Human-Imposed,Fine-Grained Patch Burning Explains the Population Stability of a Fire-Sensitive Conifer in a Frequently Burnt Northern Australia Savanna

Woody plant demographics provide important insight into ecosystem state-shifts in response to changing fire regimes. In Australian tropical savannas, the switch from patchy landscape burning by Aborigines to unmanaged wildfires within the past century has been implicated in biodiversity declines including the fire-sensitive conifer, Callitris intratropica. C. intratropica commonly forms small, closed-canopy groves that exclude fire and allow recruitment of conspecifics and other fire-sensitive woody plants. C. intratropica groves provide a useful indicator of heterogeneity and fire regime change, but the mechanisms driving the species’ persistence and decline remain poorly understood. We examined the hypothesis that C. intratropica population stability depends upon a regime of frequent, low-intensity fires maintained by Aboriginal management. We combined integral projection models of C. intratropica population behaviour with an environmental state change matrix to examine how vital rates, grove dynamics and the frequency of high- and low-intensity fires contribute to population stability. Closed-canopy C. intratropica groves contributed disproportionately to population growth by promoting recruitment, whereas singleton trees accounted for a larger proportion of adult mortality. Our patch-based population model predicted population declines under current fire frequencies and that the recruitment of new groves plays a critical role in the species’ persistence. Our results also indicated that reducing fire intensity, a key outcome of Aboriginal burning, leads to C. intratropica population persistence even at high fire frequencies. These findings provide insight into the relationship between ecosystem composition and human–fire interactions and the role of fire management in sustaining the mosaics that comprise ‘natural’ systems.     

Global ammonia emissions from synthetic nitrogen fertilizer applications in agricultural systems: Empirical and process‐based estimates and uncertainty

Excessive ammonia (NH₃) emitted from nitrogen (N) fertilizer applications in global croplands plays an important role in atmospheric aerosol production, resulting in visibility reduction and regional haze. However, large uncertainty exists in the estimates of NH₃ emissions from global and regional croplands, which utilize different data and methods. In this study, we have coupled a process‐based Dynamic Land Ecosystem Model (DLEM) with the bidirectional NH₃ exchange module in the Community Multiscale Air‐Quality (CMAQ) model (DLEM‐Bi‐NH₃) to quantify NH₃ emissions at the global and regional scale, and crop‐specific NH₃ emissions globally at a spatial resolution of 0.5° × 0.5° during 1961–2010. Results indicate that global NH₃ emissions from N fertilizer use have increased from 1.9 ± 0.03 to 16.7 ± 0.5 Tg N/year between 1961 and 2010. The annual increase of NH₃ emissions shows large spatial variations across the global land surface. Southern Asia, including China and India, has accounted for more than 50% of total global NH₃ emissions since the 1980s, followed by North America and Europe. Rice cultivation has been the largest contributor to total global NH₃ emissions since the 1990s, followed by corn and wheat. In addition, results show that empirical methods without considering environmental factors (constant emission factor in the IPCC Tier 1 guideline) could underestimate NH₃ emissions in context of climate change, with the highest difference (i.e., 6.9 Tg N/year) occurring in 2010. This study provides a robust estimate on global and regional NH₃ emissions over the past 50 years, which offers a reference for assessing air quality consequences of future nitrogen enrichment as well as nitrogen use efficiency improvement.     

Spontaneous mutations in a regulatory gene induce phenotypic heterogeneity and adaptation of Ralstonia solanacearum to changing environments

An evolution experiment with the bacterial plant pathogen Ralstonia solanacearum revealed that several adaptive mutations conferring enhanced fitness in plants arose in the efpR gene encoding a regulator of virulence and metabolic functions. In this study, we found that an efpR mutant systematically displays colonies with two morphotypes: the type S (‘smooth’, similar to the wild type) and the type EV (‘efpR variant’). We demonstrated that the efpH gene, a homologue of efpR, plays a key role in the control of phenotypic heterogeneity, the ΔefpR-ΔefpH double mutant being stably locked into the EV type. Using mixed infection assays, we demonstrated that the type EV is metabolically more proficient than the type S and displays fitness gain in specific environments, whereas the type S has a better fitness into the plant environment. We provide evidence that this efpR-dependent phenotypic heterogeneity is a general feature of strains of the R. solanacearum species complex and could occur in natural conditions. This study highlights the potential role of phenotypic heterogeneity in this plant pathogen as an adaptive trait to changing environments.     

Immunodominant Francisella tularensis antigens identified using proteome microarray

Stimulation of protective immune responses against intracellular pathogens is difficult to achieve using non-replicating vaccines. BALB/c mice immunized by intramuscular injection with killed Francisella tularensis (live vaccine strain) adjuvanted with preformed immune stimulating complexes admixed with CpG, were protected when systemically challenged with a highly virulent strain of F. tularensis (Schu S4). Serum from immunized mice was used to probe a whole proteome microarray in order to identify immunodominant antigens. Eleven out of the top 12 immunodominant antigens have been previously described as immunoreactive in F. tularensis. However, 31 previously unreported immunoreactive antigens were revealed using this approach. Twenty four (50%) of the ORFs on the immunodominant hit list belonged to the category of surface or membrane associated proteins compared to only 22% of the entire proteome. There were eight hypothetical protein hits and eight hits from proteins associated with different aspects of metabolism. The chip also allowed us to readily determine the IgG subclass bias, towards individual or multiple antigens, in protected and unprotected animals. These data give insight into the protective immune response and have potentially important implications for the rational design of non-living vaccines for tularemia and other intracellular pathogens.     

Cloning and over-expression of an alkaline protease from Bacillus licheniformis

The alkaline protease gene, apr, from Bacillus licheniformis 2709 was cloned into a Bacillus shuttle expression vector, pHL, to yield the recombinant plasmid pHL-apr. The pHL-apr was expressed in Bacillus subtilis WB600, yielding a high expression strain BW-016. The amount of alkaline protease produced in the recombinant increased by 65% relative to the original strain. SDS-PAGE analysis indicated a Mr of 30.5 kDa. The amino acid sequence deduced from the DNA sequence analysis revealed a 98% identity to that of Bacillus licheniformis 6816.     

Activities of MAP-kinase pathways in normal uroepithelial cells and urothelial carcinoma cell lines

It is often assumed that MAPK pathways drive proliferation of normal uroepithelial (UEC) and urothelial carcinoma (TCC) cells. To check this assumption, activities and inducibilities of promoters containing serum-response elements (SRE) or AP-1 binding sites were investigated in cultured UEC and seven TCC lines. Reporter plasmids dependent on SRE or AP-1 sites were highly active in UEC, but significantly less so in TCC lines. Reporter activity in TCC lines could be induced by constitutively active MEKK4 or TPA. Accordingly, phosphorylation of the MAPK pathway components MEK, ERK, and ELK1 was most pronounced in UEC and lower in TCC lines. MAPK-dependent promoter activities and bromodeoxyuridine incorporation decreased in UEC upon withdrawal of growth factors, but less so in TCC lines, in which serum diminution increased apoptosis. Likewise, E2F-dependent promoters responded to growth factors in UEC, but were more serum-independent in the TCC lines, which lack either RB1 or p16(INK4A). MEK inhibitors inhibited BrdU incorporation in UEC more strongly than in TCC lines. Thus, proliferation of normal uroepithelial cells is indeed associated with activation of MAPK pathways. However, autonomous proliferation of TCC lines--unexpectedly--appears much less dependent on MAPK activation and may rather be promoted by defects in cell cycle regulation.