Cell-to-cell transfer of bacterial outer membrane lipoproteins

Myxococcus xanthus cells can glide forward by retracting type IV pili. Tgl, an outer membrane lipoprotein, is necessary to assemble pili. Tgl mutants can be transiently "stimulated" if brought into end-to-end contact with tgl+ donor cells. By separating the stimulated recipient cells from donor cells, we found that Tgl protein was transferred from the donors to the rescued recipient cells. Mutants lacking CglB lipoprotein, which is part of a second gliding engine, could also be stimulated, and CglB protein was transferred from donor to recipient cells. The high transfer efficiency of Tgl and CglB proteins suggests that donor and recipient cells briefly fuse their outer membranes.     

Chimeras of two isoprenoid synthases catalyze all four coupling reactions in isoprenoid biosynthesis

The carbon skeletons of over 55,000 naturally occurring isoprenoid compounds are constructed from four fundamental coupling reactions: chain elongation, cyclopropanation, branching, and cyclobutanation. Enzymes that catalyze chain elongation and cyclopropanation are well studied, whereas those that catalyze branching and cyclobutanation are unknown. We have catalyzed the four reactions with chimeric proteins generated by replacing segments of a chain-elongation enzyme with corresponding sequences from a cyclopropanation enzyme. Stereochemical and mechanistic considerations suggest that the four coupling enzymes could have evolved from a common ancestor through relatively small changes in the catalytic site.     

Contributions of two types of calcium channels to synaptic transmission and plasticity

In Aplysia sensory and motor neurons in culture, the contributions of the major classes of calcium current can be selectively examined while transmitter release and its modulation are examined. A slowly inactivating, dihydropyridine-sensitive calcium current does not contribute either to normal synaptic transmission or to any of three different forms of plasticity: presynaptic inhibition, homosynaptic depression, and presynaptic facilitation. This current does contribute, however, to a fourth form of plasticity--modulation of transmitter release by tonic depolarization of the sensory neuron. By contrast, a second calcium current, which is rapidly inactivating and dihydropyridine-insensitive, contributes to release elicited by the transient depolarization of an action potential and to the other three forms of plasticity.     

Transition From Sagebrush Steppe to Annual Grass (Bromus tectorum): Influence on Belowground Carbon and Nitrogen

Vegetation changes associated with climate shifts and anthropogenic disturbance have major impacts on biogeochemical cycling. Much of the interior western United States currently is dominated by sagebrush (Artemisia tridentata Nutt.) ecosystems. At low to intermediate elevations, sagebrush ecosystems increasingly are influenced by cheatgrass (Bromus tectorum L.) invasion. Little currently is known about the distribution of belowground organic carbon (OC) on these changing landscapes, how annual grass invasion affects OC pools, or the role that nitrogen (N) plays in carbon (C) retention. As part of a Joint Fire Sciences-funded project called the Sagebrush Treatment Evaluation Project (SageSTEP), we quantified the depth distribution of soil OC and N at seven sites experiencing cheatgrass invasion. We sampled plots that retained sagebrush, but represented a continuum of cheatgrass invasion into the understory. Eighty-four soil cores were taken using a mechanically driven diamond-tipped core drill to a depth of 90 cm, or until bedrock or a restrictive layer was encountered. Samples were taken in 15-cm increments, and soil, rocks, and roots were analyzed for OC and total N. We determined that cheatgrass influences the vertical distribution of OC and N within the soil profile and might result in decreased soil OC content below 60  cm. We also found that OC and total N associated with coarse fragments accounted for at least 10% of belowground pools. This emphasizes the need for researchers to quantify nutrients in deep soil horizons and coarse fragments.     

Fertilizer-induced pathogenicity of mycorrhizal fungi to sweetgum seedlings

One isolate of Glomus clarus, two of G. etunicatus, and one of G. claroideum, obtained from plants growing on abandoned stripmine sites in Kentucky, and an isolate of G. fascicutatus known to stimulate growth of various woody plants, were evaluated for their influence on growth of sweetgum seedlings in a mixture of sand and stripmine soil. Soils were supplemented with various rates of a complete slow-release fertilizer. Throughout the growth period, G. fasciculatus, and most of the stripmine isolates, stimulated growth at low fertilizer rates. At higher fertilizer rates, including the level optimum for non-mycorrhizal plants, the stripmine isolates inhibited plant growth. After 14 weeks, plants inoculated with one of the four stripmine isolates overcame the early growth depression, and those inoculated with a second isolate appeared to be overcoming the growth depression. G. fasciculatus was not inhibitory at any fertilizer rate. Root colonization by all three isolates evaluated was inhibited by the highest fertilizer rate, but this effect was not related to growth inhibition of plants. The other two isolates colonized roots at an extremely low rate (< 1%). Sporulation of all the stripmine isolates, but not G. fasciculatus, was also inhibited by the highest fertilizer rate. The G. fasciculalus isolate used in this study may be atypical of mycorrhizal fungi occurring randomly in nature in its mutualistic or neutral effect on plants under a wide range of growth conditions.     

Effects of genotype,environment, and postharvest storage on the total ascorbate content of potato (Solanum tuberosum) tubers

The total ascorbate content of potato tubers from 33 Solanum tuberosum genotypes grown at three geographically diverse sites in Europe in each of two years was determined immediately postharvest and after approximately 4 months of storage at 4 degrees C. Statistically significant differences in total ascorbate concentration were observed between genotypes both at harvest and after storage. In all genotypes, the levels of ascorbate decreased during storage. These results are discussed in terms of their implications for diet and health as well as in terms of breeding for improved vitamin C content in potatoes.     

Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine

Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S)-labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S)-tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions.