To assess distributional shifts of species in response to recent warming, historical distribution records are the most requisite information. The surface seawater temperature (SST) of Kochi Prefecture, southwestern Japan on the western North Pacific, has significantly risen, being warmed by the Kuroshio Current. Past distributional records of subtidal canopy-forming seaweeds (Laminariales and Fucales) exist at about 10-year intervals from the 1970s, along with detailed SST datasets at several sites along Kochi's >700 km coastline. In order to provide a clear picture of distributional shifts of coastal marine organisms in response to warming SST, we observed the present distribution of seaweeds and analyzed the SST datasets to estimate spatiotemporal SST trends in this coastal region. We present a large increase of 0.3°C/decade in the annual mean SST of this area over the past 40 years. Furthermore, a comparison of the previous and present distributions clearly showed the contraction of temperate species' distributional ranges and expansion of tropical species' distributional ranges in the seaweeds. Although the main temperate kelp Ecklonia (Laminariales) had expanded their distribution during periods of cooler SST, they subsequently declined as the SST warmed. Notably, the warmest SST of the 1997–98 El Niño Southern Oscillation event was the most likely cause of a widespread destruction of the kelp populations; no recovery was found even in the present survey at the formerly habitable sites where warm SSTs have been maintained. Temperate Sargassum spp. (Fucales) that dominated widely in the 1970s also declined in accordance with recent warming SSTs. In contrast, the tropical species, S. ilicifolium, has gradually expanded its distribution to become the most conspicuously dominant among the present observations. Thermal gradients, mainly driven by the warming Kuroshio Current, are presented as an explanation for the successive changes in both temperate and tropical species' distributions.

The nuclear-encoded ITS and associated 5.8S rDNA regions were sequenced for 72 specimens of Ulva collected from 44 rivers across Japan, including U. prolifera Müller from the Shimanto River, Kochi Prefecture, as well as 26 samples originally identified as U. linza L. from 20 coastal marine areas. Sequence data revealed that the samples fall into six distinct clades: the U. flexuosa Wulfen clade (2 samples), the Ulva linza-procera-prolifera (LPP) complex clade (75 samples), Ulva sp. 1 clade (3 samples), Ulva sp. 2 clade (7 samples), Ulva sp. 3 clade (4 samples) and Ulva sp. 4 clade (7 samples). The LPP complex contained a mixture of 26 samples collected from seashores and 49 samples obtained from rivers, including U. prolifera from the Shimanto River, and GenBank data for U. linza and U. procera Ahlner. The samples of the LPP complex differed by only 0-7 substitutions (0-1.149%). Subsequent phylogeographic analyses of the LPP complex based on the 5S rDNA spacer region revealed the presence of two further groupings: a group including 22 strictly marine littoral U. linza samples and a U. prolifera group composed of a mixture of 4 marine samples and all 49 river samples. The monophyly of all river samples indicates that adaptation to low salinity might have occurred only once in the evolutionary history of the LPP complex.

SUMMARY In order to elucidate the species composition of free‐floating Ulva that cause green tide in several bays in Japan, and to clarify the generic status of Ulva and Enteromorpha (Ulvales, Ulvophyceae), the nuclear encoded internal transcribed spacer (ITS) region including the 5.8S gene and the plastid encoded large subunit of ribulose‐1, 5‐bisphosphate carboxylase/ oxgenase (rbcL) gene sequences for 15 species were determined. Both ITS and rbcL analyses indicate that free‐floating Ulva samples are divided into four different lineages that correspond to Ulva lactuca Linnaeus, U. pertusa Kjellman, U. armoricana Dion etal. and U. fasciata Delile. These four species are distinguished by cell morphology including the arrangement of cells, the shape and size of cells and the position of chloroplasts. Molecular data also indicated that Ulva and Enteromorpha are not separated as respective monophyletic groups within a large monophyletic clade and congeneric as shown by previous molecular studies using the ITS sequences alone. This strongly suggests that these genera are congeneric and Enteromorpha should be reduced to the synonym of Ulva.

Three metabolites of fucoxanthin were isolated from a brown alga, Scytosiphon lomentaria, and the structure of a new compound was determined by NMR.The content of fucoxanthin, a biologically active carotenoid, in four edible brown algae, cultivated in deep seawater, was studied.

Synchronous zooid formation in Uiva pertusa Kjellman was induced in excised disks maintained in sterilized seawater at 20X, 12:12 h L: D cycle and fluorescent light at 100 |xmol photons m ^ S"^ Zooids were reieased from mature disk tissue on the morning of the second or the third day after excision. The degree of zooid formation was found to be dependent on disk size and the region of the mother thalius from which the disk tissue was excised. Zooid formation was induced in more than 90% of small disks (0.9 mm in diameter) which were taken from the margins of the Uiva thalii. When disks were incubated together with a perforated mother thalius, the disks remained sterile. The presence of maturation inhibitors in vegetative thalli is suggested.

A new "germling cluster" method is proposed for tank cultivation of seaweed in a free-floating form. This method was applied to the tank cultivation of Ulva prolifera using deep seawater (DSW) pumped up from over 300 m depth off the cape of Muroto in southwest Japan. Numerous zoids of U. prolifera were induced by cutting thalli into 1-2 mm long pieces. Three days after fragment production, the zoids were released. The zoid suspension was concentrated to a density of more than 10 4 zoids per mL medium, and placed in a Petri dish for culture. The dense, germinating zoids began to adhere to each other and form aggregations. The germling aggregations were then removed from the bottom of the dish and torn into a large number of small "germling clusters" using an electric mixer. Each cluster contained 10-100 germlings. Once the germling clusters had attained more than 5 mm diameter in culture, they were transplanted as free-floating forms to a 500 L outdoor tank with continuous aeration, to which DSW was supplied at an exchange rate of 3 volumes per day. As a result, the average daily growth rate (DGR) in the tank throughout the year was 37%, though the DGR fluctuated with seasonal temperature changes.

ABSTRACT:Although most members of the genus Enteromorpha are important edible green algae, some species are also potentially economically valuable crops. Samples of E. prolifera were obtained from the Yoshino River estuary, Tokushima, Japan and cultured in laboratory conditions at 10°C, a salinity of 20 psu, under white light with 12 h light : 12 h dark cycle and at a photon fluence rate (PFR) of 40 mmol/m 2 per s for 30 days. In the present study, the effect of salinity and PFR on the induction of reproductive cells and rhizoid formation were investigated. Synchronous formation of swarmers by thalli was induced in excised disks of 1.2 mm diameter after 2-5 days incubation. The optimum salinity for maturation of reproductive cells was between 5.0 and 52.0 psu, and between 13.2 and 45.3 psu for swarmer release, although the lower limit for swarmer release was 5.0 psu. Maturation of reproductive cells and swarmer release required a PFR higher than 16 mmol/m 2 per s. The minimum PFR for swarmer release was 8 mmol/m 2 per s. Many rhizoids were formed between 1.6 and 52.0 psu and photon fluence rates between 8 and 320 mmol/m 2 per s. Rhizoids were formed in a polarized manner.

In order to elucidate the species composition of freefloating Ulva that cause green tide in several bays in Japan, and to clarify the generic status of Ulva and Enteromorpha (Ulvales, Ulvophyceae), the nuclear encoded internal transcribed spacer (ITS) region including the 5.8S gene and the plastid encoded large subunit of ribulose-1, 5-bisphosphate carboxylase/ oxgenase ( rbc L) gene sequences for 15 species were determined. Both ITS and rbc L analyses indicate that free-floating Ulva samples are divided into four different lineages that correspond to Ulva lactuca Linnaeus, U. pertusa Kjellman, U. armoricana Dion et al . and U. fasciata Delile. These four species are distinguished by cell morphology including the arrangement of cells, the shape and size of cells and the position of chloroplasts. Molecular data also indicated that Ulva and Enteromorpha are not separated as respective monophyletic groups within a large monophyletic clade and congeneric as shown by previous molecular studies using the ITS sequences alone. This strongly suggests that these genera are congeneric and Enteromorpha should be reduced to the synonym of Ulva .