Hybridization studies in Bostrychia. 1: B. radicans (Rhodomelaceae, Rhodophyta) from Pacific and Atlantic North America

Bostrychia radicans(Montagne) Montagne is a pantropical/temperate red alga associated with mangroves and saltmarsh plants. Collections were made from a similar north-south geographic distribution along both the Pacific and Atlantic coasts of North America. Hybridization studies were performed with cultured isolates to assess the extent of interfertility and reproductive isolation along these two coastlines. All male and female gametophytes derived from single tetrasporophytes were intercompatible. Almost all isolates extending over 1500 km of coast line from northern Pacific Mexico are compatible, forming cystocarps that released viable carpospores. Even isolates which morphologically would be placed in two species [B. radicans and B. moritziana(Sender ex Kützing) J. Agardh], based on the presence or absence of monosiphonous branches, were capable of hybridizing. Crosses of isolates from the Atlantic USA showed a greater amount of incompatibility. Certain isolates were not compatible with any other isolates including isolates collected in close proximity (North Carolina isolates), while other isolates from the same locality were compatible (South Carolina). An isolate from South Carolina formed tetrasporophytes with isolates from Pacific Mexico but tetraspores were not viable. Certain incompatible crosses formed ‘pseudocystocarps’ but viable carposporophytes did not develop. Generalizations about reproductive isolation within a species must also consider differences between populations from different biogeographic regions that may reflect different paleoclimatological histories, founder effects and unique dispersal events.     

CLONAL VARIATION IN PHOTOTAXIS AND SETTLEMENT BEHAVIORS OF HINCKSIA IRREGULARIS (PHAEOPHYCEAE) SPORES1

Hincksia irregularis (Kützing) Amsler isolates from North Carolina were used to assess clonal variations in spore swimming behavior and settlement responses within a population. These behaviors were also compared with those previously reported from a Florida panhandle clone to assess differences between geographically distinct populations. Minor variation between morphologies, reproductive responses, and settlement intervals were observed between all clones. Spores from all North Carolina clones exhibited positive phototaxis, but there was significant variation in the lower threshold irradiances that could elicit the behavior. In contrast, spores from the previously studied Florida panhandle clone exhibited negative phototaxis. Hincksia irregularis spore settlement responses to hydrophobic and hydrophilic surfaces were consistent for all clones currently and previously tested. Interactive effects of light and charge on settlement were evident in spores from North Carolina isolates, but these effects differed from the Florida panhandle isolate. Settlement preference and swimming behavior differences may have a considerable effect on an individual's ecological fitness and placement within the marine environment. However, the perceived relevance of these differences needs to be validated further in light of previously reported genetic, physiological, and behavioral variations observed in other algal clones that have called into question traditional assessments of individuality.     

Distribution and Identification of an Invasive Gracilaria Species that is Hampering Commercial Fishing Operations in Southeastern North Carolina, USA

A newly abundant Gracilaria species in the sounds of southeastern North Carolina has become a problem for commercial fishing and industries drawing water from the lower Cape Fear River. DNA sequence analyses have shown that this species is Gracilaria vermiculophylla, a taxon originally described from East Asia. Surveys for G. vermiculophylla have shown that it has a discontinuous distribution in the sounds of southeastern North Carolina, and suggest that it is spreading. Gracilaria vermiculophylla meets 6 of the 10 criteria used to help identify invasive species in that it has only recently appeared in southeastern North Carolina; is associated with human mechanisms of dispersal; has a restricted distribution; has disjunct populations in isolated oceans; no means of active dispersal, and an exotic evolutionary origin. The species may also meet two additional criteria as its local range is believed to be expanding, and it is filling a previously unoccupied seasonal niche. These factors taken together strongly suggest that G. vermiculophylla is an invasive species in southeastern North Carolina.     

Nucleotide sequences ofCyanophora paradoxa cellular and cyanelle-associated 5S ribosomal RNAs: The cyanelle as a potential intermediate in plastid evolution

Summary The 5S ribosomal RNAs from the cell cytoplasm and cyanelle (photosynthetic organelle) ofCyanophora paradoxa have been isolated and sequenced. The cellular and cyanelle 5S rRNAs were 119 and 118 nucleotides in length, respectively. Both RNAs exhibited typical 5S secondary structure, but the primary sequence of the cellular species was clearly eukaryotic in nature, while that of the organellar species was prokaryotelike. The primary sequence of the cyanellar 5S rRNA was most homologous to cyanobacterial 5S sequences, yet possessed secondary-structural features characteristic of higher-plant chloroplast 5S rRNAs. Both sequence comparison and structural analysis indicated an evolutionary position for cyanelle 5S rRNA intermediate between blue-green alga and chloroplast 5S rRNAs.Contribution from the Department of Biochemistry, School of Agriculture and Life Sciences and School of Physical and Mathematical Sciences, North Carolina State University, Raleigh, North Carolina. This is paper no. 10259 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601, USA     

Parasexual fusion products in green algae: Enteromorpha and Ulvaria (Ulvales,Chlorophyta)

Enteromorpha linza and Ulvaria oxysperma in North Carolina reproduce exclusively by asexual zoospores. Calcofluor white staining indicated that newly released zoospores lack significant cellulose cell wall material, making them suitable for treatment as protoplasts in a parasexual fusion process using high pH-Ca²⁺, PEG and centrifugation. Presumptive fusion products were identified by their larger size, twin chloroplasts and eyespots, and presence of fluorescence labelled and unlabelled portions. Parasexual fusion and karyogamy were confirmed by elevated levels of nuclear DNA in fusion cell germlings. In addition, aceto-orcein staining of fusion cell products revealed a diploid chromosome complement of 2N = 20 in Enteromorpha linza. Fusion cells were isolated by killing the more numerous adjacent unfused zoospores with 2-3 min exposure to blue light (410–490 nm). Unexposed fusion cells could be readily distinguished and recovered by micropipette at the 10-day stage.Center for Marine Science Research UNC-W Contribution No. 008.     

Ethylene-binding characteristics in Phaseolus,Citrus, and Ligustrum plants

A number of plants were tested for their ability to bind ethylene and the number of binding sites present in each was calculated. Primary leaves of laboratory-grown beans (Phaseolus vulgaris) bound 140 dpm/g fwt (1794 dpm/g dry wt) when exposed to 1.0 Ci/1 of [¹⁴C]ethylene (110 ci/mol). Phytotron-grown leaves were less succulent but only bound 90 dpm/g fwt (1046 dpm/g dry wt). Bean roots bound 30 dpm/g fwt. Citrus and Ligustrum bound 207 and 240 dpm/g fwt, respectively. The time required to achieve equilibrium of leaves with the gas phase was 15 min for bean, 30 min for Citrus, and 30–60 min for Ligustrum. The time for 1/2 of the bound ethylene to diffuse out of the leaves was 20 min for bean, 10 min for Citrus, and 30 min for Ligustrum. The amount of ethylene needed to occupy 1/2 of the binding sites was obtained from Scatchard plots. This value (K_d) was 0.2 l/1 for bean, 0.15 for Citrus, and 0.31 for Ligustrum. The quantity of binding sites in the tissues was 2.0×10^-9 mol of binding sites/kg tissue for bean leaves, 5.7×10^-9 for Citrus leaves, and 6.8×10^-9 for Ligustrum. Pretreatment with indoleacetic acid (IAA), ehtylene, and cycloheximide (1 mg/1) had little effect on the level of ethylene-binding sites in Citrus.Contribution from the Department of Biochemistry, School of Agriculture and Life Sciences and School of Physical and Mathematical Sciences, North Carolina State University. Paper No. 8445 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601.North Carolina-Israel exchange Scholar for 1981 at the Department of Biochemistry, North Carolina State University Raleigh, North Carolina, USA     

Production of zearalenone,T-2 toxin,and deoxynivalenol by Fusarium spp. isolated from plant materials grown in North Carolina

Fusarium spp. isolated from plant materials grown in the hot, humid climate of North Carolina were tested for production of mycotoxins. Isolates of F. acuminatum, F. graminearum, F. moniliforme, F. oxysporum, and F. solani produced zearalenone while isolates of F. equiseti and F. graminearum produced T-2 toxin and deoxynivalenol, respectively. This is the first report of zearalenone production by F. solani. The toxins were identified by capillary gas chromatography-mass spectrometry. These findings suggest that there are toxigenic strains of Fusarium indigenous to the warmer regions of the USA and that fasariotoxicoses of animals in this region are not necessarily the result of importing toxic grains from the cooler, upper midwestern USA.Paper No. 8953 of the Journal Series of North Carolina Agricultural Research Service, Raleigh, North Carolina. The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Research Service of the product named nor criticism of similar ones not mentioned.     

RECENT VERSUS RELIC: DISCERNING THE GENETIC SIGNATURE OF FUCUS VESICULOSUS (HETEROKONTOPHYTA; PHAEOPHYCEAE) IN THE NORTHWESTERN ATLANTIC1

Fucus vesiculosus L. is one of the most widespread macrophytes in the northwestern Atlantic, ranging from North Carolina (USA) to Greenland (DK). We investigated genetic diversity, population differentiation, patterns of isolation by distance, and putative glacial refugial populations across seven locations from North Carolina (USA) to Cape Breton Island, Nova Scotia (Canada), with microsatellite analyses. Distinct northern versus southern (Delaware–North Carolina) populations were revealed by microsatellite data. Five of six microsatellite loci were fixed in populations in North Carolina, suggesting a recent founder event or a bottleneck, and the same homozygous genotype was found in herbarium materials collected on the North Carolina coast from more than 60 years ago. An additional set of individuals from the northern limit in Greenland was included in our analysis of mitochondrial intergenic spacer (mt IGS) haplotypes in the northwestern Atlantic. Remarkably, 184 of 188 F. vesiculosus specimens from North Carolina to Greenland shared the same haplotype. Recent colonization of the North American shore from Europe is hypothesized based upon the ubiquity of this common haplotype, which was earlier reported from Europe.     

Isozyme variability in species of the genus Drosophila. VIII. The alcohol dehydrogenase polymorphism in north carolina populations of D. melanogaster

Adult Drosophila melanogaster flies collected from populations broadly dispersed over ecological and geographic strata of North Carolina, and over a period of 4 years, were analyzed for alcohol dehydrogenase phenotypes by gel electrophoresis. Gene frequencies in spring-summer-fall field collections were remarkably stable over all strata. Two winter collections exhibited contrasting gene frequency changes. In one case the results are interpreted in terms of long-distance migration from Florida, while the other is explicable by assignment of a causal role to environmental factors which accompany the winter season.This investigation was supported in part by NIH Research Grant No. GM11546 from the National Institute of General Medical Sciences and by Contract No. AT-(40-1)-3980 from the United States Atomic Energy Commission.Paper No. 4719 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, North Carolina 27609.     

FIELD AND CULTURE STUDIES ON GROWTH AND REPRODUCTION OF CALLITHAMNION BYSSOIDES (RHODOPHYTA,CERAMIALES) IN NORTH CAROLINA1,2

The life history of Callithamnion byssoides Arnott ex Harv. in Hook. from North Carolina is shown to be of the Polysiphonia-type. Cross-gradient light-temperature culture in 8:16 and 14:10 LD cycles was utilized to investigate the effects of light intensity, temperature and day-length on growth and reproduction. Carposporophyte and tetrasporangium development were controlled by total light energy rather than photoperiod. Results of these studies explain the observed seasonal periodicity of growth and reproduction on the North Carolina coast. Variations between these results and previously published data for this species in Texas are explained in terms of ecotypic differentiation.     

Rhizobium meliloti distribution in the soil following alfalfa inoculation

Summary Alfalfa seeds, inoculated with an antibiotic-resistantRhizobium meliloti strain, were planted in three replicated field plots at Clayton, N.C. Core samples were taken three times in the next year at 0, 10, and 20 cm from the edge of each plot. Soil subsamples were taken from within each core sample at 0, 6, 12, and 18 cm depths. The numbers of the inoculum Rhizobium strain in each soil subsample were determined by inoculation of alfalfa plants with diluted soil samples. In general the distribution of rhizobia showed some movement outward and downward in the soil. Lower counts were obtained at the surface during summer. The Rhizobium persistence pattern in the soil differed in the three plots which is consistent with the variability in Rhizobium numbers often observed in established alfalfa stands. Cooperative investigation of the United States Department of Agriculture, Science and Education Administration, Agricultural Research and the North Carolina Agricultural Research Service, Raleigh, North Carolina. Paper No. 6818 of the Journal Series of the North Carolina Agricultural Research Service at Raleigh.     

Inheritance,intracellular localization,and genetic variation of phosphoglucomutase isozymes in maize (Zea mays L.)

Phosphoglucomutase (PGM; EC 2.7.5.1) isozyme variants were studied in a large number of inbred lines, crosses, and races of maize (Zea mays L.). Patterns of Mendelian inheritance demonstrated for PGM isozyme variants indicated that they are encoded by nuclear genes. Two unlinked loci, Pgm1 and Pgm2, located on the long arm of chromosome 1 and the short arm of chromosome 5, respectively, specify the observed electrophoretic variation on starch gels. No intra- or interlocus hybrid bands were found, suggesting that each isozyme band consists of a single polypeptide. PGM isozymes were present in all plant parts studied and the activity specified by both loci appears to reside in the cytoplasm. In studies of 520 racial collections of maize from Latin America, a single allele at each locus predominated in most collections. Likewise, the same alleles predominated in a set of 406 inbred lines of maize from the United States and Canada.This work was supported in part by NIH Research Grant GM 11546.Paper No. 8496 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina.     

Variation in thrips species composition in field crops and implications for tomato spotted wilt epidemiology in North Carolina

Thrips were surveyed in tomato spotted wilt-susceptible crops in five areas across North Carolina. Tomato, pepper, and tobacco plants in commercial fields were sampled and 30 species of thrips were collected over a 3-year period. The most common species overall was Frankliniella tritici (Fitch). The most common thrips species that are known to vector Tomato Spotted Wilt Virus (TSWV) were F. fusca (Hinds), and F. occidentalis (Pergande). Relatively low numbers of Thrips tabaci Lindeman, another reported vector, were collected. The spatial and temporal occurrence of vectors varied with sampling method, crop species, region of North Carolina, and localized areas within each region. In a laboratory experiment, no difference was detected between the ability of F. fusca and F. occidentalis to acquire and transmit a local isolate of TSWV. Based on vector efficiency and occurrence, F. fusca is considered the most important vector of TSWV in tobacco, whereas both F. fusca and F. occidentalis are important vectors of TSWV in tomato and pepper.     

Bariatric Surgeries in North Carolina, 1990 to 2001: A Gender Comparison

Objective: The purpose of this research was to determine the number of bariatric procedures in obese men and women in a well‐defined population and to examine gender differences among bariatric patients. Research Methods and Procedures: Data on bariatric patients were taken from the North Carolina Hospital Discharge Database, which contains information on all nonfederal hospital discharges in North Carolina from 1990 to 2001. Using North Carolina Hospital Discharge Data, Census North Carolina resident estimates, and North Carolina obesity prevalence estimates, we constructed annual rates for bariatric procedures for the obese male and female population in North Carolina. Results: Overall, 2197 bariatric procedures were performed between 1990 and 2001. The annual rate of bariatric procedures in obese women increased rapidly, particularly between 1998 and 2001, whereas the increase for men was considerably less than that for women. Controlling for age and year of procedure, the odds ratio for obese female North Carolina residents of having a bariatric procedure was 4.96 (95% confidence interval: 4.39, 5.59) and of having a Roux‐en‐Y procedure was 5.57 (95% confidence interval: 4.67, 6.64) compared with obese male North Carolina residents. Controlling for age, comorbidity burden, payment source, and year of procedure, obese male North Carolina residents had a significantly greater (22%) amount of inpatient days than obese female North Carolina residents. Discussion: After controlling for population rates of obesity and year of procedure, women are more likely than men to undergo bariatric surgery, suggesting that gender‐related factors may influence use. More research is needed to determine the causes for this large gender disparity.     

NORTH CAROLINA MARINE ALGAE. IX. ONSLOWIA ENDOPHYTICA GEN. ET SP. NOV. (PHAEOPHYTA,SPHACELARIALES) AND NOTES ON OTHER NEW RECORDS FOR NORTH CAROLINA1

Four benthic algae are reported here for the first time in the North Carolina flora. The new brown algal genus and species, Onslowia endophytica Searles, is described as an endophyte of Halymenia floridana from the North Carolina continental shelf. New records of Boodleopsis pusilla and Naccaria corymbosa from North Carolina constitute range extensions of these tropical species on the American coast north from Florida. Blastophysa rhizopus, an endophyte and epiphyte known from the North Atlantic coast of Europe and America as well as the Caribbean is reported from North Carolina for the first time and in a new host, Predaea feldmannii.     

RIBOSOMAL DNA LENGTH POLYMORPHISMS WITHIN POPULATIONS OF XYLARIA MAGNOLIAE (ASCOMYCOTINA)

Xylaria magnoliae forms abundant conidial and sexual fruiting structures on the fallen strobili of Magnolia spp. throughout the southeastern United States. Populations of X. magnoliae may be identified as isolates originating from a single strobilus, among different strobili from different host trees, and different host tree localities. We examined the stratified structure of X. magnoliae populations by analysis of their ribosomal DNA restriction patterns. A sample of 48 single-conidial isolates was obtained from strobili of both Magnolia grandiflora and M. fraseri in the piedmont and mountains of North Carolina and Georgia. Southern blotting experiments revealed considerable variation in rDNA lengths between and within different isolates. Although a single rDNA phenotype usually dominates within a particular collecting locality, different phenotypes were also observed within a locality and even between isolates obtained from a single strobilus. Some rDNA restriction phenotypes are distributed over localities separated by up to 400 km. These results suggest a fine-grained population structure for X. magnoliae, together with the potential for genetic exchange and dispersal between widely separated localities.     

Biosynthesis of superoxide dismutase and catalase inSaccharomyces cerevisiae: effects of oxygen and cytochromec deficiency

Summary Two strains ofSaccharomyces cerevisiae were used to study the synthesis of superoxide dismutase. One strain (cytochromec-deficient) contained 5–10% of the normal amounts of total cytochromec, while the other strain was a wild type. The cytochromec-deficient mutant had lower specific growth rate, growth yield, and oxygen uptake than the wild type. The superoxide dismutase and catalase activities, in both strains, were significantly lower under anaerobic than under aerobic conditions. Furthermore, under aerobic conditions the mutant contained higher levels of superoxide dismutase than the wild type which may be attributed to the higher intracellular flux of superoxide radicals caused by the cytochromec deficiency. The mutant also showed a lower level of catalase which was due to glucose repression.Paper Number 10007 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695, U.S.A. The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Research Service of the products named, nor criticism of similar ones not mentioned.     

Lack of specific binding of labeledRhizobium japonicum lipopolysaccharides to wild soybean (Glycine soja) roots

Summary Lipopolysaccharides (LPS) were extracted from two strains ofRhizobium japonicum (61A76NS and 3I1b110-I). The extracted LPS was purified by gel filtration column chromatography and the amount of 2-keto-3-deoxyoctonate (KDO) was determined. Column purified LPS from both strains were conjugated to rhodamine isothiocyanate on celite to examine binding of this purified, labeled surface component to aseptically grownGlycine soja (wild soybean) seedlings as a basis for symbiotic specificity using fluorescent microscopy. Rhodamine conjugated LPS from both strains ofRhizobium japonicum did not exhibit specific binding to wild soybean seedling roots.Paper no. 8130 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27650, USA.     

Variability in sequence of Strawberry vein banding virus

The variability of the Strawberry vein banding virus (SVBV) isolates was investigated. In total 267 strawberry plants from 6 European countries and North America were tested for the presence of SVBV. Only 4 plants were positive. Partial genomic sequences of the capsid protein gene of three North American SVBV isolates were determined. Only minor sequence variability (0.7 %) was observed during a comparison with existing nucleotide data of the European and the North American isolates (9 isolates). No variability at all could be found in the annealing regions of primers and probes used for molecular detection of SVBV for these isolates. However, a comparison to a sequence of a Chinese isolate published recently revealed a much higher DNA sequence difference (9.5 %) of this isolate.     

MORPHOLOGICAL AND GENETIC VARIABILITY IN PLANTAGO CORDATA (PLANTAGINACEAE), A THREATENED AQUATIC PLANT

Morphological and genetic variation were studied in Plantago cordata Lam., an imperiled aquatic plant. The existence of distinct seasonal morphs was confirmed by numerical morphological analyses of specimens from 53 populations. Plants from two North Carolina populations possessed leaves that were significantly shorter, narrower, and fewer-veined than populations in other portions of the species range. Genetic variation within and among ten populations in seven states/provinces (Illinois, Missouri, New York, North Carolina, Ohio, Ontario, and Wisconsin) was examined electrophoretically. The species is an apparent allopolyploid with fixed heterozygosity observed at 67% of the loci surveyed. Electrophoretic variation was mostly partitioned among populations. The genetic identity among populations was high except for North Carolina populations which differed at 27% of the loci surveyed.