始红藻的类囊体膜研究方面应用的可行性。     

三株赤潮硅藻5.8S rDNA及转录间隔区(ITS)的克隆及序列分析

对引发赤潮的3株硅藻——1株尖刺拟菱形藻(Pseudo-nitzshia pungens)和2株中肋骨条藻(Skeletonema costatum)的5.8SrDNA和ITS(internal transcribed spacers)序列进行了PCR扩增、克隆和序列测定,并分析了硅藻门10株赤潮藻(7株从GenBank获得)的系统进化关系.研究结果表明,尖刺拟菱形藻的ITS和5.8SrDNA的长度为693bp,SK-1(分离自东海赤潮暴发区)测序得到715bp,除ITS和5.8SrDNA外,还包含部分18SrDNA和28SrDNA;SK-2(分离自青岛养殖场)的ITS和5.8SrDNA的长度为331bp,尖刺拟菱形藻与从GenBank中获得的2株尖刺拟菱形藻相似程度最高,为100%,与该属的多列拟菱形藻相似程度稍低,为82.9%.SK-2的ITS序列与SK-1的相似程度很低,只有51%,但与拟中肋骨条藻的ITS序列相似程度高,为95.5%.SK-1的ITS序列与拟中肋骨条藻的相似程度也低,为56.7%.系统进化树反映的结果与相似性反映的结果一致.研究的该株尖刺拟菱形藻从根据ITS序列研究的结果与形态鉴定的结果看是一致的;SK-2可能属于拟中肋骨条藻;SK-1比较特殊,有待于用其他的方法进一步研究确定其分类地位.     

龙须菜琼胶的提取方法研究

采用多种方法提取了龙须菜(Gracilaria lemaneiformis)中的琼胶,并研究了碱处理、微波处理、超声波处理等方法对龙须菜的琼胶提取率、硫酸基质量分数和多糖质量分数的影响。结果表明:当碱质量分数由2.5%增加到10%时,琼胶产率由15%降低到9.8%,硫酸基质量分数由2.66%降低到2.06%,同时,多糖质量分数由98.3%增加到102.6%;微波和超声波处理在不同程度上增加了琼胶的提取率和多糖质量分数,同时降低了硫酸基质量分数。另外,还从宏观和微观两个角度考察了龙须菜碱处理前后藻体的变化。     

生物制氢研究进展

目前, 世界上利用的能源90%以上都是“化石能源”(如石油、煤炭和天然气等)。随着人类不断利用这些不可再生的“化石能源”, 它将会逐渐枯竭, 且其燃烧形成的产物(如CO2和SO2等)造成的严重环境污染(如温室效应和酸雨), 也有害于人类健康。因此, 人类正面临着能源紧缺和环境污染的双重压力。为满足能源需求量日益增长和环境保护的需要, 我们必须寻找环保型的可再生能源来替代“化石能源” 以满足人类对能源的需求。 在各种可再生能源中氢能将有可能替代“化石能源”,成为未来能源利用的主要形式。氢能具有许多优点:①氢能燃烧时只生成水, 不产生任何污染物, 甚至连CO2也不会产生, 实现真正的“零污染” 和“零排放”;②氢能的能量密度高, 放热高达122 kJ/g, 是焦炭放热的4.5倍, 汽油的2.68倍;③氢能可经济有效的输送和储存, 可以利用已有的天然气管道输送, 输送成本低, 甚至优于输电, 因为输送氢能不会产生如电能输送过程中所产生的能量损耗;氢能的存储也比较简便, 其中储氢合金材料就是一种非常理想的储氢方法, 该方法储氢能力强、运输方便、操作容易, 且安全可靠。在利用储氢合金进行储氢时, 还可用以进行制冷或采暖(Kulkova et al., 2006), 因为储氢材料在吸氢时放热, 在放氢时吸热。正因为氢能具有如此多的优点, 许多大的跨国公司开始对制氢技术高度关注, 如主要汽车制造商(通用、福特、戴姆勒-克莱斯勒等)对开发以氢能作为燃料的汽车投入大量的人力和物力;宝马公司从20世纪70年代就开始研制以氢能为能源的汽车, 宝马公司现在已拥有了时速达240km的氢能汽车(Munro,2003)。同时, 一些发达国家的科研院所的科学家们对氢能的制备方法、制氢机制以及提高产氢量等进行了大量的研究。     

大叶藻(Zostera marina L.)PS I和PS II复合物的分离鉴定

采用蔗糖密度梯度离心法分离纯化大叶藻类囊体膜 ,经 1 0 %SDS增溶后 ,用蔗糖密度梯度超速离心分离其色素蛋白质复合物。经稳态光谱分析、DCIP光还原活性测定及P680 、P70 0 差示光谱检测结果表明 ,2 0 %蔗糖层的CP3和 40 %蔗糖层 (上 )的CP4为PSⅡ复合物 ,具有光化学活性 ;40 %蔗糖层 (下 )的CP5为PSⅠ复合物 ,其P70 0 特征吸收峰位于 695nm处。CP3和CP4的DCIP光还原活性 :CP3为 34.2 7微电子当量 /(mgchl·h) ,CP4为 7.2 9微电子当量 /(mgchl·h)。