QUANTITATIVE GENETIC ANALYSIS OF MORPHOLOGICAL VARIATION IN AN ANTARCTIC DIATOM GROWN AT TWO LIGHT INTENSITIES1

Ten clonal isolates of Thalassiosira tumida (Janisch) Hasle were grown in duplicate semi-continuous batch cultures at 116 and 11.6 μE·m⁻²·s⁻¹ acclimated cells were harvested during exponential growth and cleaned for examination by light microscopy (LM) and scanning electron microscopy (SEM). Number of strutted processes surrounding the central annulus (SP) and average number of satellite pores per process (AVSAT) were counted using SEM on 20 valves from each culture grown in high light, for a total of 400 valves examined; number of marginal labiate processes (LP) and overall diameter (DIAM) were measured using LM on 20 valves from each culture grown in both high and low light for a total of 800 valves examined. Univariate analysis of variance showed that bottle effects resulting from microenvironmental differences between replicates were a small but significant source of variation in DIAM, LP, and SP but not AVSAT. Significant differences among clones were observed for all characters. Decreased irradiance resulted in a significant decrease in valve diameter but no significant effect on LP; no light × clone interaction was observed. Significant covariance between characters among clones was also observed; since valve diameter is known to decrease during asexual growth, the correlation coefficients for SP, AVSAT, and LP with DIAM were used to correct the data for this source of nongenetic differences between clones. Analysis of the size-corrected data showed that the proportion of total phenotypic variance in SP, LP, and AVSAT caused by genetic differences among clones was 0.14, 0.14, and 0.30, respectively. This indicates that the majority of total phenotypic variance was due to environmental or developmental causes, but that sufficient genetic variability exists to support rapid phenotypic evolution in SP, LP, and AVSAT under continued directional selection. Finally, the results of the genetic analysis revealed a high (0.82) genetic correlation between SP and LP.     

AN INVESTIGATION OF DISTRIBUTIONAL PATTERNS IN THE DIATOM FLORA OF NETARTS BAY,OREGON, BY CORRESPONDENCE ANALYSIS1

Distributional patterns in assemblages of epiphytic and sediment-associated diatoms were investigated in Netarts Bay, Oregon. The method of reciprocal averaging revealed a floristic discontinuity between the epiphytic and sediment samples in ordination space. The basis for this discontinuity was the presence of a large number of sediment-associated taxa that were either very rare or not observed in the epiphytic samples. Within the sediment samples, the diatom flora formed a distributional continuum which had relatively high correlations with mean grain size, a sediment sorting coefficient, and the organic matter content of the sediment. A comparison of the flora in Netarts Bay with floras in other Oregon estuaries indicates that epiphytic, epilithic, and sediment-associated diatom assemblages do not exhibit conspicuous latitudinal changes along the coast of Oregon, and that many of the same taxa can be expected to occur in samples from comparable habitats in estuaries throughout the temperate regions of the world.     

MICROSATELLITE MARKER DEVELOPMENT AND GENETIC VARIATION IN THE TOXIC MARINE DIATOM PSEUDO‐NITZSCHIA MULTISERIES (BACILLARIOPHYCEAE)1

The genetic structure of phytoplankton populations is largely unknown. In this study we developed nine polymorphic microsatellite markers for the domoic acid–producing marine diatom Pseudo‐nitzschia multiseries (Hasle) Hasle. We then used them in the genotyping of 25 physiologically diverse field isolates and six of their descendants: 22 field isolates originated from eastern Canadian waters, two from European waters, and one from Russian waters. The numbers of alleles per locus ranged from three to seven and the observed heterozygosities from 0.39 to 0.70. A substantial degree of genetic variation was observed within the field isolates, with 23 different genotypes detected. The Russian isolate was the most genetically distinct, although there was also evidence of genetic differentiation at a more local scale. Mating experiments demonstrated that alleles were inherited in a Mendelian manner. Pseudo‐nitzschia multiseries primer pairs were tested on DNA from four congeners: P. calliantha Lundholm, Moestrup et Hasle; P. fraudulenta (P. T. Cleve) Hasle; P. pungens (Grunow ex P. T. Cleve) Hasle; and P. seriata (P. T. Cleve) H. Peragallo. Cross‐reactivity was only observed in P. pungens. Our results are a first step in understanding the genetic variation present at the Pseudo‐nitzschia“species” level and in determining the true biogeographic extent of Pseudo‐nitzschia species.     

ELECTROPHORETIC ANALYSIS OF GENETIC VARIATION IN THE CHAROPHYTA. I. GENE DUPLICATION VIA POLYPLOIDY1

Electrophoretic analysis of 12 species of Chara indicates that functional gene duplication via polyploidy has commonly occurred in the genus. Duplication has been followed by differentiation of the loci encoding phosphoglucose isomerase (PGI). This has led directly to generation of substantial levels of genie variation both within and between taxa. A simple genetic model is proposed to account for the large array of PGI phenotypes encountered in natural populations. Constraints imposed by the reproductive biology of members of the genus, such as selfing, appear to have resulted in selective premiums on intrinsic mechanisms of generating genetic variation. Levels of variation in PGI were higher than would be predicted on the basis of charophyte reproductive characteristics; haploid loci segregate approximately two alleles within each species.     

RELATIONSHIPS BETWEEN NITRATE REDUCTASE ACTIVITY AND RATES OF GROWTH AND NITRATE INCORPORATION UNDER STEADY-STATE LIGHT OR NITRATE LIMITATION IN THE MARINE DIATOM THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE)1

Although activity of the enzyme nitrate reductase (NR) can potentially be used to predict the rate of nitrate incorporation in field assemblages of marine phytoplankton, application of this index has met with little success because the relationship between the two rates is not well established under steady-state conditions. To provide a basis for using NR activity measurements, the relationships among NR activity, growth rate, cell composition, and nitrate incorporation rate were examined in cultures of Thalassiosira pseudonana (Hustedt)Hasle and Heimdal, growing a) under steady-state light limitation, b) during transitions between low and high irradiance (15 or 90 μmol quanta.m^?2.s^?1), and c) under steady-state nitrate limitation. Using a modified assay for NR involving additions of bovine serum albumin to stabilize enzyme activity, NR activity in light-limited cultures was positively and quantitatively related to calculated rates of nitrate incorporation, even in cultures that were apparently starved of selenium. During transitions in irradiance, growth rates acclimated to new conditions within 1 day; through the transition, the relationship between NR activity and nitrate incorporation rate remained quantitative. In nitrate-limited chemostat cultures, NR activity was positively correlated with growth rate and with nitrate incorporation rates, but the relationship was not quantitative. NR activity exceeded nitrate incorporation rates at lower growth rates (<25% of nutrient-replete growth rates), but chemostats operating at such low dilution rates may not represent ecologically relevant conditions for marine diatoms. The strong relationship between NR activity and nitrate incorporation provides support for the idea that NR is rate-limiting for nitrate incorporation or is closely coupled to the rate-limiting step. In an effort to determine a suitable variable for scaling NR activity, relationships between different cell components and growth rate were examined. These relationships differed depending on the limiting factor. For example, under light limitation, cell volume and cell carbon content increased significantly with increased growth rate, while under nitrate limitation cell volume and carbon content decreased as growth rates increased. Despite the differences found between cell composition and growth rate under light and nitrate limitation, the relationships between NR activity scaled to different compositional variables and growth rate did not differ between the limitations. In field situations where cell numbers are not easily determined, scaling NR activity to particulate nitrogen content may be the best alternative. These results establish a strong basis for pursuing NR activity measurements as indices of nitrate incorporation in the field.     

QUANTITATIVE GENETICS OF BRYOZOAN PHENOTYPIC EVOLUTION. II. ANALYSIS OF SELECTION AND RANDOM CHANGE IN FOSSIL SPECIES USING RECONSTRUCTED GENETIC PARAMETERS

The roles of natural selection and random genetic change in the punctuated phenotypic evolution of eight Miocene-Pliocene tropical American species of the cheilostome bryozoan Metrarabdotos are analyzed by quantitative genetic methods. Trait heritabilities and genetic covariances reconstructed by partitioning within- and among-colony phenotypic variance are similar to those previously obtained for living species of the cheilostome Stylopoma using breeding data. The hypothesis that differences in skeletal morphology between species of Metrarabdotos are entirely due to mutation and genetic drift cannot be rejected for reasonable rates of mutation maintained for periods brief enough to account for the geologically abrupt appearances of these species in the fossil record. Except for one pair of species, separated by the largest morphologic distance, directional selection acting alone would require unrealistically high rates of selective mortality to be maintained for these periods. Thus, directional selection is not strongly implicated in the divergence of Metrarabdotos species. Within species, rates of net phenotypic change are slow enough to require stabilizing selection, but mask large, relatively rapid fluctuations, all of which, however, can be attributed to chance departures from the mean phenotype by mutation and genetic drift, rather than to tracking environmental fluctuation by directional selection. The results are consistent with genetic models involving shifts between multiple adaptive peaks on which phenotypes remain more or less static through long-term stabilizing selection. Regardless of the degree to which directional selection may be involved in peak shifts, phenotypic differentiation is thus related to processes different than the pervasive stabilizing selection acting within species.     

EFFECT OF ZINC AVAILABILITY ON GROWTH,MORPHOLOGY, AND NUTRIENT INCORPORATION IN A COASTAL AND AN OCEANIC DIATOM1

We investigated the effect of Zn availability on growth rate (μ), cell morphology, and elemental stoichiometry and incorporation rate in two marine diatoms. For the coastal diatom Skeletonema costatum (Grev.) Cleve, the half‐saturation constant (K_S) for growth was 4.1 pM Zn²⁺, and growth ceased at ≤ 2.6 pM Zn²⁺, whereas for the oceanic diatom Thalassiosira oceanica Hasle, K_S was 0.5 pM Zn²⁺, and μ remained at ～40%μ_max even at 0.3 pM Zn²⁺. Under Zn‐limiting (Zn‐L) conditions, S. costatum decreased cell size significantly, leading to an 80% increase in surface area to volume ratio (SA/V) at Zn²⁺ of 3.5 pM compared to Zn‐replete (Zn‐R) conditions (at Zn²⁺ of 13.2 pM), whereas T. oceanica’s morphology did not change appreciably. Cell quotas of C, N, P, Si, and chl a significantly decreased under Zn limitation in S. costatum (at Zn²⁺ of 3.5 pM), whereas Zn limitation in T. oceanica (at Zn²⁺ of 0.3 pM) had little effect on quotas. Elemental stoichiometry was ～85C:10N:9Si:1P and 81C:9N:5Si:1P for S. costatum, and 66C:5N:2Si:1P and 52C:6N:2Si:1P for T. oceanica, under Zn‐R and Zn‐L conditions, respectively. Incorporation rates of all elements were significantly reduced under Zn limitation for both diatoms, but particularly for Si in S. costatum, and for C in T. oceanica, despite its apparent tolerance of low Zn conditions. With [Zn²⁺] in some parts of the ocean being of the same order (～0.2 to 2 pM) as our low Zn conditions for T. oceanica, our results support the hypothesis that in situ growth and C acquisition may be limited by Zn in some oceanic species.     

GENETIC AND MORPHOLOGICAL IDENTIFICATION OF FUCUS RADICANS SP. NOV. (FUCALES,PHAEOPHYCEAE) IN THE BRACKISH BALTIC SEA1

Brown seaweeds of the genus Fucus occupy a wide variety of temperate coastal habitats. The genus is evolutionary dynamic with recent radiations to form morphologically distinct taxa. In the brackish Baltic Sea, fucoids are the only perennial canopy‐forming macroalgae. The most northern populations of Fucus occur permanently submerged in extremely low salinity (3–5 psu). These are currently referred to as Fucus vesiculosus L. but are morphologically distinct with a narrow frond without bladders. We report here that a population of this unique morphotype is reproductively isolated from a truly sympatric population of common F. vesiculosus and conclude that the northern morphotype represents a previously undescribed species. We describe Fucus radicans sp. nov., which is attached and dioecious with broadly elliptic receptacles, characterized by a richly branched narrow flat frond (2–5 mm), short thallus (<26 cm), and a high capacity for vegetative recruitment of attached plants. Analysis of five highly polymorphic microsatellite DNA loci showed genetic differentiation between sympatric populations of F. radicans and F. vesiculosus, whereas allopatric populations of the same species revealed a coherent pattern of genetic variation. Sequences of the RUBISCO region in F. radicans were identical to or differing at only one to two dinucleotide positions from those of F. vesiculosus, indicating a recent common origin of the two species.     

PSEUDOCODIUM MUCRONATUM,A NEW SPECIES FROM NEW CALEDONIA,AND AN ANALYSIS OF THE EVOLUTION OF CLIMATIC PREFERENCES IN THE GENUS (BRYOPSIDALES,CHLOROPHYTA)1

A new species, Pseudocodium mucronatum, is described from the Chesterfield platform off the west coast of New Caledonia. The species differs from its congeners in having mucronate utricules. A phylogenetic analysis of rbcL and tufA sequences showed that P. mucronatum is most closely associated with P. natalense De Clerck, Coppejans et Verbruggen and P. devriesii Weber Bosse, with which it shares compressed axes, depressed apices, and plastids in the utricles and the medullar siphons. We studied the evolution of climatic and ecological preferences in the genus using an interdisciplinary approach consisting of relaxed molecular clock analysis, extraction of macroecological data from satellite imagery in a geographic information system (GIS) framework, and ancestral character state estimation. It was shown that the genus originated in tropical waters during the Early Mesozoic. Whereas the P. floridanum‐okinawense lineage remained tropical, the lineage including P. natalense, P. devriesii, and P. mucronatum gradually invaded more temperate waters during Cenozoic times. Except for P. devriesii, which occurs in shallow and intertidal habitats, all Pseudocodium species grow in deep‐water habitats, and this ecological preference appears to be ancestral.