拟衣藻系统分类新证据

拟衣藻 (Chloromonas)与衣藻属 (Chlamydomonas)的亲缘关系及分类学位置在藻类学界一直没有定论。其重要原因是单细胞鞭毛类是否具有蛋白核这一特征在系统分类学上具有重要意义 ,而光镜形态与色素体上具 1到多个蛋白核的衣藻极其相似的拟衣藻 ,其不具蛋白核这一特征的稳定性受到许多学者的怀疑。本文观察并报道了中华拟衣藻 (ChloromonasSinica)的超微结构 ,通过对拟衣藻与衣藻超微结构的探究和比对 ,发现除了从孢子时期开始的整个生活史中 ,中华拟衣藻都不具蛋白核外 ,无论显微还是超微结构 ,拟衣藻与衣藻都显示出高度的相似性。从而提出在尚未获得更多资料之前 ,将拟衣藻从衣藻属分离出来成为独立的属较为合适     

中国衣藻属研究（I）

报道了衣藻属（Chlamydomonas）的9个新种：近条纹衣藻、具孔衣藻、顶角衣藻、近环形衣藻、具粒衣藻、近具喙衣藻、不整衣藻、疣突衣藻、倒卵形衣藻,以及4个新变种：阿美衣藻大型变种、似博泡衣藻大型变种、侏儒衣藻陈氏变种和锥形衣藻胶被变种。     

衣藻的藻种保存`扩大培养和观察

衣藻属(Chlamydomonas)是常见的淡水绿藻。在生长季节里,衣藻常在营养特别是氮、磷较丰富的坑塘中有时长成纯群,但在池水或缓流中则常与其他藻类等混生,不易采集。在非生长季很难找到它们。这给教学和科研带来许多困难。几年来,我们对衣藻的藻种保存、扩大培养进行观察试验,总结出一些经验,现介绍如下:     

衣藻叶绿体表达系统的研究

真核藻类作为一种新型的蛋白表达系统,因其培养方法简单,成本低廉并且能大规模繁殖,最近几年成为人们关注的焦点.作为一种模式生物,单细胞的真核生物莱茵衣藻已经成为人们研究的重点.外源蛋白不仅能在衣藻核中进行表达而且也能在叶绿体中表达,但衣藻叶绿体的表达系统较之核表达有巨大的优越性.在到目前为止,已经有许多的药用蛋白在莱茵衣藻的叶绿体中成功表达的报道,证明了莱茵衣藻叶绿体作为生物反应器的能力.将对衣藻叶绿体的表达做详细的描述.     

中国衣藻属研究(Ⅰ)

报道了衣藻属(Chlamydomonas)的9个新种:近条纹衣藻、具孔衣藻、顶角衣藻、近环形衣藻、具粒衣藻、近具喙衣藻、不整衣藻、疣突衣藻、倒卵形衣藻,以及4个新变种:阿美衣藻大型变种、似博泡衣藻大型变种、侏儒衣藻陈氏变种和锥形衣藻胶被变种。     

衣藻生物制氢的研究进展

综述了利用衣藻生产氢气作为再生能源的研究进展。分别介绍了衣藻产氢的代谢机理、培养条件、衣藻氢化酶的特性以及利用分子生物学手段、生物信息学手段和生物工程技术提高衣藻生物制氢效率的方法,包括氢化酶的氧耐受性的改造、外源氢化酶基因的表达、影响衣藻产氢的关键基因的筛选、利用缺硫培养基和固定化培养方法提高氢气产量等。最后,还对利用衣藻生物制氢的可行性和经济性进行了分析,对其发展方向提出自己的看法。     

异形藻属一新种——扁球异形藻及其生活史的研究

本文报道了团藻目、壳衣藻科、异形藻属一新种——扁球异形藻。此种从非洲的土壤标本中分离 获得。营养细胞(囊壳、原生质体)与已知的其它8个种有明显的不同。生活史的培养观察发现有性生殖为异配,这是此属未曾被描述的一个特征。     

莱茵衣藻细胞核转化体系研究进展

莱茵衣藻(Chlamydomonas reinhardtii)是一种三套基因组都可以进行遗传转化的模式生物,具有培养条件简单、生长速度快、光合效率高等优点,细胞核转化体系相对更为成熟,将其开发为生物反应器具有广阔的应用前景。该文对衣藻核基因组特点及转化机理、转基因衣藻的筛选方法、外源基因的表达以及影响因素等方面进行了综述。     

衣藻细胞玻璃化超低温保存技术的研究

本研究以衣藻为材料,探讨其玻璃化超低温保存的条件和方法,结果表明,衣藻经含0.25mol/L蔗糖溶液的TAP培养基预培养一天后,在玻璃化冷冻保护剂中脱水5分钟,直接投稿液氮,48小时后快速化冻,去保护剂并用含0.5mol/L蔗糖溶液的TAP培养基境培养一天,再转到ATP培养基暗培养一天,最后置光照条件下恢复培养,其存活率可达31.45％,恢复培养后衣藻细胞的生长规律与未冻存的衣藻相一致。     

中国团藻目研究（I）

为了配合中国孢子植物志《团藻目》编志的需要,作者到广西壮族自治区、内蒙古自治区、湖北省等地采集标本进行分类研究。文中报道了团藻目6个属的4个新种,1个新变种和9个中国新记录,其中拟衣藻属Chloromonas和朴罗藻属Provasoliella为我国新记录属,武汉朴罗藻、嗜碱衣藻、多粒衣藻、广西拟衣藻为新种,星状四鞭藻广西变种为新变种。     

Snow algae of the Windmill Islands, continental Antarctica 3. Chloromonas polyptera (Volvocales, Chlorophyta)

A new name, Chloromonas hohamii, is proposed to accommodate a common North American snow alga previously incorrectly referred to as Chloromonas polyptera. Chloromonas hohamii differs in having the motile vegetative cells with a cup-shaped chloroplast opening in the anterior end of the cell, shorter, narrower, ellipsoidal to elongate to somewhat fusiform, sexual spores with non-spiralled wall flanges, shorter and narrower daughter cells derived from the spores, and it grows in snow of significantly lower pH and conductivity. Received: 29 August 1997 / Accepted: 24 April 1998     

Cytoplasmic inheritance of mitochondria and chloroplasts in the anisogamous brown alga Mutimo cylindricus (Phaeophyceae)

Based on the morphology of gametes, sexual reproduction in brown algae is usually classified into three types: isogamy, anisogamy, and oogamy. In isogamy, chloroplasts and chloroplast DNA (chlDNA) in the sporophyte cells are inherited biparentally, while mitochondria (or mitochondrial DNA, mtDNA) is inherited maternally. In oogamy, chloroplasts and mitochondria are inherited maternally. However, the patterns of mitochondrial and chloroplast inheritance in anisogamy have not been clarified. Here, we examined derivation of mtDNA and chlDNA in the zygotes through strain-specific PCR analysis using primers based on single nucleotide polymorphism in the anisogamous brown alga Mutimo cylindricus. In 20-day-old sporophytes after fertilization, mtDNA and chlDNA derived from female gametes were detected, thus confirming the maternal inheritance of both organelles. Additionally, the behavior of mitochondria and chloroplasts in the zygotes was analyzed by examining the consecutive serial sections using transmission electron microscopy. Male mitochondria were isolated or compartmentalized by a double-membrane and then completely digested into a multivesicular structure 2 h after fertilization. Meanwhile, male chloroplasts with eyespots were observed even in 4-day-old, seven-celled sporophytes. The final fate of male chloroplasts could not be traced. Organelle DNA copy number was also examined in female and male gametes. The DNA copy number per chloroplast and mitochondria in male gametes was lower compared with female organelles. The degree of difference is bigger in mtDNA. Thus, changes in different morphology and DNA amount indicate that maternal inheritance of mitochondria and chloroplasts in this species may be based on different processes and timing after fertilization.

     

A New Species of Dysmorphococcus (Phacotaceae,Volvocales) and Its Life Cycle

This paper deals with the morphology and life cycle of a new species of Dysmorphococcus oblatus Yu et Wei (Phacotaceae, Volvocales) from Africa. The alga seems to be morphologically rather distinct from the other members (lorica, protoplast and number of pyrenoid). Moreover, a comparison is made of overall morphological and reproductive characteristics of the present taxon with the other 8 known species of this genus. The cultural experiment of the life cycle of this alga reveals certain attributes unrecorded for early described species of this genus. The alga is homothallic, and heterogametic and sexual reproduc-tion produces 32-64 male gametes and 2 female gametes.     

Life history of Chnoospora implexa (Chnoosporaceae, Phaeophyceae) in culture

The life history of the brown alga Chnoospora implexa J. Agardh (Chnoosporaceae, Scytosiphonales) from Japan was studied in laboratory cultures. This species showed a heteromorphic and diphasic life history, alternating between erect gametophytes and discoid sporophytes. The gametophytes were dioecious and produced isogametes. The zygotes developed into sporophytes at 20°C under long‐day conditions, which formed plurilocular zoidangia. Zoids released from the plurilocular zoidangia developed again into sporophytes that always formed plurilocular zoidangia at 20°C and 25°C in long‐day conditions, and mainly unilocular zoidangia at 25°C in short‐day conditions. Zoids released from unilocular zoidangia developed into dioecious gametophytes. At 15°C zygotic erect thalli were formed and were revealed to be diploid by microspectrofluorometric measurements of nuclear DNA contents. The development and reproduction of unfused gametes were similar to those of zygotes. Some strains showed a direct‐type life history; gametophytic thalli were produced, but not via a sporophytic phase.     

Physiological and morphological processes in the Alpine snow alga Chloromonas nivalis (Chlorophyceae) during cyst formation

Amongst a specialised group of psychrophilic microalgae that have adapted to thrive exclusively in summer snow fields, Chloromonas nivalis has been reported as a species causing green, orange or pink blooms in many alpine and polar regions worldwide. Nevertheless, the cytology, ecophysiology and taxonomy of this species are still unresolved. Intracellular processes during cyst formation, which is the dominant stage on snow fields, were examined with samples from the European Alps to better understand the cellular strategies of a green alga living in this harsh habitat. We show with two different methods, i.e. oxygen optode fluorometry and by chlorophyll fluorescence, that the cysts are photosynthetically highly active, although they do not divide, and that Chloromonas nivalis can cope with low as well as high light conditions. During cyst formation, the chloroplast is fragmented into several smaller parts, enlarging the surface to volume ratio. The pool of xanthophyll-cycle pigments is significantly enlarged, which is different from other snow algae. The cytoplasm is filled with lipid bodies containing astaxanthin, a secondary carotenoid that causes the typical orange colour. The cyst wall surface possesses characteristic elongate flanges, which are assembled extracellulary by accumulation of material in the periplasmatic interspace. Comparison of Chloromonas nivalis samples from different locations (Austrian Alps, Spitsbergen) by molecular methods indicates genetic variations due to spatial isolation, while a North American strain has no close relationship to the taxon.     

Analysis of structural characteristics of alpha-tubulins in plants with enhanced cold tolerance

The uniqueness of the point substitutions in the sequences of two alpha-tubulin isotypes from psychrophilic alga Chloromonas that can determine the increased cold tolerance of this alga was analyzed. The comparison of all known amino acid sequences of plant alpha-tubulins enabled to ascertain that only M268-->V replacement is unique and may have a significant influence on spatial structure of plant alpha-tubulins. Modeling of molecular surfaces of alpha-tubulins from Chloromonas, Chalmydomonas reinhardtii and goose grass Eleusine indica showed that insertion of the amino acid replacement M268-->V into the sequence of goose grace tubulin led to the likening of this protein surface to the surface of native alpha-tubulin from Chloromonas. Alteration of local hydrophobic properties of alpha-tubulin molecular surface in interdimeric contact zone as a result of the mentioned replacement was shown that may play important role in increasing the level of cold resistance of microtubules. The crucial role of amino acid residue in 268 position for forming the interdimeric contact surface of alpha-tubulin molecule was revealed. The assumption is made about the importance of replacements at this position for plant tolerance to abiotic factors of different nature (cold, herbicides).     

ASEXUAL AND SEXUAL REPRODUCTION IN GONIUM QUADRATUM (CHLOROPHYTA) WITH A DISCUSSION OF PHYLOGENETIC RELATIONSHIP WITHIN THE GONIACEAE1

Morphological details of asexual and sexual reproduction in Gonium Quadratum Pringsheim ex Nozaki (Goniaceae, Chlorophyta) were observed by light microscopy, based on clonal cultured materials originating from Nepal. In asexual reproduction, the alga exhibited two different patterns of cell cleavage during formation of 8-and 16-celled daughter colonies. Sexual reproduction was heterothallic and isogamous. The gametes bore a tubular mating structure (bilateral mating papilla) at the base of the flagella, and the papillae of the two gametes. The germinating zygote gave rise to four biflagellate gone cells joined in a colony (germ colony). Possible phylogenetic relationships within the Goniaceae at the species level are outlined, mainly on the basis of reproduction characteristics.     

LIFE HISTORY OF COLPOMENIA SINUOSA (SYCTOSIPHONACEAE,PHAEOPHYCEAE) IN THE AZORES1

Colpomenia sinuosa (Mertens ex Roth) Derbès and Solier (Scytosiphonaceae, Phaeophyceae) is a common species on the rocky intertidal shores of the Azores, where reproductive gametophytes occur throughout the year. Life‐history studies of this species were carried out in culture, and both sexual and asexual reproduction were observed. Anisogamous gametes fused to form zygotes. The zygotes gave rise to a filamentous prostrate sporophyte generation bearing unilocular sporangia, under both short‐day and long‐day conditions at 15 and 22°C, and to both unilocular and plurilocular sporangia, under the lower temperature condition. Unispores developed into gametophytes, and plurispores gave rise to filamentous sporophytes. Asexual reproduction was carried out by unfused female gametes and asexual plurispores produced from the same gametophyte. Unfused gametes developed into filamentous prostrate sporophytes producing unilocular sporangia in both culture conditions, and unispores released from the sporangia gave rise to gametophytes. Asexual plurispores from field gametophytes, under both culture conditions, developed directly into new gametophytes. The species exhibited three types of life history: a heteromorphic, diplohaplontic; a heteromorphic, monophasic (both with alternation between the erect and filamentous prostrate thalli); and a monomorphic, monophasic.     

Unusual medium-chain polyunsaturated fatty acids from the snow alga Chloromonas brevispina

The gas chromatography-mass spectrometry (GC-MS) method was used to identify unusual medium-chain polyunsaturated fatty acids (PUFAs) in the snow alga Chloromonas brevispina collected in 2006 from surface layers of a snow field with conspicuous green patches in Bohemian Forest (Czech Republic). PUFAs formed more than 75% total fatty acids. Among them, mass spectroscopy of picolinyl esters showed sizable proportions of medium-chain PUFA, e.g., 5,8,11-tetradecatrienoic and 6,9,12-pentadecatrienoic acids. The high relative content of PUFA indicates that PUFA are an important element ensuring cell survival. Our report appears to be the first to describe the presence of short- and medium-chain PUFAs in green psychrophilic algae of the genus Chloromonas.