PHYLOGENY OF THE ULVOPHYCEAE (CHLOROPHYTA): CLADISTIC ANALYSIS OF NUCLEAR-ENCODED rRNA SEQUENCE DATA1

Cladistic analysis of nuclear-encoded rRNA sequence data provided us with the basis for some new hypotheses of relationships within the green algal class Ulvophyceae. The orders Ulotrichales and Ulvales are separated from the clade formed by the remaining orders of siphonous and siphonocladous Ulvophyceae (Caulerpales, Siphonocladales /Cladophorales [S/C] complex, and the Dasycladales), by the Chlorophyceae and Pleurastrophyceae. Our results suggest that the Ulvophyceae is not a monophyletic group. Examination of inter- and intra-ordinal relationships within the siphonous and siphonocladous ulvophycean algae revealed that Cladophora, Chaetomorpha, Anadyomene, Microdictyon, Cladophoropsis and Dictyosphaeria form a clade. Thus the hypothesis, based on ultrastructural features, that the Siphonocladales and Cladophorales are closely related is supported. Also, the Caulerpales is a monophyletic group with two lineages; Caulerpa, Halimeda, and Udotea comprise one, and Bryopsis and Codium comprise the other. The Dasycladales (Cymopolia and Batophora) also forms a clade, but this clade is not inferred to be the sister group to the S/C complex as has been proposed. Instead, it is either the sister taxon to the Caulerpales or basal to the Caulerpales and S/C clade The Trentepohliales is also included at the base of the siphonous and siphonocladous ulvophycean clade. The Pleurastrophyceae, which, like the Ulvophyceae, posses a counter-clockwise arrangement of flagellar basal bodies, are more closely related to the Chlorophyceae than to the Ulvophyceae based on rRNA sequences. Thus, the arrangement of basal bodies does not diagnose a monophyletic group. Previously reported hypotheses of phylogenetic relationships of ulvophycean algae were tested. In each case, additional evolutionary steps were required to obtain the proposed relationships. Relationships of ulvophycean algae to other classes of green algae are discussed.     

Structure,Ultrastructure and Function of the Neural Gland Complex of Ascidia interrupta (Chordata,Ascidiacea): Clarification of Hypotheses Regarding the Evolution of the Vertebrate Anterior Pituitary

Abstract The neural gland complex of Ascidia interrupta consists of three parts: dorsal tubercle, ciliated duct, and neural gland. The dorsal tubercle protrudes above the pharyngeal lining and bears a ciliated funnel. The funnel opens into a ciliated duct which opens into the neural gland, a blind sac in a blood sinus below the brain. Funnel and duct cells are joined by adhaerens junctions and, apically, by putative tight junctions. The neural gland wall is a loose, irregular, non-ciliated epithelium of phagocytes. Adhaerens, but not tight, junctions join the cells. Secretory cells were not observed. Tracers delivered onto the dorsal tubercle and dissolved in seawater around undissected animals are transported unidirectionally inward into the neural gland. The continuous ciliary incurrent moves the tracers across the wall of the neural gland and into the pharyngeal blood vessels. Small particulates and large macromolecules, however, are removed from the water stream by endocytosis on neural gland cells. Large particulates delivered onto the dorsal tubercle do not enter the system but rather are rejected by cilia on the surface of the tubercle. The neural gland complex is interpreted as an organ of blood volume regulation that consists of a pump (cilia) and coarse (tubercle) and fine (gland) filters. Analogous and homologous systems are discussed.     

PATTERNS OF PLANT ONTOGENY THAT MAY INFLUENCE GENOMIC STASIS

It is an axiom in biology that genes control the ontogeny and ultimately the final form of an organism. In plants a given morphological form can often arise through more than one ontogenetic pattern of cell divisions. Different ontogenetic patterns have different properties with regard to the final age in cell divisions of the initials in the meristems for a given morphological form. If mutation per genome per cell division is an important biological metric, then since the age of a cell in cell divisions is a function of ontogeny, the cellular ontogeny will influence the degree of mutation-loading in meristematic initials. Thus, ontogeny and form may affect the genes (by promoting or lessening genomic stasis) as well as, of course, being determined by the genes. This paper explores mathematically the relationship between different patterns of cell division and mutation-loading.     

EMBRYOLOGY OF CALYPSO BULBOSA. I. OVULE DEVELOPMENT

Calypso bulbosa is a terrestrial orchid that grows in north temperate regions. Like many orchids, the Calypso has ovules that are not fully developed at anthesis. After pollination, the ovule primordia divide several times to produce a nucellar filament which consists of five to six cells. The subterminal cell of the nucellar filament enlarges to become the archesporial cell. Through further enlargement and elongation, the archesporial cell becomes the megasporocyte. An unequal dyad results from the first meiotic division. A triad of one active chalazal megaspore and two inactive micropylar megaspores are the end products of meiotic division. Callose is present in the cell wall of the megaspore destined to degenerate. In the mature embryo sac the number of nuclei is reduced to six when the chalazal nuclei fail to divide after the first mitotic division. The chalazal nuclei join the polar nucleus and the male nucleus near the center of the embryo sac subsequent to fertilization.     

RESTRICTION FRAGMENT ANALYSIS OF CHLOROPLAST DNA AND THE SYSTEMATICS OF VIGUIERA AND RELATED GENERA (ASTERACEAE: HELIANTHEAE)

Phylogenetic relationships among 31 species representing variously Helianthus, Helianthopsis, Heliomeris, Simsia, Viguiera, and Tithonia were assessed by chloroplast DNA restriction site mapping. A total of 147 mutations including 92 informative ones was detected using 16 restriction enzymes, with an estimated sequence divergence within the group of 1.4%. Parsimony analysis produced a single most parsimonious tree with a length of 161 steps and a consistency index of 0.91. Statistically significant clades, as assessed by the bootstrap method, correspond to a number of taxa recognized previously, including Helianthus (3 species), Helianthopsis (5 species), and several groups within Viguiera, including sect. Maculatae (4 species), the Baja California group (6 species), sect. Paradosa (2 species) and V. dentata (3 samples). However, species of Viguiera collectively form a highly paraphyletic group relative to species of other genera. Helianthus and Helianthopsis were separated into different clades, supporting their recent segregation. Placement of H. porteri in Helianthus rather than Heliomeris was confirmed; the single sample of the latter genus was most similar to the Baja California group of Viguiera. An expected relationship between Simsia (2 species) and one member of Viguiera ser. Grammatoglossae was confirmed (although not with two other putatively related members of Viguiera) and an unexpected relationship between Simsia and Tithonia was suggested. The presence of Mexican taxa as the more basal groups in the tree points toward a possible Mexican origin for Viguiera and related genera. A molecular clock hypothesis is rejected in many pairwise comparisons involving woody taxa with herbaceous ones, although it could not be rejected in most pairwise comparisons involving taxa of similar habit.     

Contribution of pilA to Competitive Colonization of the Squid Euprymna scolopes by Vibrio fischeri

Vibrio fischeri colonizes the squid Euprymna scolopes in a mutualistic symbiosis. Hatchling squid lack these bacterial symbionts, and V. fischeri strains must compete to occupy this privileged niche. We cloned a V. fischeri gene, designated pilA, that contributes to colonization competitiveness and encodes a protein similar to type IV-A pilins. Unlike its closest known relatives, Vibrio cholerae mshA and vcfA, pilA is monocistronic and not clustered with genes associated with pilin export or assembly. Using wild-type strain ES114 as the parent, we generated an in-frame pilA deletion mutant, as well as pilA mutants marked with a kanamycin resistance gene. In mixed inocula, marked mutants were repeatedly outcompeted by ES114 (P < 0.05) but not by an unmarked pilA mutant, for squid colonization. In contrast, the ratio of mutant to ES114 CFUs did not change during 70 generations of coculturing. The competitive defect of pilA mutants ranged from 1.7- to 10-fold and was more pronounced when inocula were within the range estimated for V. fischeri populations in Hawaiian seawater (200 to 2,000 cells/ml) than when higher densities were used. ES114 also outcompeted a pilA mutant by an average of twofold at lower inoculum densities, when only a fraction of the squid became infected, most by only one strain. V. fischeri strain ET101, which was isolated from Euprymna tasmanica and is outcompeted by ES114, lacks pilA; however, 11 other diverse V. fischeri isolates apparently possess pilA. The competitive defect of pilA mutants suggests that cell surface molecules may play important roles in the initiation of beneficial symbioses in which animals must acquire symbionts from a mixed community of environmental bacteria.     

Primate terminal complement inhibitor homologues of human CD59

The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession numbers L22862 (African green monkey CD59) and L22863 (baboon CD59)     

Growth and bioluminescence of Noctiluca scintillans on varying algal diets

The effects of food concentration and food quality on the growthrate and bioluminescence potential of the heterotrophic dinoflagellateNoctiluca scintillans were examined. The growth rate of N. scinullansvaried greatly with the phytoplankton food it was provided,and those foods yielding higher growth rates also produced cellsthat were smaller in diameter, but with a greater bioluminescencepotential per unit volume. Small phytoplankton species, includingIsochrysis galbana and an unidentified chrysophyte responsiblefor the Texas brown tide, supported the lowest growth rates.Other small flagellates (Dunaliella tertiolecta) and dinoflagellates(Gyrodinium dorsum, Prorocentrum minimum) supported moderategrowth rates. The highest growth rates were supported by diatomsand prasinophytes. Maximum growth rates (0.5 day^-1) were obtainedwith the diatom Thalassiosira sp. at concentrations     

Radiative properties of hardwood leaves to ultraviolet irradiation

Spectral reflectance and transmittance of leaves to ultraviolet irradiation were determined under laboratory conditions for seven species of hardwood trees, namely red oak (Quercus rubra, L), black oak (Q. velutina, Lamarch), white oak (Q. alba, L.), sugar maple (Acer saccharum), Norway maple (A. plantanoides), hickory (Carya tomemtosa), sweetgum (Liquidambar styraciflua), and black oak litter. The experimental system consisted of a solar simulator, an integrating sphere, and a spectroradiometer. Experiments were repeated three to five times for both adaxial and abaxial surfaces of fresh leaves chosen at randomly. The spectral distributions and simple averages of the radiative properties in the wavelength ranges of ultraviolet-B (UV-B, 280–320 nm) and ultraviolet-A (UV-A, 320–400 nm) were determined. The spectral distributions of reflectance were similar between adaxial and abaxial surfaces, although the magnitude varied among tree species. Leaf reflectance was very low for the ultraviolet spectrum in general and varied among species and between adaxial and abaxial surfaces. It was generally higher over the UV-A waveband compared to UV-B, and higher on the abaxial than adaxial surface. The broadband reflectance in the UV-A range (over all species) was 5.0 and 3.9% for abaxial and adaxial surface, respectively, compared to 3.5 and 2.8% in UV-B. The transmittance through leaves was extremely small in the UV-B (<0.1%) and nearly zero in the UV-A spectral range. Consequently, the absorptance of ultraviolet radiation by leaves, as determined from the measured reflectance and transmittance, was quite high, being more than 90% for all the combinations of species and wavebands examined. The reported results are useful for studies requiring spectral radiative properties of the examined leaves with respect to ultraviolet irradiation.     

Population genetic response to microsite ecological stress in wild barley, Hordeum spontaneum

Genetic diversity was studied in six subpopulations (a total of 60 individuals) of wild barley, Hordeum spontaneum , the progenitor of cultivated barley, sampled from six stations located along a transect of 300 m across the two opposing slopes of 'Evolution Canyon', a Mediterranean microsite at Lower Nahal Oren, Mt Carmel. The two opposing slopes are separated by between 100 and 400 m and designated SFS (South-Facing Slope) and NFS (North-Facing Slope) with each having three equidistant test stations. The SFS, which receives up to 300% more solar radiation, is drier, ecologically more heterogeneous, fluctuating, and more stressful than the NFS. Analysis of 12 RAPD primers, representing a total of 51 putative loci, revealed a significant inter- and intraslope variation in RAPD band polymorphism. A significantly higher proportion of polymorphic RAPD loci was found amongst the subpopulations on the SFS (mean P = 0.909) than on the NFS (mean P = 0.682), on the basis of the presence and absence of 22 strong bands. Polymorphism generally increased upwards from the bottom to the top of the SFS (0.636, 0.773, 0.955) and NFS (0.409, 0.500, 0.545), respectively. Gametic phase disequilibria estimates, D, revealed SFS and NFS unique predominant combinations which sharply differentiated the two slopes and indicated that there is differential interslope selection favouring slope-specific multilocus combinations of alleles, or blocks of genes over tens to hundreds of meters. This suggests that selection overrides migration. RAPD polymorphism appears to parallel allozyme diversity which is climatically adaptive and driven by natural selection in the same subpopulations at the microsite.