CONTROL OF PLASMODIAL FUSION IN A PANAMANIAN ISOLATE OF DIDYMIUM IRIDIS

Plasmodial fusion in a Panamanian isolate (Pan. 1) of Didymium iridis is controlled by at least six fusion loci. Only plasmodia with identical phenotypes for these six loci are capable of fusion. No evidence was obtained in this study to indicate that any one fusion locus more strongly affects the fusion process than any of the other loci. Control of plasmodial fusion in the Pan. 1 strain of D. iridis is similar to that found earlier for a Honduran strain (Hon. 1).     

Infections of Plasmodium vivax in Saguinus geoffroyi*

SYNOPSIS Panamanian marmosets Saguinus geoffroyi can be infected with Plasmodium vivax. Infections resulting from inoculation of vivax blood from man are generally low-grade but, in many cases, significant infections result from the inoculation of blood from experimentally infected monkeys. Nine of 29 marmosets were infected from man; 9 of 25 from the night monkey Aotus trivirgatus; 13 of 31 from other Panamanian marmosets; and 1 of 1 from a black spider monkey Ateles fusciceps. Average maximum parasitemias for the 4 donor species were, respectively; 2,320; 24,410; 19,380; and 12,160 per cmm. Splenectomy, administration of Imuran, and a combination of splenectomy and administration of Imuran conditioned Panamanian marmosets for infection. Significant parasitemias resulted from infections with the Emperador, Santa Rosa 1, and Achiote strains of vivax malaria. The Emperador strain was passed serially thru 3 marmoset-to-marmoset passages.     

Sigmodontine rodents diversified in South America prior to the complete rise of the Panamanian Isthmus

The extant distribution of sigmodontine rodents encompasses most of the New World, and the majority of the species in this subfamily inhabit South America. Nevertheless, the basal lineages of the Sigmodontinae are distributed in North and Central America, and the fossil record indicates a North American origin. This evidence has produced contentious theories concerning the evolution of these rodents. The dispute usually stems from a disagreement about the way in which sigmodontines reached South America, which was an isolated landmass during most of the Cenozoic. Fundamentally, the debate is associated with the role of Panamanian Isthmus formation and the Great American Biotic Interchange (GABI) in the diversification of the clade. An early hypothesis implies that sigmodontines arrived in South America before the complete rise of the Panamanian Isthmus, whereas a late hypothesis directly correlates the diversification of the lineage with this event. To address this question, we have sequenced nuclear and mitochondrial sequences, as well as the first Sigmodontinae mitochondrial genomes (Akodon montensis and Wiedomys cerradensis) and performed a Bayesian dating analysis. Our results showed that the most recent common ancestor of the subfamily lived at approximately 15 Ma. Although the diversification of sigmodontines was not associated with the complete rise of the Panamanian Isthmus, we cannot exclude the hypothesis that this event played a relevant role in the evolution of the lineage during the Miocene.     

NUCLEAR DNA CONTENT OF MYXAMOEBAE AND PLASMODIA IN SIX NON-HETEROTHALLIC ISOLATES OF A MYXOMYCETE,DIDYMIUM IRIDIS

The nuclear DNA content of six non-heterothallic isolates of the myxomycete Didymium iridis was measured by combining the Feulgen reaction with absorption microspectrophotometry. This allowed us to distinguish between homothallic (sexual) and apogamic (non-sexual) isolates. Four of the isolates studied, Panamanian 4 and 5, California 1, and Missouri 1 are homothallic. Moreover, the average DNA content of the myxamoebal and plasmodial nuclei (0.32 and 0.61 respectively) does not differ significantly from the calculated haploid and diploid values for heterothallic isolates of D. iridis (0.34 and 0.63). Hence, it is concluded that in each of these isolates the myxamoebae are haploid and the plasmodia diploid. In two of the isolates investigated, Georgia 1 and Hawaii 1, the DNA content of the myxamoebal and plasmodial nuclei did not differ significantly. Therefore, in both of these isolates the plasmodia appear to develop apogamically. In addition the mean DNA values recorded for the Ha-1 isolate suggest that it is aneuploid.     

DIFFERENCES IN NITROGEN REQUIREMENTS OF TWO CLONES OF CONVOLVULUS ARVENSIS IN VITRO

Various nitrogen sources were shown to alter the growth and modify nitrate reductase activities of stem callus tissue derived from two clones of Convolvulus arvensis L. (field bindweed). Callus from a Washington (S) clone grew better and had a higher level of nitrate reductase activity than callus from a New Mexico (R) clone when nitrate was the only source of nitrogen available in the culture medium. The addition of glycine to the culture medium decreased growth of the R clone and increased growth of the S clone, but glutamic acid repressed growth of both clones. An amide source of nitrogen such as glutamine or asparagine, or ammonium was required to produce maximum growth of both bindweed clones. Glutamine increased nitrate reductase activity in the two clones, and glycine increased nitrate reductase activity in the S clone but decreased it in the R clone. Glutamic acid decreased nitrate reductase activities in both the R and S tissues.     

BIOCHEMICAL TAXONOMY OF MARINE PHYTOPLANKTON BY ELECTROPHORESIS OF ENZYMES. II. LOSS OF HETEROZYGOSITY IN CLONAL CULTURES OF THE CENTRIC DIATOMS SKELETONEMA COSTATUM AND THALASSIOSIRA PSEUDONANA1, 2

An electrophoretic survey of 12 new isolates of Thalassiosira pseudonana Hasle & Heimdal and 25 new isolates of Skeletonema costatum (Grev.) Cleve revealed several heterozygote genotypes at malate dehydrogenase (MDH) and phosphohexose isomerase (PHI) loci. The new clones were maintained in culture for 6 mo and then reasayed at these two loci. All MDH heterozygotes and halj of the PHI heterozygotes had become homozygous. This resulted in a collection of clones that are largely homozygous but that are samples of polymorphic species. The physlogical implications of this loss of heterozygosity in clonal cultures has not been analyzed. Hawever, any change in a clone that is the result of culturing conditions reduces the usefulness of that clone as a laboratory test organism for ecological correlations.     

Phylogeography of the harlequin beetle-riding pseudoscorpion and the rise of the Isthmus of Panamá

Molecular and geological evidence indicates that the emergence of the Isthmus of Panamá influenced the historical biogeography of the Neotropics in a complex, staggered manner dating back at least 9 Myr bp. To assess the influence of Isthmus formation on the biogeography of the harlequin beetle-riding pseudoscorpion, Cordylochernes scorpioides, we analysed mitochondrial COI sequence data from 71 individuals from 13 locations in Panamá and northern South America. Parsimony and likelihood-based phylogenies identified deep divergence between South American and Panamanian clades. In contrast to low haplotype diversity in South America, the Panamanian Cordylochernes clade is comprised of three highly divergent lineages: one clade consisting predominantly of individuals from central Panamá (PAN A), and two sister clades (PAN B1 and PAN B2) of western Panamanian pseudoscorpions. Breeding experiments demonstrated a strictly maternal mode of inheritance, indicating that our analyses were not confounded by nuclear-mitochondrial pseudogenes. Haplotype diversity is striking in western Atlantic Panamá, where all three Panamanian clades can occur in a single host tree. This sympatry points to the existence of a cryptic species hybrid zone in western Panamá, a conclusion supported by interclade crosses and coalescence-based migration rates. Molecular clock estimates yield a divergence time of approximately 3 Myr between the central and western Panamanian clades. Taken together, these results are consistent with a recent model in which a transitory proto-Isthmus enabled an early wave of colonization out of South America at the close of the Miocene, followed by sea level rise, inundation of the terrestrial corridor and then a second wave of colonization that occurred when the Isthmus was completed approximately 3 Myr bp.     

UPTAKE OF ORGANIC,COMPOUNDS BY TWO FACULTATIVELY HETEROTROPHIC MARINE CENTRIC DIATOMS1,2

Glucose and galactose uptake ability in Cyclotella cryptica (clone WT-1-8) and glucose uptake ability in Coscinodiscus sp. develop rapidly in the dark. Induction of sugar uptake ability in the dark does not require the presence of sugar in the medium. The sugars are taken up by carrier-mediated systems. In C. cryptica glucose and galactose are probably taken up by the same system. The capacity of glucose uptake in this recently isolated clone of C. cryptica is nearly 5 times that of a previously studied clone (0-3A). Other organic compounds, which by themselves do not support heterotrophic growth, can be taken up and respired by both diatoms at considerable rates compared, to glucose and galactose. Therefore, in nature, these diatoms may be able to utilize a variety of dissolved organic compounds as sources of intermediary metabolites and as respiratory substrates.     

COMPARATIVE MEASUREMENTS OF NUCLEAR DNA IN A HETEROTHALLIC AND A SELF-FERTILE ISOLATE OF THE MYXOMYCETE,DIDYMIUM IRIDIS

Comparative measurements were made of the nuclear Feulgen-DNA content of a heterothallic and a self-fertile isolate of the myxomycete Didymium iridis. Plasmodial nuclei of both isolates contain the diploid amount of DNA. The replicated diploid (4C) values for the heterothallic and the self-fertile isolates are 5.66 and 5.95, respectively. Myxamoebae, however, are quite dissimilar in their nuclear DNA content. Those of the heterothallic isolates, Honduran 1–2 (A¹) and Panamanian 2–4 (A⁷), have mean values of 3.81 and 3.69, whereas myxamoebae of the self-fertile Philippine-1 isolate were found to have a mean value of 6.07. Myxamoebae of the Ph-1 isolate are, therefore, at the same ploidy level as the Ph-1 Plasmodium. Mean DNA values for Ph-1 sporangial nuclei were in category 4C. Measurement of the DNA content of mitotic metaphases in sporangia at T = 6 hr confirmed that the mean DNA content of both Ph-1 myxamoebae and plasmodial nuclei is equivalent to 4C. It is concluded that nuclear phase alternance is lacking in the Ph-1 isolate and that the Plasmodium of this isolate develops by apogamy.     

Malaria Parasites of the “Borriguerro'’Lizard,Ameiva ameiva (Sauria: Teiidae) in Panama*

SYNOPSIS. The teiid lizard Ameiva ameiva praesignis in Panama is parasitized by 2 species of Plasmodium: P. cnemidophori Carini and P. diminutivum sp. n. The Panamanian strain of P. cnemidophori has thick macrogametocytes which are larger than the oval microgametocytes, and schizonts which contain 42–119 nuclei. P. diminutivum is characterized by round to oval gametocytes which are usually smaller than the host cell nucleus; fan-shaped schizonts with 4–6 nuclei; and prominently vacuolated young stages, with a thick band of chromatin along the periphery of the vacuole.     

Phylogenetic distribution of symbiotic bacteria from Panamanian amphibians that inhibit growth of the lethal fungal pathogen Batrachochytrium dendrobatidis

The introduction of next‐generation sequencing has allowed for greater understanding of community composition of symbiotic microbial communities. However, determining the function of individual members of these microbial communities still largely relies on culture‐based methods. Here, we present results on the phylogenetic distribution of a defensive functional trait of cultured symbiotic bacteria associated with amphibians. Amphibians are host to a diverse community of cutaneous bacteria and some of these bacteria protect their host from the lethal fungal pathogen Batrachochytrium dendrobatidis (Bd) by secreting antifungal metabolites. We cultured over 450 bacterial isolates from the skins of Panamanian amphibian species and tested their interactions with Bd using an in vitro challenge assay. For a subset of isolates, we also completed coculture experiments and found that culturing isolates with Bd had no effect on inhibitory properties of the bacteria, but it significantly decreased metabolite secretion. In challenge assays, approximately 75% of the bacterial isolates inhibited Bd to some extent and these inhibitory isolates were widely distributed among all bacterial phyla. Although there was no clear phylogenetic signal of inhibition, three genera, Stenotrophomonas, Aeromonas and Pseudomonas, had a high proportion of inhibitory isolates (100%, 77% and 73%, respectively). Overall, our results demonstrate that antifungal properties are phylogenetically widespread in symbiotic microbial communities of Panamanian amphibians and that some functional redundancy for fungal inhibition occurs in these communities. We hope that these findings contribute to the discovery and development of probiotics for amphibians that can mitigate the threat of chytridiomycosis.     

QUANTITATIVE DETERMINATION OF CANNABINOIDS IN INDIVIDUAL GLANDULAR TRICHOMES OF CANNABIS SATIVA L. (CANNABACEAE)

Cannabinoid levels of individual mature glandular trichomes from two clones and two strains of Cannabis sativa L., which included both drug and fiber phenotypes, were investigated by gas-liquid chromatographic analyses. Capitate-stalked glands were selectively harvested from vein and nonvein areas of pistillate bracts while capitate-sessile glands were harvested from these areas of leaves. The qualitative cannabinoid profile characteristic of the strain or clone was maintained in the individual capitate-stalked glands while the quantitative cannabinoid profiles varied with each strain or clone and between vein and nonvein areas as well. Capitate-sessile glands were found to contain conspicuously lower levels of cannabinoids than capitate-stalked glands. This study emphasizes that glands of Cannabis represent a dynamic system within the cannabinoid synthesizing activities of this plant.     

ASEXUAL DEVELOPMENTAL PATTERNS OF THE DIATOM STAURONEIS ANCEPS IN CULTURE123

During several months under controlled growth conditions, cell division in the diatom Stauroneis anceps resulted in a gradual decrease in length as predicted by the Macdonald-Pfitzer hypothesis. Cells of a clone decreased in average length from 35 to 17 μ. The smallest cell observed measured 12 μ. Cells longer than 26 μ typically had capitate ends. As the cells became shorter, there was a gradual decrease in this capitate configuration with the smallest cells being oval in outline. The estimated increment of size reduction due to cell division in the clone was 0.1 μ per division. This value is dependent on the assumption of random division. Populations with cells less than 28 μ in length showed an increase in valve aberrations, but these cells continued to reproduce. A cell 13 μ long and similar to the smallest specimen of the clone was observed from a core sample with fossil specimens of S. anceps.     

STRUCTURE OF CLONES OF NORTHERN PRICKLY ASH (XANTHOXYLUM AMERICANUM)

Clones of northern prickly ash (Xanthoxylum americanum) were excavated to expose their horizontal root structure and pattern of vegetative spread; stems and horizontal roots were mapped. Most of the root system was very shallow (< 10 cm deep). Stems and roots were aged from annual rings. The four clones studied ranged in age from 26 to 33 yrs. All root connections between shoots were maintained for the life of the clone. Clones were found to be single individuals in a physical as well as a genetic sense. There was virtually no overlap of adjacent clones; horizontal roots turned away or stopped growing when the border of another clone was encountered. No differences were found between male and female clones in above- or below-ground morphology. There was, however, a great deal of intrasexual variation among clones.     

B12 SPECIFICITY OF MARINE CENTRIC DIATOMS1, 2

Twenty-one species (23 isolates) of marine diatoms were examined for their capacity to utilize analogs of cyanocobalamin for growth at the ecologically significant concentration of 4 ng 1-¹. Yields due to the analogs were compared to those produced by B₁₂. Responses of the various clones to the analogs were not all-or-none, but varied continuously; thus, assigning the clones to the conventional B₁₂ specificity types is a convenient but arbitrary classification. The use of 10 and 1% levels of response is suggested for such classification. At the 10% level of response, 11 clones had coliform, 4 lactobacillus, and 8 mammalian specificity patterns. At the 1% response level, 14 had coliform, 5 lactobacillus, and 4 mammalian specificities. All clones exceed the 10% response level on all benzimi-dazole-containine analogs tested. Few clones showed definite enough patterns of response to make them potentially useful for differential bioassay. The clones suggested are clone 675-D (Bidclulphia sp.?), clone F^;-3 (Fragilaria sp.?), and the estuarine clone of Cyclotella nana (3H).     

A STATISTICAL CHARACTERIZATION OF GROWTH AMONG CLONES OF SYNURA PETERSENII (SYNUROPHYCEAE)1

Each of four clones from the Synura petersenii complex was grown at different pHs (5.5, 6.5, 7.5, 8.5) in batch culture experiments. Growth response curves and exponential growth rates were compared among clones and pH treatments in order to examine growth trend variation among the clonal groups. The clones were isolated from geographically distant North American localities. The clonal groups represented distinct mating types, an isolate and its subisolate, and S. petersenii- and S. glabra-like scale morphologies. No consistent relationship existed between growth response curve, and culture medium pH. Additionally, the trends across time differed according to clone and pH combination. Pairwise comparisons of linear trends from transformed growth response curves indicated two distinct clonal associations. Although the clonal associations corresponded with the final cell density of the cultures, growth response curves did not correspond with mating type, the parent-isolate and subisolate, or scale morphology. Clones with glabra-like scales had greater growth rates than the clone with petersenii-like scales. The conflicting results generated from growth response curve and growth rate analyses support the concept that S. petersenii and S. glabra form a highly variable, homogeneous grouping.     

KINETICS OF SILICON-LIMITED GROWTH IN THE FRESHWATER DIATOM ASTERIONELLA FORMOSA1,2

Growth rates of two clones of the freshwater planktonic diatom Asterionella formosa Hass. were measured under conditions in which external silicon concentrations controlled growth. Clone AfOH2 from Lake Ohrid, Yugoslavia, had a higher maximum growth rate (μ_max= 1.11 doublings/day) and apparent half-saturation constant (K_si] + Si_o= 1.93 μM Si) than clone L262 from Lake Windermere, England. (μ_max= 0.61 doublings/day; K_si+ Si_o= 1.09 μM Si). K_lim, the silicon concentration at μ= 0.9 μ_max, is 13.8 μM Si for clone AfOH2 and 6.5 μM Si for clone L262. These values agree well with published field observations showing A. formosa populations decreasing below 0.5 mg/l SiO₂ (= 8.4 μM Si). Calculations of yield gave a range of 0.5–1.5 μM Si/10⁶ cells for clone AfOH2 and 0.6–1.9 μM Si/10⁶ cells for clone L262.     

SURFACE CONFIGURATION OF THE LORICA OF THE EUGLENOID TRACHELOMONAS AS REVEALED WITH SCANNING ELECTRON MICROSCOPY

The surface of Trachelomonas was examined readily with SEM at magnifications up to X 20,000 in unfixed or fixed material. Cells were placed directly on specimen holders without washing, air dried, and coated with aluminum or gold. Clones of T. grandis, T. hispida var. coronata, and T. oblonga var. punctata and three species from a natural collection showed the following features: the lorica was punctate or solid, and the surface projections, when present, consisted of minute papillae (0.1 μm), larger, often globose papillae (up to 0.8 μm), and spines which tapered to a point or which had parallel sides and ended bluntly; spines appeared hollow or solid. Pringsheim's clone of T. oblonga var. punctata, which he described as a new spineless variety, possessed short tapering spines and globose papillae. Such observations suggest that a major problem in delimiting species may be in discovering the ornamentation potential of a clone. The discovery of new features of the lorica (e.g., minute papillae and hollow projections), the clarification of shapes of surface features, and, in general, the excellent resolution at 10 times the usable magnification of light microscopy dictate a reexamination of known species with SEM. We suggest that a SEM study of the variation in the lorica surface in clonal material will lead to a recognition of polymorphism and eventually to a clearer understanding of taxonomic entities described from natural collections.     

Proposed nutritional importance of plant exudates in the diet of the Panamanian Tamarin,Saguinus oedipus geoffroyi

Data collected on the feeding behavior of Saguinus oedipus geoffroyi(Panamanian tamarin) in Panama show that plant exudates, as well as insects and fruits, are a major component of the diet. During the months of May, June, and July, tamarins were observed clinging to the trunks of Anacardium excelsum(Anacardiaceae) while feeding on exudates. These observations accounted for greater than 14% of the total feeding time of this primate. Nutritional analyses indicate that Anacardiumexudates provide these tamarins with a rich source of the mineral calcium as well as protein, carbohydrates, and water. The seasonal pattern of exudate feeding observed in S. oedipusappears to coincide with certain nutritionally sensitive periods of the reproductive cycle and may serve to offset a potential mineral imbalance incurred during the terminal phase of gestation and lactation.     

ABUNDANCE OF A NOVEL DINOFLAGELLATE PHYLOTYPE IN THE ROSS SEA,ANTARCTICA1

The biodiversity of protistan assemblages present in microhabitats of the Ross Sea, Antarctica, was examined using molecular biological methods to obtain a greater understanding of the genetic diversity present. Sequencing of 18S clone libraries indicated genetically diverse collections of organisms in the water column, ice, and meltwater layer (slush), but a single small subunit ribosomal DNA (srDNA) sequence type dominated clone libraries (>30%) from seawater and slush samples taken within the ice pack of this ecosystem. The BLAST searches indicated that this dominant clone was derived from a dinoflagellate, and that it shared sequence similarity (97.6%–98.3%) with both Karenia and Karlodinium species. Phylogenetic analyses based on small subunit ribosomal gene sequences supported its placement as a sister group to these taxa, and suggested that it represented a novel genus. The dinoflagellate was successfully recovered in culture, and morphological analyses have shown that it contains chloroplasts, is gymnodinoid, appears not to have thecal plates, and has an apical groove and sulcal structure that confirm its placement as a relative of the Karenia/Karlodinium group. The abundance of this phylotype in natural samples was confirmed by quantitative PCR analyses of water and slush communities, and suggests that this dinoflagellate can be a major constituent of the protistan assemblages of some Antarctic microhabitats of the Ross Sea.