Post-cultivation Secondary Succession in a Venezuelan Lower Montane Rain Forest

Since tropical rain forests are widely threatened by conversion to agriculture, even within protected areas, an understanding of recovery processes is important for restoration of forest ecosystems and thus conservation of their biodiversity. Secondary succession following land clearance and crop cultivation was studied in a lower montane rain forest in a protected area of the Venezuelan Cordillera de la Costa Central. Forest recovery was studied using a chronosequence of eight 20 × 20 m plots which represented four forest types ca.10 year-old Secondary Forest, ca. 20 year-old Secondary Forest, ca. 35 year-old (uncultivated) secondary forest and mature forest. Species richness and structural complexity increased during succession, with the oldest secondary forest having a physiognomy comparable to the mature forest. Species diversity was lower in the secondary forests than the mature forest, and their floristic composition was distinct. Four phases are hypothesized to occur in the succession process, each with a distinctive species assemblage: initial colonisation by non-woody vegetation; establishment and canopy closure by short-lived small-seeded woody pioneer species; replacement by longer-lived secondary species; and gradual replacement by mature forest large-seeded climax species. Full recovery of the forests in the protected area is likely to take many years, although it may be assisted through conservation management measures.     

Modeling of drug delivery into tissues with a microneedle array using mixture theory

In this paper, we apply mixture theory to quantitatively predict the transient behavior of drug delivery by using a microneedle array inserted into tissue. In the framework of mixture theory, biological tissue is treated as a multi-phase fluid saturated porous medium, where the mathematical behavior of the tissue is characterized by the conservation equations of multi-phase models. Drug delivery by microneedle array imposes additional requirements on the simulation procedures, including drug absorption by the blood capillaries and tissue cells, as well as a moving interface along its flowing pathway. The contribution of this paper is to combine mixture theory with the moving mesh methods in modeling the transient behavior of drug delivery into tissue. Numerical simulations are provided to obtain drug concentration distributions into tissues and capillaries.     

Growth response of barley and tomato to nitrogen stress and its control by abscisic acid,water relations and photosynthesis

Barley (Hordeum vulgare L.) and tomato Lycopersicon esculentum Mill.) were grown hydroponically and examined 2, 5, and 10 d after being deprived of nitrogen (N) supply. Leaf elongation rate declined in both species in response to N stress before there was any reduction in rate of dryweight accumulation. Changes in water transport to the shoot could not explain reduced leaf elongation in tomato because leaf water content and water potential were unaffected by N stress at the time leaf elongation began to decline. Tomato maintained its shoot water status in N-stressed plants, despite reduced water absorption per gram root, because the decline in root hydraulic conductance with N stress was matched by a decline in stomatal conductance. In barley the decline in leaf elongation coincided with a small (8%) decline in water content per unit area of young leaves; this decline occurred because root hydraulic conductance was reduced more strongly by N stress than was stomatal conductance. Nitrogen stress caused a rapid decline in tissue NO ₃ ^- pools and in NO ₃ ^- flux to the xylem, particularly in tomato which had smaller tissue NO ₃ ^- reserves. Even in barley, tissue NO ₃ ^- reserves were too small and were mobilized too slowly (60% in 2 d) to support maximal growth for more than a few hours. Organic N mobilized from old leaves provided an additional N source to support continued growth of N-stressed plants. Abscisic acid (ABA) levels increased in leaves of both species within 2 d in response to N stress. Addition of ABA to roots caused an increase in volume of xylem exudate but had no effect upon NO ₃ ^- flux to the xylem. After leaf-elongation rate had been reduced by N stress, photosynthesis declined in both barley and tomato. This decline was associated with increased leaf ABA content, reduced stomatal conductance and a decrease in organic N content. We suggest that N stress reduces growth by several mechanisms operating on different time scales: (1) increased leaf ABA content causing reduced cell-wall extensibility and leaf elongation and (2) a more gradual decline in photosynthesis caused by ABA-induced stomatal closure and by a decrease in leaf organic N.Abbreviation and symbols ABA abscisic acid - c_i leaf internal CO₂ concentration - L_p root hydraulic conductance     

Plant rhizosphere influence on microbial C metabolism: the role of elevated CO2, N availability and root stoichiometry

Microbial decomposer C metabolism is considered a factor controlling soil C stability, a key regulator of global climate. The plant rhizosphere is now recognized as a crucial driver of soil C dynamics but specific mechanisms by which it can affect C processing are unclear. Climate change could affect microbial C metabolism via impacts on the plant rhizosphere. Using continuous ¹³C labelling under controlled conditions that allowed us to quantify SOM derived-C in all pools and fluxes, we evaluated the microbial metabolism of soil C in the rhizosphere of a C4 native grass exposed to elevated CO₂ and under variation in N concentrations in soil and in plant root C:N stoichiometry. Our results demonstrated that this plant can influence soil C metabolism and further, that elevated CO₂ conditions can alter this role by increasing microbial C efficiency as indicated by a reduction in soil-derived C respiration per unit of soil C-derived microbial biomass. Moreover, under elevated CO₂ increases in soil N, and notably, root tissue N concentration increased C efficiency, suggesting elevated CO₂ shifted the stoichiometric balance so N availability was a more critical factor regulating efficiency than under ambient conditions. The root C:N stoichiometry effect indicates that plant chemical traits such as root N concentration are able to influence the metabolism of soil C and that elevated CO₂ conditions can modulate this role. Increased efficiency in soil C use was associated with negative rhizosphere priming and we hypothesize that the widely observed phenomenon of rhizosphere priming may result, at least in part, from changes in the metabolic efficiency of microbial populations. Observed changes in the microbial community support that shifting microbial populations were a contributing factor to the observed metabolic responses. Our case study points at greater efficiency of the SOM-degrading populations in a high CO₂, high N world, potentially leading to greater C storage of microbially assimilated C in soil.     

Detection of methanotrophs with highly divergent pmoA genes from Arctic soils

Tundra soil samples from the Canadian Arctic community, Kuujjuaq, were analyzed for the presence of the soluble (sMMO) and particulate (pMMO) methane monooxygenase genes. Total genomic DNA extracted from these soils was used as template for PCR using sMMO- and pMMO-specific primers, mmoX1-mmoX2 and A189-A682, respectively. pMMO and sMMO genes were detected in the Kuujjuaq soil samples. Isolation of sMMO-possessing methanotrophic microorganisms from the three soils, as determined by the colony naphthalene oxidation assay, was carried out using direct plating (5 degrees C) and methane enrichment studies (5 degrees C and 25 degrees C). Direct plating did not yield sMMO-possessing methanotrophic bacteria, whereas methane enrichments yielded isolates possessing and expressing sMMO activity. Analysis of derived amino acid sequences of pmoA genes and partial 16S rRNA genes obtained by PCR, using DNA isolated directly from this environment and from isolates, revealed the presence of highly divergent PmoA/AmoA sequences and 16S rRNA sequences that cluster closely with but are distinct from the genes from the genera Methylosinus and Methylocystis.     

Omega-3 biotechnology: Thraustochytrids as a novel source of omega-3 oils

Thraustochytrids are large-celled marine heterokonts and classified as oleaginous microorganisms due to their production of docosahexaenoic (DHA) and eicosapentaenoic (EPA) ω-3-fatty acids. The applications of microbial DHA and EPA for human health are rapidly expanding, and a large number of clinical trials have been carried out to verify their efficacy. The development of refined isolation and identification techniques is important for the cultivation of thraustochytrids. With a high proportion of lipid biomass, thraustochytrids are also amenable to various production strategies which increase omega-3 oil output. Modifications to the existing lipid extraction methods and utilisation of sophisticated analytical instruments have increased extraction yields of DHA and EPA. Other metabolites such as enzymes, carotenoids and extracellular polysaccharides can also be obtained from these marine protists. Approaches such as the exploration for more diverse isolates having fast growth rates, metabolic engineering including gene cloning, and growing thraustochytrids on alternate low cost carbon source, will further enhance the biotechnological potential of thraustochytrids.     

The entire N-terminal half of TatC is involved in twin-arginine precursor binding

Translocation of twin-arginine precursor proteins across the cytoplasmic membrane of Escherichia coli requires the three membrane proteins TatA, TatB, and TatC. TatC and TatB were shown to be involved in precursor binding. We have analyzed in vitro a number of single alanine substitutions in tatC that were previously shown to compromise in vivo the function of the Tat translocase. All tatC mutants that were defective in precursor translocation into cytoplasmic membrane vesicles concomitantly interfered with precursor binding not only to TatC but also to TatB. Hence structural changes of TatC that affect precursor targeting simultaneously abolish engagement of the twin-arginine signal sequence with TatB and block the formation of a functional Tat translocase. Since these phenotypes were observed for tatC mutations spread over the first half of TatC, this entire part of the molecule must globally be involved in precursor binding.     

Recent Advances in Understanding Proteins Involved in Lipid Droplet Formation,Growth and Fusion

Lipid droplets （LDs） were once viewed as simple, inert lipid micelles. However, they are now known to be organelles with a rich proteome involved in a myriad of cellular processes. LDs are heterogeneous in nature with different sizes and compositions of phospholipids, neutral lipids and proteins. This review takes a focused look at the roles of proteins involved in the regulation of LD formation, expansion, and morphology. The related proteins are summarized such as the fat-specific protein （Fsp27）, fat storage-inducing trans- membrane （FIT） proteins, seipin and ADP-ribosylation factor 1-coat protein complex I （Arf-COPI）. Finally, we present important challenges in LD biology for a deeper understanding of this dynamic organelle to be achieved.     

Quantitative evaluation of siRNA delivery in vivo

Effective small interfering RNA (siRNA)-mediated therapeutics require the siRNA to be delivered into the cellular RNA-induced silencing complex (RISC). Quantitative information of this essential delivery step is currently inferred from the efficacy of gene silencing and siRNA uptake in the tissue. Here we report an approach to directly quantify siRNA in the RISC in rodents and monkey. This is achieved by specific immunoprecipitation of the RISC from tissue lysates and quantification of small RNAs in the immunoprecipitates by stem-loop PCR. The method, expected to be independent of delivery vehicle and target, is label-free, and the throughput is acceptable for preclinical animal studies. We characterized a lipid-formulated siRNA by integrating these approaches and obtained a quantitative perspective on siRNA tissue accumulation, RISC loading, and gene silencing. The described methodologies have utility for the study of silencing mechanism, the development of siRNA therapeutics, and clinical trial design.