PHYCOGENETIC COMPARISON OF THE SMALL-SUBUNIT RIBOSOMAL DNA SEQUENCE OF COSTARIA COSTATA (PHAEOPHYTA) WITH THOSE OF OTHER ALGAE,VASCULAR PLANTS AND OOMYCETES1

The small-subunit ribosomal DNA (rDNA) coding sequence was determined for the help Costaris Costata (Phaeophyta) and compared to those of chlorophyll a + b- and chlorophyll a + c-containing vascular and nonvascular plants. Phylogenetic comparison of all sequences indicated a common ancestor for phaeophytes, chrysophytes and oomycetes. Phylogenies based on rDNA sequence data and those based on plastid characteristics were compared. Relative evolutionary distances between some taxa, derived from rDNA sequence data, conflicted with findings of plastid-based phylogenies.     

37 Production, survival and morphology of kelp (protista) "sporophytes" arising from selfings, apogamy and hybridizations

The production and survival of sporophyte morphologies that arose from selfings, parthenogenesis, male apogamy and reciprocal hybridizations of Costaria costata (C.A. Agardh) Saunders, Laminaria saccharina (L) Lamouroux, Alaria marginata Postels and Ruprecht, Eisenia arborea Areschoug, Lessoniopsis littoralis (Tilden) Reinke, Macrocystis integrifolia Bory, and Nereocystis leutkeana (Mertens) Postels and Ruprecht was tracked for 215 hybridization trials for 30 weeks. Each trial consisted of selfings for each species, solo cultures of males and females for each species and reciprocal crossings. At the end of 30 weeks, "sporophyte" survival in the 215 cases was 13% for selfings, 10% for parthenogenesis, 6% for apogamy, and 16% for the reciprocal hybridizations. The morphologies of sporophytes resulting from selfings were typical of the selfed species. Parthenosporophytes and reciprocal hybrids usually had the distinctive morphology of the female parent. Male, apogamously derived "sporophytes" often had reduced morphological features. For example, A. marginata male apogamous sporophytes often lacked a midrib. The sporophyte morphologies that arose in the reciprocal crossings are considered to be putative hybrids. Molecular evidence is required to establish their genetic origins.     

When is a family not a family?

Recent molecular investigations of the small-subunit rRNA gene sequences have indicated that established taxonomic hierarchies can be clearly at odds with the degree of evolutionary divergence (as inferred from molecular divergence) between supposedly equivalent taxa at every rank (i.e., species through class), both within and across biological divisions. This is particularly true between the Rhodophyta and the Phaeophyta, whose higher-order taxa appear under- and overinflated, respectively. We present two plausible alternatives that algal taxonomists might adopt in order to invoke discussion on their relative merits.     

AN EVALUATION OF METHODS USED TO ASSESS INTERGENERIC HYBRIDIZATION IN KELP USING PACIFIC LAMINARIALES (PHAEOPHYCEAE)1

Kelp intergeneric laminarialean hybridizations and hybridization protocol were assessed using seven northeast Pacific kelp species: Alaria marginata Postels and Ruprecht, Costaria costata (C. A. Agardh) Saunders, Eisenia arborea Areschoug, Laminaria saccharina (L) Lamouroux, Lessoniopsis littoralis (Tilden) Reinke, Macrocystis integrifolia Bory, and Nereocystis leutkeana (Mertens) Postels and Ruprecht. Survival and development of sporophyte morphologies derived from selfings, separate males and females, and reciprocal crosses were evaluated over 30 weeks of cultivation. All cultures were initiated from cloned gametophytes. Two closely related species, Laminaria angustata Kjellman and L. japonica Areschoug, demonstrated the efficacy of long‐term (up to 30 years) cloned gametophytes in hybridization studies. Sporophyte morphologies appeared in 34%–69% of control and hybridization trials, and 6%–16% of all trials produced sporophytes in control and hybridization conditions that persisted through 30 weeks of cultivation. Sporophytes in control and hybridization conditions could appear normal or abnormal. Usually, the morphology of sporophytes in hybridizations and female controls resembled the female parent, whereas the sporophytes in male controls often had an abbreviated morphology, lacking definitive generic features. Species‐specific rDNA internal transcribed spacer molecular primers were used to determine the parentage of five putative hybrids. Only the L. japonica♀/L. angustata♂ hybrid bore both parental genomes. That negative controls could produce persistent and normal‐appearing sporophytes negates their value and emphasizes the importance of molecular confirmation in hybridization studies. These findings were applied to critique the only known wild intergeneric hybrid, Pelagophycus/Macrocystis.     

Mathematical model ofLaminaria production near a British Columbian salmon sea cage farm

The technical and economical feasibility of farmingLaminaria saccharina for a food base product near a salmon sea cage farm was evaluated. Suitability of kelp for nutrient removal was also analyzed. A computer model of a conceptualized system was developed in order to make the assessments. Kelp growth was modelled as a linear function of temperature and background dissolved inorganic nitrogen concentration, and it was partially experimentally validated. Based on model simulations, aLaminaria farm containing 10,60 m ropes on each end of a salmon sea cage farm is fertilized by the salmon farm and yields annually 1600 kg of dried kelp. The payback period for the initial investment of $61 × 10³ is 6 years after which an annual net profit of 20 × 10³ Canadian dollars ($16.68 × 10³ US) can be achieved. The net present worth of the kelp farm was positive for a rate of return up to 25%. Kelp production on multiple salmon farms or at a higher kelp density could increase the overall revenue.The kelp farm does not appreciably affect background nutrient or oxygen levels. With a few modifications in the model,Nereocystis andMacrocystis farming can be substituted and evaluated for feasibility and nutrient removal efficiency.     

Mitochondrial DNA sequence variation of four <Emphasis Type="Italic">Saccharina</Emphasis> species (Laminariales,Phaeophyceae) growing in Japan

The complete mitochondrial DNA (mtDNA) was sequenced for four Saccharina species including three varieties of Saccharina japonica in Hokkaido: S. japonica; S. japonica var. religiosa; S. japonica var. ochotensis; S. japonica var. diabolica; Saccharina longipedalis; Saccharina angustata; and Saccharina coriacea. Furthermore, the structure and the sequence were compared among them. The total nucleotide length was 37,500–37,657 bp. All mtDNAs were mapped and no differences in the organization of the coding region were found. From the total alignment of S. japonica including the three varieties and S. longipedalis, nucleotide substitutions were detected at 68 sites, and a nucleotide insertion/deletion was detected at one site on rps19 for the whole genome. Variable regions useful for varieties distinction of S. japonica were trnI, trnM, rps19, ORF41, and seven spacers. The pairwise distance between S. japonica, S. angustata, and S. coraiacea was 0.000–0.116. Sequencing comparisons of rps7, rps11, rpl5, and tatC were thought to be useful tools for discrimination and phylogenetic analysis of Saccharina species having similar morphologies.     

RESTRICTION ENDONUCLEASE ANALYSIS OF RIBOSOMAL DNA SEQUENCE VARIATION IN LAMINARIA (PHAEOPHYTA)1

The taxonomy and evolutionary relationships of species in the genus Laminaria are poorly understood. Previous studies have demonstrated significant plasticity of morphological characters used to describe taxa, and interfertility has been reported among putative species. We analyzed nuclear ribosomal DNA (rDNA) sequence variation in eight species of Laminaria (L. agardhii Kjell., L. digitata (Huds.) Lamour., L. groenlandica Rosenv. [sensu Druehl 1968], L. longicruris De la Pyl., L. longipes Bory, L. saccharina (L.) Lamour., L. setchellii Silva, and L. yezoensis Miyabe) to elucidate evolutionary relationships in this genus. Restriction maps were constructed using a small subunit rDNA probe from Costaria costata (Turn.) Saunders, an rDNA repeat from the nematode Caenorhabditis elegans, and 11 hexameric restriction endonucleases in an annealing analysis of genomic DNA. Laminaria rDNA restriction maps were compared to each other and to that of the outgroup taxon, C. costata. rDNA restriction maps of Laminaria species and C. costata were similar. Restriction fragment length polymorphisms mapped to both the coding regions and the nontranscribed spacer of rDNA. Laminaria species were distinguished with this method. The restriction maps of L. agardhii, L. saccharina, and L. longicruris were identical, supporting a previous hypothesis that these species are conspecific. Comparison of restriction maps of Laminaria species suggested that the generic subdivision of Sections Simplices and Digitatae may be invalid.     

NUCLEOTIDE SEQUENCES OF THE SMALL-SUBUNIT RIBOSOMAL RNA GENES FROM SELECTED LAMINARIALES (PHAEOPHYTA): IMPLICATIONS FOR KELP EVOLUTION1

The entire small-subunit (SSU) ribosomal ribonucleic acid sequence was inferred for kelp representing seven genera: Alaria marginata Postels and Ruprecht (1824 bp), Egregia menziesii (Turner) Areschoug (1825 bp), Lessoniopsis littoralis (Tilden) Reinke (1825 bp), Macrocystis integrifolia Bory (1825 bp), Nereocystis leutkeana (Mertens) Postels and Ruprecht (1824 bp), Postelsia palmaeformis Ruprecht (1826 bp), and Pterygophora californica Ruprecht (1825 bp). We obtained a partial sequence for Eisenia arborea Areschoug (1669 bp) from a single clone of polymerase chain reaction-amplified product. The SSU sequence was too conserved among these morphologically distinct taxa to permit phylogenetic analysis. The divergence between the most distant taxa was only 0.66%. This value was used in a SSU molecular clock to suggest that the most distantly related kelp investigated in this study diverged between 16 and 30 (more probably 16 and 20) million years ago.     

A MOLECULAR ANALYSIS OF ANALIPUS AND RALFSIA (PHAEOPHYCEAE) SUGGESTS THE ORDER ECTOCARPALES IS POLYPHYLETIC1

We report partial 18S ribosomal DNA sequences of Analipus japonicus (Harvey) Wynne and Ralfsia fungiformis (Grunnerus) Setchell et Gardner. These sequences were compared with the corresponding sequences of 13 brown algae representing six phaeophycean orders: Dictyotales, Ectocarpales, Fucales, Laminariales, Sphacelariales, and Syringodermatales. These 15 brown algae included 10 ectocarpoids representing nine ectocarpacean (sensu Gabrielson et al. 1989) families: Chordariaceae, Dictyosiphonaceae, Ectocarpaceae, Elachistaceae, Heterochordariaceae, Leathesiaceae, Punctariaceae, Ralfsiaceae, and Scytosiphonaceae. We addressed the controversial taxonomic placement of A. japonicus and R. fungiformis in the Ectocarpales by analysis of DNA sequences. Neighbor-joining and maximum parsimony-inferred phylogenies provided evidence that A. japonicus and R. fungiformis are not closely associated with the other representatives of the Ectocarpales. Bootstrap analyses suggest polyphyly of the order Ectocarpales (sensu Gabrielson et al.). However, our analysis failed to resolve the phylogenetic relationship between A. japonicus and R. fungiformis. Our results suggest that the ectocarpoids are just as distantly related to A. japonicus and R. fungiformis as they are to members of the advanced orders Desmarestiales, Dictyotales, Fucales, Laminariales, Sphacelariales, and Syringodermatales.