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单细胞多核管状绿藻原生质团聚及发育研究
其他题名The aggregation and development of the protoplasm of the siphonaceous Chlorophyta
李德茂
学位类型博士
2007-06-13
学位授予单位中国科学院海洋研究所
学位授予地点海洋研究所
关键词藓羽藻 刺松藻 齿形蕨藻 原生质 团聚 发育
摘要在实验室条件下研究了三种单细胞多核管状绿藻(藓羽藻、刺松藻和齿形蕨藻)原生质的团聚和发育过程。结果表明: 藓羽藻原生质团聚过程中,Na+或Ca2+浓度分别为0.5M和0.2 M时形成的藓羽藻亚原生质体数量最多,在原生质体团聚过程中细胞核发生了形变;藓羽藻无成熟叶绿体原生质能够在海水中团聚,并形成无成熟叶绿体的亚原生质体,该亚原生质体有细胞膜形成,但是无细胞壁形成,72小时后会死亡,分离纯化的线粒体在海水中不能团聚,分离纯化的叶绿体在海水中的团聚不是很明显,只是有2-3个叶绿体形成团聚体,而且数量也很少;在藓羽藻团聚过程中外源壳孢子和藓羽藻原生质会很快分开,这样壳孢子就不会被包围到藓羽藻原生质团聚体中,即使有的壳孢子被团聚进藓羽藻亚原生质体中,最后也是以藓羽藻亚原生质体的破裂而壳孢子发育而告终,同时加入壳孢子后还导致藓羽藻亚原生质体的发育方式发生了一定的变化,从以前的单极萌发变成二极萌发;在实验室条件下继代培养了藓羽藻原生质体三代,而且在团聚数量、大小和发育能力方面能够稳定遗传;野生藓羽藻原生质在6-12月份能够团聚,但是只能在8-12月份间发育; 刺松藻原生质体在碱性人工海水中会团聚形成亚原生质体,但是亚原生质体在72h内全部破裂,酶学和染色实验显示刺松藻原生质团聚过程中的絮凝胶状物质含有蛋白质和多糖类物质,松藻是一种非常合适的研究亚原生质体形成的实验模式材料;刺松藻的营养芽、游动细胞、丝状体和果胞都能够再生,刺松藻片断生长的最佳碳和氮浓度分别为:HCO3—C 3mM和NO3—N 70μM,刺松藻对所研究的不同氮源吸收能力基本相同; 齿形蕨藻的光饱和点为200 μmol/m2 s,最佳生长温度为25-30℃;齿形蕨藻片断的再生与光照强度成反比,而且只能在20-35℃间再生。齿形蕨藻原生质在碱性海水中能够团聚,但是与藓羽藻相比较数量非常的少。
其他摘要The aggregation of the protoplasm of Bryopsis hypnoides, Codium fragile and Caulerpa serrulata, which were coenocytic Siphonales, formation and development of the newly formed protoplasts were studied in laboratory conditions. The results indicated that: The number of the protoplasts formed under Na+ concentration of 0.5 M was the most, for the Ca2+ was under 0.2 M. The nuclei of B. hypnoides were changed in appearance from spherical to triangle or long strands during protoplasm aggregation. The protoplasm without mature chloroplasts (PMC) of B. hypnoides aggregated together in artificial seawater and protoplasts without mature chloroplasts (PTCs) were formed. Cell membrane was regenerated from the PTCs, however, the cell wall was not regenerated and the PTCs died in 72 h. For the isolated chloroplasts, when they were transferred into seawater, there were only two or three chloroplasts aggregated together, which were very small. In comparison with the PMC and the chloroplasts, the isolated mitochondria and the mixed six layers cell organelles (separated by the PMC with sucrose density centrifugation) could not aggregate in the artificial seawater. An incompatibility barrier was present during protoplast formation in B. hypnoides to exclude foreign conchospores. Even when the conchospores were incorporated into protoplasts in some experimental condition, they were expelled from the protoplasts or the protoplasts were degenerated within several days. However, the regeneration polarity was changed from one direction to two or three directions when conchospores were added to the protoplasm. There were three generations regenerated from the protoplasts of B. hypnoides. The number, diameter and regeneration of the protoplasts trait were inherited stably for 3 generations in succession. The protoplasm of B. hypnoides could be aggregated and new protoplasts from June to December; however, the newly formed protoplasts were regenerated from August to December. The protoplasm of C. fragile could be aggregated in artificial seawater. The newly formed sub-protoplasts of C. fragile disintegrated in 72h. The enzyme and staining experiments showed that the long and gelatinous thread involved in the formation of the protoplasts was composed of protein and saccharide. The experiments indicated that C. fragile would be a suitable model alga for studying the formation of protoplasts. The propagation bud, swimming cell, filamentous thalli and utricle could be regenerated and developed to mature individuals which would be propagation methods in C. fragile life history. The optimum concentration of HCO3—C and NO3—N were 3mM and 70μM. No significant differences in growth of thalli were detected when the nitrate that was normally used in the culture medium was replaced by nitrite, ammonium or urea. The light saturation point of C. serrulata was 200 μmol/(m2 s) and the optimum growth temperature was 25-30°C. The recruitment of the fragmented C. serrulata showed a negative correlation with the light intensity and it was only recruited at 20-35°C. The protoplasm of C. serrulata could be aggregated and new protoplasts could be formed in the artificial seawater. However, compared with the number of formed by B. hypnoides in the artificial seawater, it formed by C. serrulata was very little.
页数147
语种中文
文献类型学位论文
条目标识符http://ir.qdio.ac.cn/handle/337002/323
专题海洋环流与波动重点实验室
推荐引用方式
GB/T 7714
李德茂. 单细胞多核管状绿藻原生质团聚及发育研究[D]. 海洋研究所. 中国科学院海洋研究所,2007.
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