Pb^2＋对衣藻和微囊藻种间竞争的影响

以衣藻和微囊藻的混合培养来模拟水体生态系统的浮游植物群落，研究了Pb^2＋不同质量浓度对水体生态系统中衣藻和微囊藻种问竞争的影响．结果显示，其优势种随Pb^2＋质量浓度的变化而变化，在乙酸铅低质量浓度下铜绿微囊藻为优势种，而在乙酸铅高质量浓度下衣藻变为优势种．测定了Pb^2＋不同质量浓度对衣藻和微囊藻的生长、叶绿素a含量以及蛋白质的影响，结果表明：微囊藻在乙酸铅低质量浓度污染下生长速度较快，而衣藻对Pb^2＋高质量浓度污染的耐受能力较微囊藻高，这可能是导致衣藻和微囊藻混合培养的培养体系结构随环境Pb^2＋污染强度而变化的重要原因．     

水葫芦根厌氧发酵与沙角衣藻生长偶联研究

本文对水葫芦采用不同浓度的NaOH预处理,研究其对水葫芦厌氧发酵产气量的影响. 在培养基中分别加入乙酸钠、碳酸氢钠和发酵上清液培养沙角衣藻,讨论对沙角衣藻生长的影响. 结果发现,2% 浓度的NaOH预处理对根发酵产气最为有利,根叶混合物经预处理后可获得802.6mL/100g新鲜底物的最佳产气量;以乙酸钠作培养基碳源,沙角衣藻生物量能得到明显积累,可从初始的0.4×106个/mL达到10d后的2.84×106个/mL.     

莱茵衣藻鞭毛内运输蛋白IFT81调控鞭毛组装

利用插入突变的方式获得了一株衣藻不运动突变体ift81,该突变体表现出鞭毛缺失或者短鞭毛的性状。基因序列分析表明,外源基因aphⅧ插入了突变体中IFT81( intraflagellar transport, IFT)基因的第五个外显子内,并导致该外显子原有的52个碱基对被替换。把含有完整IFT81基因的重组质粒导入突变体ift81后,其鞭毛恢复为野生型且可以检测到IFT81-HA融合蛋白的表达,这证明突变体的鞭毛缺陷确实是由于IFT81基因突变所导致。电镜观察显示突变体中鞭毛的显微结构发生改变,免疫荧光实验证实IFT81蛋白主要定位于基体和鞭毛部位。上述结果表明：IFT81蛋白缺失会导致衣藻鞭毛组装缺陷,在鞭毛组装所需蛋白的运输过程中,IFT81蛋白是必不可少的。     

Transfer of a eubacteria-type cell division site-determining factor CrMinD gene to the nucleus from the chloroplast genome in Chlamydomonas reinhardtii

MinD is a ubiquitous ATPase that plays a crucial role in selection of the division site in eubacteria, chloroplasts, and probably Archaea. In four green algae, Mesostigma viride, Nephroselmis olivacea, Chlorella vulgaris and Prototheca wickerhamii, MinD homologues are encoded in the plastid genome. However, in Arabidopsis, MinD is a nucleus-encoded, chloroplast-targeted protein involved in chloro- plast division, which suggests that MinD has been transferred to the nucleus in higher land plants. Yet the lateral gene transfer (LGT) of MinD from plastid to nucleus during plastid evolution remains poorly understood. Here, we identified a nucleus-encoded MinD homologue from unicellular green alga Chlamydomonas reinhardtii, a basal species in the green plant lineage. Overexpression of CrMinD in wild type E. coli inhibited cell division and resulted in the filamentous cell formation, clearly demon- strated the conservation of the MinD protein during the evolution of photosynthetic eukaryotes. The transient expression of CrMinD-egfp confirmed the role of CrMinD protein in the regulation of plastid division. Searching all the published plastid genomic sequences of land plants, no MinD homologues were found, which suggests that the transfer of MinD from plastid to nucleus might have occurred be- fore the evolution of land plants.     

Transfer of a eubacteria-type cell division site-determining factor CrldfnD 8ene to the nucleus from the chloroplast genome in Chlamydomonas reinhardtii

MinD is a ubiquitous ATPase that plays a crucial role in selection of the division site in eubacteria, chloroplasts, and probably Archaea. In four green algae, MesosUgma viride, Nephroselmis olivacea, Chlorella vulgaris and Prototheca wickerhamii, MinD homologues are encoded in the plastid genome. However, in Arabidopsis, MinD is a nucleus-encoded, chloroplast-targeted protein involved in chloroplast division, which suggests that MinD has been transferred to the nucleus in higher land plants. Yet the lateral gene transfer （LGT） of MinD from plastid to nucleus during plastid evolution remains poorly understood. Here, we identified a nucleus-encoded MinD homologue from unicellular green alga Chlamydomonas reinhardtii, a basal species in the green plant lineage. Overexpression of CrMinD in wild type E. coil inhibited cell division and resulted in the filamentous cell formation, clearly demonstrated the conservation of the MinD protein during the evolution of photosynthetic eukaryotes. The transient expression of CrMinD-egfp confirmed the role of CrMinD protein in the regulation of plastid division. Searching all the published plastid genomic sequences of land plants, no MinD homologues were found, which suggests that the transfer of MinD from plastid to nucleus might have occurred before the evolution of land plants.     

Mutagenesis of Ser24 of Cytochrome b559 α Subunit Affects PSII Acitivies in Chlamydomonas reinhardtii

In order to study the functions of cytochrome b559 (Cyt b559) in photosystem two (PSII) activity, mutant S24F of Chlamydomonas reinhardtii was constructed using site directed mutagenesis, in which Serine24 (Ser24) locating downstream of Histidine23 (His23) in α subunit of Cyt b559 was replaced by Phenylalanine (Phe). Physiological and biochemical analysis showed that mutant S24F could be grown photoautotrophically or photoheterotrophically. However, their growth rate was slower either on HSM or TAP medium than that of the control; Analysis of PSII activity revealed that its oxygen evolution was about 71% of wild type (WT); The Photochemical efficiency of PSII (Fv/Fm) of S24F was reduced 0.23 compared with WT; S24F was more sensitive to strong light irradiance than the wild type; Furthermore, SDS-PAGE and Western-blotting analysis indicated that the expression levels of α subunit of Cyt b559, LHCII and PsbO of S24F were a little less than those of the wild type. Overall, these data suggests that Ser24 plays a significant role in making Cyt b559 structure maintain PSII complex activity of oxygen evolution although it is not directly bound to heme group.     

莱茵衣藻-se两步法产氢用培养基的研究

分析了TAP(Tris-Acet-Phosphate)、seTA(se-Tris-Acet)、seP(se-Phosphate)、seTH(se-Tris-HCl)和se这5种培养基对莱茵衣藻-se生长产氢的影响,并进行了培养基的含量分析.结果发现,培养基的pH和氮素的含量是影响莱茵衣藻生长的重要因素;而培养基中醋酸根的含量则有助于提高厌氧产氢过程中衣藻的氢酶活性,从而提高其产氢量.综合考虑两步法产氢对细胞生长周期和细胞密度的要求以及最终厌氧诱导获得的氢酶活性,最终选择seTA培养基作为莱茵衣藻-se产氢用培养基.     

沙角衣藻转化难溶性磷的研究

以磷矿石粉为沙角衣藻（Chlamydomonas sajao Lwein）的唯一生长磷源，研究沙角衣藻对难溶性磷源（矿石磷）的转化利用作用，结果表明：沙角衣藻对矿石磷的转化强度可达40.56%-63.49%，以有效磷mg/g磷源为单位可达41－64，是一般溶磷细菌溶磷强度的2－4倍，相当于每产1g鲜藻体可得磷酸1.52g，该藻不仅能快速地转化利用磷矿石粉中的难溶性磷源以满足其自身的生理需要，而且能迅速提高介质中的有效磷浓度，为环境中其它生物的生活提供磷源；而且这种转化作用的强度与其对磷的需要相协调，可确保体系供磷和需磷的和谐同步进行。