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题名: 空间飞行及回转器模拟微重力对微藻的生物学效应研究
作者: 陈浩峰
答辩日期: 1999
导师: 刘永定 ; 俞敏娟
专业: 水生生物学
授予单位: 中国科学院水生生物研究所
授予地点: 中国科学院水生生物研究所
学位: 博士
关键词: 暹罗鱼腥藻 ; 突变株 ; 二元生命系统 ; 封闭系统 ; 空间飞行 ; 微重力 ; 空间辐射 ; 回转器 ; 模拟微重力
摘要: 在系统阐述了空间生物学的发展、空间环境条件对生物(尤其是藻类和高等植物)的生物学效应及空间受控生态生命保障系统(Controlled Ecological Life Support System,CELSS)的基础上,本研究利用我国第17颗返回式科学实验卫星提供空间飞行条件,应用由中国科学院水生生物研究所与中国科学院上海技术物理研究所共同研制的“空间通用生物培养箱”提供基本生保条件,共进行了12种微藻的空间飞行,探索了空间环境条件对微藻的存活、种群增长、生理生化、遗传特性的影响及水生生物二元生命微型生态系统的影响。卫星在轨飞行l5天。运用遥测技术对暹罗鱼腥藻(Anabaena siamensis)种群在空间的生长进行了实时观测,在我国首次获得了微藻种群在空间飞行期间的实时生长曲线。卫星返地后,发现培养物的气体组成比例与地面对照有较大差别,且微藻返地后的生长比对照明显减慢,但经继代培养后两者生长速率趋同;返地样品固氮酶活性明显高于对照,但光合放氧速率与光合荧光效率与对照差异不显著。对经过空间飞行的暹罗鱼腥藻进行了普通光镜和荧光标记观察,并测定了返地鱼腥藻与对照的DNA含量,结果发现搭载返地样品与地面对照在藻体在细胞学水平上有较明显差异。基于此发现,在经空间飞行的暹罗鱼腥藻中分离出一株在形态和生理特性上均与出发株不同的突变株。并在实验室中利用回转器(clinostat)提供模拟微重力环境,研究了模拟微重力对暹罗鱼腥藻的生物学效应并与空间飞行真实微重力的作用作了比较。研究发现,该突变株与其野生型及经回转器模拟微重力条件培养的藻株三者的形态、生长、光合作用特性等方面存在着较大差异,突变株藻丝加长、细胞数增多,其异形胞的分布方式及与营养细胞的比率也与野生型有区别;生长速度与光合放氧、固氮酶活性明显高于野生型,而回转器培养的藻株则在形态上与野生型没有差异,其生长速度、光合放氧与固氮酶活性低于野生型藻株。透射电镜观察显示:突变株藻细胞类囊体片层很密集,胞内羧体较多而且体积较大,细胞外物质少,细胞壁相对较薄:对照藻细胞内类囊体片层较为稀疏,胞内羧体较少而且体积较小,且胞外有较多的胞外物质积累,细胞壁相对较厚;经回转器处理的藻细胞则与对照藻细胞相似。比较了三者的色素吸收光谱和光合作用荧光发射光谱,结果表明,突变株在藻胆蛋白向光系统I及光系统II传递能量的效率方面比野生型高,其两个光系统的活性也高于野生型。回转器的模拟微重力则使暹罗鱼腥藻能量传递效率降低,而且抑制了两个光系统尤其是光系统I的活性。运用200个10核苷酸随机引物对暹罗鱼腥藻突变株与野生型的基因组DNA进行了RAPD分析,发现在扩增出的1000余条清晰可辨的条带中,在两者之间有8个引物的扩增产物中出现21个多态性差异条带。从而证实了空间突变株在遗传水平上与出发株存在差异。研究结果说明,空间环境存在使生物产生变异的因素,产生突变的原因可能是空间环境各种因素如宇宙辐射、微重力等等因素的综合作用,而微重力本身的作用只改变微藻的生长状况、生理特性及代谢状况。利用返回式卫星搭载的“空间通用生物培养箱”进行了水生生物二元微生态系统(异养藻-自养藻系统)的空间结构与功能的实验研究,采用二个类型的微藻混合生长封闭系统。第1种组合Poterioochromonas sp.(兼性营养型)+Chlorella pyrenoidosa(自养型)运行良好,系统中的气体组分与大气气体组分相近:第2种组合Collodictyon sp.(异养型)+Microcystis sp.,Synechococuus sp.,Carteria sp.,Oscillatoria sp.(均为自养型)基本运行正常,其中的异养藻仍具有捕食性。该微生态系统的成功搭载为空间CELSS的研究提供了基础资料。
英文摘要: A review on the development and outlook of space biology and controlled ecological life support system (CELSS), the biological effects of space environmental elements such as microgravity, space radiation etc. on microalgae and higher plants was presented. 12 strains of microalgae were chosen to be carded in the 17~(th) retrievable scientific experimental satellite of China, and traveled in space for 15 days to study the influences of space environment to their survival, population growth, physiological and biochemical characters, heredity characters and the structure and functions of artificial hydro-biological micro-ecosystems. The tested organisms were loaded in a newly developed instrument to provide basic life support conditions for the organisms (temperature control and illumination), the "multipurpose biological incubator", which was invented and made by the Institute of Hydrobiology and Shanghai Institute of Technological Physics, Chinese Academy of Sciences. The instrument was installed in the retrievable cabin of the satellite during spaceflight and retrieved when the satellite came back to earth. By means of the remote sensing equipments installed in the satellite, we gained the instant growth curve on Anabaena siamensis (Blue-green alga or cyanobacterium) population during spaceflight, which was the first time in the studies of space biology in China. The curve has its own characters and the growth rate was slower than the ground control. After retrieval, the gas composition of the spaceflight culture system has high 0_2 and high CO_2 radio, it was quite different from the ground control; Inoculation of the spaceflight microalga showed that the growth rate was distinctively slower than the ground control, but after 6 days of culture, using the cultured microalga again as inoculation, indicated the growth rates were similar. The activity of nitrogenase of the spaceflight A. siamensis was about 2 times higher than the control, but the rate of oxygen evolution and photosynthesis fluoresce were of no difference. The comparative observation of cytological structure of A. siamensis was made between spaceflight and control. It has been studied by the methods of light microscopic observation, electron microscopic observation and fluorescence dyeing observation, and the DNA contents of retrieved A. siamensis and control was also measured. Obvious difference was observed between the spaceflight strain and the ground control. And a strain of mutant (AsSR10) was selected from A. siamensis after spaceflight, which has different morphological and physiological characters with the wild type strain. The morphological character and some physiological characters such as growth rate, nitrogenase activity, photosynthetic O_2 evolution and fluorescent efficiency of photosynthesis were studied between the mutant, the wild type cells and the cells cultured by clinostat (under the condition of simulated microgravity). Morphologically, The mutant's filament is much longer than the control and has different heterocyst distributing pattern. Electron microscopic observation showed that the layers of thylakoid are denser in the mutant than in the wild type, there is more outer cellular substances at the outside of the wild type cell wall than the mutant, and the cell wall of mutant is thinner than that of control. The spaceflight mutant is much higher than the control in growth rate, photosynthetic 0_2 evolution and nitrogenase activity. But the Anabaena strain cultured in clinostat has no difference in morphological character, and it is lower than the control in growth rate, photosynthetic 0_2 evolution. The absorption spectra and low temperature fluorescence emission spectra of mutant, wild type and clinostat cultured strain of A. siamensis were also studied, the absorption spectra showed that there is no difference in the absorbing peaks of absorption spectra between the mutant and the wild type cells, but the strain cultured in clinostat had relative low chlorophyll absorbing peak. The emission spectra showed that the energy transmission efficient from phycobiliprotein to photosystem I and photosystem II was higher in mutant A. siamensis cells than that of control, and the activity of the 2 photosystems are also higher than the other two. And alga cells cultured in clinostat showed low energy transmission efficiency from phycobiliprotein to the 2 photosystems and the activity of PSI and PSII was lower than control, especially PSI. The polymorphism of genomic DNA between the mutant and the wild type was studied by RAPD analysis, 200 random primers were used. In the total 1016 amplified electrophoresis bands, 21 polymorphic bands were discernible between the mutant and wild type DNA, the polymorphism was around 2%, that is to say, there was changes at the genetic level in Anabaena siamensis after space flight. A conclusion may be drown from former phenomena that there are environmental elements in space which changed the microalgae at physiological and genetic level. After exclude other elements, the 2 main elements are microgravity and space radiation. After the comparative study between real and simulated microgravity, it can be inferred that microgravity can only change the situation of growth and metabolic characters, it is the comprehensive effect of the radiation and microgravity who changed the genetic character of Anabaena siamensis. Two artificial micro-systems (heterotrophical microalgae and autotrophical algae) were chosen to conduct a study on their structure and function in Space, they were divided into two groups: 1. Poterioochromonas sp.+ Chlorella pyrenoidosa, and 2. Collodictyon sp. + Synechococus sp., Microcystis sp., Carteria sp., Oscillatoria sp. the first group nm very well, its gas compositions were similar to that of atmosphere on earth, the 2~(nd) one was also normal in space. The food-chain relationship between the heterotrophical and the autotrophical algae was set-up well.
语种: 中文
内容类型: 学位论文
URI标识: http://ir.ihb.ac.cn/handle/342005/12526
Appears in Collections:中科院水生所知识产出(2009年前)_学位论文

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Recommended Citation:
空间飞行及回转器模拟微重力对微藻的生物学效应研究.陈浩峰[d].中国科学院水生生物研究所,1999.20-25
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