Distribution of Purple Photosynthetic Bacteria in Wetland and Woodland Habitats of Central and Northern Minnesota

Enrichment cultures for purple nonsulfur and sulfur photosynthetic bacteria were prepared from soil samples collected in central and northern Minnesota. The purple nonsulfur bacteria were found in most wetland soils sampled but were uncommon in woodland and grassland soils. The pH range of the soils in which these bacteria occurred was 3.8 to 7.8, and the oxidation-reduction potential (E(h)) range was +510 to -65 mV. Soils with a pH below 5.0 or an E(h) above +370 mV had few purple nonsulfur bacteria (<10/g of soil). Rhodopseudomonas viridis, a photosynthetic bacterium containing bacteriochlorophyll b, and the purple sulfur bacteria were common only in low-acidity wetland soils that were usually being reduced.     

The chemistry of vitamin B12. The coordination of biologically important molecules

The following equilibrium constants (given as logK in units of m⁻¹) were determined for the substitution of co-ordinated H₂O in aquocobalamin by glycine (bound through N) 5.8, cysteine (bound through S) 6.0 or 8.3, depending on the value chosen for the pK of the thiol group, and phenolate 2.9. The spectrum of the phenolate cobalamin shows an additional intense absorption band at 468nm with a molar extinction coefficient of 1.1×10⁴, which is assigned to a charge transfer from the phenolate to the cobalt ion. Equilibrium constants have also been determined for the equilibria between adenylcobamide cyanide and CN⁻, HO⁻ and H⁺, which show that the adenine is more easily displaced by CN⁻ and HO⁻ than is 5,6-dimethylbenziminazole in vitamin B₁₂, but can be protonated by acid while still remaining co-ordinated to the cobalt. It is shown that in the binding of corrinoids to proteins and polypeptides the formation of hydrogen bonds is far more important than co-ordination by the metal.     

An electron microscope study of nebenkern formation and differentiation in spermatids of Murgantia histrionica (Hemiptera, Pentatomidae)

Mitochondria in early spermatids of many insects aggregate and form a round body, the nebenkern. The nebenkern undergoes a structural differentiation and then divides into two separate equal-sized bodies. In the present study, nebenkerns of Murgantia histrionica, a Hemipteran insect, were reconstructed using electron micrographs of serial sections to determine how the mitochondria transform into the two separate bodies. Newly formed nebenkerns are made of one piece, an anastomosis of rod-like segments. Some segments interconnect to join networks of rings. Each network interlocks with another similar network, but networks which interlock are connected with each other by other segments of the nebenkern. Later, the entire nebenkern is made of two unconnected and interlocked networks of rings. The nebenkern appears to remain bipartite during subsequent differentiation. Since the two pieces are interlocked, breaks must occur before the pieces can separate. As breaks occur, each network transforms into a set of curved sheets, producing a nebenkern made of four concentric layers. The three outer layers are each made of two curved sheets which surround a bipartite central core. The surface sheets meet at a furrow in the surface of the nebenkern; segments in each layer are roughly symmetrical with each other about the plane in which the furrow lies. Rod-like segments join alternate segments. The number of layers then decreases to three, and later, to two. These nebenkerns resemble four-layered nebenkerns, but fewer connections between alternate segments are present. The two pieces constituting the nebenkern probably separate after most of the latter connections disappear. Hypotheses to account for the observed changes in nebenkern structure are presented.     

Fusion protein-based epitope mapping of phytochrome. Precise identification of an evolutionarily conserved domain

Fusion proteins are used to define with precision an evolutionarily conserved domain on the carboxyl-terminal portion of the chromoprotein phytochrome. Simultaneously, assignments of two other epitopes are made with significantly greater precision, while the location of a fourth is confirmed. The epitope-mapping method that is described here is systematic, using complementary, overlapping nested sets of fusion proteins of predefined sequence rather than randomly generated peptides. Moreover, in contrast to previous methods, this approach yields rigorous and unambiguous assignments because it relies solely upon the ability of an antibody to detect a given polypeptide. A cDNA fragment encoding phytochrome amino acids 464-1129, which is its carboxyl terminus, was identified in lambda gt11 and subcloned in frame into the lacZ alpha sequence of pUC18. Four nested sets of subclones in pUC18 were created by digestion with selected restriction endonucleases and with the exonuclease Bal31. Fusion proteins were analyzed by immunoblotting following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The epitope for monoclonal antibody Oat-13 was confirmed to be between residues 551 and 617, while the epitopes for Oat-8 and Oat-28 were narrowed to 624-686 and 624-747, respectively. The epitope recognized by Pea-25, Pea-2, and Oat-15 was resolved unequivocally to a sequence of only seven residues (residues 765-771): N-Pro-Ile-Phe-Gly-Ala-Asp-Glu-C.     

Detecting non-neutral heterogeneity across a region of DNA sequence in the ratio of polymorphism to divergence

Natural selection, in the form of balancing selection or selective sweeps, can result in a decoupling of the amounts of molecular polymorphism and divergence. Thus natural selection can cause some areas of DNA sequence to have greater silent polymorphism, relative to divergence between species, than other areas. It would be useful to have a statistical test for heterogeneity in the polymorphism to divergence ratio across a region of DNA sequence, one that could identify heterogeneity greater than that expected from the neutral processes of mutation, drift, and recombination. The only currently available test requires that a region be arbitrarily divided into sections that are compared with each other, and the subjectivity of this division could be problematic. Here a test is proposed in which runs of polymorphic and fixed sites are counted, where a "run" is a set of one or more sites of one type preceded and followed by the other type. The number of runs is smaller than otherwise expected if polymorphisms are clumped together. By simulating neutral evolution and comparing the observed number of runs to the simulations, a statistical test is possible which does not require any a priori decisions about subdivision.