Exploring Environmental Factors That Drive Diel Variations in Tree Water Storage Using Wavelet Analysis

Internal water storage within trees can be a critical reservoir that helps trees overcome both short- and long-duration environmental stresses. We monitored changes in internal tree water storage in a ponderosa pine on daily and seasonal scales using moisture probes, a dendrometer, and time-lapse electrical resistivity imaging (ERI). These data were used to investigate how patterns of in-tree water storage are affected by changes in sapflow rates, soil moisture, and meteorologic factors such as vapor pressure deficit. Measurements of xylem fluid electrical conductivity were constant in the early growing season while inverted sapwood electrical conductivity steadily increased, suggesting that increases in sapwood electrical conductivity did not result from an increase in xylem fluid electrical conductivity. Seasonal increases in stem electrical conductivity corresponded with seasonal increases in trunk diameter, suggesting that increased electrical conductivity may result from new growth. On the daily scale, changes in inverted sapwood electrical conductivity correspond to changes in sapwood moisture. Wavelet analyses indicated that lag times between inverted electrical conductivity and sapflow increased after storm events, suggesting that as soils wetted, reliance on internal water storage decreased, as did the time required to refill daily deficits in internal water storage. We found short time lags between sapflow and inverted electrical conductivity with dry conditions, when ponderosa pine are known to reduce stomatal conductance to avoid xylem cavitation. A decrease in diel amplitudes of inverted sapwood electrical conductivity during dry periods suggest that the ponderosa pine relied on internal water storage to supplement transpiration demands, but as drought conditions progressed, tree water storage contributions to transpiration decreased. Time-lapse ERI- and wavelet-analysis results highlight the important role internal tree water storage plays in supporting transpiration throughout a day and during periods of declining subsurface moisture.

Biomolecular corona formation on CuO nanoparticles in plant xylem fluid

CuO nanoparticles selectively remove proteins from Cucurbita pepo (pumpkin) xylem to form a nanoparticle corona.

Ralstonia solanacearum Uses Inorganic Nitrogen Metabolism for Virulence, ATP Production, and Detoxification in the Oxygen-Limited Host Xylem Environment

ABSTRACTGenomic data predict that, in addition to oxygen, the bacterial plant pathogenRalstonia solanacearumcan use nitrate (NO3−), nitrite (NO2−), nitric oxide (NO), and nitrous oxide (N2O) as terminal electron acceptors (TEAs). Genes encoding inorganic nitrogen reduction were highly expressed during tomato bacterial wilt disease, when the pathogen grows in xylem vessels. Direct measurements found that tomato xylem fluid was low in oxygen, especially in plants infected by R. solanacearum. Xylem fluid contained ~25 mM NO3−, corresponding to R. solanacearum's optimal NO3−concentration for anaerobic growthin vitro. We tested the hypothesis that R. solanacearum uses inorganic nitrogen species to respire and grow during pathogenesis by making deletion mutants that each lacked a step in nitrate respiration (ΔnarG), denitrification (ΔaniA, ΔnorB, and ΔnosZ), or NO detoxification (ΔhmpX). TheΔnarG,ΔaniA, andΔnorBmutants grew poorly on NO3−compared to the wild type, and they had reduced adenylate energy charge levels under anaerobiosis. While NarG-dependent NO3−respiration directly enhanced growth, AniA-dependent NO2−reduction did not. NO2−and NO inhibited growth in culture, and their removal depended on denitrification and NO detoxification. Thus, NO3−acts as a TEA, but the resulting NO2−and NO likely do not. None of the mutants grew as well as the wild typein planta, and strains lacking AniA (NO2−reductase) or HmpX (NO detoxification) had reduced virulence on tomato. Thus, R. solanacearum exploits host NO3−to respire, grow, and cause disease. Degradation of NO2−and NO is also important for successful infection and depends on denitrification and NO detoxification systems.IMPORTANCEThe plant-pathogenic bacteriumRalstonia solanacearumcauses bacterial wilt, one of the world's most destructive crop diseases. This pathogen's explosive growth in plant vascular xylem is poorly understood. We used biochemical and genetic approaches to show that R. solanacearum rapidly depletes oxygen in host xylem but can then respire using host nitrate as a terminal electron acceptor. The microbe uses its denitrification pathway to detoxify the reactive nitrogen species nitrite (a product of nitrate respiration) and nitric oxide (a plant defense signal). Detoxification may play synergistic roles in bacterial wilt virulence by converting the host's chemical weapon into an energy source. Mutant bacterial strains lacking elements of the denitrification pathway could not grow as well as the wild type in tomato plants, and some mutants were also reduced in virulence. Our results show how a pathogen's metabolic activity can alter the host environment in ways that increase pathogen success.

The In Planta Transcriptome of Ralstonia solanacearum: Conserved Physiological and Virulence Strategies during Bacterial Wilt of Tomato

ABSTRACTPlant xylem fluid is considered a nutrient-poor environment, but the bacterial wilt pathogenRalstonia solanacearumis well adapted to it, growing to 108to 109 CFU/g tomato stem. To better understand howR. solanacearumsucceeds in this habitat, we analyzed the transcriptomes of two phylogenetically distinctR. solanacearumstrains that both wilt tomato, strains UW551 (phylotype II) and GMI1000 (phylotype I). We profiled bacterial gene expression at ~6 × 108 CFU/ml in culture or in plant xylem during early tomato bacterial wilt pathogenesis. Despite phylogenetic differences, these two strains expressed their 3,477 common orthologous genes in generally similar patterns, with about 12% of their transcriptomes significantly alteredin plantaversus in rich medium. Several primary metabolic pathways were highly expressed during pathogenesis. These pathways included sucrose uptake and catabolism, and components of these pathways were encoded by genes in thescrABYcluster. A UW551scrAmutant was significantly reduced in virulence on resistant and susceptible tomato as well as on potato and the epidemiologically important weed hostSolanum dulcamara. FunctionalscrAcontributed to pathogen competitive fitness during colonization of tomato xylem, which contained ~300 µM sucrose.scrAexpression was induced by sucrose, but to a much greater degree by growthin planta. Unexpectedly, 45% of the genes directly regulated by HrpB, the transcriptional activator of the type 3 secretion system (T3SS), were upregulatedin plantaat high cell densities. This result modifies a regulatory model based on bacterial behavior in culture, where this key virulence factor is repressed at high cell densities. The active transcription of these genes in wilting plants suggests that T3SS has a biological role throughout the disease cycle.IMPORTANCERalstonia solanacearumis a widespread plant pathogen that causes bacterial wilt disease. It inflicts serious crop losses on tropical farmers, with major economic and human consequences. It is also a model for the many destructive microbes that colonize the water-conducting plant xylem tissue, which is low in nutrients and oxygen. We extracted bacteria from infected tomato plants and globally identified the biological functions thatR. solanacearumexpresses during plant pathogenesis. This revealed the unexpected presence of sucrose in tomato xylem fluid and the pathogen’s dependence on host sucrose for virulence on tomato, potato, and the common weed bittersweet nightshade. Further,R. solanacearumwas highly responsive to the plant environment, expressing several metabolic and virulence functions quite differently in the plant than in pure culture. These results reinforce the utility of studying pathogens in interaction with hosts and suggest that selecting for reduced sucrose levels could generate wilt-resistant crops.

Analysis of Xylem Fluid Components in Almond Cultivars Differing in Resistance to Almond Leaf Scorch Disease

Almond leaf scorch (ALS) is caused by the pathogenic bacterium Xylella fastidiosa and poses a threat to the California almond industry. Almond cultivars are differentially resistant or susceptible to ALS. X. fastidiosa can infect but does not overwinter in resistant cultivars in sufficient numbers to cause symptoms or be detected by polymerase chain reaction. To better understand the biochemical or morphological factors mediating resistance, we extracted and analyzed almond xylem fluid from four almond cultivars differing in ALS susceptibility, including Butte and Carmel cultivars that are field resistant and Peerless and Sonora that are ALS susceptible. Xylem fluid was collected over winter months in 2007 to 2009, as well as July 2008 and April 2009, and analyzed for the following: pH, osmolarity, concentrations of sugars, calcium, magnesium, organic acids, and total phenolics. For most of these analyses, we found no clear differences in xylem fluid from resistant and susceptible almond cultivars. However, during the winter months, resistant cultivars tended to have higher concentrations of total phenolic compounds compared with susceptible cultivars (P = 0.05). In February 2009, Carmel had the highest total phenolic concentration measured, 233 μg/ml of gallic acid equivalents. The lowest phenolic concentrations occurred in April 2009. The cross-sectional areas of xylem vessels in Butte (resistant) and Peerless (susceptible) trees were not significantly different between cultivars.