Physical map and gene organization of the mitochondrial genome from the unicellular green alga Platymonas (Tetraselmis) subcordiformis (Prasinophyceae)

the entire mitochondrial genome (mt genome) of the unicellular green alga Platymonas subcordiformis (synonym Tetraselmis subcordiformis; Prasinophyceae) was cloned and a physical map for the four restriction enzymes Hind III, Eco RI, Bgl II and Xba I was constructed. The mt genome of P. subcordiformis is a 42.8 kb circular molecule, coding for at least 23 genes. Hybridization and sequence analysis revealed the presence of a ca. 1.5 kb inverted repeat on the mt genome of P. subcordiformis. Phylogenetic analyses based on sequences of several coxI genes were carried out. Our data indicate that mitochondria from P. subcordiformis and from land plants form a natural, monophyletic group.     

METABOLIC SEGREGATION OF INTRACELLULAR FREE AMINO ACIDS IN PLATYMONAS (CHLOROPHYTA)1

¹⁴C-amino acids were supplied to Platymonas subcordiformis (Wille) Hazen and the incorporation of radioactivity into protein and other compounds was followed. Alanine was rapidly metabolized by both N-limited and N-sufficient cells. Arginine and lysine were metabolized rapidly by N-limited cells, but were sequestered from metabolism in N-sufficient cells. This suggests the existence of two functionally distinct pools; a “metabolic” pool that is rapidly metabolized and preferentially used for incorporation into protein, and a “storage” pool rich in basic amino acids that is sequestered from metabolism.     

A Diurnal Settling Rhythm in Platymonas subcordiformis Hazen*

SYNOPSIS. In cultures of Platymonas subcordiformis Hazen, grown in appropriate light-dark cycles, as many as 75% of the cells adhered to the surface of the glass culture vessel toward the end of the light period of each day. Cell division occurred primarily while the cells were attached. Subsequently, motile daughter cells were released into the growth medium by the rupture of the mother cell theca. The settling behavior appears to be an integral part of the life cycle being synchronized to the same extent as cell division.     

CELL DIVISION IN THE FILAMENTOUS PLEURASTRUM AND ITS COMPARISON WITH THE UNICELLULAR PLATYMONAS (CHLOROPHYCEAE)1

At prophase in Pleurastrum, extranuclear spindle microtubules develop from the region of centrioles, which lie lateral to the nucleus midway between the future sites of the metaphase spindle poles. The microtubules then move laterally to overarch the nucleus and finally become incorporated into the spindle. The centrioles do not migrate and therefore lie in the same plane as the chromosomes at metaphase. At telophase, 2, more different systems of microtubules develop from the vicinity of the centrioles—a phycoplast and extensive arrays of microtubules that ensheath the daughter nuclei. Cell division in the filamentous Pleurastrum is compared to that in the green flagellate, Platymonas. The similarities between cell division in the 2 algae are interpreted as evidence: (i) that rhizoplasts (which in Platymonas resemble myofibrils) are somehow homologous to microtubules; and, (ii) that cell division in Pleurastrum differs from cell division in other examined filamentous chlorophycean genera because Pleurastrum has an independent evolutionary origin from a monad with Platymonas-like characteristics.     

Starch in prasinophycean algae

Reserve products isolated from three species of prasinophycean algae: Pyramimonas parkeae Norris and Pearson, Pyramimonas amylifera Conrad and Platymonas tetrathele West were compared. Three pieces of experimental evidence indicate that the reserve product of each of the species analyzed is a true starch. (I) There is no essential difference between the light absorption curve for the complex formed between corn starch and iodine and the corresponding curves for the algal products. (2) The β-amylase breakdown limit of these compounds is very close to that of corn starch. (3) The X-ray diffraction diagrams produced by the reserve material from Pyramimonas parkeae and Platymonas tetrathele is a so-called A-spectrum characteristic of cereal starches while the diagram produced by grains from Pyramimonas amylifera corresponds to the B-spectrum typical for tuber starches. The dissimilarity between the X-ray diagrams produced by the starches from the Pyramimonas species suggests that a re-examination of the taxonomy of these species is warranted.     

利用外源γ-氨基丁酸调控海洋绿藻亚心形四爿藻淀粉积累

微藻被看作第三代生物质能源的来源。微藻淀粉结构与高等植物的高度相似性使其可以作为粮食作物的替代,在生物能源领域有广泛的应用。γ-氨基丁酸(GABA)被认为是一种信号分子,可以调节植物细胞的生长代谢。本研究在缺氮培养条件下添加外源GABA调控海洋绿藻亚心形四爿藻生理代谢和淀粉积累。结果表明,添加外源GABA可以抑制细胞生长,降低光合作用效率;OJIP实验显示,GABA的添加增强了光合器官能量耗散,降低了光能利用效率,阻碍了电子传递,造成额外胁迫,从而促使细胞将碳流更多地分配到淀粉积累,导致藻细胞的淀粉含量、淀粉产量和淀粉产率提高。添加10 mmol/L GABA获得最大淀粉含量39%DW,比未添加GABA的对照组淀粉含量提高39%;同时获得最大淀粉产量和产率为1.72 g·L^-1和0.36 g·L^-1·d^-1,分别比未添加GABA的对照组提高39%和50%。以上结果表明在缺氮条件下添加外源GABA是一种调控亚心形四爿藻细胞代谢并提高其淀粉生产的有效方法。     

Mixotrophic cultivation of <Emphasis Type="Italic">Platymonas subcordiformis</Emphasis>

The effects of glucose concentration and irradiance on mixotrophiccultivation of Platymonas subcordiformis were investigated in flaskcultures. The optimal glucose concentration was 24 g L^-1, andirradiance was 95 mol photon m^-2 s^-1. Based on theoptimization of culture conditions and a modified SK(IA) medium, themaximum biomass was 3.68 g L^-1 after 14 days, which was about6 times that in autotrophic culture, and the specific growth rate reached0.62 d^-1, twice that in autotrophic culture. Using the optimalconditions established in flasks, mixotrophic culture in a 5.0-Lphotobioreactor was achieved, and the yield after 8 days was 3.30 gL^-1.     

应用免疫和质谱法对亚心形扁藻氢酶蛋白进行亚细胞定位和分类

亚心形扁藻(Platymonas subcordiformis)是新发现的一株产氢海洋单细胞绿藻,经过胁迫调控可实现一定时间的持续产氢。氢酶是亚心形扁藻在胁迫条件下进行光合产氢的一个关键酶。但到目前为止,亚心形扁藻氢酶相关信息仍不清楚。利用蛋白合成抑制剂氯霉素和放线菌酮对亚心形扁藻氢酶活性进行考察,同时利用免疫印迹技术和免疫胶体金电镜对亚心形扁藻氢酶蛋白进行亚细胞定位分析。结果表明:亚心形扁藻氢酶蛋白可能由胞浆内合成,在叶绿体行使功能。采用免疫共沉淀技术富集亚心形扁藻细胞氢酶蛋白,SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)对免疫共沉淀复合物进行分离,从胶中切取目的蛋白条带,胶内酶解后进行基质辅助激光解吸飞行时间质谱(MALDI-TOF-MS)分析,得到相应的肽指纹图谱,通过搜索数据库检索初步断定亚心形扁藻氢酶蛋白为铁氢酶。     

扁藻多糖的性质及结构特征的研究

利用不同浓度范围的乙醇从培养至平衡期后的扁藻细胞内沉淀分离出不同组分的扁藻多糖级分,对其理化性质和结构特征进行了相应的测定和对比分析。结果表明,经过凝胶层析分离后,乙醇浓度在0%~50%范围时沉淀的扁藻多糖与乙醇浓度为50%~75%范围时沉淀的扁藻多糖相比,前者分子量大、糖含量高、其结合的蛋白质也多,但前者的溶解度和粘度小于后者。两个级分的扁藻多糖的官能团相似,均含有硫酸基和氨基,但其含量不同。所得结果为深入研究扁藻多糖的结构及其实际应用提供了实验依据。     

扁藻培养条件初探

在250 mL三角瓶中对扁藻的生长条件作了初步探索。在f/2培养基的基础上,通过正交实验确定主要营养盐的浓度分别为:NaNO30.08 g/L,KH2PO40.006 g/L,NaHCO30.8 g/L,可以使扁藻的倍增时间由原来的1.34 d缩短为0.64 d,并得出了主要营养盐含量与扁藻生物量的关系。研究表明,人尿对扁藻的生长有良好的促进作用,流加营养盐和人尿可使藻液浓度达4.25×106个/mL,比对照提高了36%。