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题名: 蓝藻的碳、氮代谢-多变鱼腥藻(Anabaena variabilis)的固氮酶诱导合成、活性变化与细胞内含糖原和碳代谢的关系
作者: 刘永定
答辩日期: 1989-03-18
导师: 黎尚豪
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
摘要: 在细胞生物学的基础上综述了蓝藻(蓝细菌)碳和氮的代谢生理。采用BioFlo型恒化器和小型全玻璃发酵罐两种类型的连续培养装置成功地建立了异形胞蓝藻多变鱼腥藻(Anabaena variabilis ATCC 29413)的半稳定状态连续培养,并用于生理学研究。在光一暗周期(12:12)条件下,初始培养物经5-6天达到半稳定状态。此时,培养物的生理活性保持稳定,生长参数符合方程X = X-0e~((μ-D)t)所表述的关系。由于光一暗周期交替的影响,种的生长速度和细胞组分的增长都呈周期变化,而且变化速度各异。在光照时间内,由于CO_2同化和固氮作用,生物量增长,在黑暗时间内,由于连续稀释和呼吸作用,生物量下降。生物量在光照时间内增加的速度与黑暗时间内下降的速度在数值上几乎相等。生物量在光照时间内的增加主要是由于糖原的合成和累积。光照时间内糖原可累积到干重的20%,而后在黑暗中消耗到占干重的1-2%。糖原在光照时间过程内合成的速度是不一致的,光照开始时最高(2.6 μg.μg -1 chl . h~(-1))。供给通常空气浓度(0.03%)的CO_2,实验条件下的培养物受CO_2限制;提高CO_2供应浓度到1%,生物量增加到原来的1.6-1.8倍,培养物的糖原浓度相增加。在从稳定的低CO_2培养到高CO_2培养之间的过渡期内,糖原的合成速度增加很快,开始达6.1μg.ug-1chl.h~(-1),含量达干重的56%,以后逐日下降,直到新的半稳定状态生长。据此,推断了细胞对无机碳的亲和力,讨论了细胞从外向内转运CO_2的可能的结构模式。外源供铵,加速培养物生物量的增长,但糖原累积减少,同时外源供给NH_4~+和加富CO_2,使培养物的生物量和糖原总量提高,但细胞内糖原累积不能提高。光照开始前固氮酶活性很低,随着光照开始,固氮酶活性迅速增加。光诱导的固氮酶活性在光照开始后第4至第6小时达到最大值,然后由于酶的周转而活性下降。异形胞蓝藻多变鱼腥藻只是在光照时间刚开始的一段时间内合成固氮酶。光照开始时加入铵或氯霉强烈地抑制固氮酶活性。在整个光一暗周期内,样品于无光条件下测试到的固氮酶活性只有照光测到的数值的约五分之一。没有发现黑暗时间内固氮酶活性与糖原含量的直接关系,光合作用产生的糖原不决定黑暗时间内的固氮酶活性。固氮酶活性也根据全氮量的增加进行了计算,由形成铵所需的电子数说明没有游离氢产生。90%以上的固氮发生在光照时间内,多变鱼腥藻的氮同化基本上是与光照过程相关连的。在整个光一暗周期内没有发现生物量的含氮量有显著变化,可是提取的蛋白质的含氮量在光照时间内有增加,在黑暗时间减少即光照时间内同化的氮暂时以含氮较多的蛋白质形式贮存起来。本文所报道的半稳定状态连续培养在德意志联邦共和国康斯坦茨大学是首次建立,在我国是新的方法。有关异形胞的多变鱼腥藻在不一暗周期条件下糖原合成、累积、降解固氮酶合成与活性变化及蛋白质含氮量与氮同化的关系的结果是首次报导。对于藻类生理学、生理生态学和藻类培养与利用生物技术具有意义。
英文摘要: A brief review of the carbon and nitrogen metabolisms in blue-green algae (cyanobacteria) had is been presented on the basis of cell biology. Semi-steady state continuous culture of Anabaena variabilis ATCC 29413 (a heterocystous blue-green alga) had been was set up and was applied to the physiological research using two types of apparatus, i.e., a BioFlo chemostat and a small whole-glass fermentor. Under a light-dark (12:12) regime the culture reached to the semi-steady state in 5-6 days. Physiological activities of the cells maintained on a stable level, growth parameters of the culture met the relationship expressed in the equation X = X_oe~(μ-D)t. Specific growth rate and the change rates of cell components followed the light-dark cycle and were different from each other. During the light period biomass increased due to CO_2-assimilation and nitrogen-fixation, due to the continuous dilution and respiration it decreased approximately at the same rate in the dark. The increase of biomass in the light was mainly due to the synthesis and accumulation of glycogen. Glycogen accumulated up to 20% of biomass at the end of the light period, then degraded down to 1-2% of dry weight in the dark. The synthesis rate of glycogen during light period was not uniform being highest at the onset of illumination (2.6 μg.μg~(-1) chl.h~(-1)). The culture was CO_2 - limited when supplied with normal air (0.03%. CO_2). Higher CO_2 (1%) supply caused that the biomass increased to 1.6-1.8 times and the concentration of glycogen in culture also increased correspondly. Synthesis rate of glycogen increased rapidly in the transitional phase from low CO_2 to high CO_2 supply, it was 6.1 μg.μg~(-1) chl.h~(-1) and the glycogen per dry weight was 56% at the beginning of the phase. Afterwards it decreased gradually day and day to a new semi-steady state. According to the data the affinity of inorganic carbon to the cell had been discussed and a possible model for the CO_ and carbonate transport system of the cell was proposed. Adding ammonium to the culture stimulated the increase of biomass but the accumulation of glycogen decreased. Simultaneously supplying NH_4~+ and enriched CO_2 to the culture led to a enhancement of biomass but not the accumulation of glycogen in cells. Nitrogenase activity was low before the onset of light period, then it increased rapidly in the following hours. Maxium of light-induced nitrogenase activity was reached 4 to 6 hours after light on followed by a decrease due to enzyme turnover. Nitrogenase synthesis in the heterocystous blue-green alga, A. variabilis, only occurred in the first hours of light period. Supplying ammonium and chloramphenicol to the culture at the onset of light, the nitrogen activity was inhibited strongly. Throughout the diurnal cycle nitrogenase activity determined in dark assay was one fifth of light-stimulated activity. No correlation was found of dark activity with the glycogen content of whole filaments. Glycogen derived from photosynthesis did not determine the nitrogenase activity in the dark. Also nitrogenaso activity was calculated from the increase of total nitrogen measured. The number of electrons for ammonium formation indicated that no free hydrogen was produced. More than 90% of nitrogen-fixation occurred during the light period, nitrogen-assimilation of A. variabilis was essentially bound to the light period. No significant change in the nitrogen content of the biomass was observed during the light-dark cycle, but the nitrogen content of extracted proteins increased in the light and decreased in the dark indicating that nitrogen assimilation in the light was reserved transiently in nitrogen-rich proteins. The semi-steady state of continuous culture of a heterocystous blue-green alga was successfully set up for the first time in the University Constance, F. R. Germany (Prof. Boger's statement, 1987). It is a new method for the study of algal physiology in our country as well. The results of glycogen synthesis, accumulation, degradation and the nitrogenase synthesis, activity as well as the nitrogen content of proteins related with nitrogen assimilation under light-dark cycle were reported here for the first time. Interest from these results would be expected in algal physiology, eco-physiology and algal biotechnology.
语种: 中文
内容类型: 学位论文
URI标识: http://ir.ihb.ac.cn/handle/342005/12804
Appears in Collections:中科院水生所知识产出(2009年前)_学位论文

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蓝藻的碳、氮代谢-多变鱼腥藻(Anabaenavariabilis)的固氮酶诱导、合成、活性变化与细胞内含糖原和碳代谢的关系.pdf(6030KB)----暂不开放-- 联系获取全文

Recommended Citation:
蓝藻的碳、氮代谢-多变鱼腥藻(Anabaena variabilis)的固氮酶诱导合成、活性变化与细胞内含糖原和碳代谢的关系.刘永定[d].中国科学院水生生物研究所,1989.20-25
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