利用盐藻（Dunaliella）培养生产β—胡萝卜素

盐藻是一种高含β－胡萝卜素的广盐性微藻。本文介绍了β－胡萝卜的功能、生产方法、协菏的生物学特征,研究了利用盐藻生产β－胡萝卜素的优化工艺条件和新型光生物反应器的开发应用。     

杜氏盐藻核基质附着区的分离及特征性分析

采用0.5%TritonX 100破碎细胞,15%Percoll分离盐藻细胞核,25mM二碘水杨酸锂(lithiumdi iodosalicylate,LIS)抽提核蛋白,限制酶消化除去结合松弛的DNA,蛋白酶K SDS处理,酚/氯仿抽提,乙醇 沉淀提取核基质附着DNA,限制酶酶切连至pUC18载体上构建MARs文库。随机挑选6个克隆进行体外结 合实验筛选,筛选出一能与核基质结合的克隆,测序分析结果表明该序列具明显的MAR序列特征。     

杜氏盐藻光系统Ⅰ反应中心psaB基因cDNA克隆及系统进化分析

杜氏盐藻是一种抗逆性很强的单细胞真核绿藻,能在010525mol/L 的盐水中生长,繁殖1。杜氏盐藻的突出优点在于培养条件简单,光合自养,抗逆性极佳,这些特性为利用其进行实验提供了便利。本研究通过比较现有已知物种的psaB 基因的氨基酸序列,利用 psaB 基因的氨基酸高度保守序列设计一对简并引物,采用 RT2PCR 从杜氏盐藻中克隆了psaB cDNA 片段,为进一步研究杜氏盐藻A2 亚基的结构与功能以及它的光合机理打下基础,同时通过 psaB 基因的进化分析可以更加深刻地了解杜氏盐藻与其他高等植物以及真核藻类之间的亲缘关系.       

盐藻小G蛋白基因(DsRab)的克隆及在高盐胁迫下的表达分析

采用RT-PCR和RACE技术扩增了杜氏盐藻小G蛋白基因cDNA全长序列(GenBank Accession No.JN989548),命名为DsRab,对其进行生物信息学分析,并通过实时荧光定量PCR方法检测盐胁迫下该基因的表达情况。结果表明,DsRab基因的cDNA全长为1 299 bp,开放阅读框(ORF)为612 bp,编码203个氨基酸,5’非编码区78 bp,3’非编码区609 bp;保守性结构域分析可知编码的小G蛋白有4个GTP/GDP保守结构域,1个效应区、1个羧基端的半胱氨酸结构域和5个Rab亚家族共有的结构域;二级结构预测表明该蛋白有32.02%的α-螺旋,23.65%的伸展片段,44.33%的自由卷曲,三维建模成功;比对分析发现DsRab蛋白与多种生物的Ypt/Rab的氨基酸序列具有较高的同源性。荧光定量PCR结果表明,盐藻在高盐(3.0 mol/L)胁迫下,DsRab基因表达量显著上调,1 h后表达量达到最大值,为正常培养下对照组(0 h)的4.9倍,差异极显著(P<0.01)。     

重金属及微量元素诱导蓝藻形成液泡*

鱼腥藻595(Anabaena sp.595)、织线藻246(Plectonema boryanum 246)和伪枝藻248(Scynetonema hofmanni 248)在两种重金属汞、镉和两种微量元素铜、锌诱导下,藻丝细胞均能发生膨大、液泡化现象。不同重金属及微量元素对3种蓝藻液泡化诱导作用强弱程度不同。汞的诱导作用明显强于镉、铜和锌,0．1μmol／L汞可诱导3种蓝藻发生液泡化。采用压片法均观察到诱导形成的液泡,液泡在相差显微镜下显示为圆球形,基本透明。     

RESPONSE TO SALINITY AND HEAT STRESS IN TWO HALOTOLERANT CHLOROPHYTE ALGAE1

Two new isolates of halotolerant chlorophyte algae from the Salt Plains National Wildlife Refuge in Oklahoma, USA, tentatively identified as Dunaliella sp. Teodoresco and Nannochloris sp. Naumann, were characterized with respect to interaction between growth salinity and short‐term heat tolerance. Cells were cultured at 23–25° C over a wide range of salinity. In both species, salinity alone had little effect on maximum photochemical yield (measured by pulse modulated fluorescence) and integrity of the light harvesting system (77 K fluorescence emission spectra). In contrast, Nannochloris exhibited decreasing growth rate (μ), light‐saturated photosynthetic capacity (P^cell_max), respiration (R_d), light‐harvesting efficiency (α^cell), and chl content with increasing salinity. Cultures were heated for 2 h near their upper temperature limits (41.5° C for Dunaliella and 45° C for Nannochloris grown at 50 psu). Dunaliella was progressively more heat‐tolerant with increasing salinity. Photochemical yield of cells at 100 and 50 psu was inhibited by about 15% and 40%, respectively, and largely recovered within 30 min after return to 23° C. Thermal inhibition of photochemical yield in Nannochloris was about 45% at both 50 and 100 psu, but recovery was slower at 100 psu. At 20 psu, both species were almost 90% inhibited by high temperature and required more than a day to recover. In both species, 2 h of heating increased the PSI:PSII fluorescence emission ratio (714:690 nm) at all salinities. This ratio largely recovered within 24 h in Dunaliella at 50 and 100 psu and partially recovered in Nannochloris at 100 psu, but cells of both species heated at 20 psu were chlorotic the next day.     

海涂围滩养鱼生态系统中藻类种群和生物量的研究

研究了滩涂养鱼生态系统中藻类的种群结构等,结果表明共有7门127种,年平均143.92×106个/L、平均生物量105.60mg/L;藻类种群结构、数量、生物量等存在动态变化;结构上以蓝、绿、硅藻门为主,裸、隐、黄、甲藻门为次要门类,绿藻门的种数最多并且占绝对优势;春季绿、硅藻门的数量和生物量大于蓝藻门,夏季蓝藻门藻数远多于其他藻类,秋季蓝、绿、硅藻门生物量接近;藻类组成与异龙湖接近、生物量较其小,但远超过洞庭湖区高产塘的生物量上限。     

铅锌矿废水中重金属在宽叶香蒲（Typha latifolia）的积累与分布

 本文通过对净化塘系统和对照点的水、土和宽叶香蒲组织的分析结果指出：宽叶香蒲具有较强的忍耐、吸收和积累Pb、Zn等重金属的能力。Pb、Zn等重金属在宽叶香蒲体内的积累数量与分布规律是根>地下茎>叶;老地下茎>嫩地下茎;Pb在叶的分布是老叶>成熟叶>嫩叶。在净化塘系统内,各种重金属在宽叶香蒲根部积累比例最大的是Pb,其次是Zn、Cu、Cd、Ni、Mn。     

WATER USE AND SALT BALANCE IN THREE SALT MARSH SUCCULENTS

Water-use characteristics and potential salt accumulation rates were studied in three halophytes, Salicornia virginica, Balis marítima and Borrichia frutescens, inhabiting a salinity gradient in the high marsh. Xylem pressure potential (ψ_ρ), leaf osmotic potential (ψ_π) and leaf relative water content were measured seasonally in the three species. Species growing on the high end of the salinity gradient developed more negative xylem pressure potentials compared to species growing at lower soil salinities. This trend was also observed for leaf osmotic potentials. Low mean leaf ψ_π (below –15 to –36 bars) and high ash contents (0.27–0.48 g NaCl/g DW) indicated salt accumulation in transpiring tissues. However, calculations of potential salt accumulation, based on rates of transpiration and substrate salinity, suggest that some mechanism of salt exclusion at the roots may be operating.     

用藻类监测和评价图们江的水污染

在图们江上、中游进行了人工基质着生藻类调查和室内栅藻检测,以监测和评价水污染状况。在所采用的野外监测指标中,以着生藻类总数与污染程度的关系最为明显。为便于比较和综合,采用了相对藻类总数(PN) 根据调查结果,提出了可作为图们江污染的指示种类。多样性指数与污染的关系,因受到其他非污染因子的影响,并不稳定。研究结果表明:图们江的主要污染源是尾矿废水(悬浮物)和纸浆废水(悬浮物、色度和有毒物质)。最严重污染江段为菜队到图们江段,次之为南坪江段。       

ION FLUXES AND MODIFICATION OF THE EXTRACELLULAR MATRIX DURING GAMETE RELEASE IN FUCOID ALGAE

A rapid phase transition in the extracellular matrix (ECM) of the reproductive tissues (= receptacles) has been proposed to cause gamete release in fucoid algae. We tested this model with cryoanalytical energy dispersive x-ray microanalysis (EDX) scanning electron microscopy (SEM) of rapidly frozen hydrated receptacles of Silvetia compressa Serrão, Cho, Boo et Brawley that were planed to smooth faces using a cryoultramicrotome. Every receptacle typically contained a region(s) of intracellular accumulation of K and Cl and a region(s) of efflux of K and Cl to the ECMs, regardless of treatment (e.g. calm vs. shaken conditions in the light, anthracene-9-carboxylic acid). Although longitudinal variations in [K] and [Cl] between ECM and cells were common, a tissue within any transverse plane of the receptacle was in the same state of efflux or accumulation of K and Cl. The strongest and most extensive effluxes occurred during the time of irreversible commitment to gamete release (i.e. 2–4 min darkness after light potentiation). A receptacle may oscillate rapidly between global states of accumulation and efflux at this time. The polysaccharide collar between the oogonium and stalk cell expanded during Cl and K efflux into the ECM of the conceptacle and condensed when [K] and [Cl] were low in the conceptacle ECM. During periods of strong efflux associated with gamete release, the collar and other components of the exochiton ruptured. Detached gametangia were guided to the pore by the paraphyses. The ECM of the conceptacle is sulfur rich (226 mM) compared with the ECM of the medulla (8 mM). This analysis supports the importance of osmotic modification of the ECM during gamete release and demonstrates that the receptacle is a dynamic signaling organ.