Photosynthetic inorganic carbon utilization of gametophytes and sporophytes of Undaria pinnatifida (Phaeophyceae)

The characteristics of inorganic carbon assimilation by photosynthesis were investigated in male and female gametophytes and juvenile sporophytes of Undaria pinnatifida. Gametophytes and sporophytes have detectable extracellular and intracellular carbonic anhydrase (CA) activity, and the CA inhibitor, acetazolamide (AZ), significantly inhibited their photosynthesis O-2 evolution. In pH-drift experiments, it was found that gametophytes did not raise the final pH of seawater above 9.00 (CO2 concentrations of about 2.2 mu M), indicating a low ability to utilize inorganic carbon. In contrast, sporophytes rapidly raised pH to over 9.53 and depleted the free CO2 Concentration to less than 0.16 mu M. The apparent photosynthetic affinity for CO2 was almost the same for gametophytes and sporophytes, whereas gametophytes had a much lower affinity for HCO3- than sporophytes. Two inhibitors of band 3 anion exchange protein (DIDS and SITS) inhibited the photosynthesis of gametophytes but not that of sporophytes. It was indicated that both gametophytes and sporophytes were capable of using HCO3-, which involved the external CA activity, and a direct HCO3- use also occurred in the former, but the latter showed a greater capacity of HCO3- use than the former. In addition, male and female gametophytes did not show great differences in the inorganic carbon uptake mechanism underlying photosynthesis.

Expression of the lacZ reporter gene in sporophytes of the seaweed Laminaria japonica (Phaeophyceae) by gametophyte-targeted transformation

The seaweed Laminaria japonica (Phaeophyceae) has a two-generation life cycle consisting of haploid gametophytes and diploid sporophytes. Female and/or male gametophytes were transformed using particle bombardment and the histological LacZ assay was performed on sporophytes generated by either parthenogenesis or inbreeding. Female gametophyte-targeted transformation resulted in similar lower efficiencies in both parthenogenetic and zygotic sporophytes, and only a chimeric expression pattern was observed. Male gametophyte-targeted transformation led to a higher efficiency, with 3.5% of the zygotic sporophytes stained completely blue (all-blue), implying the integration of lacZ at the one-cell stage. Polymerase chain reaction analysis using primers specific for a lacZ-vector juncture fragment and subsequent blotting indicated the presence of the introduced gene in the sporophytes. The method reported here has a potential for seaweed transformation using spore-based bombardment followed by the developmental process.

Genetic analysis of the gametophytes of Undaria pinnatifida (Phaeophyceae) with ISSR method

Inter-simple sequence repeat (ISSR) analysis was used to assess eleven pairs of Undaria pinnatifida (Harv.) Suringar male and female gametophytes. After screening fifty primers, 18 ISSR primers were selected for final analysis. A total of 104 loci were obtained, of which 77 were polymorphic, among the gametophytes studied. Genetic relationships were analyzed with simple matching (S), Jaccard's (J) and Dice's (D) distance coefficients. Little genetic variations were found among the selected Undaria gametophytes, for instance, the genetic distances ranging from 0.010 to 0.125 with Dice coefficients. UPGMA dendrograms showed that 11 pairs of Undaria gametophytes were distributed into five groups. Most Undaria strains cultivated in China exhibited closely genetic relationships with the strains from Japan. However, gametophytes from Qingdao appeared as distinct clades from other Undaria strains with all three distance coefficients used. Mantel test showed that the three distance measurements generated congruent clustering patterns on the same data. Our results demonstrated the feasibility of applying ISSR markers for genetic analysis of Undaria gametophytes. (c) 2006 Elsevier B.V. All rights reserved.

The genetic analysis and germplasm identification of the gametophytes of Undaria pinnatifida (Phaeophyceae) with RAPD method

Eleven pairs of Undaria pinnatifida (Harv.) Suringar gametophytes were identified with random amplified polymorphic DNA (RAPD) technique. After screening 100 primers, 20 ten-base primers were determined for the RAPD analysis. A total of 312 polymorphic loci were obtained, of which 97.7% were polymorphic. The primer S198 was found to distinguish all the selected Undaria pinnatifida gametophytes. The genetic distances between each two of the twenty-two U. pinnatifida gametophytes ranged from 0.080 to 0.428, while the distances to the Laminaria was 0.497 on average. After reexamination, two sequences characterized amplification region (SCAR) markers were successfully converted, which could be applied to U. pinnatifida germplasm identification. All these results demonstrated the feasibility of applying RAPD markers to germplasm characterization and identification of U. pinnatifida gametophytes, and to provide a molecular basis for Undaria breeding.

Identification and assessing the cultivars of Laminaria Lamx. (phaeophyceae) with molecular markers

Molecular markers were used to identify and assess cultivars of Laminaria Lamx. and to delineate their phylogenetic relationships. Random amplified polymorphic DNA (RAPD) analysis was used for detection. After screening, 11 primers were selected and they yielded 133 bands in all, of which approximately 99.2% were polymorphic. The genetic distances between gametophytes ranged from 0.412 to 0.956. Two clusters were formed with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on the simple matching coefficient. All cultivars of Laminaria japonica Aresch. used for breeding in China fell into one cluster. L. japonica from Japan, L. saccharina (L.) Lam., and L. angustata Kjellm. formed the other cluster and showed higher genetic variation than L. japonica from China. Nuclear ribosomal DNA (rDNA) sequences, including internal transcribed spacers (ITS1 and ITS2) were studied and aligned. The nucleotides of the sequences ranged from 634 to 668, with a total of 692 positions including TTS1, ITS2, and the 5.8S coding region. The phylogenetic tree obtained by the neighbor-joining method favored, to some extent, the results revealed by RAPD analysis. The present study indicates that RAPD and ITS analyses could be used to identify and assess Laminaria germplasm and to distinguish some species and, even intraspecies, in Laminaria.

Effect of alginic acid decomposing bacterium on the growth of Laminaria japonica (Phaeophyceae)

We collected the diseased blades of Laminaria japonica from Yantai Sea Farm from October to December 2002, and the alginic acid decomposing bacterium on the diseased blade was isolated and purified, and was identified as Alterornonas espejiana. This bacterium was applied as the causative pathogen to infect the blades of L. japonica under laboratory conditions. The aim of the present study was to identify the effects of the bacterium on the growth of L. japonica, and to find the possibly effective mechanism. Results showed that: (1) The blades of L. japonica exhibited symptoms of lesion, bleaching and deterioration when infected by the bacterium, and their growth and photosynthesis were dramatically suppressed. At the same time, the reactive oxygen species (ROS) generation enhanced obviously, and the relative membrane permeability increased significantly. The contents of malonaldehyde (MDA) and free fatty acid in the microsomol membrane greatly elevated, but the phospholipid content decreased. Result suggested an obvious peroxidation and deesterrification in the blades of L. japonica when infected by the bacterium. (2) The simultaneous assay on the antioxidant enzyme activities demonstrated that superoxide dismutase (SOD) and catalase (CAT) increased greatly when infected by the bacterium, but glutathione peroxidase (Gpx) and ascorbate peroxidase (APX) did not exhibit active responses to the bacterium throughout the experiment. (3) The histomorphological observations gave a distinctive evidence of the severity of the lesions as well as the relative abundance in the bacterial population on the blades after infection. The bacterium firstly invaded into the endodermis of L. japonica and gathered around there, and then resulted in the membrane damage, cells corruption and ultimately, the death of L. japonica.

The extent of grazing release from epiphytism for Sargassum muticum (Phaeophyceae) within the invaded range

The overall biotic pressure on a newly introduced species may be less than that experienced within its native range, facilitating invasion. The brown alga Sargassum muticum (Yendo) Fensholt is a conspicuous and successful invasive species originally from Japan and China. We compared S. muticum and native macroalgae with respect to the biotic pressures of mesoherbivore grazing and ectocarpoid fouling. In Strangford Lough, Northern Ireland, S. muticum thalli were as heavily overgrown with seasonal blooms of epiphytic algae as native macroalgal species were. The herbivorous amphipod Dexamine spinosa was much more abundant on S. muticum than on any native macroalga. When cultured with this amphipod, S. muticum lost more tissue than three native macroalgae, Saccharina latissima (Linnaeus) Lane et al., Halidrys siliquosa (Linnaeus) Lyngbye and Fucus serratus Linnaeus. Sargassum muticum cultured with both ectocarpoid fouling and amphipods showed a severe impact, consistent with our previous findings of large declines in the density of S. muticum observed in the field during the peak of fouling. Despite being a recent introduction into the macroalgal community in Strangford Lough, S. muticum appears to be under biotic pressure at least equal to that on native species, suggesting that release from grazing and epiphytism does not contribute to the invasiveness of this species in Strangford Lough.

Seasonal differences in the effects of oscillatory and uni-directional flow on the growth and nitrate-uptake rates of juvenile Laminaria digitata (Phaeophyceae)

The influence of oscillatory versus unidirectional flow on the growth and nitrate-uptake rates of juvenile kelp, Laminaria digitata, was determined seasonally in experimental treatments that simulated as closely as possible natural environmental conditions. In winter, regardless of flow condition (oscillatory and unidirectional) or water velocity, no influence of water motion was observed on the growth rate of L. digitata. In summer, when ambient nitrate concentrations were low, increased water motion enhanced macroalgal growth, which is assumed to be related to an increase in the rate of supply of nutrients to the blade surface. Nitrate-uptake rates were significantly influenced by water motion and season. Lowest nitrate-uptake rates were observed for velocities <5 cm · s−1 and nitrate-uptake rates increased by 20%–50% under oscillatory motion compared to unidirectional flow at the same average speed. These data further suggested that the diffusion boundary layer played a significant role in influencing nitrate-uptake rates. However, while increased nitrate-uptake in oscillatory flow was clear, this was not reflected in growth rates and further work is required to understand the disconnection of nitrate-uptake and growth by L. digitata in oscillatory flow. The data obtained support those from related field-based studies, which suggest that in summer, when insufficient nitrogen is available in the water to saturate metabolic demand, the growth rate of kelps will be influenced by water motion restricting mass transfer of nitrogen.

Reproduction in Sphacelaria biradiata Askenasy (Sphacelariales, Phaeophyceae) in Southern Australia

Sphacelaria biradiata Askenasy is a little known but common brown alga of southern Australian coasts. This paper describes its reproduction. Populations of S. biradiata reproduced asexually by vegetative propagules in all localities examined. The propagules were produced throughout the year and did not appear to be affected by season, although, if sexual structures occurred, propagule numbers declined. Sexual reproduction can occur and involves an alternation of generations, but it is a rare event. Gametophytes may produce male gametangia, female gametangia or both. Male gametangia, however, are extremely rare and are described for the first time. Production of female gametangia was influenced by season and occurred at all areas studied. The sexual behaviour of the male and female gametes is described.

Dispersal of Hormosira banksii (phaeophyceae) via detached fragments : reproductive viability and longevity

Drifting, fertile thalli are well documented to be the primary long-distance dispersal vector for many marine macroalgae, but little information about reproductive viability of drift is known. This study examined the reproductive viability and longevity of floating fragments of the intertidal Australasian fucoid Hormosira banksii (Turner) Decne. Beach wrack surveys and field experiments were conducted to test the model that long-distance dispersal is achieved in H. banksii via floating, fertile fronds. High densities of beach wrack fragments were evident during summer compared to autumn. The majority of beach wrack occurred on sandy beaches rather than rocky shores. Both male and female fragments were present in the beach wrack. Detached fronds were capable of releasing gametes up to 8 weeks after detachment. Beach wrack produced high fertilization rates and recruited successfully onto artificial panels. Results suggest that detached fragments are reproductively viable and that floating, fertile fronds may be an important mechanism for facilitating long-distance dispersal in this species. Nevertheless, the frequency of fronds reaching a suitable habitat and contributing to gene flow between populations, or colonizing new populations, may not be proportional to the total density of beach wrack.

Dislodgment and attachment strength of the intertidal macroalga Hormosira banksii (Fucales, Phaeophyceae)

Hormosira banksii is a dominant intertidal alga characterised by limited dispersal of propagules, yet it has a broad distribution throughout temperate Australasia. However, the high abundance of beach wrack of H. banksii on sandy beaches along southwest Victoria, the frequency of frond dislodgment and the force required to break it from the substratum remain unknown. Dislodgment of macroalgae has been shown to influence dispersal processes; therefore, we tested the model that long-distance dispersal of H. banksii is facilitated by weak attachment to the substratum and frequent dislodgment of fertile fronds. We monitored the dislodgment of H. banksii individuals and conducted in situ pull tests to determine the attachment strength of fronds of H. banksii. We further tested whether thallus size or pull direction influenced attachment strength of H. banksii. We found that breakage of vesicles and fronds was a regular event during the survey period and that the mean attachment strength of fronds of H. banksii was weak compared to other intertidal algae. Furthermore, we found that thallus size did not influence the force required to break fronds of H.banksii from the substratum but that the direction that fronds were pulled from the substratum did. We suggest that fronds of H. banksii do drift frequently and that long-distance dispersal is likely to be an important mechanism for the distributional success of this species.

Divergence patterns in Fucus seaweeds (Phaeophyceae) in the northern Baltic Sea and in the Tjongspollen area

To be able to interpret patterns of biodiversity it is important to understand the processes by which new species evolve and how closely related species remain reproductively isolated and ecologically differentiated. Divergence and differentiation can vary during speciation and it can be seen in different stages. Groups of closely related taxa constitute important case studies to understand species and new biodiversity formation. However, it is important to assess the divergence among them at different organismal levels and from an integrative perspective. For this purpose, this study used the brown seaweeds genus Fucus as a model to study speciation, as they constitute a good opportunity to study divergence at different stages. We investigated the divergence patterns in Fucus species from two marginal areas (northern Baltic Sea and the Tjongspollen area), based on phenetic, phylogenetic and biological taxonomical criteria that are respectively characterised by algal morphology, allele frequencies of five microsatellite loci and levels of secondary polyphenolic compounds called phlorotannins. The results from this study showed divergence at morphological and genetic levels to certain extent but complete lack of divergence at biochemical level (i.e. constitutive phlorotannin production) in the Baltic Sea or Norway. Morphological divergence was clearly evident in Tjongspollen (Norway) among putative taxa as they were identified in the field and this divergence corresponds with their neutral genetic divergence. In the Baltic, there are some distinguishable patterns in the morphology of the swedish and finnish individuals according to locality to certain extent but not among putative taxa within localities. Likewise, these morphological patterns have genetic correspondence among localities but not within each locality. At the biochemical level, measured by the phlorotannin contents there were neither evidence of divergence in Norway or the Baltic Sea nor any discernable aggregation pattern among or within localities. Our study have contributed with further understanding of the Baltic Sea Fucus system and its intriguingly rapid and recent divergence as well as of the Tjongspollen area systems where formally undescribed individuals have been observed for the first time; in fact they appear largely differentiated and they may well warrant a new species status. In current times, climate change threatens, peripheral ecosystems, biodiversity, and increased knowledge of processes generating and maintaining biodiversity in those ecosystems seem particularly important and needed.