The phylogenetic relationship between the Chlamydomonadales and Chlorococcales inferred from 18SrDNA sequence data

Nuclear-encoded small subunit ribosomal RNA gene (185rDNA) sequences were determined for Chlamydomonas moewusii Gerloff and five chlorococcalean algae (Chlorococcum hypnosporum Starr; Chlorococcum oleofaciens Trainor et Bold; Chlorococcum sp.; Tetracystis aeria Brown et Bold; Protosiphon botryoides (Kützingl Klebs). All these algae are characterized by a clockwise CCW) flagellar apparatus. Phylogenetic trees were constructed from sequences from these algae together with 20 green algae. All algae with a CW flagellar apparatus form a monophyletic clade (CW group). Three principal clades can be recognized in the CW group, although no morphological character supports monophyly of any of these three clades. The 185rDNA trees clearly demonstrate the non-monophyly of the Chlamydomonadales and Chlorococcales, suggesting that vegetative morphology does not reflect phylogenetic relationships in the CW group. The paraphyly or polyphyly of the genus Chlamydomonas and Chlorococcum are also revealed. Present analysis suggests that the presence or absence of a zoospore's cell wall and the multinucleate condition have limited taxonomic values at higher taxonomic ranks.     

Role of pyrenoids in the CO2-concentrating mechanism: comparative morphology, physiology and molecular phylogenetic analysis of closely related strains of Chlamydomonas and Chloromonas (Volvocales)

The morphology of the pyrenoid and the physiology of the CO₂-concentrating mechanism (CCM) were investigated in Chlamydomonas (Cd.) mutabilis Gerloff UTEX 578, Cd. radiata Deason et Bold UTEX 966, Cd. augustae Skuja UTEX 1969, Cd. macrostellata Lund SAG 72.81, Cd. bipapillata Bourrelly SAG 11-47, and Chloromonas (Cr.) insignis Gerloff et Ettl NIES-447, all of which are closely related phylogenetically to the pyrenoid-less strains of Chloromonas. In the chloroplasts of Cd. mutabilis UTEX 578, Cd. radiata UTEX 966, Cd. augustae UTEX 1969, and Cd. macrostellata SAG 72.81, a typical, spheroidal, electron-dense pyrenoid matrix surrounded by starch granules was present, and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) molecules were highly concentrated in the pyrenoid matrix. On the other hand, while the pyrenoid matrix of Cr. insignis NIES-447 was electron-dense that of Cd. bipapillata SAG 11-47 was not, and neither was surrounded by starch granules. The pyrenoid matrices of these two species exhibited a higher concentration of Rubisco molecules than the thylakoid region (thylakoid and stroma) of the chloroplasts; however, the densities of Rubisco molecules in these pyrenoid matrices were low compared with those of the other four Chlamydomonas strains examined in this study and that of Cd. reinhardtii Dangeard. In all six strains examined, the presence of the CCM was indicated by relatively high photosynthetic affinities for CO₂ (low values of K_0.5(CO₂)). However, differences in the inorganic carbon (Ci) pools were recognized in relation to the differences in pyrenoid morphology among the strains. In the typical pyrenoid-containing strains. Cd. mutabilis UTEX 578 and Cd. radiata UTEX 966, the ratio of internal to external inorganic carbon was about 20, while in Cr. insignis NIES-447 and Cd. bipapillata SAG 11-47 the ratio was only 2–3 similar to the two pyrenoid-less, CCM-containing strains of Chloromonas previously examined (E. Morita et al., 1998, Planta 204: 269–276). It is thus speculated that the presence of typical pyrenoids with a high concentration of Rubisco molecules is related to the formation of large Ci pools in the CCM. Detailed phylogenetic relationships among these Chlamydomonas/Chloromonas strains and the pyrenoid-less Chloromonas strains previously investigated were inferred based on the sequence of rbcL, the gene for the large subunit of Rubisco. Two monophyletic groups were resolved with high bootstrap values. Based on the tree topology resolved, it was inferred that loss of the typical pyrenoids accompanied by a decrease in intracellular Ci pools might have taken place independently in the two groups. Received: 21 August 1998 / Accepted: 30 November 1998     

Immunological Characterization of Gamete Autolysins in Chlamydomonas reinhardtii

Polyclonal antibodies were raised in rabbits against the C. reinhardtii cell wall lytic enzyme, autolysin, which dissolves the cell wall of gametes prior to cell fusion. The purified immunoglobulins react with both the native and the deglycosylated forms of this gametic autolysin and specifically inhibit enzyme activity. In addition, the immunoglobulins selectively detect the gametic autolysin in immunoblots of crude extracts and do not cross-react with the autolysin of the vegetative cells. The antibodies have been used to study the time of synthesis of the enzyme during gametogenesis and to compare gametic autolysins of different strains of Chlamydomonas.     

MITOSIS AND CLEAVAGE DURING ZOOSPOROGENESIS IN SEVERAL COCCOID GREEN ALGAE1

The fine structure of mitosis and cleavage in Axilosphaera vegetata Cox & Deason, Chlorococcum echinozygotum Starr, Chlorosarcinopsis eremi Chantanachat & Bold, Nautococcus mammilatus Korshikoff, N. terrestris Archibald, N. soluta Archibald, Neospongiococcum solitarium Deason, and Tetracystis aeria Brown & Bold was observed. All possess a phycoplast, but differences in basal body position during mitosis were observed. Nautococcus mammilatus differs from the others as protoplast rotation occurs in many cells prior to cleavage.     

COMPARATIVE ULTRASTRUCTURE OF THREE SPECIES OF CHLOROSARCINA (CHLOROSARCINACEAE,CHLOROPHYTA) 1,2

Ultrastructural studies revealed that Chlorosarcina stigmatica Deason differs from C. brevispinosa and C. longispinosa Chantanachat and Bold because the vegetative cell possess a pyrenoid, and the zoospores have chlorophycean characteristics. Zoospores of the latter species exhibit pleurastrophycean characteristics including counterclockwise absolute orientation of the flagellar apparatus components. Nuclear division in C. longispinosa, like that of Friedmannia israelensis Chantanachat and Bold, is metacentric. It is suggested that a new genus, Desmotetra, be erected for Chlorosarcina stigmatica, and that it, along with Chlorosarcinopsis Herndon, be placed in the Chlorococcales, Chlorophyceae. Chlorosarcina brevispinosa and C. longispinosa belong in the Pleurastrales, Pleurastrophyceae.     

PHYLOGENYOF THE VLE-14 CHLAMYOMONAS (CHLOROPHYCEAE) GROUP: A STUDY OF 18S rRNA GENE SEQUENCES1

Comparative specificity of sporangial wall autolysins (i.e. vegetative lytic enzymes [VLE]) derived from sporulating cultures has been used to group Chlamydomonas taxa into 15 different VLE types. The VLE-14 group, including isolates of C. geitleri, C. noctigama, C. monoica, C. pinicola, C. terricoia, and C. hindakii, is one of the largest of these VLE groups. Genetic studies have shown that a number of the VLE-14 taxa are interfertile, albeit with little or reduced viability of progeny. A reevaluation of the VLE-14 group suggested that all members should be regarded as distinct isolates of C. noctigama. The present investigation tests the phylogenetic implications of the VLE evidence and examines the validity of the taxonomic reevaluation in a phylogenetic context by analysis of 18S rRNA gene sequence data. Results from analyses of the sequence data are consistent with an interpretation of the VLE evidence as indicative of monophyletic taxa. Phylogenetic analyses of the sequence data are also consistent with the taxonomic reevaluation and reidentification of the group. However, at least some of the VLE-14 isolates studied in this investigation fit criteria for distinct biologic or phylogenetic species. It is concluded that the VLE-14 taxa represent a very closely allied group that includes some isolates that are in the early stages of speciation ly reproductive isolation.     

A marine Chlamydomonas sp. emerging as an algal model

The freshwater microalga Chlamydomonas reinhardtii, which lives in wet soil, has served for decades as a model for numerous biological processes, and many tools have been introduced for this organism. Here, we have established a stable nuclear transformation for its marine counterpart, Chlamydomonas sp. SAG25.89, by fusing specific cis‐acting elements from its Actin gene with the gene providing hygromycin resistance and using an elaborated electroporation protocol. Like C. reinhardtii, Chlamydomonas sp. has a high GC content, allowing reporter genes and selection markers to be applicable in both organisms. Chlamydomonas sp. grows purely photoautotrophically and requires ammonia as a nitrogen source because its nuclear genome lacks some of the genes required for nitrogen metabolism. Interestingly, it can grow well under both low and very high salinities (up to 50 g · L^‐1) rendering it as a model for osmotolerance. We further show that Chlamydomonas sp. grows well from 15 to 28°C, but halts its growth at 32°C. The genome of Chlamydomonas sp. contains some gene homologs the expression of which is regulated according to the ambient temperatures and/or confer thermal acclimation in C. reinhardtii. Thus, knowledge of temperature acclimation can now be compared to the marine species. Furthermore, Chlamydomonas sp. can serve as a model for studying marine microbial interactions and for comparing mechanisms in freshwater and marine environments. Chlamydomonas sp. was previously shown to be immobilized rapidly by a cyclic lipopeptide secreted from the antagonistic bacterium Pseudomonas protegens PF‐5, which deflagellates C. reinhardtii.     

MERCURY AND TEMPERATURE INTERACTIONS ON THE GROWTH RATES OF THREE SPECIES OF FRESHWATER PHYTOPLANKTON1,2

Cultures of freshwater algae, representing three algal divisions, Synura petersenii Korshikov; Chlamydomonas sp.: and Nitzschia sp., were subjected to four different mercury-temperature shock interactions to demonstrate synergistic effects between mercury and temperature. Algal growth, measured by temporal changes in vivo fluorescence of chlorophyll was used to ascertain the effects. Mercury addition and temperature shock had various inhibitory effects on algal growth. Prior environmental conditions influence the effect of subsequent treatments. Growth of S. petersenii was severely inhibited by mercury in all experiments. Control cultures would not grow at 30 C and died when shacked at this temperature. Chlamydomonas sp. cultures initially inhibited by mercury were able to recover under most conditions after a period of reduced growth. Nitzschia sp. was resistant to mercury except when simultaneously shocked with temperature. Mercury analyses showed that Nitzschia cells at 25 C and 30 C contained a high percentage of the mercury initially added to the cultures. There was a significant loss of mercury at the end of each experiment from all cultures, probably due to volatilization.     

Zoospore production in selected xanthophycean algae

The effects of incubation time and deficiencies of calcium and nitrate on zoospore production were investigated in Botrydiopsis arhiza Borzi, B. intercedens Vischer et Pascher, Bumilleriopsis peterseniana Vischer, Pseudobumilleriopsis pyrenoidosa Deason et Bold, and Ophiocytium maius Naegeli. Release of zoospores occurred after the transfer of stationary phase, vegetative cells into fresh Bold's basal medium. Maximum yields of zoospores were produced on the second day after transfer, 0·5–1·0 h after the start of the light period of a 12–12 h light-dark cycle. Lack of calcium or nitrate reduced the yield of zoospores in each taxon. Only O. maius produced large numbers of motile cells in nitrate-free medium.     

FATTY ACID AND LIPID COMPOSITION OF THE ACIDOPHILIC GREEN ALGA CHLAMYDOMONAS SP.1

The lipid and fatty acid compositions of Chlamydomonas sp. isolated from a volcanic acidic lake and C. reinhardtii were compared, and the effects of pH of the medium on lipid and fatty acid components of Chlamydomonas sp. were studied. The fatty acids in polar lipids from Chlamydomonas sp. were more saturated than those of C. reinhardtii. The relative percentage of triacylglycerol to the total lipid content in Chlamydomonas sp. grown in medium at pH 1 was higher than that in other cells grown at higher pH. A probable explanation might be that Chlamydomonas sp. has two low pH adaptation mechanisms. One mechanism is the saturation of fatty acids in membrane lipids to decrease membrane lipid fluidity, and the other is the accumulation of triacylglycerol, as a storage lipid, to prevent the osmotic imbalance caused by high concentrations of H₂SO₄.     

Generic and species concepts in Microglena (previously the Chlamydomonas monadina group) revised using an integrative approach

Traditionally the genus Microglena Ehrenberg has been used to contain species that belong to the Chrysophyceae; however, the type species of Microglena, M. monadina, represents a green alga, which was later transferred to the genus Chlamydomonas. The taxonomic status of the genus has therefore remained unclear. We investigated 15 strains previously assigned to C. monadina and two marine species (C. reginae and C. uva-maris) using an integrative approach. Phylogenetic analyses of SSU and ITS rDNA sequences revealed that all strains form a monophyletic lineage within the Chlorophyceae containing species from different habitats. The strains studied showed similar morphology with respect to cell shape and size, but showed differences in chloroplast and pyrenoid structures. Some representatives of this group have the same type of sexual reproduction (homothallic advanced anisogamy). Three different morphotypes could be recognized. Strains belonging to type I have a cup-shaped chloroplast with a massive basal part, in which a large, single, ellipsoidal pyrenoid is located. The members of type II also have a cup-shaped chloroplast, which is partly lobed and has a thinner basal part than type I; here the pyrenoid is half-ring or horseshoe-shaped and occupies different positions in the chloroplast depending on the strain. The strains of type III have multiple pyrenoids, which appear to have developed from the subdivision of a single ring-shaped pyrenoid into several parts. We compared the results of our morphological investigations with the literature and found that 15 strains could be identified with existing species. Two strains did not fit with any described species. As a result of our study, we transfer all strains to the genus Microglena, propose 11 new combinations, and describe two new species. Comparison of the ITS-1 and ITS-2 secondary structures confirmed the species delineations. All species have characteristic compensatory base changes in their ITS secondary structures and are supported by ITS-2 DNA barcodes.     

PHYLOGENY OF CHLAMYDOMONAS (CHLOROPHYTA) BASED ON CLADISTIC ANALYSIS OF NUCLEAR 18S rRNA SEQUENCE DATA1

The genus Chlamydomonas Ehrenberg may contain as many as 450 described species. Morphological, physiological and molecular data show that variation among some Chlamydomonas species can he great, leading to speculation that multiple, generic-level lineages exist within this genus. The most recent systematic studies of Chlamydomonas have led to proposals of nine distinct morphological and 15 distinct sporangial autolysin groups. Partial sequences from the nuclear small subunit rRNAs from 14 Chlamydomonas species representing 12 autolysin and four morphological groups, and from three flagellates thought to he related to Chlamydomonas were determined in a phylogenetic study of relationships among these algae. Sequence comparisons among some Chlamydomonas species revealed differences comparable to the sequence divergence between soybeans and cycads. Cladistic analysis of the sequence data suggests that multiple lineages exist among species of Chlamydomonas. Some of these lineages represent alliances of both Chlamydomonas and non-Chlamydomonas taxa; thus, the current taxonomy does not reflect natural, or monophyletic, groups. Collectively, these lineages may represent distinct families or even orders.     

ISOLATION AND CHARACTERIZATION OF A CELL WALL‐DEFECTIVE MUTANT OF CHLAMYDOMONAS MONOICA (CHLOROPHYTA)1

Cell wall–defective strains of Chlamydomonas have played an important role in the development of transformation protocols for introducing exogenous DNA (foreign genes or cloned Chlamydomonas genes) into C. reinhardtii. To promote the development of similar protocols for transformation of the distantly related homothallic species, C. monoica, we used UV mutagenesis to obtain a mutant strain with a defective cell wall. The mutant, cw‐1, was first identified on the basis of irregular colony shape and was subsequently shown to have reduced plating efficiency and increased sensitivity to lysis by a non‐ionic detergent as compared with wild‐type cells. Tetrad analysis of crosses involving the cw‐1 mutant confirmed 2:2 segregation of the cw:cw⁺ phenotypes, indicating that the wall defect resulted from mutation of a single nuclear gene. The phenotype showed incomplete penetrance and variable expressivity. Although some cells had apparently normal cell walls as viewed by TEM, many cells of the cw‐1 strain had broken cell walls and others were protoplasts completely devoid of a cell wall. Several cw‐1 isolates obtained from crosses involving the original mutant strain showed a marked enhancement of the mutant phenotype and may prove especially useful for future work involving somatic cell fusions or development of transformation protocols.     

Cellular aggregation in Chlamydomonas (Chlorophyceae) is chimaeric and depends on traits like cell size and motility

How the variation in phenotypic traits like cell size and motility impacts predator-induced cellular aggregation is not known. Furthermore the genetic composition of cell groups in mixed populations of Chlamydomonas has not been investigated. An examination of these two questions will not only enhance our understanding of Chlamydomonas ecology, but also shed light on the primordial steps before integrated multicellular groups were established. Group living comes with viability and reproductive costs and it is not known how these are shared if groups are genetically heterogeneous. We observed that the natural predator Peranema trichophorum (Euglenoidea) induced clumping in Chlamydomonas. When co-cultured with P. trichophorum cells protected themselves by forming facultative groups (reverting back to a unicellular lifestyle once predators were removed). The dynamics of group formation in different Chlamydomonas species and strains correlated with cell size and swimming speed. Small or less motile strains aggregated more readily than large, fast-swimming ones. Interestingly, Chlamydomonas groups were both intra-species and inter-species chimaeric. This suggests that the predator-induced group formation in Chlamydomonas involved cells coming together rather than staying together and during aggregation cells showed little or no discrimination between self and non-self. These data demonstrate that the dynamics of cell aggregation, in unicellular volvocines at least, depends on phenotypic traits like cell size and motility and high genetic relatedness is not mandatory at this initial stage. These findings further our understanding of aggregation in mixed Chlamydomonas populations and have implications for understanding the very first steps on the road to simple multicellularity.     

A COMBINED 18S rDNA AND rbcL PHYLOGENETIC ANALYSIS OF CHLOROMONAS AND CHLAMYDOMONAS (CHLOROPHYCEAE,VOLVOCALES) EMPHASIZING SNOW AND OTHER COLD‐TEMPERATURE HABITATS1

An extensive phylogenetic analysis of the biflagellate genera, Chlamydomonas Ehrenberg and Chloromonas Gobi emend. Wille, was undertaken using 18S rDNA and rbcL gene sequence analysis. Emphasis was placed on 21 cold‐tolerant taxa of which 10 are from snow. These taxa occurred in four distinct clades each in the 18S rDNA and rbcL phylogenies, and when taken together suggest at least five distinct origins in cold habitats. Most of these taxa occur in a single clade (A), and all snow species occurred in this clade. In the rbcL and combined rbcL–18S rDNA analyses, the snow taxa fell into three groups. Two groups occurred in subclade 1: Chlamydomonas augustae Skuja CU, Chlamydomonas augustae UTEX, and Chlamydomonas sp.‐A and Chloromonas clathrata Korshikov, Chloromonas rosae Ettl CU, and Chloromonas rosae v. psychrophila var. nov. The third snow group, subclade 2, included three species with unique cell divisions, Chloromonas brevispina (Fritsch) Hoham, Roemer et Mullet, Chloromonas pichinchae (Lagerheim) Wille, and Chloromonas sp.‐D, and the basal Chloromonas nivalis (Chodat) Hoham et Mullet with normal cell divisions. This suggests that the snow habitat has been colonized at least twice and possibly three times in the history of these biflagellates. In the 18S rDNA tree, one cold‐tolerant Chloromonas species fell outside clade A: Chloromonas subdivisa (Pascher et Jahoda) Gerloff et Ettl. In the rbcL tree, three cold‐tolerant Chloromonas species fell outside clade A: Chloromonas subdivisa, Chloromonas sp.‐ANT1, and Chloromonas sp.‐ANT3. These results support previous findings that pyrenoids have been gained and lost several times within this complex.     

Chlamydomonas neoplanoconvexa nom. nov. and its unique phylogenetic position within Volvocales (Chlorophyceae)

Chlamydomonas Ehrenb. is a unicellular volvocalean genus consisting of 400–600 species, most of which are known solely based on microscopy. In this study, a newly isolated strain of Chlamydomonas neoplanoconvexa (M. O. P. Iyengar) Nakada nom. nov. (≡Chlamydomonas planoconvexa M. O. P. Iyengar non J. W. G. Lund) was examined using light microscopy and combined 18S rRNA, rbcL and psaB gene phylogeny. The C. neoplanoconvexa strain was quite similar to Chlamydomonas perpusilla (Korshikov) Gerloff in terms of its small fusiform vegetative cells (～10 µm in length), parietal chloroplast containing a single pyrenoid, and nucleus posterior to the pyrenoid. However, C. neoplanoconvexa was distinguished from C. perpusilla based on the morphology of the papillae and pyrenoids. The two species belong to the clade Caudivolvoxa in the order Volvocales, but are distantly related to each other. Chlamydomonas perpusilla was shown to be sister to Chlorogonium Ehrenb. (clade Chlorogonia, within Caudivolvoxa), while C. neoplanoconvexa represented a second basal lineage within Caudivolvoxa, next to the clade Characiosiphonia. Although the morphology of C. neoplanoconvexa was not particularly outstanding, its unique phylogenetic position will encourage further investigation of this species and its uncultivated relatives.     

PLOIDY ANALYSIS OF THE TWO MOTILE FORMS OF CHRYSOCHROMULINA POLYLEPIS (PRYMNESIOPHYCEAE)1

In some cultures of the flagellate Chrysochromulina polylepis Manton et Parke, established from cells isolated from the massive bloom in Skagerrak and Kattegat in 1988, we observed, two motile cell types. They were termed authentic and alternate cells and differed with respect to scale morphology. To investigate whether or not the two cell forms were joined in a sexual life cycle, the relative DNA content per cell and relative size of cells of several clonal cultures of C. polylepis were determined by flow cytometry. Percentages of authentic and alternate cells in the cultures were estimated by transmission electron microscopy. Pure authentic cultures (α) contained cells with the lowest level of DNA and were termed haploid. Two pure alternate cultures (β) contained cells with double the DNA content of authentic cells and were termed diploid. Other pure alternate cultures contained haploid cells only, or both haploid and diploid cells. Three cell types were observed, each capable of vegetative propagation: authentic haploid, alternate haploid, and alternate diploid cells. Both the haploid and diploid alternate cells were larger than the haploid authentic cells. Cultures containing diploid cells appeared unstable: cell type ratio and ploidy ratio changed during the experiment where this cell type was present, particularly when grown in continuous light. In contrast, cultures with only haploid cells remained unchanged at all growth conditions tested. Light condition may influence cell type ratio and ploidy ratio. Our attempt to induce syngamy by mixing different authentic haploid clones did not result in mating. Assuming that the authentic and alternate cell types are of the same species, the life cycle of C. polylepis includes three flagellated scale-covered cell forms. Two of the cell types are haploid and may function as gametes, and the third is diploid, possibly being the result of syngamy.     

ASSESSING THE RELATIONSHIPS OF SOME COCCOID GREEN LICHEN ALGAE AND THE MICROTHAMNIALES (CHLOROPHYTA) WITH 18S RIBOSOMAL RNA GENE SEQUENCE COMPARISONS1

Eight complete nuclear-encoded small-subunit ribosomal RNA (18S rRNA) gene sequences were determined for four genera of the Microthamniales (Pleurastrophyceae) and for Gloeotilopsis planctonica Iyengar & Philipose (Ulvophyceae, Ulotrichales) to investigate evolutionary relationships within the Microthamniales and the taxonomic position of this order within the green algae. Phylogenies inferred from these data revealed specific relationships at the level of genera and species that disagree with those inferred from vegetative cell morphology but agree with those inferred from motile cell characters. The rRNA phylogenies provide even better resolution than that gained from morphology alone. The coccoid lichen alga Trebouxia spp. is specifically related to other coccoid lichen and soil algae (i.e. Myrmecia biatorellae Boye-Petersen and Friedmannia israelensis Chantanachat & Bold), forming the “Lichen Algae Group,” an evolutionarily distinct lineage within the Microthamniales. Trebouxia is a paraphyletic and Pleurastrum a polyphyletic genus in rRNA phylogenies. In contrast to previous hypotheses based on morphology, Pleurastrum terrestre Fritsch & John is not closely related to Trebouxia but occupies an isolated position within the Microthamniales. The filamentous alga Microthamnion kuetzingianum is not ancestral to coccoid members of the Microthamniales but is closely related with the coccoid Fusochloris perforata (Lee & Bold) Floyd, Watanabe & Deason. The Microthamniales are inferred to be an array of independent lineages that radiate nearly simultaneously and may also include some autosporic coccoid taxa previously classified in the Chlorophyceae. Monophyly of the Microthamniales could not be demonstrated unequivocally. In contrast to a hypothesis based on ultrastructure, the Microthamniales are evolutionarily distinct from the Ulotrichales. The latter are ancestral to the radiation of the Microthamniales and the Chlorophyceae in the rRNA phylogenies.     

Calmodulin Sensitivity of the Flagellar Membrane Adenylate Cyclase and Signaling of Motile Responses by cAMP in Gametes of Chlamydomonas reinhardtii

Cyclic AMP (cAMP) has been shown to be a primary signal of the agglutination-induced mating events of flagellar tip activation, cell wall loss, and mating structure activation in the unicellular alga Chlamydomonas reinhardtii (Pasquale and Goodenough, Cell Biol. 105 (1987), 2279–2293). The flagellar membrane adenylate cyclase of Chlamydomonas is here shown to be inhibited in vitro by EGTA, La³⁺, and trifluoperazine, and to be stimulated in the presence of calcium by incubation with exogenous calmodulin. Also, the motility of detergent-extracted models of Chlamydomonas is shown to be enhanced by cAMP. These observations suggest the hypothesis that the twitching motility characteristic of agglutinating Chlamydomonas gametes may be signaled by cAMP produced locally within the flagella by a calmodulin-sensitive adenylate cyclase.