Effect of wind speed on loss of water from Nostoc flagelliforme colonies

The effects of wind speed on loss of water from N. flagelliforme colonies were investigated indoors in an attempt to assess its ecological significance in field. Wind enhanced the process of waterloss; the half-time of desiccation at wind speeds of 2.0 and 3.4 m s^-1 was, respectively, shortened to one-third and one-fifth at 20°C and, to one-sixth and one-eighth at 27°C that of still air. Photosynthetic efficiency was not affected before the wet alga lost about 50% water. This revised version was published online in June 2006 with corrections to the Cover Date.     

PHOTOSYNTHETIC BICARBONATE UTILIZATION BY A TERRESTRIAL CYANOBACTERIUM, NOSTOC FLAGELLIFORME (CYANOPHYCEAE)

The photosynthetic characteristics of the terrestrial cyanobacterium, Nostoc flagelliforme , after complete recovery by rewetting, was investigated to see whether it could use bicarbonate as the external inorganic carbon source when submerged. The photosynthesis–pH relationship and high pH compensation point suggested that the terrestrial alga could use bicarbonate to photosynthesize when submerged. The photosynthetic oxygen evolution rates were significantly inhibited in Na ⁺ -free and Na ⁺ + Li ⁺ media but were not affected by the absence of Cl ⁻ , implying that the bicarbonate uptake was associated with Na ⁺ / HCO₃ ⁻ symport rather than Cl ⁻ /HCO₃ ⁻ exchange system.     

发状念珠藻藻殖段的分化及其光合特性的研究

发状念珠藻 (NostocflagelliformeBorn .etFlah .)存在着两个重要而明显的个体发育阶段 ,即营养藻丝体和藻殖段。采用弱光 (铺垫砂粒遮光 ) ,红光或在白光下向培养基中加入DCMU (3,4_dichlorophenyl_1,1_dimethylurea)等方法 ,可促进营养藻丝体转变成藻殖段。用可见光吸收光谱、低温荧光发射光谱和光合放氧活性表示发状念珠藻藻丝体与藻殖段的光合特性 ,表明营养藻丝体和藻殖段的可见光吸收光谱和色素含量差别不大。而两者在不同光强范围 (110～ 12 0 0 μmol·m-2 ·s-1)和不同温度 (15～ 45℃ )下的光合放氧活性 ,表明发状念珠藻的藻殖段比营养藻丝体可能更适合在低光强下和较高的温度下生长。从荧光发射光谱可以看出 ,在光合能量传递中营养藻丝体比藻殖段在两个光系统之间的光能分配上更加均衡 ;但是藻殖段中藻胆体吸收光能向两个光系统的传递比营养藻丝体的更加有效。可以认为藻殖段的形成对光合作用的结构与功能产生影响。     

发状念珠藻藻殖段的分化及其光合特性的研究

Hormogonia of Nostoc flagelliforme is one of the developmental stages in the life cycle of cyanobacterium. High yields of pure hormogonia were obtained by weak light (the filaments were covered by sterilized sand for blocking light), red light, white light plus DCMU (3, 4-dichlorophenyl-1, 1-dimethylurea) in the culture. These pure fractions of hormogonia allowed the study of physiological measurements in comparison to vegetative filaments. The photosynthesis in the hormogonia and the vegetative filaments was characterized by fluorescence emission spectra at 77 K, absorption spectrum and oxygen evolution. Absorption spectrum of the hormogoia and vegetative filaments did not reveal difference. The data indicated the similarity of pigment contents between hormogonia and vegetative filaments. Some differences were observed in oxygen evolution of vegetative filaments and hormogonia in the temperature range of 15 ℃ to 45 ℃ and light intensity around 110 μmol·m-2·s-1 to 1200 μmol·m-2·s-1. The fluorescence emission spectra showed that energy distribution between the two photosystems in mature colonies was more balance than in hormogonia. The absorption of light energy in phycobilisomes and the transfer to the two photosystems in the hormogonia were more effective than in the mature colonies. It may be concluded that the formation of hormogonia affected on the structure and function of phytosynthesis.     

Dried field populations of Nostoc flagelliforme (Cyanophyceae) require exogenous nutrients for their photosynthetic recovery

The effects of nutrients on the photosynthetic recovery of Nostoc flagelliforme during re-hydration were investigated in order to see if their addition was necessary. Net photosynthesis was negligible in distilled water without nutrient-enrichment. Addition of K⁺ resulted in significant enhancement of net photosynthesis, whereas other nutrients (Fe³⁺, Mg²⁺, Na⁺, NO₃ ^-, PO₄ ^3-, Cl^-) and trace-metals (A₅) showed little effect. The recovered net photosynthetic activity increased with the increased K⁺, and reached the maximum at concentrations above 230 μM. Desiccation and re-hydration did not affect the dependence of photosynthetic recovery on K⁺. It was concluded that dried field populations of N. flagelliforme require exogenous addition of potassium for photosynthetic recovery and that growth may be potassium-limited in nature. This revised version was published online in August 2006 with corrections to the Cover Date.     

发菜细胞培养物对盐胁迫的响应

用不同浓度(0、0.1、0.2、0.4 m o l.L-1)的N aC l处理BG 110培养的发菜细胞,结果显示,发菜光合速率与叶绿素荧光强度随N aC l浓度的升高先增加后降低,当N aC l浓度为0.1 m o l.L-1时光合速率与叶绿素荧光具有最大值,表明发菜细胞培养物能耐受一定浓度的盐胁迫.以BG 110+0.4 m o l.L-1N aC l为对照,在BG 11+0.4m o l.L-1N aC l的胁迫实验中,光合速率与叶绿素荧光强度下降较慢;丙二醛、脯氨酸含量较低;类胡萝卜素含量较高,表明在培养液中添加外源硝酸盐后可以缓解N aC l对发菜细胞培养物的生理胁迫效应,增强其抗盐性.     

Paraphysomonas corbidifera sp. nov., a marine,colourless, scale-bearing member of the Chrysophyceae

Paraphysomonas corbidifera sp. nov. has a spherical body bearing two flagella of unequal length, the longer possessing two rows of mastigonemes, the shorter smooth. The body is covered in scales of one basic form, open crown-like structures. The organism is compared with other members of the genus.     

发菜光合特性与水分关系的研究

处于自然光照与室内散射光照和2种不同供水时间 8∶00与20∶00 条件下的发菜藻殖段,用Li-6200光合仪作其光合速率日变化测试,结果得出:自然光照与室内散射光照及2种供水时间对发菜光合速率、蒸腾速率、温度、水汽压亏缺等因子之间的影响关系.自然光照条件下,对发菜20∶00供水,在次日的发菜光合速率日变化中,光合速率与水汽压亏缺呈显著性正相关关系 r=0.6216 .发菜光合速率日变化曲线与水汽压亏缺日变化曲线趋于一致,当光照强度为1324μmol·m-2·s-1时,水汽压亏缺达到44.5mb,发菜光合速率日变化出现峰值 1.68mg·g-1·s-1 .对发菜8∶00供水,其光合速率日变化在供水5h后出现峰值;在室内散射光照条件下,在8∶00和20∶00供水都使发菜在光合速率日变化中,保持了较高的相对含水量,光合速率降低,并处于较长时间的呼吸作用状态.     

Effect of Temperature Water Content and Light Intensity on Nitrogenase Activity of Nostoc flagelliforme

The optimal temperature for the nitrogenase activity in the terrestrial cyanobacterium N. flagelliforme was 21–28℃; the optimal water content in thallus was 1000--1500%; the light saturation was between 150–200 J·m-2·s-1. The thallus of N. flagelliforme is extremely sensitive to higher temperature in wet. Long-term exposure of wetted thallus to high temperature at 45℃ causes rapid declination of its nitr0genase activity to zero. Under dry condition, N. flagelliforme is extremely resistant to extensive desiccation and heat exposure. Dry thalli exposed to 55℃, 5 hours daily for 21 days, show no marked change in its nitrogenase activity. The thalli preincubated in wet condition for 4–5 days, are highly sensitive against desication. However, repeated drying/wetting cycles induce a slow and gradual increase of its nitrogenase activity and improve the resistance of its nitrogenase activity against desiccation. High concentrated NaC1 salt solution (0.17–0.43 mol/L) depletes nitrogenase activity of the thalli quickly. Above result shows that N. flagelliforme is not able to resist against salt. The physiological characteristics of nitrogen fixation of cyanobacterium N. flagelliforme may be eonsidered as a result of drought adaptation of the terrestrial ecological condition aad the drying westting cycle is perhaps a necessary factor to maintain its growth.     

发菜(Nostoc flagelliforme)培养条件的研究

研究了光照强度、日供水次数、CO2浓度和培养基成分对发菜生长的影响。结果显示,中度光强(114μmol.m-2.s-1)下发菜生长最快;发菜的生长基本同供水次数成正相关系;CO2浓度的升高并没有显著促进发菜生长,低光条件下(57μmol.m-2.s-1),高浓度的CO2(2800μL/L)抑制了发菜的生长;用BG11培养的发菜生物量的增长显著高于用BG110培养的;BG11培养基中K+和CO32-的缺失并没有显著影响发菜的生长。     

Growth characteristics of Nostoc flagelliforme at intermittent elevated CO2 concentrations

Algal cultivation is a potential candidate for CO₂ mitigation. CO₂ plays important roles in mass cultivation of algae, including supplying carbon source and adjusting medium pH. To assess the possibility of using edible cyanobacterium Nostoc flagelliforme as carbon storage device, the growth characteristics of N. flagelliforme batch cultured under elevated CO₂ concentrations (0, 2.5, 5, 20, and 40%) were investigated in this study. Results showed that the net photosynthetic rate, efficiency and carbon sequestration rate at 20% CO₂ were increased at a maximum of 121 μmol O₂ (mg chla)^?1 h^?1 8.40% and 0.17 g CO₂ L^?1 day^?1, and increased by 0.42, 1.03 and 1.13 folds compared with that of the control, respectively. Higher CO₂ concentration resulted in the declines in photosynthetic rate, efficiency and carbon sequestration rate because of medium pH reduction. Accordingly, the dry cell weight, amount of exopolysaccharides and protein content of N. flagelliforme cells at 20% CO₂ were obtained at a maximum of 1.45 g L^?1, 54.98 mg L^?1 and 57.75%, increased by 0.93, 0.29 and 0.8 folds compared with that of the control, respectively. These results provided important information for CO₂ mitigation by N. flagelliforme and would shed more light on elucidating the mechanisms of CO₂ tolerance in cyanobacterium.     

Studies on Photosynthesis,Respiration and Morphology of Nostoc flagelliforme

Nostoc flagelliforme Born. et Flah. is one of the terrestrial cyanobacteria naturally distributed in arid and semi-arid areas in the Northern and the North-western parts of China. The cyanobacterium is an edible delicacy with special medical value. However, commercial N. flagelliforme has nov been collected from the field only. For cultivation of this valuable cyanobacterium, it is necessary to understand how it grows and how it adapts to the environment.The experimental material was collected in Siziwangqi of Nei Monggol. The effects of light intensity, temperature, pH, salinity, length of thallus and rewetting on photosynthesis and respiration of N. fiagelli[orme were measured using an oxygen electrode. The results were as follows: The photosynthetic light compensation point was around 40–90μmol photons·m-2·s-1, the light saturation point was 1200μmol photons·m-2·s-1, and no photoinhibition appeared when the light intensity was increased to 1800μmol photons· m-2·s-1. N. fiagelliforme exhibited its photosynthetic and respiratory activities in the temperature range of 5–45℃. The optimum temperature for its photosynthesis was 25℃ and that for respiration was 35--40℃. Between pH range of 4.5–9.5 N. flagelliforme had photosynthetic activity and respiratory activity at pH range of 4-10, with optimum pH for photosynthesis at 7.5 and for respiration at 7.5–8.0. N. flagelliforme exhibited maximum net photosynthesis in 0.15mol/L of NaC1 in BG-11 medium. When the salinity was increased to 0.9 mol/L the net photosynthesis dropped down to zero. Respiration decreased concordantly with the increasing salinity as well. Maxima photosynthesis and respiration was also detected when the thallus of N. flagelliforme reached a length of 0.5cm and aftewords the more the length the less the activities. The recovery time attaining to the maximum photosynthesis and respiration activities after rewetting was dependent on storage time in dryness. The cyanobacterial mats after being reserved for 3 months, attained its maximum photosynthesis by 0.5h after rewetting, and that being reserved for 18 months needed 3.5h after rewetting. For respiration, the mats reserved for 3 months and 18 months required 5 minutes and lh after rewetting, respectively to attain its maximum. Under scanning electron microscope, cells of N. flagelliforme were wrapped up within a gluey sheath, and usually attached closely to each other in pairs and the filaments were uni-trichome with branches in some cases. The surface of thallus tip was rougher than other parts which meant that the tip portion had greater surface area beneficial to water absorption and cell growth.     

念珠藻发菜的水分生理特性

对野生风干发菜水分生理指标进行了测定与分析,结果表明:风干含水量为7.3%～11.2%,饱和含水量为616.0%～1258.4%,相对含水量1%左右,水分饱和亏为99%左右,水势为-8.2MPa;最大吸水速率为8.029gH2O·g-1·min-1,饱和吸水后其体积膨大10～14倍,平均吸胀率为1286.9%,其中伸长率为23.5%,增粗率为235.0%;导水力极弱;干燥发菜具有超强的吸水力,能在相对湿度>28.4%的空气中吸取水分;发菜的保水力随环境而变,在阴暗或弱光照的环境中具有较强的保水力.另外,探讨了不同种源发菜的水分生理指标与环境因子的关系.     

Energy Transfer from Phycobilisomes to Photosystems of Nostoc flagelliforme Born. et Flah. During the Rewetting Course and Its Physiological Significance

During the non-frost season, the condensation of dew makes Nostocflagelliforme Born. et Flah., a highly drought-tolerant terrestrial cyanobacterium, frequently undergo rehydration-dehydration.Rehydration begins in the dark at night. After rewetting in the dark, photochemical activity and the structure of photosystem (PS)II were not recovered at all; the structure of PSI, energy transfer in phycobilisomes, and energy transfer from phycobilisomes to PSI were recovered within 5 min, as in the light. The recovery of energy transfer from phycobilisomes to PSII was light dependent and energy transfer from phycobilisomes to PSII was only partially recovered in the dark. These results suggest that the two-trigger control (water and light) of photosynthetic recovery may make IV, flagelliforme avoid unnecessary energy consumption and, at the same time, the partial recovery of energy transfer from phycobilisomes to PSII in the dark could help N. flagelliforme accumulate more photosynthetic products during the transient period of rehydration-dehydration.     

水分对发状念珠藻生理活性的影响

研究了水分对发状念珠藻(Nostor flagelliforme Born.et Flah.)生理活性的影响作用。结果表明：干藻体在湿润的过程中,呼吸、光合和固氮活性依次恢复;且随水分含量的增加,光合活性和固氮活性逐渐增强,呼吸作用缓慢减弱并在一定水平上保持相对稳定,自由水是束缚水8倍左右时发菜生理活性全面恢复。吸水饱和的藻体在干燥过程中,光合、呼吸、固氮作用依次停止;呼吸作用随水分的丧失逐渐下降;固氮活性、光合活性在水分丧失20％～40％时有一定程度的增强,出现活性高峰;此后,生理活性下降,水分完全丧失时,光合作用终止,呼吸和固氮作用极其微弱。试验证明,水分是发状念珠藻生理活性的重要限制因子,适宜的水分有助于发菜维持正常的生理代谢和生长。     

发状念珠藻胞外多糖的纯化与性质分析

采用DEAE阴离子交换层析和Sephadex G100凝胶层析对液体悬浮培养发状念珠藻胞外多糖进行纯化, 得到两个组分NFPS1和NFPS2。对组分NFPS2进行理化性质分析, 并与野生发状念珠藻多糖NFPS0的性质进行对比。结果表明二者具有相似的单糖组成, 均为葡萄糖、木糖、半乳糖、甘露糖; 表观分子量分别为2.79×105、2.26×105; 均不含核酸、蛋白质等物质, 是非硫酸化多糖; 有较高的热稳定性, 其降解温度在245oC左右。但在微观结构上, 两者存在一定差别。     

发状念珠藻胞外多糖的纯化与性质分析

采用DEAE阴离子交换层析和Sephadex G100凝胶层析对液体悬浮培养发状念珠藻胞外多糖进行纯化, 得到两个组分NFPS1和NFPS2。对组分NFPS2进行理化性质分析, 并与野生发状念珠藻多糖NFPS0的性质进行对比。结果表明二者具有相似的单糖组成, 均为葡萄糖、木糖、半乳糖、甘露糖; 表观分子量分别为2.79×105、2.26×105; 均不含核酸、蛋白质等物质, 是非硫酸化多糖; 有较高的热稳定性, 其降解温度在245oC左右。但在微观结构上, 两者存在一定差别。     

Observations on Microscopic Structure and Ultrastructure of Nostoc flagelliforme

Using the Iight mieroseope, scanning and transmission eleetron microscopes, it is found that each individuaI Nostoe flagelliforme is composed of hundreds and thousands torulose cells. The filaments are uni-triehome with branches in some cases. The thallus is surrounded by a mueoid sheath with different thicknesses, rough surface, and numerous cracky appearance. The Nostoc flagelliforme cells are typieal prokaryote eells ill which there is no discrete nuclear body. However nueleoplasm exists. occupying the center of the ceils. The cell wall of this prokaryote ceil is three-layered. Bet, when the nueleoplasm and the wall, a great quantity of thylakoids disperse. Moreover, structured granules, polyhedral and polyphosphate bodies intersperse. All of vegetative eells seem to be able to divide. The reproductive manner is of horizontal split, i.e. the regenerated new cell walls expand from periphery towards the center, then the mother cells eonstriet and finally split into two daughter cells. In our opinion, the function of this structural characteristic of heteroeysts is to endure extreme envi-ronment, so that they are able to maintain their species progeny.     

发菜NdhK的抗体制备与蛋白表达

发菜是一种陆生蓝藻,分布于一些干旱和半干旱区域。其NADPH脱氢酶(NDH-1)是一种重要的光合膜蛋白复合体,参与CO₂吸收、围绕光系统Ⅰ的循环电子传递和细胞呼吸。为研究该物种中ndhK基因的功能,本研究利用特异性引物,通过PCR方法从发菜中扩增ndhK基因并克隆到原核表达载体pET-32a上,得到表达载体pET-32a-ndhK,将其转入大肠杆菌BL21(DE3),经异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,得到分子量大小为43 kDa的融合蛋白NdhK。随后,采用亲和层析,对融合蛋白进行纯化回收,并以此回收蛋白作为抗原进行免疫,制备NdhK的多克隆抗体。最后,利用Western blot蛋白免疫印迹对所得抗体的特异性进行验证。从而为进一步探索发菜ndhK基因的功能以及发菜中NDH-1复合体各亚基的作用进行前期准备。     

发菜蛋白质组双向电泳技术的建立及优化

为建立适用于发菜(Nostoc flagelliforme)蛋白质组研究的双向电泳技术,对发菜蛋白质的提取、裂解、上样量、IEF及SDS-PAGE电泳等关键步骤进行了优化,结果显示:发菜蛋白质主要分布在pH 4～7范围内,采用改良TCA法可提高提取液中蛋白质的含量和双向电泳图谱的分辨率,裂解液含60 mmol/L DTT,24 cm IPG胶条上样量1.5 mg时不仅提高了蛋白质的溶解性,而且改善了双向电泳的分离效果,得到近800个蛋白点,且蛋白点清晰,图谱分辨率较好.采用优化后的双向电泳体系提高了发菜蛋白质双向电泳的分辨率和重复性,建立起一套适用于发菜蛋白质组分析的双向电泳方法.