Investigation of a freshwater acrochaetioid alga (Rhodophyta) with molecular and morphological methods

An enigmatic acrochaetioid alga was collected from Niangziguan spring in Shanxi Province, northern China. Morphological data indicated that this alga reproduces exclusively asexually by monosporangia and its morphological characteristics suggested that it might be referred to Audouinella heterospora. To ascertain its phylogenetic position, phylogenetic trees were reconstructed using partial sequences of the plastid‐encoded gene (rbcL) and the nuclear‐encoded gene (SSU rDNA) applying Bayesian inference (BI), maximum parsimony (MP) and maximum likelihood (ML). However, phylogenetic reconstructions showed that this acrochaetioid alga does not belong in a clade with the genus Audouinella, but forms a clade with Thorea hispida (Thoreales). Based on this analysis it is concluded that A. heterospora represents the ‘chantransia’ stage of T. hispida.     

Three gene phylogeny of the Thoreales (Rhodophyta) reveals high species diversity

The freshwater red algal order Thoreales has triphasic life history composed of a diminutive diploid “Chantransia” stage, a distinctive macroscopic gametophyte with multi‐axial growth and carposporophytes that develop on the gametophyte thallus. This order is comprised of two genera, Thorea and Nemalionopsis. Thorea has been widely reported with numerous species, whereas Nemalionopsis has been more rarely observed with only a few species described. DNA sequences from three loci (rbcL, cox1, and LSU) were used to examine the phylogenetic affinity of specimens collected from geographically distant locations including North America, South America, Europe, Pacific Islands, Southeast Asia, China, and India. Sixteen species of Thorea and two species of Nemalionopsis were recognized. Morphological observations confirmed the distinctness of the two genera and also provided some characters to distinguish species. However, many of the collections were in “Chantransia” stage rather than gametophyte stage, meaning that key diagnostic morphological characters were unavailable. Three new species are proposed primarily based on the DNA sequence data generated in this study, Thorea kokosinga‐pueschelii, T. mauitukitukii, and T. quisqueyana. In addition to these newly described species, one DNA sequence from GenBank was not closely associated with other Thorea clades and may represent further diversity in the genus. Two species in Nemalionopsis are recognized, N. shawii and N. parkeri nom. et stat. nov. Thorea harbors more diversity than had been recognized by morphological data alone. Distribution data indicated that Nemalionopsis is common in the Pacific region, whereas Thorea is more globally distributed. Most species of Thorea have a regional distribution, but Thorea hispida appears to be cosmopolitan.     

UV‐radiation effects on photosynthesis and photoprotection in gametophytic and sporophytic stages of the freshwater red alga Kumanoa ambigua (Rhodophyta,Batrachospermales)

The aim of this study was to analyze the photosynthetic performance of gametophytic and sporophytic (‘Chantransia’) stages of Kumanoa ambigua in culture under UV radiation. We hypothesized that both life history stages of K. ambigua would exhibit different photosynthetic responses to UVR exposure. Experiments were performed under three conditions: (i) photosynthetically active radiation (PAR) only (400–700 nm), P control; (ii) PAR + UVA (320–700 nm), PA treatment; and (iii) PAR + UVA + UVB (280–700 nm), PAB treatment. The photosynthetic parameters were measured as in vivo chlorophyll a fluorescence. Differences were found between life stages, observing higher values of NPQ and effective quantum yields (ΔF/F_m′) under UVA and PAR in gametophytes compared to sporophytes. One type of mycosporine‐like amino acid (MAA) was detected in the gametophyte in all treatments, but not in the ‘Chantransia’ stage. The increased photosynthetic performance for some parameters and the presence of MAA in gametophyte suggest that it is less sensitive to UV radiation, particularly UVA, in comparison to sporophyte under culture conditions. This approach is relevant for a better understanding of the adaptation and physiological acclimation of freshwater Rhodophyta to varying light climates in terms of global changes.     

The marine algae of the Hebridean Island of Colonsay

Four specimens attributable to the freshwater red algal taxon, Ptilothamnion richardsii, were examined from locations in Australia and French Guiana, and partial rbcL gene sequence data were obtained. Phylogenetic analyses showed these specimens to be allied to taxa in the Batrachospermales. The two specimens from Australia were determined to be Batrachospermum antipodites, differing from that taxon by only 15?bp (1.2%). The two specimens from French Guiana were within the Batrachospermum section Contorta clade, but differed considerably (≥8%) from the taxa for which there are presently rbcL gene data. The sequence data clearly show these specimens to be chantransia stages of Batrachospermum taxa. Recently, other described freshwater Audouinella taxa have also been shown to be chantransia stages. Like previous authors, we argue that it is useful to continue to recognize entities with particular morphological characters and propose the form taxa Chantransia richardsii, C. macrospora and C. pygmaea. A key to these is provided.     

Systematics and distribution of Thorea (Thoreaceae, Rhodophyta) from central Mexico and south-eastern Brazil

Thirteen populations of Thorea were analyzed from central Mexico and south‐eastern Brazil. All populations were considered as belonging to a single species [Thorea hispida (Thore) Desvaux], with wide variation of morphological features. Secondary branches varying in frequency were observed in several populations with an overlapping in the range of branch density for Thorea violacea Bory and T hispida (0–9 and 11–41 per 30 mm, respectively). As this is the most distinguishing character and on the basis of the overlapping (within a same population or even a single plant), we regarded T. violacea as a synonym of T hispida.‘Chantransia’ stage in culture, as well as gametophyte and carposporophyte were described in detail. We confirmed tine coexistence of asexual monosporangia with sexual reproductive structures (carpogonia and spermatangia) and carposporangia. Size, content, arrangement and chromosome number were the most distinctive characteristics among spermatangia, carposporangia and monosporangia. Monosporangia can be promptly differentiated from spermatangia by their granulated content and larger size but they are similar to carposporangia in shape and size; however, monosporangia are not arranged in fascicles. Structures resembling bisporangia were observed in female plants of some populations. Chromosome numbers were n = 4 for spermatangia and fascicle cells, and 2n ca 8 for gonimoblast filaments, carpospores and the ‘Chantransia’ stage cells. The populations of Thorea from central Mexico and south‐eastern Brazil corroborated the known world distribution for T. hispida, consisting dommantly of tropical to subtropical rainforests, sometimes extending into warm temperate areas. Thorea hispida occurred in warm (temperature 17.6–28.O°C), neutral to alkaline (pH 7.0–8.0), high ion content (specific conductance 59–2140 μS cm^?1), moderate flowing (current velocity 17–43 cm/s) and shallow waters (depth <50cm); these data are essentially similar to previous reports.     

A conserved, Mg²+-dependent exonuclease degrades organelle DNA during Arabidopsis pollen development

In plant cells, mitochondria and plastids contain their own genomes derived from the ancestral bacteria endosymbiont. Despite their limited genetic capacity, these multicopy organelle genomes account for a substantial fraction of total cellular DNA, raising the question of whether organelle DNA quantity is controlled spatially or temporally. In this study, we genetically dissected the organelle DNA decrease in pollen, a phenomenon that appears to be common in most angiosperm species. By staining mature pollen grains with fluorescent DNA dye, we screened Arabidopsis thaliana for mutants in which extrachromosomal DNAs had accumulated. Such a recessive mutant, termed defective in pollen organelle DNA degradation1 (dpd1), showing elevated levels of DNAs in both plastids and mitochondria, was isolated and characterized. DPD1 encodes a protein belonging to the exonuclease family, whose homologs appear to be found in angiosperms. Indeed, DPD1 has Mg²⁺-dependent exonuclease activity when expressed as a fusion protein and when assayed in vitro and is highly active in developing pollen. Consistent with the dpd phenotype, DPD1 is dual-targeted to plastids and mitochondria. Therefore, we provide evidence of active organelle DNA degradation in the angiosperm male gametophyte, primarily independent of maternal inheritance; the biological function of organellar DNA degradation in pollen is currently unclear.     

A PROPOSAL FOR A NEW RED ALGAL ORDER,THE THOREALES1

Representatives of the freshwater red algal family Thoreaceae were studied to resolve their taxonomic and phylogenetic status. Three specimens of Nemalionopsis and five collections of Thorea were examined for pit plug ultrastructure and analyzed for the sequences of the genes coding for the large subunit of RUBISCO (rbcL) and the small subunit of rRNA (18S rRNA). The phylogenetic trees generated from the two genes, and a combined tree all showed the Thoreaceae to be contained in a well‐supported monophyletic clade that is separate from the other two families currently classified in the Batrachospermales, the Batrachospermaceae and the Lemaneaceae. In addition, secondary structure elements of the 18S rRNA gene were observed at positions 650 and 1145 (Escherichia coli numbering system) that are not present in other members of the Rhodophyta. The pit plugs of the gametophytic and chantransia stages of the Thoreaceae contain two cap layers, the outer one of which is typically plate‐like, though occasionally inflated ones have been seen. No pit plug cap membrane has been observed. These findings indicate the Thoreaceae has been misclassified in the Batrachospermales and should be placed in its own order, the Thoreales. This order is characterized by having freshwater representatives with multiaxial gametophytes, a uniaxial chantransia stage, and pit plugs with two cap layers, the outer one of which is usually plate‐like.     

Different feeding behaviours in a single predatory mite species. 2. Responses of two populations of Phytoseiulus longipes (Acari: Phytoseiidae) to various prey species,prey stages and plant substrates

Tritrophic studies involving several populations of the predatory mite Phytoseiulus longipes showed distinct life history traits depending on the prey offered and/or the plant substrate. In order to better understand the biology of this predator, the response to several combinations of prey species (Tetranychus evansi and Tetranychus urticae), prey stages (eggs and mobile stages) and plant substrates (bean and tomato leaf discs) has been assessed for two populations of P. longipes. Unlike what was found for life history traits, both populations displayed similar behaviour: they went and stayed more on tomato than on bean, they preferred T. urticae over T. evansi and mobile stages over eggs. Combining the previous life history data with the present results suggests that P. longipes may display host-plant mediated specialization on a prey species, T. evansi. Possible underlying mechanisms are discussed, as well as directions for future work.     

Variation of thymidine incorporation patterns in the alternating vegetative and sexual life cycles of Chlamydomonas reinhardtii

The cellular distribution of thymidine kinase activities in Chlamydomonas reinhardtii, as manifested by the in vivo incorporation of exogenous thymidine and 5-bromodeoxyuridine into different DNA species, appeared to be organelle specific and varied with different developmental stages in the life cycle of this organism. During vegetative growth and gametogenic differentiation, thymidine and 5-bromodeoxyuridine were shown to be selectively incorporated into chloroplast but not nuclear DNA. On the other hand, during zygotic germination in which meiosis occurs and the ensuing vegetative divisions of meiotic products, thymidine as well as 5-bromodeoxyuridine were incorporated into both nuclear and chloroplast DNA. These results suggest that, in addition to the thymidine kinase activity that is constantly present in the chloroplast, a cytoplasmic thymidine kinase is derepressed only during the sexual reproductive cycle of C. reinhardtii.     

LIGHT AND ELECTRON MICROSCOPIC OBSERVATIONS OF PREFERENTIAL DESTRUCTION OF CHLOROPLAST AND MITOCHONDRIAL DNA AT EARLY MALE GAMETOGENESIS OF THE ANISOGAMOUS GREEN ALGA DERBESIA TENUISSIMA (CHLOROPHYTA)1

Gametogenesis in male and female gametophytes was studied by light microscopy and EM in the dioecious multinucleate green alga Derbesia tenuissima (Moris & De Notaris) P. Crouan & H. Crouan, where male and female gametes differ in size. Gametogenesis was divided into five stages: 32 h (stage 1), 24 h (stage 2), 16 h (stage 3), 8 h (stage 4), and 0.5 h (stage 5) before gamete release. At stage 1, the first sign of gametogenesis observed was the aggregation of gametophyte protoplasm to form putative gametangia. At stage 2, gametangia were separated from the vegetative protoplasm of gametophytes. Morphological changes of nuclei and organelles occurred at this early stage of male gametogenesis, and organelle DNA degenerated. At stage 3, male organelle DNA had completely degenerated, whereas in female gametangia, organelle DNA continued to exist in both chloroplasts and mitochondria. Gametogenesis was almost completed at stage 4 and fully at stage 5. Small male gametes had a DNA‐containing nucleus and a large mitochondrion and one or several degenerated chloroplasts. The mitochondria and plastids were devoid of DNA. The large female gametes had a nucleus and multiple organelles, all of which contained their own DNA. Thus, degeneration of chloroplast DNA along with morphological changes of organelles occurred at male gametogenesis in anisogamous green algae (Bryopsis and D. tenuissima), in contrast with previous studies in isogamous green algae (Chlamydomonas, Acetabularia caliculus, and Dictyosphaeria cavernosa) in which degeneration of chloroplast DNA occurred after zygote formation.     

Mitochondrial and Nucleolar Localization of Cysteine Desulfurase Nfs and the Scaffold Protein Isu in Trypanosoma brucei

Trypanosoma brucei has a complex life cycle during which its single mitochondrion is subjected to major metabolic and morphological changes. While the procyclic stage (PS) of the insect vector contains a large and reticulated mitochondrion, its counterpart in the bloodstream stage (BS) parasitizing mammals is highly reduced and seems to be devoid of most functions. We show here that key Fe-S cluster assembly proteins are still present and active in this organelle and that produced clusters are incorporated into overexpressed enzymes. Importantly, the cysteine desulfurase Nfs, equipped with the nuclear localization signal, was detected in the nucleolus of both T. brucei life stages. The scaffold protein Isu, an interacting partner of Nfs, was also found to have a dual localization in the mitochondrion and the nucleolus, while frataxin and both ferredoxins are confined to the mitochondrion. Moreover, upon depletion of Isu, cytosolic tRNA thiolation dropped in the PS but not BS parasites.     

Cytological studies on meiosis and male gametophyte development in autotetraploid cucumber

With improved staining and chromosome preparation techniques, meiosis of pollen mother cells (PMCs) and male gametophyte development in autotetraploid cucumber (Cucumis sativus L.) was studied to understand the correlation between chromosomes behaviour and fertility. Various chromosome configurations, e.g. multivalent, quadrivalents, trivalents, bivalents and univalents were observed in most PMCs at metaphase I. Lagging chromosomes were frequently observed at anaphase in both meiotic divisions. In addition, chromosomes segregations were not synchronous and equal in some PMCs during anaphase II and telophase II. Dyads, triads, tetrads with micronuclei and polyads were observed at tetrad stage, and the frequencies of normal tetrad with four microcytes were only 55.4 %. The frequency of abnormal behaviour in each stage of meiosis was counted, and the average value was 37.2 %. The normal meiotic process could be accomplished to form the microspore tetrads via simultaneous cytokinesis. Most microspores could develop into fertile gametophytes with 2 cells and 3 germ pores through the following stages: single-nucleus early stage, single-nucleus late stage and 2-celled stage. The frequency of abnormalities was low during the process of male gametophyte development. The germination rate of pollen grains was 46.9 %. These results suggested that abnormal meiosis in PMCs was the reason for low pollen fertility in the autotetraploid cucumber.