发状含珠藻藻殖段的分化及其光合特性的研究

发状念球藻（Ｎｏｓｔｏｃ ｆｌａｇｅｌｌｉｆｏｒｍｅ Ｂｏｒｎ．ｅｔ Ｆｌａｈ．）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（铺垫闰遮光）,红光或在白光下向培养基中加入ＤＣＭＵ（３,４－ｄｉｃｈｌｏｒｏｐｈｅｎｙｌ－１,１－ｄｉｍｅｔｈｙｌｕｒｅａ）等藻丝体与藻殖段的光合特性,表明营养营丝体和藻殖段的可见光吸收光变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     

念珠藻葛仙米的藻殖段分化——信号转导参与过程的初步研究

钙调素拮抗剂W_7和FPZ均可抑制葛仙米藻殖段的分化。利用FPZ作为荧光染色剂发现 ,经过钙调素拮抗剂处理的藻体有较低的钙调素分布。在达到一定浓度后 ,金属离子螯合剂EGTA也抑制藻殖段分化。金霉素 (CTC)荧光显示 ,经EGTA处理的藻细胞其膜钙分布大大减少。但实验表明 ,藻殖段得以正常分化却是多种金属离子共同参与的结果。一氧化氮 (NO)可以大大降低葛仙米藻殖段的分化率 ,一氧化氮合成酶 (NOS)的抑制剂NNA也可以抑制藻殖段的分化。外源地加入氧自由基抑制了藻殖段的分化 ,但是氧自由基的清除物过氧化氢酶 (CA)也使藻殖段分化完全受抑制。以上结果初步表明 ,葛仙米藻殖段分化是个复杂的、信号转导参与的过程     

念珠藻葛仙米的藻殖段分化——信号转导参与过程的初步研究

钙调素拮抗剂W-7和FPZ均可抑制葛仙米藻殖段的分化。利用FPZ作为荧光染色剂发现,经过钙调素拮抗剂处理的藻体有较低的钙调素分布。在达到一定浓度后,金属离子螯合剂EGTA也抑制藻殖段分化。金霉素（CTC）荧光显示,经EGTA处理的藻细胞其膜钙分布大大减少。但实验表明,藻殖段得以正常分化却是多种金属离子共同参与的结果。一氧化氮（NO）可以大大降低葛仙米藻殖段的分化率,一氧化氮合成酶（NOS）的抑制剂NNA也可以抑制藻殖段的分化。外源地加入氧自由基抑制了藻殖段的分化,但是氧自由基的清除物过氧化氢酶（CA）也使藻殖段分化完全受抑制。以上结果初步表明,葛仙米藻殖段分化是个复杂的、信号转导参与的过程。     

发状念珠藻对盐胁迫的响应

探讨了发状念珠藻(NostocflagelliformeBornetFlah)对盐胁迫的耐受适应机制,采用含不同浓度NaCl(0、01、02、04、06、08、10mol/L)的BG110培养液处理具有正常生理活性的丝状体,25±05℃,40μmol/m2/s下照光培养12h,测定藻体光合作用、呼吸作用等生理活性以及体内一些物质的含量,结果表明:随培养液中NaCl浓度的升高藻体光合作用、呼吸作用以及PSⅡ活性(Fv/Fm)降低;质膜透性不断增大,丙二醛含量升高,自由水含量、自由水/束缚水比值下降,类胡萝卜素、可溶性糖含量增加,脯氨酸含量变化不大。由此可知,盐胁迫下发状念珠藻正常生理活性受到抑制而表现出一定的抗逆能力;该藻对盐胁迫具有一定的耐受能力,类胡萝卜素的增加有助于清除藻体内的氧自由基,可溶性糖可能是其主要渗透调节物质之一,脯氨酸在盐胁迫中的渗透调节作用不大。       

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.     

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。     

发状念珠藻干旱胁迫响应基因NfGR的克隆与鉴定

该研究采用PCR技术,从发状念珠藻细胞中克隆了谷胱甘肽还原酶(glutathione reductase,GR)基因,命名为NfGR,其开放阅读框长1 374bp,编码458个氨基酸,蛋白相对分子量为49.42kD,理论等电点为5.49。氨基酸序列分析表明,NfGR蛋白具有NADPH结合位点超家族(NADB-Rossmann superfamily)和吡啶氧化还原酶二聚体超家族(Pyr_redox_dim superfamily)2个结构域,与点形念珠藻(Nostoc punctiforme)的相似性达93%。系统进化树分析表明,NfGR与点形念珠藻处在同一进化枝上,亲缘关系较近。qRT-PCR表达分析表明,在不同浓度PEG-6000处理下,NfGR基因均保持上调表达,其中,PEG-6000浓度为8%时,NfGR基因的相对表达量达到峰值(32.69)。研究推测,谷胱甘肽还原酶可能参与了发状念珠藻对干旱胁迫过程的响应。     

丝状体蓝藻藻殖段的分化及其调节机制

本文介绍了丝状体蓝藻(亦称蓝细菌) 的藻殖段的分化及其调节机制。藻殖段与正常藻丝体的区别在于细胞形状、细胞内存有气囊和可移动的短而直的藻丝链等。本文对许多环境因子包括光和营养因素等促进或抑制藻殖段的分化进行了讨论;还介绍了念珠藻(Nostoc) ,单歧藻(Tolypothrix) 和眉藻(Calothrix)所具有复杂的细胞发育过程,即具气囊又可移动的藻殖段分化,异形胞分化以及营养细胞的补偿性色适应。这三种细胞类型的适应形成取决于两种不同的光受体系统。藻殖段和异形胞两者的分化可能取决于光合电子传递链;而营养细胞的补偿性色适应则受光敏色素的调节。此外,谷酰胺合成酶合成和活性调节的PII蛋白,在协同藻殖段分化、异形胞分化及营养细胞的补偿色适应中起重要作用。由于蓝藻藻殖段分化及其调节机制是一个新的研究领域,关于它的知识尚不完整,亟待人们加强研究。     

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.     

不同培养基对发菜细胞生长和光合活性的影响

研究测定了发菜(NostocflagelliformeBorn.etFlah.)细胞在不同培养基中的生长速率、光合作用和叶绿素荧光活性。结果显示培养11d后:Detmer培养基中叶绿素a的含量为1.08mg/L,Kratz-Myers培养基中叶绿素a的含量为1.87mg/L,水生104号培养基中叶绿素a的含量为1.21mg/L,BG11培养基中叶绿素a的含量为2.18mg/L,表明在BG11培养基中培养的细胞具有最高生长速率;与另外4种不同浓度的BG11培养基相比,上述BG11培养基培养的发菜具有最大的光合速率Pm(218.1μmolO2.mg-1chla.h-1)和最高的PSII光化学效率(Fv/Fm=0.349)。实验结果表明,BG11是适合发菜生长的培养基,对其光合作用和叶绿素荧光活性具有显著促进作用。     

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.     

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.     

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.     

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.