Predicting leaf gravimetric water content from foliar reflectance across a range of plant species using continuous wavelet analysis

Leaf water content is an important variable for understanding plant physiological properties. This study evaluates a spectral analysis approach, continuous wavelet analysis (CWA), for the spectroscopic estimation of leaf gravimetric water content (GWC, %) and determines robust spectral indicators of GWC across a wide range of plant species from different ecosystems. CWA is both applied to the Leaf Optical Properties Experiment (LOPEX) data set and a synthetic data set consisting of leaf reflectance spectra simulated using the leaf optical properties spectra (PROSPECT) model. The results for the two data sets, including wavelet feature selection and GWC prediction derived using those features, are compared to the results obtained from a previous study for leaf samples collected in the Republic of Panamá (PANAMA), to assess the predictive capabilities and robustness of CWA across species. Furthermore, predictive models of GWC using wavelet features derived from PROSPECT simulations are examined to assess their applicability to measured data. The two measured data sets (LOPEX and PANAMA) reveal five common wavelet feature regions that correlate well with leaf GWC. All three data sets display common wavelet features in three wavelength regions that span 1732-1736 nm at scale 4, 1874-1878 nm at scale 6, and 1338-1341 nm at scale 7 and produce accurate estimates of leaf GWC. This confirms the applicability of the wavelet-based methodology for estimating leaf GWC for leaves representative of various ecosystems. The PROSPECT-derived predictive models perform well on the LOPEX data set but are less successful on the PANAMA data set. The selection of high-scale and low-scale features emphasizes significant changes in both overall amplitude over broad spectral regions and local spectral shape over narrower regions in response to changes in leaf GWC. The wavelet-based spectral analysis tool adds a new dimension to the modeling of plant physiological properties with spectroscopy data.     

Cloning and initial characterization of a new subunit for mammalian serine-palmitoyltransferase

Serine-palmitoyltransferase (SPT) catalyzes the rate-limiting step of the de novo synthesis of sphingolipids. SPT is considered to be a heterodimer composed of two subunits, SPTLC1 and SPTLC2. Here we report the identification of a novel, third, SPT subunit (SPTLC3) that shows 68% homology to the SPTLC2 subunit. Quantitative real-time PCR revealed that SPTLC3 expression is highly variable between different human tissues and cell lines. The highest expression was observed in placenta tissue and human trophoblast cell lines. The overexpression of SPTLC3 in Hek293 cells, which otherwise have very little endogenous SPTLC3, led to a 2- to 3-fold increase in cellular SPT activity. Silencing of SPTLC3 expression in HepG2 cells or human trophoblast cells by transfecting SPTLC3-specific siRNA resulted in a significant reduction of cellular SPT activity. The expression of two SPT isoforms could be a cellular mechanism to adjust SPT activity to tissue-specific requirements of sphingolipid synthesis.     

Function and evolution of plasmid-borne genes for pyrimidine biosynthesis in Borrelia spp

The thyX gene for thymidylate synthase of the Lyme borreliosis (LB) agent Borrelia burgdorferi is located in a 54-kb linear plasmid. In the present study, we identified an orthologous thymidylate synthase gene in the relapsing fever (RF) agent Borrelia hermsii, located it in a 180-kb linear plasmid, and demonstrated its expression. The functions of the B. hermsii and B. burgdorferi thyX gene products were evaluated both in vivo, by complementation of a thymidylate synthase-deficient Escherichia coli mutant, and in vitro, by testing their activities after purification. The B. hermsii thyX gene complemented the thyA mutation in E. coli, and purified B. hermsii ThyX protein catalyzed the conversion of dTMP from dUMP. In contrast, the B. burgdorferi ThyX protein had only weakly detectable activity in vitro, and the B. burgdorferi thyX gene did not provide complementation in vivo. The lack of activity of B. burgdorferi's ThyX protein was associated with the substitution of a cysteine for a highly conserved arginine at position 91. The B. hermsii thyX locus was further distinguished by the downstream presence in the plasmid of orthologues of nrdI, nrdE, and nrdF, which encode the subunits of ribonucleoside diphosphate reductase and which are not present in the LB agents B. burgdorferi and Borrelia garinii. Phylogenetic analysis suggested that the nrdIEF cluster of B. hermsii was acquired by horizontal gene transfer. These findings indicate that Borrelia spp. causing RF have a greater capability for de novo pyrimidine synthesis than those causing LB, thus providing a basis for some of the biological differences between the two groups of pathogens.     

Revisiting the localisation of Zn cations sorbed on pathological apatite calcifications made through X-ray absorption spectroscopy

The role of oligo-elements such as Zn in the genesis of pathological calcifications is widely debated in the literature. An essential element of discussion is given by their localisation either at the surface or within the Ca apatite crystalline network. To determine the localisation, X-ray absorption experiments have been performed at SOLEIL. The Exafs results suggest that Zn atoms, present in the Zn²⁺ form, are bound to about 4 O atoms at a distance of 2.00 Å, while the interatomic distance R_CaO ranges between 2.35 Å and 2.71 Å. Taking into account the content of Zn (around 1000 ppm) and the difference in ionic radius between Zn²⁺ (0.074 nm) and Ca²⁺ (0.099 nm), a significant longer interatomic distance would be expected in the case of Zn replacing Ca within the apatite crystalline network. We thus conclude that Zn atoms are localised at the surface and not in the apatite nanocrystal structure. Such structural result has essential biological implications for at least two reasons. Some oligoelements have a marked effect on the transformation of chemical phases, and may modify the morphology of crystals. These are both major issues because, in the case of kidney stones, the medical treatment depends strongly on the precise chemical phase and on the morphology of the biological entities at both macroscopic and mesoscopic scales.     

Biofilm Formation and Cell Surface Properties among Pathogenic and Nonpathogenic Strains of the Bacillus cereus Group

Biofilm formation by 102 Bacillus cereus and B. thuringiensis strains was determined. Strains isolated from soil or involved in digestive tract infections were efficient biofilm formers, whereas strains isolated from other diseases were poor biofilm formers. Cell surface hydrophobicity, the presence of an S layer, and adhesion to epithelial cells were also examined.The Bacillus cereus group includes B. cereus sensu stricto, B. anthracis, and B. thuringiensis, three genetically close pathogenic species. Based on genetic evidence, it has been suggested that they could represent one species (7). B. cereus sensu stricto is itself an opportunistic human pathogen occasionally found to cause various diseases such as endophthalmitis or periodontitis but is more frequently involved in gastrointestinal diseases with diarrheal or emetic syndromes (4, 12). Emetic syndromes result from the presence of cereulide, a heat-stable toxin produced in food before ingestion, whereas diarrheal syndromes require survival of the bacterium in the host digestive tract. B. thuringiensis is an insect pathogen, and B. anthracis causes anthrax, a lethal human disease.The persistent contamination of industrial food processing systems by B. cereus (12) may facilitate its involvement in gastroenteritis. This persistence is due to spores, which may survive pasteurization, heating, and gamma-ray irradiation (9, 13), and to biofilms, which have been shown to be highly resistant to cleaning procedures (18). Biofilms are also suspected to be involved in bacterial pathogenicity, as they may form on host epithelia (15).In this study, we wanted to test whether biofilm formation by species of the B. cereus group could be connected to the pathogenicity of the bacterium. For this purpose, we screened a collection of 102 pathogenic (diarrheal, emetic, and oral diseases) and nonpathogenic strains of B. cereus and B. thuringiensis for their capability to form biofilms. As adhesion to inert or living surfaces is a prerequisite for biofilm formation, we have investigated relationships within our collection of strains between biofilm formation and cell surface hydrophobicity, the presence of an S-layer, or adhesion to epithelial cells.     

Changes in the Treatment Responses to Artesunate-Mefloquine on the Northwestern Border of Thailand during 13 Years of Continuous Deployment

Background

Artemisinin combination treatments (ACT) are recommended as first line treatment for falciparum malaria throughout the malaria affected world. We reviewed the efficacy of a 3-day regimen of mefloquine and artesunate regimen (MAS₃), over a 13 year period of continuous deployment as first-line treatment in camps for displaced persons and in clinics for migrant population along the Thai-Myanmar border.

Methods and Findings

3,264 patients were enrolled in prospective treatment trials between 1995 and 2007 and treated with MAS₃. The proportion of patients with parasitaemia persisting on day-2 increased significantly from 4.5% before 2001 to 21.9% since 2002 (p<0.001). Delayed parasite clearance was associated with increased risk of developing gametocytaemia (AOR = 2.29; 95% CI, 2.00–2.69, p = 0.002). Gametocytaemia on admission and carriage also increased over the years (p = 0.001, test for trend, for both). MAS₃ efficacy has declined slightly but significantly (Hazards ratio 1.13; 95% CI, 1.07–1.19, p<0.001), although efficacy in 2007 remained well within acceptable limits: 96.5% (95% CI, 91.0–98.7). The in vitro susceptibility of P. falciparum to artesunate increased significantly until 2002, but thereafter declined to levels close to those of 13 years ago (geometric mean in 2007: 4.2 nM/l; 95% CI, 3.2–5.5). The proportion of infections caused by parasites with increased pfmdr1 copy number rose from 30% (12/40) in 1996 to 53% (24/45) in 2006 (p = 0.012, test for trend).

Conclusion

Artesunate-mefloquine remains a highly efficacious antimalarial treatment in this area despite 13 years of widespread intense deployment, but there is evidence of a modest increase in resistance. Of particular concern is the slowing of parasitological response to artesunate and the associated increase in gametocyte carriage.     

Calcium Is a Major Determinant of Xylem Vulnerability to Cavitation

Xylem vulnerability to cavitation is a key parameter in the drought tolerance of trees, but little is known about the control mechanisms involved. Cavitation is thought to occur when an air bubble penetrates through a pit wall, and would hence be influenced by the wall''s porosity. We first tested the role of wall-bound calcium in vulnerability to cavitation in Fagus sylvatica. Stems perfused with solutions of oxalic acid, EGTA, or sodium phosphate (NaPO₄) were found to be more vulnerable to cavitation. The NaPO₄-induced increase in vulnerability to cavitation was linked to calcium removal from the wall. In contrast, xylem hydraulic conductance was unaffected by the chemical treatments, demonstrating that the mechanisms controlling vulnerability to cavitation and hydraulic resistance are uncoupled. The NaPO₄ solution was then perfused into stems from 13 tree species possessing highly contrasted vulnerability to cavitation. Calcium was found to be a major determinant of between-species differences in vulnerability to cavitation. This was evidenced in angiosperms as well as conifer species, thus supporting the hypothesis of a common mechanism in drought-induced cavitation.In plants, long-distance sap transport occurs under negative pressures in xylem conduits. Sap flows between adjoining conduits through pits that form pores in the walls, and that facilitate the flow of water while preventing the passage of air bubbles. Under water stress conditions, xylem tensions increase and the conduits become vulnerable to cavitation. Cavitation provokes an air embolism that leads to a loss of hydraulic conductance, thus exacerbating plant water deficit.Species resistance to cavitation has been intensively studied over the past two decades, and is now ranked among the traits with the highest functional and ecological significance. In woody species for instance, xylem vulnerability to cavitation correlates tightly with species-specific drought tolerance (Pockman and Sperry, 2000; Tyree et al., 2003; Maherali et al., 2004), with more xerophilous species proving less vulnerable to cavitation. Substantial variations have also been found between genotypes of a different species (Cochard et al., 2007; Dalla-Salda et al., 2009). This implies that this trait could potentially be used in breeding programs to identify more drought-tolerant species or genotypes. However, efforts in this direction are still strongly impeded by a lack of understanding of the molecular and genetic basis of cavitation resistance. Our work represents a first significant step toward resolving this challenging issue.Understanding the fine mechanism of cavitation formation is a pivotal step toward identifying the key structures and the key genes coding for these structures, yet we currently have only partial insights. According to a hypothesis first formulated by Zimmermann (1983), water stress-induced cavitation is thought to occur when a tiny air bubble penetrates through a pit membrane, and would consequently be strongly influenced by the porosity of the membrane (Tyree and Sperry, 1988; Cochard, 2006). There is also experimental evidence for a role of the mechanical properties of the pit membrane in this cavitation process (Choat et al., 2004; Sperry and Hacke, 2004). Clearly, the structural, physical, and chemical properties of pit membranes are central to the determinism of cavitation.Pit membranes are modified primary cell walls made of tightly interwoven cellulose microfibrils in a matrix of hydrated hemicelluloses and pectins. Pectins consist of a complex set of GalUA (GalA)-rich polysaccharides, and four pectic domains can be distinguished: homogalacturonan (HG), rhamnogalacturonan I, rhamnogalacturonan II, and xylogalacturonan (Willats et al., 2001). The high degree of structural complexity and heterogeneity across the pectin family is the result of both biosynthesis in the endomembrane system and the action of an array of wall-based pectin-modifying enzymes (Willats et al., 2001). HG units are synthesized in the Golgi apparatus and deposited in the cell wall in a form containing 70% to 80% methyl-esterified GalA residues (O''Neill et al., 1990; Mohnen, 1999). The removal of methyl ester groups by pectin methyl esterase (PME) within the cell wall matrix produces free carboxyl groups capable of being cross-linked by calcium cations in an “egg-box” structure (Grant et al., 1973; Pelloux et al., 2007). These calcium-dependent cross-linkages are dependent both on the degree and the distribution of methyl-esterified GalA units through the HG network (Willats et al., 2001). Calcium therefore plays a central role as it determines the supramolecular assembly of the pectic chains and the formation of a pectate gel.Pectins capable of Ca²⁺ cross-linking are particularly common in bordered pit membranes (Chaffey et al., 1997; Hafren et al., 2000). Moreover, pectin-bound calcium influences wall elasticity (Ezaki et al., 2005; Proseus and Boyer, 2006; Derbyshire et al., 2007), and could therefore influence the stretching properties of the pit membranes and, consequently, the mechanism of cavitation. We tested the hypothesis that calcium plays a major role in the determinism of cavitation. This hypothesis was formulated long ago (Sperry and Tyree, 1988) but has not yet been thoroughly tested. We designed a series of experiments to demonstrate the specific role of calcium in this mechanism, and analyzed a large number of woody species to establish the role of calcium cross-linkage in across- and within-species variation in cavitation resistance. The data strongly support our hypothesis.