Genetic Regulation of Development in Sorghum bicolor (VIII. Shoot Growth,Tillering, Flowering,Gibberellin Biosynthesis,and Phytochrome Levels Are Differentially Affected by Dosage of the ma3R Allele

Sorghum [Sorghum bicolor (L.) Moench] homozygous for ma3R lacks a type II, light-stable phytochrome of 123 kD and has a number of phenotypic characteristics consistent with the absence of functional phytochrome B. We have used plants heterozygous at Ma3 (Ma3/ma3R and ma3/ma3R) to determine the effect of dosage of ma3R on plant growth, flowering, gibberellin (GA) levels, and content of the 123-kD phytochrome. Both Ma3/ma3R and ma3/ma3R produced the same number of tillers per plant as their respective homozygous non-ma3R parents. Height of the heterozygotes was intermediate between the homozygous parents, although it was more similar to the non-ma3R genotypes. In both field and growth-chamber environments, the timing of floral initiation and anthesis in the heterozygotes also was intermediate, again more similar to non-ma3R plants. In Ma3/ma3R, levels of GA53, GA19, GA20, and GA1 were almost exactly intermediate between levels detected in Ma3/Ma3 and ma3R/ma3R plants. Immunoblot analysis indicated that there was less of the 123-kD phytochrome in Ma3/ma3R than in homozygous Ma3, whereas none was detected in ma3R/ma3R. The degree of dominance of Ma3 and ma3 over ma3R varies with phenotypic trait, indicating that mechanisms of activity of the 123-kD phytochrome vary among the biochemical processes involved in each phenotypic character. Although the heterozygotes were similar to homozygous Ma3 and ma3 plants in growth and flowering behavior, Ma3/ma3R contained 50% less of the bioactive GA (GA1) than non-ma3R genotypes. Thus, sensitivity to endogenous GAs also may be regulated by the 123-kD phytochrome. To fully regulate plant growth and development, two copies of Ma3 or ma3 are required to produce sufficient quantities of the light-stable, 123-kD phytochrome.     

Probing protein-cofactor interactions in the terminal oxidases by second derivative spectroscopy: study of bacterial enzymes with cofactor substitutions and heme A model compounds.

Second derivative absorption spectra are reported for the aa3-cytochrome c oxidase from bovine cardiac mitochondria, the aa3-600 ubiquinol oxidase from Bacillus subtilis, the ba3-cytochrome c oxidase from Thermus thermophilis, and the aco-cytochrome c oxidase from Bacillus YN-2000. Together these enzymes provide a range of cofactor combinations that allow us to unequivocally identify the origin of the 450-nm absorption band of the terminal oxidases as the 6-coordinate low-spin heme, cytochrome a. The spectrum of the aco-cytochrome c oxidase further establishes that the split Soret band of cytochrome a, with features at 443 and 450 nm, is common to all forms of the enzyme containing ferrocytochrome a and does not depend on ligand occupancy at the other heme cofactor as previously suggested. To test the universality of this Soret band splitting for 6-coordinate low-spin heme A systems, we have reconstituted purified heme A with the apo forms of the heme binding proteins, hemopexin, histidine-proline-rich glycoprotein and the H64V/V68H double mutant of human myoglobin. All 3 proteins bound the heme A as a (bis)histidine complex, as judged by optical and resonance Raman spectroscopy. In the ferroheme A forms, none of these proteins displayed evidence of Soret band splitting. Heme A-(bis)imidazole in aqueous detergent solution likewise failed to display Soret band splitting. When the cyanide-inhibited mixed-valence form of the bovine enzyme was partially denatured by chemical or thermal means, the split Soret transition of cytochrome a collapsed into a single band at 443 nm.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of chelators on iron uptake and release by the brain in the rat

The iron chelators desferrioxamine (DFO), pyridoxal isonicotinoyl hydrazone (PIH), 2,2-bipyridine, diethylenetriamine penta-acetic acid (DTPA) and 1,2 dimethyl-3-hydroxy pyrid-4-one (CP20) were analysed for their ability to change⁵⁹Fe uptake and release from the brain of 15- and 63-day rats either during or after intravenous injection of⁵⁹Fe-¹²⁵I-transferrin. DTPA was the only chelator unable to significantly reduce iron uptake into the brain of 15-day rats. This indicates that iron is not released from transferrin at the luminal surface of brain capillary endothelial cells. CP20 was able to reduce iron uptake in the brain by 85% compared to 28% with DFO. Only CP20 was able to significantly reduce brain iron uptake in 63 day rats. Once⁵⁹Fe had entered the brain no chelator used was able to mediate its release. All of the chelators except CP20 had similar effects on femur iron uptake as they did on brain uptake, suggesting similar iron uptake mechanisms. It is concluded that during the passage of transferrin-bound iron into the brain the iron is released from transferrin within endothelial cells after endocytosis of transferrin.     

Effect of ovarian antral follicle cauterization on the interestrus interval of the gilt

The objective of the present study was to determine if destruction of ovarian antral follicles by laser-cauterization affects CL lifespan during the estrous cycle of the gilt. Cyclic gilts were randomly assigned to either SHAM, laser (L) or laser-estradiol (L-E2) treatment groups, with the L-E2 group receiving a 5-mg intramuscular (i.m.) injection of estradiol-17beta cypionate at the time of the first surgery. Ovarian antral follicles were laser-cauterized on either Days 12 and 14 (L12) or Days 14 and 17 (L14) of the estrous cycle. In the L12-E2 group, 3 of 4 gilts had extended mean interestrus intervals of more than 22 days compared with 0 of 4, 0 of 6, 0 of 7 and 1 of 5 gilts in the SHAM, L12, L14 and L14-E2 groups, respectively. The L12-E2 gilts had a longer (P<0.05) mean interestrus interval (23.5+/-1.3 days) than the L12 (20.0+/-1.1 days), L14 (20.7+/-1.0 days) and SHAM (20.5+/-1.3 days). The mean interestrus interval of L14-E2 gilts (21.8+/-1.2 days) did not differ from those of the L12-E2 group or the L12, L14 and SHAM group gilts. Six additional gilts were injected with 5 mg estradiol cypionate-17beta to serve as nonsurgical controls for E2 treatment. Gilts (3 of 3) given an E2 injection on Day 12 had extended mean interestrus interval (26.0+/-2.6 days), while 2 of 3 gilts injected with E2 on day 14 had extended mean interestrus intervals (27.7+/-2.1 days). These results indicate that in cyclic gilts destruction of ovarian follicles by laser-cauterization did not affect CL lifespan, and that luteolysis is not dependent on the presence of antral follicles.     

Description and SEM Observations of Meloidogyne sasseri n. sp. (Nematoda: Meloidogynidae), Parasitizing Beachgrasses

Meloidogyne sasseri n. sp. is described and illustrated from American beachgrass (Ammophila breviliffulata) originally collected from Henlopen State Park and Fenwick Island near the Maryland state line in Delaware, United States (6). Its relationship to M. graminis, M. spartinae, and M. californiensis is discussed. Primary distinctive characters of the female perineal pattern were a high to rounded arch with shoulders, widely spaced lateral lines interrupting transverse striations, a sunken vulva and anus, and coarse broken striae around the anal area. Second-stage juvenile body length was 554 μm (470-550), stylet length 14 μm (13-14.5), tail length 93 μm (83-115), tapering to a finely rounded terminus. Male stylet length 20 μm (19-21.5), spicule length 33 μm (30-36). Scanning electron microscope observations provided additional details of perineal patterns and face views of the female, male, and J2 head. Wheat, rice, oat, Ammophila sp., Panicum sp., bermudagrass, zoysiagrass and St. Augustinegrass were tested as hosts. Distribution of the species was the coasts of Delaware and Maryland. The common name "beachgrass root-knot" is proposed for M. sasseri n. sp.     

DISTRIBUTION AND SYSTEMATICS OF FRESHWATER CERAMIALES (RHODOPHYTA) IN NORTH AMERICA1

Twenty-five freshwater populations of Ceramiales were collected in North America, 24 of which were from the tropical rainforest region of Central America and the Caribbean. The streams tended to be moderate in mean current velocity (X?= 23.3 cm·S^?1) and maximum width (X?= 6.3 m) but high in temperature (X?= 23.1°C), pH (X?= 7.9), and specific conductance (X?= 320 μS·cm^?1). Three Bostrychia species were restricted to the Caribbean islands: B. moritziana (Sonder ex Kütz.) J. Ag. (syn. B. cornigera Mont. and B. radicans f. moliforme Post), with ecorticate indeterminate axes, monosiphonous ultimate branches, and cladohaptera; B. radicans (Mont.) Mont. (syn. B. leprieurii Mont and B. rivularis Harv.), with ecorticate and polysiphonous axes throughout and cladophaptera; and B. tenella (Lamour.) J. Ag., with corticate indeterminate axes, monosiphonous ultimate branches, and peripherohaptera. Ballia prieurii Kütz. was found in Belize and Costa Rica and was characterized by rebranched determinate laterals, opposite branching, and long apical cells (X?= 61 μm) and axial cells (X?= 43 μm). Caloglossa leprieurii (Mont.) J. Ag. was localized in Puerto Rico while. C. ogasawaerensis Okam. was collected only in Costa Rica. The two species were separated by site of branching (midrib vs. margin) and blade width (X?= 384 vs. 861 μm). Polysiphonia subtilissima Mont. from Florida and Jamaica had four pericentral cells, no cortication, rhizoids arising from pericentral cells, and branches initiated at trichoblast scars.     

The Effect of Deleterious Mutations on Neutral Molecular Variation

Selection against deleterious alleles maintained by mutation may cause a reduction in the amount of genetic variability at linked neutral sites. This is because a new neutral variant can only remain in a large population for a long period of time if it is maintained in gametes that are free of deleterious alleles, and hence are not destined for rapid elimination from the population by selection. Approximate formulas are derived for the reduction below classical neutral values resulting from such background selection against deleterious mutations, for the mean times to fixation and loss of new mutations, nucleotide site diversity, and number of segregating sites. These formulas apply to random-mating populations with no genetic recombination, and to populations reproducing exclusively asexually or by self-fertilization. For a given selection regime and mating system, the reduction is an exponential function of the total mutation rate to deleterious mutations for the section of the genome involved. Simulations show that the effect decreases rapidly with increasing recombination frequency or rate of outcrossing. The mean time to loss of new neutral mutations and the total number of segregating neutral sites are less sensitive to background selection than the other statistics, unless the population size is of the order of a hundred thousand or more. The stationary distribution of allele frequencies at the neutral sites is correspondingly skewed in favor of rare alleles, compared with the classical neutral result. Observed reductions in molecular variation in low recombination genomic regions of sufficiently large size, for instance in the centromere-proximal regions of Drosophila autosomes or in highly selfing plant populations, may be partly due to background selection against deleterious mutations.     

Base composition analysis of oligonucleotides containing apurinic sites.

Upon base composition analysis, oligonucleotides which are labeled at the 3'-terminus with fluorescein or biotin generate an additional, late eluting peak in the HPLC chromatogram. Investigation of this effect revealed the haptens acted as apurinic sites, and phosphodiesterase cleavage of the phosphate bond between the upstream nucleotide and apurinic site is inhibited. Extension of this work with a base-stable apurinic site inserted into all possible junctures of 5'-TGAC-3' tetramers showed this to be a general effect. As a consequence of this work, acid-catalyzed depurination resulting in apurinic sites can be monitored in oligonucleotide synthesis.     

Characterization and classification of virulent lactococcal bacteriophages isolated from a Cheddar cheese plant

C.N. CASEY, E. MORGAN, C. DALY AND G.F. FITZGERALD, 1993. Twenty-two bacteriophages, isolated from cheese-vat whey samples over a period of 4 years, were found to be active against one or more of four different strains of Lactococcus lactis subsp. cremoris used in a defined strain starter system in an Irish Cheddar cheese factory. All the phages were small isometric-headed with non-contractile tails, a baseplate and a collar; they had genome sizes of approximately 30 kb, and belonged to a single DNA hybridization group. All 22 phages could be classified into four distinct groups based on restriction analysis which overlapped perfectly with those based on host range. Each group of phage examined showed cross-reactive host ranges. None of the phage DNAs hybridized to the chromosomes of any of the seven cultures used in the factory during the four cheesemaking seasons, and neither could phages be induced from any of the strains by mitomycin-C or ultraviolet light treatment.