OBSERVATIONS ON THE FINE STRUCTURE OF OEDOGONIUM. V. EVIDENCE FOR THE DE NOVO FORMATION OF PYRENOIDS IN ZOOSPORES OF OE. CARDIACUM123

Ultrastructural studies of the chloroplasts of zoospores and developing zoospores of Oedogonium carcliacum have disclosed the occurrence of numerous incipient pyrenoids. A single developing zoospore may possess several score of these structures which appear to arise de novo in the chloroplast stroma and seem to lack any direct association with mature pyrenoids which are also present in the cells. The incipient pyrenoids lack the associated starch grains and the membrane-limited channels characteristic of mature pyrenoids, but they are readily recognized in the chloroplasts since they demonstrate a greater granularity and electron density than the surrounding chloroplast stroma. The granularity and electron density of the incipient pyrenoids match the ultra-structural appearance of the matrix of mature pyrenoids. The smallest of the incipient pyrenoids examined from serial sections had a maximum diameter of less than 0.3 μ. This may be compared with the size of mature pyrenoids, many with a maximum diameter of over 5.0 μ. In all the zoospores and developing zoospores examined, only one mature pyrenoid was observed in an apparent stage of division.     

NEW ULTRASTRUCTURAL ASPECTS OF PYRENOIDS OF THE LICHEN PHOTOBIONT TREBOUXZA (MICROTHAMNIALES,CHLOROPHYTA)1

The pyrenoid structure of Trebouxia, a photobiont of two lichen species, Umbilicaria cinereorufescens (Schaer.) Frey and Parmelia sulcata Taylor, was investigated. In both lichen species, the pyrenoid of the photobiont exhibited straight, unbranched, long or short tubules. In the first lichen species, multiple pyrenoids were observed occasionally, while in the second one, homogeneous masses, called protein bodies, appeared between the thylakoids. These protein bodies were previously observed in some other species of the family Umbilicariaceae. Serial sections from single pyrenoids showed that tubules of the Impressa-type pyrenoid were closely associated with pyrenoglobuli. The three-dimensional reconstruction of a complete chloroplast of a P. sulcata algal cell showed that the protein bodies were spatially separate structures. Immunolocalization techniques to detect the presence of ribulose-bisphosphate carboxylase (Rubisco) in the chloroplast showed that this enzyme was present primarily in the pyrenoid matrix. When protein bodies were present in the chloroplast, Rubisco appeared to be localized in these structures. The presence of pyrenoid satellites and protein bodies with reactivity to anti-Rubisco may be related to the nutritional conditions of the thalli.     

Immunocytochemical Localization of Phosphoribulokinase in Microalgae

Employing immunogold electron microscopy, the subcellular location of the Calvin cycle enzyme phosphoribulokinase (PRK) was determined for two diverse species of microalgae. In both the red alga Porphyridium cruentum and the green alga Chlamydomonas reinhardtii, PRK was distributed throughout the thylakoid-containing chloroplast stroma. In contrast, the next enzyme in the pathway, ribulose 1,5-bisphosphate carboxylase/oxygenase, was predominantly pyrenoid-localized in both species. In Porphyridium, the chloroplast stroma abuts the pyrenoid but in Chlamydomonas and other green algae, the pyrenoid appears encased in a starch sheath. Unique inclusions found in the pyrenoid of Chlamydomonas were immunolabelled by anti-PRK and thus identified as regions of chloroplast stroma. It is postulated that such PRK-containing stromal inclusions in the pyrenoids of Chlamydomonas and perhaps other green algae provide a means for exchange of Calvin cycle metabolites between pyrenoid and stroma.     

FINE STRUCTURE OF THE ZOOSPORE OF OEDOCLADIUM CAROLINIANUM (CHLOROPHYTA) WITH SPECIAL REFERENCE TO THE FLAGELLAR APPARATUS1, 2

Ultrastructure of the motile zoospore has been investigated in Oedocladium catolinianum & Hoffman. An unwalled zoospore is usually produced from the contents of a terminal vegetative cell and consists of two principal regions: a small anterior dome and a larger body region; a ring of flagella marks the juncture of these two areas. Chloroplast inclusions consist of thylakoids, mature and incipient pyrenoids, starch and striated microtubules; no eyespot has been observed. Zoospores appear to possess permanent contractile vacuoles with numerous accessory vacuoles, coated vesicles and occasionally coated tubules. The cytoplasm of the dome contains numerous mitochondria ER and golgi bodies, as well as two distinct types of vesicles. The first contains an electron-dense; granular core and is surrounded by a loose, sinuate membrane. The second vesicle is electron-opaque and is found at the apex of the dome: it contains mucopolysaccharides employed during zoospore adhesion. A complex flagellar apparatus encircles the lower region of the dome. It consists of ca. 30–65 flagella, a ring-shaped fibrous band, flagella roots and additional supporting material. The flagella and roots alternate with one another beneath the fibrous band. The compound flagellar roots consist of two superimposed components: an outer ribbon-like unit composed of three microtubular elements and a single striated inner component. A band of support material lies beneath the proximal end of the basal bodies. It is a continuous fibrous band, although it often appears as three distinct, repetitive units.     

COMPARATIVE ULTRASTRUCTURE OF CULTURED SPECIES OF TREBOUXIA1,2

Five species of cultured Trebouxia—T. anticipata, T. decolorans, T. erici, T. gelatinosa, and T. impressa—were examined with the electron microscope. A comparative examination of their pyrenoids revealed pyrenoglobuli associated with single pyrenoid thylakoids. The pyrenoids of T. decolorans, T. erici, and T. gelatinosa possess single thylakoids that cross or deeply penetrate the pyrenoid matrix and are often disposed in parallel arrays. T. anticipata possesses both single and double pyrenoid thylakoids within the matrix. T. impressa possesses vesiculate invaginations of thylakoid membranes into the pyrenoid matrix. The phycobiont. T. erici was examined in detail at the light and electron microscopic levels for pyrenoid alterations associated, with varied environmental regimes and with cell division. A greater amount of starch is present in cells grown in organic culture at 215 lux light intensity than in cells of similar size grown at 1075 or 3600 lux. Pyrenoglobuli are present throughout the life cycle and occur both in aplanospores and in zoospores.     

STRUCTURE AND FUNCTION OF THE ALGAL PYRENOID. I. ULTRASTRUCTURE AND CYTOCHEMISTRY DURING ZOOSPOROGENESIS OF TETRACYSTIS EXCENTRICA1

The fine structure of the pyrenoid in the mature vegetative cell of Tetracystis excentrica Brown and Bold is described. During zoosporogenesis, the pyrenoid undergoes regression, and the ultrastructure of this process is described in detail. The ground substance undergoes dissolution, and reticulate fibrillar structures appear as well as intruding chloroplast thylakoids. Pyrenoid-associated starch plates diminish, and quantities of starch not associated with the pyrenoid are produced. New pyrenoids appear late in the division cycle after all other major organelles associated with the motile cell have been formed. Zoospore pyrenoids develop in thylakoid-free spaces of the chloroplast which are similar to the DNA-containing regions. The new pyrenoid ground substance, which is loosely fibrillar, arises in close proximity to starch grains which may be formed in the stroma. Then the zoospore pyrenoid produces 2 hemispherical starch plates identical to those in the mature vegetative cell. Zoospore pyrenoids lack the 2 convoluted thylakoids between the starch plates and the ground substance characteristic of those in the mature vegetative cell. Instead, the thylakoids are identical to those of the chloroplast at first, and then develop into a convoluted state in the vegetative cell. Cytochemical tests for DNA, RNA, and protein were made for the cytoplasm, nucleus, nucleolus, and pyrenoid. Conclusive evidence is presented for the presence of RNA in the cytoplasm and nucleolus, DNA in the nucleus, and protein in the pyrenoid. The tests did not conclusively demonstrate the presence or absence of DNA and RNA in the pyrenoid; however, they suggested that small amounts of both DNA and RNA may be present.     

FLAGELLAR APPARATUS OF THE BIFLAGELLATE ZOOSPORES OF THE ENIGMATIC MARINE GREEN ALGA BLASTOPHYSA RHIZOPUS1

The flagellar apparatus of the biflagellate zoospores from Blastophysa rhizopus Reinke has 180° rotational symmetry of the major components and counterclockwise absolute orientation. The basal bodies are connected anteriorly by a prominent striated distal fiber and posteriorly by two proximal striated bands. The C microtubules in the basal bodies terminate proximal to the transition region. Terminal caps and well-defined proximal sheaths are absent. The four microtubular rootlets diverge at a very small angle from the basal bodies. Six to eight (usually seven) microtubules are present in the s rootlets and two microtubules in the d rootlets. Rootlet 1s is associated with the eyespot. Each d rootlet is subtended by a coarsely striated fiber. Rootlet Id also has a finely striated fiber, roughly opposite the coarsely striated fiber, associated with it. Rhizoplasts and mating structures were not observed. Ultrastructural features of B. rhizopus zoospores are essentially identical with those found in examined members of the Siphonocladales sensu lato (= Siphonocladadales/Cladophorales complex) and Dasycladales, and have relatively few features in common with motile cells of caulerpalean algae. Blastophysa rhizopus probably does not represent an intermediate between the Siphonocladadales and the Caulerpales. Its evolutionary history is different from that of other algae placed in the siphonocladalean family Chaetosiphonaceae. Whether or not Blastophysa is representative of the ancestor to the Siphonodadales and Dasycladales is unclear.     

LIGHT AND ELECTRON MICROSCOPIC CHARACTERIZATION OF TWO TYPES OF PYRENOIDS IN GONIUM (GONIACEAE,CHLOROPHYTA)1

The single, basal pyrenoids of Gonium quadratum Pringsheim ex Nozaki and G. pectorale Müller (Goniaceae, Chlorophyta) differed in appearance when vegetative colonies were cultured photoheterotrophically in medium containing sodium acetate. Chloroplasts of G. quadratum had distinct pyrenoids when grown in medium without major carbon compounds. However, the pyrenoids degenerated and were markedly reduced in size when such cells were inoculated into a medium containing 400 mg·L^?1 of sodium acetate. No pyrenoids were visible under the light microscope; however, with electron microscopy small pyrenoids and electron-dense bodies were visible within the degenerating chloroplasts, which had only single layers of thylakoid lamellae at the periphery. The chloroplasts subsequently developed distinct pyrenoids and several layers of thylakoid lamellae as the culture aged. In contrast, vegetative cells of G. pectorale always showed distinct pyrenoids when cells were inoculated into medium containing sodium acetate, sodium pyruvic acid, sodium lactate, and/or yeast extract. Therefore, we propose two terms, “unstable pyrenoids” and “stable pyrenoids,” for pyrenoids of G. quadratum and G. pectorale, respectively. Chloroplasts of the colonial green flagellates should thus be examined under various culture conditions in order to determine whether their pyrenoids are unstable or stable when pyrenoids are used as taxonomic indicators. Immunogold electron microscopy showed that the ratios of gold particle density of ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCO) between pyrenoid matrix and chloroplast stroma in G. quadratum grown in medium with or without sodium acetate were lower than those of G. pectorale. Heavy labeling by anti-RuBisCO was observed in both the electron-dense bodies and pyrenoid matrix of G. quadratum. This is the first electron microscopic demonstration of degeneration and development of both pyrenoids and thylakoid lamellae in the chloroplast as a function of culture condition in green algae.     

NEW PYRENOID FORMATION IN THE BROWN ALGA,SCYTOSIPHON LOMENTARIA (SCYTOSIPHONALES,PHAEOPHYCEAE)1

The Scytosiphon lomentaria (Lyngbye) Link cell characteristically has only one chloroplast with a prominent protruding pyrenoid. We observed the appearance of a new pyrenoid in each chloroplast during first mitosis in zygotes of S. lomentaria, using the freeze substitution technique. At first, a pyrenoid matrix appeared within the outermost stroma, in which thylakoid triplets and ribosomes were absent. At this time, the surface of this part remained smooth. The old pyrenoid was covered with a pyrenoid cap on the cytoplasmic side, whereas there was no pyrenoid cap on the new pyrenoid before protrusion. Irregularly shaped membranous sacs containing fine granular materials associated with the cytoplasmic side of the new pyrenoid. The sacs fused with each other and changed conformation and finally transformed into the pyrenoid cap. The new pyrenoid gradually protruded toward the cytoplasm, and the new pyrenoid cap became curved along the surface of pyrenoid. Cytokinesis occurred, and each chloroplast had two prominent protruding pyrenoids in two‐celled zygotes. We examined immunolocalization of β‐1,3‐glucans within the pyrenoid cap with a monoclonal antibody, using EM. Gold particles indicating localization of β‐1,3‐glucans were detected in vacuoles but never in the pyrenoid cap. This observation suggests that the pyrenoid cap in brown algae contains no photosynthetic products such as polysaccharide.     

Re-examination of ultrastructures of the stellate chloroplast organization in brown algae: Structure and development of pyrenoids

Some taxa of brown algae have a so‐called ‘stellate’ chloroplast arrangement composed of multiple chloroplasts arranged in a stellate configuration, or else a single chloroplast with radiating lobes. The fine structures of chloroplasts and pyrenoids have been studied, but the details of their membrane configurations as well as pyrenoid ontogeny have not been well understood. The ultrastructure of the single stellate chloroplast in Splachnidium rugosum and Scytothamnus australis were re‐examined in the present study, as well as the stellate arrangement of chloroplasts in Asteronema ferruginea and Asterocladon interjectum, using freeze‐substitution fixation. It was confirmed that the chloroplast envelope invaginated into the pyrenoid in Splachnidium rugosum, Scytothamnus australis and Asteronema ferruginea, but chloroplast endoplasmic reticulum (CER) remained on the surface of the chloroplast. The space between the invaginated chloroplast envelope and CER was filled with electron‐dense material. In Asteronema ferruginea, CER surrounding each pyrenoid was closely appressed to the neighboring CER over the pyrenoids, so that the chloroplasts formed a stellate configuration; however, in the apical cells chloroplasts formed two or more loose groups, or were completely dispersed. The pyrenoids of Asterocladon interjectum did not have any invagination of the chloroplast envelope, but a unique membranous sac surrounded the pyrenoid complex and occasionally other organelles (e.g. mitochondria). Immunolocalization of β‐1,3‐glucans showed that the membranous sac in Asterocladon interjectum did not contain photosynthetic products such as chrysolaminaran. Observations in the dividing cells of Splachnidium rugosum and Scytothamnus australis indicated that the pyrenoid in the center of the chloroplast enlarged and divided into two before or during chloroplast division.     

LIGHT AND ELECTRON MICROSCOPY OF PYRENOIDS AND SPECIES DELIMITATION IN VOLVULINA (CHLOROPHYTA,VOLVOCACEAE)1

The appearances of pyrenoids in the vegetative cells of Volvulina steinii Playfair and V. pringsheimii Starr were observed in detail by light and electron microscopy in relation to the culture age to clarify the taxonomic relationship between the two species. In V. pringsheimii, the pyrenoids were always present in the bottom of the cupshaped chloroplasts and their gross morphology did not vary in relation to the culture age, while those of V. steinii appeared de novo and developed as the culture aged. In 24-h cultures of V. steinii, pyrenoids were not observed in the chloroplasts. In 48-h cultures, a pyrenoid matrix developed apparently de novo in the brim of the cupshaped chloroplast. Subsequently, starch grains appeared around the pyrenoid matrix in 72-h cultures. The volume of the matrix and the associated starch grains increased and tubular channels entered into the pyrenoid matrix in 96-h cultures. In addition, the pyrenoid in the parental chloroplast of V. pringsheimii divided and was distributed to each daughter cell during cell divisions in daughter colony formation, while the parental pyrenoid of V. steinii did not divide and went to one of the daughter cells. Therefore, these two species can be clearly distinguished by the differences in the position of pyrenoids in the cupshaped chloroplasts and stability of pyrenoid appearance in relation to the culture age, as well as in the fate of parental pyrenoids during daughter colony formation.     

CHLOROPLAST DIVISION AND PYRENOID FORMATION IN CHLAMYDOMONAS REINHARDI1

The process of chloroplast division during mitosis in Chlamydomonas reinhardi is followed with the electron microscope. The pyrenoid and the chloroplast reproduce by fission. The dividing chloroplast contains regions of dense material that superficially resemble pyrenoids, but it is concluded that the formation of the dense material is not related to pyrenoid formation in C. reinhardi. The dense material appears to be localized over regions of chloroplast DNA.     

CYTOLOGY AND ULTRASTRUCTURE OF CHLORARACHNION REPTANS (CHLORARACHNIOPHYTA DIVISIO NOVA,CHLORARACHNIOPHYCEAE CLASSIS NOVA)1

The green amoeboid cells of Chlorarachnion reptans Geitler are completely naked and each contains a central nucleus, several bilobed chloroplasts each with a central projecting pyrenoid enveloped by a capping vesicle, several Golgi bodies, mitochondria with tubular cristae, extensive rough ER, and a distinct layer of peripheral vesicles. Complex extrusome-like organelles occur rarely in both the amoeboid and flagellate stages. The only organelles entering the reticulopodia are mitochondria, but microtubules are also present. The chloroplasts contain chlorophylls a and b, but histochemical tests suggest that the carbohydrate storage product probably is not a starch. The chloroplast lamellae are composed of one to three thylakoids or form deep stacks. A girdle lamella and interlamellar partitions are absent. Each chloroplast is bounded by either four separate membranes, a pair of membranes with vesicular profiles between them, or three membranes; all three arrangements may occur in the same chloroplast. A periplastidal compartment occurs near the base of the pyrenoid where there are always four surrounding membranes. The compartment has a relatively dense matrix and contains ribosome-like particles and small dense spheres; it extends over and into a deep invagination in the pyrenoid where its contents are enclosed in a double-membraned envelope which is penetrated by wide pores. The zoospores are ovoid and each bears a single laterally inserted flagellum which appears to be wrapped helically around the cell body during swimming. The flagellum lies in a groove in the cell surface and bears fine lateral hairs. Neither a second flagellum or vestige of one, nor an eyespot, is present. A single microtubular root and a larger homogeneous root run from the flagellar base parallel to the emerging flagellum, between the nuclear envelope and the plasmalemma. In the simple flagellar transition region, fine filaments connect adjacent axonemal doublets. A detailed comparison of C. reptans with all other algal taxa results in the conclusion that it must be segregated in the new class Chlorarachniophyceae, the only class in the new division Chlorarachniophyta. The possibility that C. reptans evolved from a symbiosis between a colorless amoeboid cell and a chlorophyll b- containing eukaryote is considered, but the possible affinities of the symbiont remain enigmatic. The implications of the unique chloroplast structure of C. reptans for current hypotheses concerning the origin of chloroplasts are discussed.     

Comparative ultrastructure of the zoospores of nine species ofNeochloris (Chlorophyta)

Nine species ofNeochloris can be divided into three groups on the basis of comparative ultrastructure of the flagellar apparatus, the cell wall and the pyrenoid of zoospores. In Group I,N. wimmeri andN. minuta, zoospores are thin-walled, pyrenoids are penetrated by stromal channels, and the basal bodies are in the clockwise absolute orientation and connected by the distal and two proximal fibers. In Group II,N. aquatica, N. vigenis, N. terrestris, N. pyenoidosa, andN. pseudostigmatica, zoospores are naked or covered by fuzzy material, pyrenoids are covered by a continuous starch sheath or invaginated by cytoplasmic channels, basal bodies are directly opposed, the distal fiber is differentiated into a ribbed structure at the central region, a striated microtubule-associated component (SMAC) is continuous between opposite two-membered rootlets and connected to the ribbed structure, proximal ends of basal bodies are covered by partial caps, each two-membered rootlet and a basal body are connected by a striated fiber to the X-membered rootlet associated with the opposite basal body, and the basal bodies, when oriented at wide angles, are joined at their proximal ends by core extensions. In Group III,N. pseudoalveolaris andN. cohaerens, zoospores are naked, pyrenoids are traversed by parallel thylakoids, basal bodies are in the counterclockwise absolute orientation and overlapped, and each X-membered rootlet is connected to the end of the opposite basal body by a terminal cap. It is suggested that the genusChlorococcopsis gen. nov. be erected for the Group I species. Group II, which includes the type species,N. aquatica, should be preserved asNeochloris. The group appears to be closely related to the coenobial generaPediastrum, Hydrodictyon, andSorastrum, and to have affinities with the coenocytic generaSphaeroplea andAtractomorpha as well. It is also suggested that the genusParietochloris gen. nov. be erected in thePleurastrophyceae for the species of Group III.     

PHYLOGENETIC TRANSITIONS IN THE CHLOROPLASTS OF THE ANTHOCEROTALES I. THE NUMBER AND ULTRASTRUCTURE OF THE MATURE PLASTIDS

Representatives of three genera of anthooerotes were examined: Phaeoceros, Notothylas, and Megaceros. Species of the first two genera were found to exemplify the typical anthocerote plastid condition. This condition is characterized by the presence in each cell of the gametophyte of only a single large chloroplast containing a “multiple” pyrenoid. The genus Megaceros, however, proved to be quite different. In two species of Megaceros the pyrenoid was observed to be composed of a highly subdivided thylakoid system of even greater complexity than the “multiple” pyrenoids of Phaeoceros. In another species only an indistinct “pyrenoid-like” area was noted while in a fourth species no evidence was found for any internal differentiation. Associated with these changes in plastid structure there are corresponding alterations in the number and the size of the chloroplasts. Together they indicate an evolutionary trend away from a primitive, algal-like condition to a more advanced land plant form.     

Observations on chloroplast growth and pyrenoid formation inSpirogyra. A study by means of uncoiled picture of chloroplast

By a newly developed method for recording a circumferential view of a cylinder cell, growth characteristics of the chloroplast and pyrenoid formation inSpirogyra were studied. Because of no active migration of pyrenoids in the chloroplast, they were used as indices for local growth of the chloroplast. The chloroplast ribbon grew diffusively and evenly in the helical direction over its entire length. Pyrenoids multiplied only throughde novo formation, but not through division. Formation of a new pyrenoid occurred after the distance between two adjacent pyrenoids exceeded a critical length. The formation was independent of the cell cycle and did not occur at specific region of the chloroplast.     

AXILOSPHAERA AND HETEROTETRACYSTIS,NEW CHLOROSPHAERACEAN GENERA FROM TENNESSEE SOIL12

Two new chlorosphaeracean genera were isolated into axenic culture from soil collected in cedar glades in Cedars of Lebanon State Forest, Wilson County, Tennessee. The distinguishing characteristics of the new monotypic genus Axilosphaera include an axile (asymmetric) ckloroplast with at least 1 pyrenoid and Chlamydomonas-type (walled) zoospores. A. vegetata is the type species. Reproduction is by dissociation of daughter cells following vegetative cell division, by zoospores, and by aplanospores. The new polytypic genus Heterotetracystis, comprising 3 species, H. akinetos, H. macrogranulosa, and H. intermedia, is characterized by a parietal chloroplast with at least 1 pyrenoid and walled zoospores with flagella of unequal length. Reproduction is by dissociation of daughter cells following vegetative cell division and by zoospores. H. akinetos is designated as the type species.     

EVOLUTIONARY ORIGIN OF GLOEOMONAS (VOLVOCALES,CHLOROPHYCEAE), BASED ON ULTRASTRUCTURE OF CHLOROPLASTS AND MOLECULAR PHYLOGENY1

Gloeomonas is a peculiar unicellular volvocalean genus because it lacks pyrenoids in the chloroplasts under the light microscope and has two flagellar bases that are remote from each other. However, ultrastructural features of chloroplasts are very limited, and no molecular phylogenetic analyses have been carried out in Gloeomonas. In this study, we observed ultrastructural features of chloroplasts of three species of Gloeomonas and Chloromonas rubrifilum (Korshikov ex Pascher) Pröschold, B. Marin, U. Schlösser et Melkonian SAG 3.85, and phylogenetic analyses were carried out based on the combined data set from 18S rRNA, ATP synthase beta‐subunit, and P700 chl a–apoprotein A2 gene sequences to deduce the natural phylogenetic positions of the genus Gloeomonas. The present EM demonstrated that the chloroplasts of the three Gloeomonas species and C. rubrifilum SAG 3.85 did not have typical pyrenoids with associated starch grains, but they possessed pyrenoid matrices that protruded interiorly within the stroma regions of the chloroplast. The pyrenoid matrices were large and broad in C. rubrifilum, whereas those of the three Gloeomonas species were recognized in only the small protruded regions of the chloroplast lobes. The present multigene phylogenetic analyses resolved that the three species of Gloeomonas belong to the Chloromonas lineage or Chloromonadinia of the Volvocales, and Chloromonas insignis (Anakhin) Gerloff et H. Ettl NIES‐447 and C. rubrifilum SAG 3.85, both of which have pyrenoids without associated starch grains, were positioned basally to the clade composed of the three species of Gloeomonas. Therefore, Gloeomonas might have evolved from such a Chloromonas species through reduction in pyrenoid matrix size within the chloroplast and by separating their two flagellar bases.