Protein composition of the carboxysomes of Thiobacillus neapolitanus

For purifying carboxysomes of Thiobacillus neapolitanus an isolation procedure was developed which resulted in carboxysomes free from whole cells, protoplasts and cell fragments. These purified carboxysomes are composed of 8 proteins and at the most of 13 polypeptides. The two most abundant proteins which make up more than 60% of the carboxysomes, are ribulose-1,5-bisphosphate carboxylase and a glycoprotein with a molecular weight of 54,000. The shell of the carboxysomes consists of four glycoproteins, one also with a molecular weight of 54,000. The other proteins are present in minor quantities. Ribulose-1,5-bisphosphate carboxylase is the only enzyme which could be detected in the carboxysomes and 3-phosphoglycerate was the only product formed during incubation with ribulose-1,5-diphosphate and bicarbonate. The supernatant of a broken and centrifuged carboxysome suspension contained the large subunit of ribulose-1,5-bisphosphate carboxylase. The small subunit of ribulose-1,5-bisphosphate carboxylase was found in the pellet together with the shell proteins which indicates that the small subunit of ribulose-1,5-bisphosphate carboxylase is connected to the shell.Abbreviations RuBisCO ribulose-1,5-bisphosphate carboxylase - PMSF phenylmethylsulfonyl fluoride - PAA gelectrophoresis, polyacrylamide gelelectrophoresis - SDS sodium dodecyl sulphate - CIE crossed immunoelectrophoresis - IEF isoelectric focusing     

Grass-Shrub Associations over a Precipitation Gradient and Their Implications for Restoration in the Great Basin,USA

As environmental stress increases positive (facilitative) plant interactions often predominate. Plant-plant associations (or lack thereof) can indicate whether certain plant species favor particular types of microsites (e.g., shrub canopies or plant-free interspaces) and can provide valuable insights into whether “nurse plants” will contribute to seeding or planting success during ecological restoration. It can be difficult, however, to anticipate how relationships between nurse plants and plants used for restoration may change over large-ranging, regional stress gradients. We investigated associations between the shrub, Wyoming big sagebrush (Artemisia tridentata ssp. wyomingensis), and three common native grasses (Poa secunda, Elymus elymoides, and Pseudoroegneria spicata), representing short-, medium-, and deep-rooted growth forms, respectively, across an annual rainfall gradient (220–350 mm) in the Great Basin, USA. We hypothesized that positive shrub-grass relationships would become more frequent at lower rainfall levels, as indicated by greater cover of grasses in shrub canopies than vegetation-free interspaces. We sampled aerial cover, density, height, basal width, grazing status, and reproductive status of perennial grasses in canopies and interspaces of 25–33 sagebrush individuals at 32 sites along a rainfall gradient. We found that aerial cover of the shallow rooted grass, P. secunda, was higher in sagebrush canopy than interspace microsites at lower levels of rainfall. Cover and density of the medium-rooted grass, E. elymoides were higher in sagebrush canopies than interspaces at all but the highest rainfall levels. Neither annual rainfall nor sagebrush canopy microsite significantly affected P. spicata cover. E. elymoides and P. spicata plants were taller, narrower, and less likely to be grazed in shrub canopy microsites than interspaces. Our results suggest that exploring sagebrush canopy microsites for restoration of native perennial grasses might improve plant establishment, growth, or survival (or some combination thereof), particularly in drier areas. We suggest that land managers consider the nurse plant approach as a way to increase perennial grass abundance in the Great Basin. Controlled experimentation will provide further insights into the life stage-specific effectiveness and practicality of a nurse plant approach for ecological restoration in this region.     

Assessing changes to ecosystem structure and function following invasion by Spartina alterniflora and Phragmites australis: a meta-analysis

Biological invasions resulting from anthropogenic activities are one of the greatest threats to maintaining ecosystem functioning and native biodiversity. Invasions are especially problematic when the invading species behaves as an ecosystem engineer that is capable of transforming ecosystem structure, function, and community dynamics. Of particular concern is the spread of emergent wetland grasses whose root systems alter hydrology and structural stability of soils, modify ecosystem functions, and change community dynamics and species richness. To address the threats posed to ecosystems across the globe, management practices focus on the control and removal of invasive grasses. However, it remains unclear how severely invasive grasses alter ecosystem functions and whether alterations persist after invasive grass removal, limiting our ability to determine if management practices are truly sufficient to fully restore ecosystems. Here, we conducted a meta-analysis to quantify ecological alterations and the efficacy of management following the invasion of Spartina alterniflora and Phragmites australis, two common and pervasive invaders in coastal wetlands. Our results indicate that S. alterniflora and P. australis significantly alter measures of ecosystem functioning and organismal abundance. Invaded ecosystems had significant elevations in abiotic carbon and nitrogen fixation and uptake in areas with invasive grasses, with differential photosynthetic pathways of these two grass species further explaining carbon fluxes. Moreover, evidence from our analyses indicates that management practices may not adequately promote recovery from invasion, but more data are needed to fully assess management efficacy. We call for future studies to conduct pairwise comparisons between uninvaded, invaded, and managed systems and provide research priorities.

     

Energetic aspects of CO2 uptake in Thiobacillus neapolitanus

Uptake of inorganic carbon (C_i) in the form of CO₂ and/or HCO ₃ ^- was studied in the chemolithoautotroph Thiobacillus neapolitanus under energy (thiosulphate) or carbon (CO₂) limitation. Uptake of C₁ was found to be a metabolic energy dependent process since in the presence of uncouplers no uptake was observed. The accumulation level of C_i was higher in the CO₂-limited cells (1000-to 1500-fold) in comparison to the thiosulphate-limited cells (500-to 800-fold). The process of uptake could be influenced by addition of ionophores. Inhibition of uptake and accumulation of C_i was found after addition of valinomycin which completely dissipated the electrical potential (). After addition of nigericin an increase in the uptake and accumulation of C_i was observed with a concomitant increase of the . These results suggest that the is the main driving force for uptake of C_i. However, uptake of C_i could never be found in the absence of electron transfer, or in cells in which the electron transfer chain was blocked by potassium cyanide. Electron transfer therefore appears to be an additional requirement for C_i uptake. Kinetic experiment on the uptake of inorganic carbon at different pH values suggest that CO₂ is the carbon species taken up by T. neapolitanus.Abbreviations RuBisCO ribulose-1,5-bisphosphate carboxylase - DCCD N,N¹-dicyclohexylcarbodiimide - CCCP carbonyl cyanide m-chlorophenyl hydrazone - FCCP carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone - EDTA sodium ethylene diamine tetraacetate     

Electron microscopic studies of carboxysomes of Thiobacillus neapolitanus

The outer part of the carboxysomes of Thiobacillus neapolitanus was examined by electron microscopy using negatively stained, cryo-treated, frozen hydrated and freeze dried specimens. From stereo-micrographs of freeze dried and fixated carboxysomes the three dimensional structure of the carboxysomes was elucidated. The carboxysomes always appear as hexagonal bodies, which possess twelve pentameric planes. This indicates that carboxysomes have the form of a pentagonal dodecahedron. Inside the carboxysomes the ribulose-1,5-bisphosphate carboxylase molecules are arranged in rows and concentric rings. Negatively stained and cryo-treated carboxysomes do not differ significantly in size. The mean size of these carboxysomes is 117.3±6.9 nm (n=782)