Life history and photoperiodic responses in Chondrus giganteus forma flabellatus (Gigartinaceae,Rhodophyta) from Japan

The life history and photoperiodic responses of the carrageenophyte Chondrus giganteus forma flabellatus (Rhodophyta) from Japan were investigated in culture. This entity has a Polysiphonia-type life history and the formation of both tetrasporangia and cystocarps is controlled by daylength. A night-break of 1 h in the middle of a 16 h night was effective in inhibiting reproduction, confirming the classical photoperiodic nature of the response. Tetrasporangia formed at daylengths of 12 h or less, whereas cystocarps only developed at daylengths of 10 h or less. The formation of spermatangia required daylengths of 12 h or less, but production was not inhibited by a night-break of 1 h in the middle of a 16 h night.     

TWO MORPHOLOGICALLY SIMILAR BIOLOGICAL SPECIES: CHONDRUS PINNULATUS AND C. ARMATUS (GIGARTINACEAE,RHODOPHYTA)1

The nature of the relationship between Chondrus pinnulatus (Harvey) Okamura f. pinnulatus and C. pinnulatus f. armatus (Harvey) Yamada et Mikami (Gigartinaceae, Rhodophyta) was investigated by comparative analysis of their biogeography, phenologies, life histories, gross and vegetative morphology, crossability, and upper thermal tolerance. The forma pinnulatus has a more northerly distribution in Japan and adjacent waters, exhibiting adaptation to the cooler regions, whereas the forma armatus has a more southerly range. The latter may be the result of a higher thermal tolerance. Both formae have a Polysiphonia-type life history and are similar in their internal vegetative morphology. They can, however, be distinguished by gross morphology: forma pinnulatus has wide, flattened axes, compressed to flattened ultimate segments and proliferations, while forma armatus has narrow, compressed to subterete axes and subterete to terete ultimate segments and proliferations. These differences persist in laboratory culture. All intraformae crosses were positive, with carpospores from the cross developing into fertile F₁ tetrasporophytes releasing tetraspores that developed into dioecious F₁ gametophytes, the female gametophytes of which formed normal cystocarps. This suggests that members of populations of each forma freely interbreed. Among interformae crosses, only some offspring derived from geographically distant strains bore normal cystocarps in the F₁ female gametophytes. Other crosses showed that interbreeding between populations of these two formae was blocked by various isolating mechanisms: incompatibility, hybrid inviability, and hybrid sterility. Reproductive isolation between f. pinnulatus and f. armatus is virtually complete in wild populations, because hybrid populations have not been found in the wild. In addition, these two entities can be considered biological species that are also referred to the taxonomic species, C. armatus and C. pinnulatus, because they do not overlap with regard to the morphology of the ultimate segments and proliferations. Subtle (but significant) gross morphological differences, partial interfertility between the two species, and deleterious hybridization in the area in which they occur sympatrically suggest that their evolutionary divergence was relatively recent.     

Phenology of Chondrus ocellatus in Cheongsapo Near Busan, Korea

The reproductive phenology of Chondrus ocellatus and the effects of temperature and light on its growth were examined in Cheongsapo near Busan, Korea, from September 1994 to August 1995. The vegetative plants dominated over the year, with a peak occurrence in January. Gameto- and tetrasporophytes were most abundant in November and August. All vegetative and reproductive plants had a peak both in length and weight in October, when seawater temperature was highest (24°C). In laboratory culture, the maximum relative growth rate (RGR) of 2.94% day⁻¹ was obtained at 20°C and 100 μmol photons m⁻² s⁻¹, whereas the lowest value was recorded at 25°C and 100 μmol photons m⁻² s⁻¹ in a 12: 12 h LD photoperiod regime. Among the three photoperiod regimes (8:16 h, 12:12 h, 16:8 h LD) tested, there was evidence of a higher RGR in the 12:12 h LD cycle. This result suggests that the growth and reproduction of C. ocellatus are correlated with the seawater temperature based on laboratory culture and field observations.     

Phylogeographic structure and deep lineage diversification of the red alga Chondrus ocellatus Holmes in the Northwest Pacific

A major goal of phylogeographic analysis using molecular markers is to understand the ecological and historical variables that influence genetic diversity within a species. Here, we used sequences of the mitochondrial Cox1 gene and nuclear internal transcribed spacer to reconstruct its phylogeography and demographic history of the intertidal red seaweed Chondrus ocellatus over most of its geographical range in the Northwest Pacific. We found three deeply separated lineages A, B and C, which diverged from one another in the early Pliocene–late Miocene (c. 4.5–7.7 Ma). The remarkably deep divergences, both within and between lineages, appear to have resulted from ancient isolations, accelerated by random drift and limited genetic exchange between regions. The disjunct distributions of lineages A and C along the coasts of Japan may reflect divergence during isolation in scattered refugia. The distribution of lineage B, from the South China Sea to the Korean Peninsula, appears to reflect postglacial recolonizations of coastal habitats. These three lineages do not coincide with the three documented morphological formae in C. ocellatus, suggesting that additional cryptic species may exist in this taxon. Our study illustrates the interaction of environmental variability and demographic processes in producing lineage diversification in an intertidal seaweed and highlights the importance of phylogeographic approaches for discovering cryptic marine biodiversity.     

Genetic variation of the cytochrome b gene in the rosy bitterling, Rhodeus ocellatus (Cyprinidae) in Japan

According to conventional views, the rosy bitterling, Rhodeus ocellatus, comprises two subspecies, R. ocellatus kurumeus and R. ocellatus ocellatus, the former being native to Japan whereas the latter was introduced into Japan from China during World War II. To examine the genetic structure of Japanese R. ocellatus, part of the mitochondrial cytochrome b gene from 48 individuals collected from various locations in Japan was sequenced. Three major mitochondrial lineages were found. Based on historical evidence, two of these represent R. ocellatus ocellatus and the third R. ocellatus kurumeus. The existence of two distinct lineages of R. ocellatus ocellatus in Japan suggests at least two colonizations. Some local populations comprised purely R. ocellatus kurumeus, but those from Kashima and Ogori included both subspecies. Because the proportion of R. ocellatus ocellatus in Kashima increased from 1994 to 1995, invasion by R. ocellatus ocellatus into R. ocellatus kurumeus habitats is apparently in progress. Received: April 30, 1999 / Revised: March 22, 2000 / Accepted: December 20, 2000     

中国杉藻科(Gigartinaceae)两个新记录种的研究

报道中国产杉藻科(Gigartinaceae)角叉菜(Chondrus ocellatus)和日本马泽藻(Mazzaella japonica)两个种的分类研究,均为中国新记录。     

A molecular investigation of polymorphism in the North Atlantic red alga Chondrus crispus (Gigartinales)*

Seven samples of Chondrus crispus Stackhouse, representing widely contrasting forms from both sides of the North Atlantic, were compared by restriction digestion of their plastid DNA. The similar banding patterns confirmed that the seven forms were conspecific and distinct from Chondrus ocellatus Holmes f. ocellatus from Japan, used as an outgroup. Nucleotide sequences of the internal transcribed spacers (ITS1 and ITS21 and the intervening 5.8S rRNA gene of the nuclear rRNA operon were investigated as a potential indicator of genetic divergence among morphological variants of C. crispus. The combined ITS regions were relatively short in Chondrus (between 719 and 731 base pairs [bp] in C. crispus and 724 bp in C. ocellatus f. ocellatus), and the sequence of the 5.8s rDNA fragment (152 bp) was identical in both species. In the aligned ITS regions, there were 0–18 base pair differences (0–2.18% divergence) in pairwise comparisons of the seven forms of C. crispus but no consistent pattern of variation according to gross morphology or geographic origin. However, the ITS sequence differed at 41–54 sites (6.22-7.56%) between C. crispus and C. ocellatus f. ocellatus, again illustrating the genetic distinctiveness of the latter species.     

Phylogeography of the Northern Atlantic species Chondrus crispus (Gigartinales, Rhodophyta) inferred from nuclear rDNA internal transcribed spacer sequences

Eighteen isolates of the red algae Chondrus crispus were collected from Northern Atlantic sites, together with C. ocellatus, C. yendoi and C. pinnulatus from the North Pacific. The nuclear rDNA internal transcribed spacer (ITS) was sequenced and compared, spanning both the ITS regions and the 5.8S rRNA gene. Percentage of nucleotide variation for C. crispus ranged from 0.3% to 4.0%. Phylogenetic analyses were performed using maximum parsimony (MP), neighbor-joining (NJ) and minimum evolution methods. They showed that two main clades existed within the C. crispus samples examined and that suggested C. crispus had a single Atlantic origin. The clustering however did not follow the geographic origin. We hypothesized that the current distribution of C. crispus populations might be a result of three main factors: temperature boundaries, paleoclimate and paleoceanography. ITS data exhibited abundant molecular information not only for phylogeographical investigation but also for systematics studies. Handling editor: C. Sturmbauer     

ATP-linked citrate lyase activity in the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum

Cell-free extracts of the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum strains 1C and L have been shown to cleave citrate with the formation of oxaloacetate and acetyl-CoA. This capacity was found in autotrophically grown cells as well as in the cells grown on media with acetate or L-glutamate. Citrate lyase activity in cell-free extracts is only measurable in the presence of citrate, adenosine-5-triphosphate, coenzyme A and Mg²⁺ or Mn²⁺. It is concluded on the basis of the obtained data that C. limicola f. thiosulfatophilum contains adenosine-5-triphosphate-linked citrate lyase (E.C.4.1.3.8). In contrast to green bacteria in the purple bacteria Ectothiorhodospira shaposhnikovii, Rhodospirillum rubrum and Thiocapsa roseopersicina citrate lyase activity was not found.     

Isolation and characterization of new microsatellite markers for rose bitterlings, Rhodeus ocellatus

The Japanese rose bitterling (Rhodeus ocellatus kurumeus) is facing imminent extinction because of hybridization and competition from an invasive alien subspecies (Rhodeus ocellatus ocellatus). Eleven new microsatellite markers for the two subspecies were developed using dinucleotide repeat specific polymerase chain reaction. The number of alleles per locus and the heterozygosity in R. o. kurumeus were lower than those in R. o. ocellatus. Most of these microsatellite markers were successfully cross‐amplified in three Acheilognathinae species.     

Taxonomic reassessment of Chondrus verrucosus (Rhodophyta,Gigartinales), with a description of Chondrus retortus sp. nov

Phylogenetic and morphological reinvestigation of Chondrus verrucosus Mikami was conducted in the Pacific coast of southeastern Honshu, Japan, including the holotype specimen and samples near the type locality. Two entities were recognized by phylogenetic analyses of plastid rbcL and nuclear EF2 sequences. In morphological analyses, the two entities were distinguished by: (i) thallus size and form, and (ii) cell shape of the medullary layer surrounding young carposporangia: one has larger thalli, is regularly canaliculated, with linear medullary cells (large‐type); the other is smaller, canaliculated with crispation and has roundish medullary cells (small‐type). The large‐type was common in southern to eastern Boso Peninsula including near the type locality; the small‐type was typical in the southwest Boso Peninsula and Sagami Bay. The large‐type was shown to be equivalent to C. verrucosus by examination of the holotype specimen. The small‐type is described as a new species, C. retortus Matsumoto et Shimada.     

List of members August, 1962

We investigated the acclimation of Chondrus crispus to growth at 5°C and 20°C in the laboratory. We were specifically interested in the responses of light-limited photosynthesis to temperature and the effects of short-term thermal changes (of the order of minutes). Thermal acclimation to constant temperatures over 3–4 weeks had significant effects on the light-use characteristics of this species such that in comparison with those grown at 5°C, 20°C-grown plants had higher concentrations of chlorophyll a and total phycobilins, which were associated with larger photosynthetic unit sizes. Plants grown at the higher temperature had greater photosynthetic efficiencies (α) and higher rates of light-limited photosynthesis at a given photon flux density than did plants acclimated to 5°C. Plants acclimated to 20°C were less sensitive to short-term temperature changes than were 5°C-acclimated plants. These results are discussed in terms of (1) the effects of growth temperature on light harvesting and (2) the implications of exposure to constant temperature for short-term thermal responses.     

Seasonal Variations in Clock‐Gene Expression in Atlantic Salmon (Salmo salar)

In homeothermic vertebrates inhabiting temperate latitudes, it is clear that the seasonal changes in daylength are decoded by the master circadian clock, which through secondary messengers (like pineal melatonin secretion) entrains rhythmic physiology to local conditions. In contrast, the entrainment and neuroendocrine regulation of rhythmic physiology in temperate teleosts is not as clear, primarily due to the lack of understanding of the clock gene system in these species. In this study, we analyzed the diel expression of the clock‐genes in brains of Atlantic salmon, a species that is both highly photoperiodic and displays robust clock‐controlled behavior. Atlantic salmon parr were acclimated to either long‐day (LD) or short‐day (SD) photoperiods for one month and thereafter sampled at 4 h intervals over a 24 h cycle. Clock, Bmal1, Per2, and Cry2 were all actively expressed in salmon brain homogenates and, with the exception of Per2, all displayed rhythmic expression under SD photoperiods that parallels that reported in zebrafish. Interestingly, daylength significantly altered the mRNA expression of all clock genes studied, with Clock, Bmal1, and Per2 all becoming arrhythmic under the LD compared to SD photoperiod, while Cry2 expression was phase delayed under LD. It is thus proposed that the clock‐gene system is actively expressed in Atlantic salmon, and, furthermore, as has been reported in homeothermic vertebrates, it appears that clock expression is daylength‐dependent.     

中国东北川续断科两个新变型

发表了东北产川续断科蓝盆花属两个新变型,即白花窄叶蓝盆花(Scabiosa comosa Fisch. ex Roem. et Schult. f. albiflora S. H. Liet S. Z. Liu)和白花华北蓝盆花(S. tschiliensis Grun. f. albiflora S. H. Liet S. Z. Liu).两个变型与原变型的主要区别为花为白色。     

Rhodopseudomonas sphaeroides forma sp. denitrificans,a denitrifying strain as a subspecies of Rhodopseudomonas sphaeroides

From polluted water of a lagoon pond a new type of denitrifying photosynthetic purple bacteria was isolated. With respect to morphology, fine structure, photopigments, requirement for growth factors, the range of utilization of organic substrates for phototrophic growth and DNA base ratio, the denitrifying strains show the closest resemblance to Rhodopseudomonas sphaeroides and were therefore described as a subspecies named R. sphaeroides forma sp. denitrificans. The new isolates grow well with nitrate anaerobically in the dark accompanying the evolution of nitrogen gas. They cannot assimilate nitrate as the nitrogen source for growth.     

Gibberellins and the photoperiodic control of leaf growth in Poa pratensis

The role of gibberellins (GAs) in photoperiodic control of leaf elongation in Poa pratensis was studied by both application of exogenous GAs and analysis of endogenous GAs. Leaf elongation was strongly increased under long day (LD, 24 h) conditions at both 9 and 21°C, leaf length at 9°C LD being similar to that in plants grown in short days (SD, 8 h) at 21°C. However, even at 21°C leaf elongation was enhanced by LD. Exogenous GA₁ could completely compensate for LD at both 9 and 21°C. Gibberellins A₂₀, A₁₉ and A₄₄ could also partly replace LD, but they were significantly less active than GA₁, GA₅₃ was inactive when applied to plants grown at 9°C in SD and exhibited only marginal activity at 9°C LD and 21°C SD. The total level of GAs of the early 13-hydroxylation pathway (A₅₃, A₄₄, A₁₉, A₂₀ and A₁) increased rapidly when plants were transferred from SD to LD at 9°C. After transfer from 9 to 21°C, there was an increase in GA levels at both LD and SD, followed by a decrease under LD conditions. In all cases, GA₁₉ was the predominant GA, accounting for 60 to 80% of the analysed GAs. Levels of the bioactive GA₁ were low and increased transiently by LD four days after transfer from SD to LD. At both temperatures, the ratio GA₁₉ to GA₂₀ and GA₂₀ to GA₁ at 9°C was enhanced by LD compared with SD. Taken together, these results support the hypothesis that photoperiodic regulation of leaf elongation in Poa pratensis is GA-mediated, and they indicate a photoperiodic control of oxidation of GA₅₃ to GA₄₄ and GA₁₉ to GA₂₀, and perhaps also of 3β-hydroxylation of GA₂₀ to GA₁.     

Upper temperature tolerance of North Atlantic and North Pacific geographical isolates ofChondrus species (Rhodophyta)

The upper survival temperature for most isolates ofChondrus crispus from localities ranging from northern Norway and Iceland to Spain, and for an isolate from Nova Scotia, was 28 °C after 2 weeks of exposure to temperatures of 20–31 °C at intervals of 1 °C. An upper survival limit of 29 °C was exhibited by a few European isolates from the English Channel, the North Sea, and one Irish isolate from the upper intertidal. The warm-temperate Japanese speciesC. nipponicus andC. giganteus formaflabellatus survived 30 °C, whereas 29 °C was the upper survival limit for the coldtemperatureC. pinnulatus formapinnulatus from northern Japan. A possible origin ofC. crispus in the north Pacific is discussed.     

Genetic markers distinguishing between the two subspecies of the rosy bitterling,Rhodeus ocellatus (Cyprinidae)

Eleven populations of the rosy bitterling,Rhodeus ocellatus, from different localities in Japan, were genetically compared at 16 protein-coding loci using starch-gel electrophoresis. Two loci,Ldh-2 andPgdh, were demonstrated as diagnostic markers for the identification of two subspecies;R. ocellatus kurumeus, which is native to Japan, andR. ocellatus ocellatus, which was introduced from China. The two subspecies were distinguished by the complete substitution of different alleles between them. Population ofR. ocellatus kurumeus occurring in Yao City, Osaka, and in Kanzaki, Saga Prefecture were genetically closely related to each other (genetic distance: D=0.056) but distantly so toR. ocellatus ocellatus from Saitama Prefecture (D=0.202 or 0.265). Electrophoretic analyses also elucidated the existence of hybrid populations of the two subspecies. The populations ofR. ocellatus kurumeus in Yao City had lower genetic variability and a lower incidence of white coloration on the ventral fins than populations of the same in Saga Prefecture. The present study strongly implies that the introduction of the foreign freshwater fishes with subspecific differentiation, into the original range of indigenous subspecies, should be averted not to bring the genetic pollution.     

Comparison of three techniques for identifying isomorphic phases of <Emphasis Type="BoldItalic">Chondrus crispus</Emphasis> (Gigartinaceae)

Chondrus crispus, a member of the economically important family Gigartinaceae, alternates between two free-living isomorphic life history phases. The carrageenan composition of the gametophyte and tetrasporophyte phases has been used to identify non-reproductive material, and thus provide a better understanding of the ecology of the species. The aim of this study was to compare three methods for identifying the isomorphic phases. The results obtained by the widely used resorcinol test and an iridescence method are compared with those obtained by a technique that involves the acquisition of mid-infrared absorbance spectra from dried samples by Fourier transform infrared spectroscopy (FT-IR). Both the resorcinol test and FT-IR produced similar results with a calculated gametophyte to tetrasporophyte ratio of 2.1 and 2.3:1, respectively. In contrast the iridescence method was less reliable, with a calculated gametophyte to tetrasporophyte ratio of 1.3:1 and a low predictive value (0.70) for selecting tetrasporophytes. The FT-IR technique provides a rapid assessment, does not involve the use of hazardous chemicals, has the potential to be applied to members of the Gigartinaceae for which the resorcinol and iridescence tests are less applicable and it has greater flexibility in that it can identify additional carrageenan fractions.