Yoshiaki Hara

An allelopathic fatty acid from the brown alga Cladosiphon okamuranus

Abstract An allelopathic substance was isolated from the methanol extract of the brown alga, Cladosiphon okamuranus . Extraction and purification of the active substance was monitored by bioassays against the conchospores of Porphyra yezoensis and Heterosigma akashiwo . The compound was identified as 6 Z ,9 Z ,12 Z ,15 Z -octadecatetraenoic acid and its activity was then tested against 36 species of microalgae. Many of eukaryotes tested were affected at 5 ppm, but prokaryotes were unaffected at 25 ppm. The tetraunsaturated fatty acid exterminated toxic red tide planktons such as Chattonella antiqua at 1 ppm concentration.

Morphology, ultrastructure and taxonomy of the raphidophycean alga Heterosigma akashiwo

Heterosigma akashiwo and its related algae were re-examined by light and electron microscopes using cultured materials including type cultures of bothH. akashiwo andH. inlandica as well as specimens referred to asOlisthodiscus luteus maintained in CCAP and UTEX. All the strains examined were similar to one another in appearance and ultrastructural features. They can be accommodated in a single species,H. akashiwo, which has been invalidly published.In this paper, the genus is validly desribed by providing a Latin diagnosis and designating a type species. Ultrastructural characteristics are also given for the genus.

The Origin of Chlorarachniophyte Plastids, as Inferred from Phylogenetic Comparisons of Amino Acid Sequences of EF-Tu

Abstract. A molecular phylogenetic analysis of elongation factor Tu (EF-Tu) proteins from plastids was performed in an attempt to identify the origin of chlorarachniophyte plastids, which are considered to have evolved from the endosymbiont of a photosynthetic eukaryote. Partial sequences of the genes for plastid EF-Tu proteins (1,080–1,089 bp) were determined for three algae that contain chlorophyll b, namely, Gymnochlora stellata (Chlorarachniophyceae), Bryopsis maxima (Ulvophyceae), and Pyramimonas disomata (Prasinophyceae). The deduced amino acid sequences were used to construct phylogenetic trees of the plastid and bacterial EF-Tu proteins by the maximum likelihood, the maximum parsimony, and the neighbor joining methods.The trees obtained in the present analysis suggest that all plastids that contain chlorophyll b are monophyletic and that the chlorarachniophyte plastids are closely related to those of the Ulvophyceae. The phylogenetic trees also suggest that euglenophyte plastids are closely related to prasinophycean plastids. The results indicate that the chlorarachniophyte plastids evolved from a green algal endosymbiont that was closely related to the Ulvophyceae and that at least two secondary endosymbiotic events have occurred in the lineage of algae with plastids that contain chlorophyll b.

Low molecular weight carbohydrates of the Bangiophycidae (Rhodophyta)

In the order Porphyridiales there are three clades based on molecular evidence. These show parallels with the low molecular weight carbohydrate (LMWCs) in different genera. Clade Porphyridiales 1 includes Dixoniella, Glaucosphaera, Rhodella, and one undescribed genus (3987) that all contain mannitol. Clade Porphyridiales 2 comprises taxa of the Stylonematales Rhodosorus and Stylonema species and contains digeneaside and sorbitol, whereas Chroodactylon has only sorbitol. In clade Porphyridiales 3 Flintiella, Porphyridium, and the undescribed genus (3797) all possess only floridoside. In the Erythropeltidales Rhodochaete contains floridoside and digeneaside, Erythrotrichia species contain only floridoside, Sahlingia subintegra has floridoside and traces of D‐floridoside, and Smithora has L‐isofloridoside plus floridoside. In the Compsopogonales Boldia and Compsopogon have only floridoside. Within these genera as presently circumscribed, the LMWCs appear to be a reliable character to supplement the usual cytological characters.

Taxonomic re-examination of Nitella (Charales, Charophyceae) from Japan, based on microscopical studies of oospore wall ornamentation and rbcL gene sequences

Abstract Nine taxa of Nitella (Charales, Charophyceae) from Japan, including five Japanese or east Asian endemics, were examined to improve understanding of their taxonomic status. The approaches used were light and scanning electron microscopy (SEM) of the oospores and sequencing of the gene (rbcL) encoding the large subunit of Rubisco. The species delineated were N. pulchella (= N. dualis f. pulchella), N. furcala, N. inversa (= N. furcata f. inversa), N. gracilens (= N. furcata f. gracilens), N. pseudoflabellata, N. hyalina, N. spieiformis (= N. gracilis f. spiciformis), N. moriokae (= N. rigida f. moriokae) and N. axillaris (= N. translucens var. axillaris). SEM observations showed that the oospore morphology of four taxa – N. inversa, N. gracilens, N. spiciformis and N. axillaris – was distinctly different from that of the species (N. furcata, N. furcata, N. gracilis and N. translucens, respectively) to which R.D. Wood assigned them as infraspecific taxa. Furthermore, the rbcL data showed that N. gracilens was separated phylogenetically from N. furcata and N. axillaris from N. translucens. This is the first integration of SEM oospore morphology and molecular phylogenetics in charalean taxonomy, demonstrating the efficiency of both approaches to address problems at lower taxonomic levels.

The Complete Plastid Genome Sequence of the Secondarily Nonphotosynthetic Alga Cryptomonas paramecium: Reduction, Compaction, and Accelerated Evolutionary Rate

The cryptomonads are a group of unicellular algae that acquired photosynthesis through the engulfment of a red algal cell, a process called secondary endosymbiosis. Here, we present the complete plastid genome sequence of the secondarily nonphotosynthetic species Cryptomonas paramecium CCAP977/2a. The ∼78 kilobase pair (Kbp) C. paramecium genome contains 82 predicted protein genes, 29 transfer RNA genes, and a single pseudogene (atpF). The C. paramecium plastid genome is approximately 50 Kbp smaller than those of the photosynthetic cryptomonads Guillardia theta and Rhodomonas salina; 71 genes present in the G. theta and/or R. salina plastid genomes are missing in C. paramecium. The pet, psa, and psb photosynthetic gene families are almost entirely absent. Interestingly, the ribosomal RNA operon, present as inverted repeats in most plastid genomes (including G. theta and R. salina), exists as a single copy in C. paramecium. The G + C content (38%) is higher in C. paramecium than in other cryptomonad plastid genomes, and C. paramecium plastid genes are characterized by significantly different codon usage patterns and increased evolutionary rates. The content and structure of the C. paramecium plastid genome provides insight into the changes associated with recent loss of photosynthesis in a predominantly photosynthetic group of algae and reveals features shared with the plastid genomes of other secondarily nonphotosynthetic eukaryotes.

EVOLUTIONARY DIVERGENCE IN THE RED ALGAE CALOGLOSSA LEPRIEURII AND C. APOMEIOTICA

Morphological comparisons, hybridization experiments, and molecular phylogenetic analyses using the RUBISCO spacer region were undertaken on 12 populations of Caloglossa leprieurii (Montagne) J. Agardh in order to clarify their relationships. In addition, data from one population of the morphologically similar but asexual species, C. apomeiotica (West et Zuccarello), were included in the assessment. Three morphological types were recognized on the basis of the number of cell rows at nodes of the main axis opposite to the lateral branch and blade width: single/ broad (with three mating groups), multiple/broad (three mating groups) and multiple/slender (one mating group). In the molecular analyses, C. leprieurii was resolved as two clusters that correspond phenetically to the single and multiple cell row types. Both the morphological and molecular data indicate that the asexual species was derived from sexual plants of the multiple cell row type. The reproductive compatibility correlates with genetic distance rather than geographical distance. Sympatric mating groups are completely incompatible and have 10–21 nucleotide changes in the examined region, whereas mating groups that produce abnormal progeny or pseudocystocarps are allopatrically distributed with 5–7 nucleotide changes. The present data suggest that the two populations, one with single and the other with multiple cell rows, which are sympatrically distributed in southeastern Japan, have probably evolved by allopatric speciation. The single/broad type that is restricted to the western Pacific, may have diverged genetically between eastern and western Australia, with subsequent dispersal from the western population as far as Japan.

Taxonomic re‐examination of 17 species of Nitella subgenus tieffallenia (Charales, Charophyceae) based on internal morphology of the oospore wall and multiple DNA marker sequences (Vol. 41:195–211)

In an attempt to reconstruct the natural taxonomic system for Nitella, 17 species of Nitella subgenus Tieffallenia were reexamined using SEM observations of the internal morphology of the oospore wall (IMOW) and phylogenetic analyses of 4553 base pairs from multiple DNA markers (atpB, rbcL, psaB, and ITS‐5.8S rRNA genes). Our SEM observations identified three types of IMOW: homogeneous (HG), weakly spongy (W‐SG), and strongly spongy (S‐SG) types. Based on differences in the IMOW, species with reticulate or tuberculate oospore wall ornamentation in the external morphology of the oospore wall (EMOW) were subdivided into two distinct groups (characterized by the HG or S‐SG types of IMOW, respectively), which were robustly separated from each other in our molecular phylogenetic analyses. In our molecular phylogeny, the subgenus Tieffallenia consisted of four robust monophyletic groups—three clades of the HG type and a spongy (S‐SG and W‐SG) type clade—that were characterized by differences in the IMOW and EMOW. In addition, our SEM observations and sequence data verified the distinct status of five species (N. japonica Allen, N. oligospira A. Braun, N. vieillardii stat. nov., N. imperialis stat. nov., and N. morongii Allen) that R. D. Wood had assigned as infraspecific taxa. Moreover, our SEM observations of the IMOW also suggested that N. megaspora (J. Groves) Sakayama originally identified by LM includes at least two distinct species, characterized by W‐SG and S‐SG types of IMOW, respectively.

Isolation, characterization and chromosomal mapping of an actin gene from the primitive red alga Cyanidioschyzon merolae

Based on the results of cytological studies, it has been assumed that Cyanidioschyzon merolae does not contain actin genes. However, Southern hybridization of C. merolae cell-nuclear DNA with a yeast actin-gene probe has suggested the presence of an actin gene in the C. merolae genome. In the present study, an actin gene was isolated from a C. merolae genomic library using a yeast actin-gene probe. The C. merolae actin gene has no intron. The predicted actin is composed of 377 amino acids and has an estimated molecular mass of 42003 Da. Southern hybridization indicated that the C. merolae genome contains only one actin gene. This gene is transcribed at a size of 2.4 kb. When Southern hybridization was performed with C. merolae chromosomes separated by pulsed-field gel electrophoresis, a band appeared on unseparated chromosomes XI and XII. A phylogenetic tree based on known eucaryote actin-gene sequences revealed that C. merolae diverged after the division of Protozoa, but before the division of Fungi, Animalia and Chlorophyta.

Pigments and fatty acids of marine raphidophytes: A chemotaxonomic re-evaluation

The patterns of occurrence of photosynthetic pigments and fatty acids among seven available species (11 strains) of marine raphidophytes were determined and used as chemotaxonomic markers. All currently recognized genera of marine raphidophytes were included for analysis: that is, Chattonella, Fibrocapsa, Heterosigma, Olisthodiscus and Haramonas. The characteristic pigment composition was shown to be chlorophyll a, chlorophylls c1 and/or c2, fucoxanthin as the major carot‐enoid, β,β‐carotene and any or all of zeaxanthin, violaxanthin and an auroxanthin‐like pigment as the minor carotenoids. The carotenoid composition of all marine raphidophyte genera investigated was virtually the same, except in Fibrocapsa and Haramonas, which differed due to the occurrence of fucoxanthinol and 19′‐butanoyloxyfucoxanthin, respectively. These fucoxanthin derivatives, in addition to fucoxanthin, have potential chemotaxonomic use for differentiating the two species. In all 11 strains, 15 fatty acids (saturated, mono‐unsaturated and polyunsaturated) were determined. Significant taxonomic distinctions between genera were reflected by their fatty acid profiles. A rapid key for the differentiation of genera, in addition to morphological features, may be the absence of the 18:4 fatty acid in Olisthodiscus; presence of 18:5 in Heterosigma; the presence of fucoxanthinol in Fibrocapsa and presence of 19′‐butanoyloxyfucoxanthin in Haramonas.