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Effects of Mercuric Chloride on the Hydraulic Conductivity of Tomato Root Systems (Evidence for a Channel-Mediated Water Pathway).

A pressure-flux approach was used to evaluate the effects of HgCl2 on water transport in tomato (Lycopersicon esculentum) roots. Addition of HgCl2 to a root-bathing solution caused a large and rapid reduction in pressure-induced root water flux; the inhibition was largely reversible upon addition of [beta]-mercaptoethanol. Root system hydraulic conductivity was reduced by 57%. There was no difference between treatments in the K+ concentration in xylem exudate. The results are consistent with the presence of a protein-mediated path for transmembrane water flow in tomato roots.

Journal Article↗

A plant flavone, luteolin, induces expression of Rhizobium meliloti nodulation genes.

The symbiotic interaction of Rhizobium meliloti and alfalfa results in the formation of nitrogen-fixing root nodules. Rhizobium meliloti nodABC genes are required for the early host responses of cortical cell divisions and root hair curling. The induction of nodABC expression by alfalfa exudates demonstrates host-symbiont signaling at an early stage in nodule development. The inducer molecule for nodABC expression was isolated from plant exudate by constructing a nodABC-lacZ fusion to monitor the inducing activity. From ultraviolet-visible absorption spectra, proton nuclear magnetic resonance, and mass spectrometry, the inducer was determined to be 3',4', 5,7-tetrahydroxyflavone (luteolin). Luteolin is a normal secondary plant metabolite found throughout the plant kingdom that may serve to control nodABC expression during nodule development. This regulatory role for a flavone contrasts with the function of some flavonoids as defense compounds.

Flavonoids↗

Effect of crop rotation and soil cover on alteration of the soil microflora generated by the culture of transgenic plants producing opines.

The culture of transgenic Lotus corniculatus plants producing opines, which are bacterial growth substrates, leads to the selection of rhizospheric bacteria able to utilize these substrates. We have investigated the fate of the opine-utilizing community over time under different experimental conditions following elimination of selective pressure exerted by the transgenic plants. These plants were removed from the soil, which was either left unplanted or replanted with wild-type L. corniculatus or wheat plants. The density of opine-utilizing bacteria in the fallow soils remained essentially unchanged throughout the experiment, regardless of the soil of origin (soil planted with wild-type or transgenic plants). When wild-type Lotus plants were used to replace their transgenic counterparts, only the bacterial populations able to utilize the opines were affected. Long-term changes affecting the opine-utilizing bacterial community on Lotus roots was dependent upon the opine studied. The concentration of nopaline utilizers decreased, upon replacement of the transgenic plants, to a level similar to that of normal plants, while the concentration of mannopine utilizers decreased to levels intermediate between transgenic and normal plants. These data indicate that: (i) the opine-utilizing bacterial populations can be controlled in the rhizosphere via plant-exudate engineering; (ii) the interaction between the engineered plants and their root-associated micro-organisms is transgene specific; and (iii) alterations induced by the cultivation of transgenic plants may sometimes be persistent. Furthermore, opine-utilizing bacterial populations can be controlled by crop rotation. Therefore, favouring the growth of a rhizobacterium of agronomic interest via an opine-based strategy appears feasible.

Arginine↗

Improvement by soil yeasts of arbuscular mycorrhizal symbiosis of soybean (Glycine max) colonized by Glomus mosseae.

The effects of the soil yeasts Rhodotorula mucilaginosa, Cryptococcus laurentii and Saccharomyces kunashirensis on the arbuscular mycorrhizal (AM) fungus Glomus mosseae (BEG 12) was studied in vitro and in greenhouse trials. The presence of yeasts or their soluble and volatile exudates stimulated the percentage spore germination and hyphal growth of G. mosseae. Percentage root length colonized by G. mosseae and plant dry matter of soybean (Glycine maxL. Merill) were increased only when the soil yeasts were inoculated prior to the AM fungus. Higher beneficial effects on AM colonization and plant dry matter were found when the soil yeasts were inoculated as an aqueous solution rather than as a thin agar slice. Although soluble and volatile exudates of yeasts benefited the AM symbiosis, their modes of action were different.

Cryptococcus↗

Xylem sap proteins.

Xylem sap from apple (Malus domestica Borkh), peach (Prunus persica Batsch), and pear (Pyrus communis L.) twigs was collected by means of pressure extrusion. This sap contained a number of acidic peroxidases and other proteins. Two other sources of xylem sap used in this study were stem exudates and guttation fluid. Similar peroxidases were also found in stem exudates and guttation fluids of strawberry (Fragaria x ananassa Duch.), tomato (Lycopersicum esculentum L.), and cucumber (Cucumis sativus L.). Isoelectric focusing activity gels showed that two peroxidases (isoelectric point [pl] 9 and pl 4.6) were present in initial stem exudates collected in the first 30 minutes after excision. Subsequent samples of stem exudate collected contained only the pl 4.6 isozyme. The pl 4.6 peroxidase isozyme was also found in root tissue and guttation fluid. These observations suggest that roots produce and secrete the pl 4.6 peroxidase into xylem sap. Cucumber seedlings were treated with 100 microliters per liter ethylene for 16 hours and the exudate from decapitated hypocotyl stumps was collected over a 3 hour period. Ethylene increased the peroxidase activity of stem exudates and inhibited the amount of exudate released. These observations suggest that xylem sap peroxidase may play a role in plugging damaged vascular tissue.

Journal Article↗

Changes in root water flow properties of solution culture-grown trembling aspen (Populus tremuloides) seedlings under different intensities of water-deficit stress.

To study the effects of water-deficit stress on root water flow properties in trembling aspen (Populus tremuloides Michx.), seedlings were grown in solution culture and subjected to water-deficit stress by placing their roots in sealed high humidity chambers. After 17 h of stress treatment, seedlings showed mild stress (MS) symptoms with a decline in shoot water potentials. Within 20 h, shoot water potentials rapidly declined, and severe stress (SS) symptoms were present. Root hydraulic conductivity (L(pr)) increased more than two-fold and the relative concentration of apoplastic tracer dye trisodium 3-hydroxy-5, 8, 10-pyrenetrisulphonate (PTS(3)) in xylem exudate decreased by 73.6% in MS seedlings. Conversely, L(pr) decreased (55.3%) and PTS(3) increased (28.6%) in SS seedlings. Treatment of roots with 0.1 mM mercuric chloride decreased root volume flow density (J(v)) by about 29.0% in control and MS plants with no decrease measured in SS seedlings. Mercuric chloride also increased PTS(3) concentration in xylem exudate of control (59%) and MS (86%) seedlings with no change observed in SS plants. The results suggest that aquaporin-mediated transport is important in the regulation of root water flow under drought stress and that root water flow properties are strongly affected by the stress level. Regulation of root water flow may represent an important drought-stress resistance mechanism.

Journal Article↗

Chemical behavior of Cd in rice rhizosphere.

Chemical behavior of Cd in rice rhizosphere as affected or not by Pb was investigated. The NH4OAc extractable Cd in the rhizosphere was distinctly lower than that in bulk soil. The depletion of Cd in the rhizosphere could not be simply attributed to Cd uptake by rice. The observed phenomena could be attributed to the decreasing pH in the rhizosphere and the complexing capabilities of soluble exudates for Cd. Extractable Cd increased in both the rhizosphere and bulk soil after the addition of Pb, which might be caused by the replacement of Pb for Cd. The extractable Cd in the non-rhizosphere varied with the distance from the root surface, especially within 0-1 mm, which was greatly affected by the combined effects of mass flow, activation and fixation, and had the lowest extractable Cd. Pb addition affected the distribution of extractable Cd in the non-rhizosphere, implying that the affinity of Pb for organic matter was greater than that of Cd. The difference of Cd species between rhizosphere and bulk soil demonstrated that the transformation of exchangeable Cd (EXC-Cd) to OM-Cd (bound to organic matter) and FMO-Cd (bound to iron and manganese oxide) occurred in the rice rhizosphere due to the exudations from the rice root, the activity of microorganisms on the root surface and the activation of Fe and Mn oxides. The interaction between Pb and Cd resulted in the content of EXC-Cd being higher in the presence of Pb, whereas the OM-Cd content was lower in the presence of Pb.

Cadmium↗

Scale-up production of puerarin from hairy roots of Pueraria phaseoloides in an airlift bioreactor.

Hairy roots of the Chinese herb, Pueraria phaseoloides, obtained from leaf explants and transformed with the Agrobacterium rhizogenes, were cultured in 2.5 l airlift bioreactors for three weeks. Puerarin accumulated at 5,570 microg g(-1) dry wt, which is near 200 times as much as in 250 ml flask cultures. In addition, puearin was exuded into the nutrient medium at final concentrations higher than in the hairy roots themselves.

Bioreactors↗

Studies of Root Function in Zea mays: III. Xylem Sap Composition at Maximum Root Pressure Provides Evidence of Active Transport into the Xylem and a Measurement of the Reflection Coefficient of the Root.

The cut ends of excised Zea mays roots were sealed to a pressure transducer and their root pressures recorded. These rose approximately hyperbolically to a maximum value of 4.21 +/- 0.34 bar after 30 to 40 minutes. Xylem exudate could not be collected at this pressure since the flow rate was zero. Samples of exudate were collected at lower applied pressures (DeltaP), however, and Deltapi, the osmotic pressure difference between them and the solution bathing the root, was measured by freezing point depression. A plot of DeltaP/Deltapi against J(v)/Deltapi, where J(v) is the volume flux, proved to be a straight line whose intercept, equal to sigma, the reflection coefficient, was 0.853 +/- 0.016. The maximum xylem concentrations of various chemical species were found by a similar extrapolative method and compared with those in the cell sap. This indicated that (a) Ca(2+), Mg(2+), NO(3) (2-), SO(4) (2-), and most amino acids move from the cells to the xylem down an electrochemical potential gradient; (b) relative to these ions H(+), NH(4) (+), glutamine and asparagine are actively transported into the xylem; and (c) H(2)PO(4) (-), and K(+) are actively retained in the symplasm.

Journal Article↗

Metabolome-driven rhizosphere microbiome assembly determining the health of medicinal herb (Angelica sinensis) against root rot.

BACKGROUND: The rhizosphere-associated microbiota plays a crucial role in plant responses to disease stress. Plant secondary metabolites are recognized as crucial mediators in the assembly of rhizosphere microbial communities, particularly by enhancing the colonization of beneficial microorganisms. Despite this recognized importance, a deeper understanding of how such metabolome-driven microbiome assembly specifically determines plant resistance against soil-borne diseases is still lacking. RESULTS: Here, we focused on the widely planted medicinal plant Angelica sinensis and demonstrated that root rot-diseased rhizosphere soils (DRS) exhibited a higher relative abundance of Fusarium and a lower relative abundance of Streptomyces compared to healthy rhizosphere soils (HRS). Shotgun metagenomic sequencing revealed that metabolism-associated genes, particularly those related to steroid degradation, are significantly enriched in HRS samples. Subsequent genome and functional gene analysis of Streptomyces revealed that the steroid degradation-related genes are associated with rhizosphere colonization in hosts. Rhizosphere Streptomyces S15 directly antagonized Fusarium and enhanced the root resistance of A. sinensis. Comparative metabolomics showed that A. sinensis plants from HRS secreted more lipid and lipid-like molecules than those from DRS, especially sterol lipids and long-chain fatty acids, which promoted the growth of Streptomyces S15 isolates. Transcriptome analysis validated that the lipid hormones are essential for sporulation, biofilm formation, and streptomycin biosynthesis of S15 strain. Finally, exogenous application of synbiotics (lipid prebiotics and S15) to A. sinensis resulted in the enrichment of S15-homologous Streptomyces amplicon sequence variant (ASV), further establishing beneficial bacterial communities in Fusarium-stressed rhizospheres. CONCLUSIONS: Our study proposes that A. sinensis recruits steroid-metabolizing Streptomyces species by exuding key lipid compounds (i.e., methyl jasmonate and brassinolide) to combat Fusarium root rot. This study provides novel insights into using functional synbiotics as a promising strategy for manipulating plant-microbiome interactions to promote sustainable agriculture. Video Abstract.

Rhizosphere↗

In vivo and in vitro evaluations of water-absorption properties of various ointments.

The water-absorption properties of various vehicles were evaluated in rats whose stratum corneum had been stripped off over the application site. Polyethylene glycol (PEG) applied to the site effectively absorbed the exudate, but other applied ointments absorbed little exudate or water. By the PEG ointment, the exudate was absorbed continuously and extensively, and a linear relationship was obtained between the amount of water absorbed and the square root of time. To establish an acceptable alternative in vitro method for the design of a suitable ointment for treating skin ulcers with exudate, in vitro evaluations were performed by three methods: the Franz cell method, the cellulose membrane diffusion method and the spontaneous water uptake method. The membrane diffusion method, and the spontaneous water uptake method appeared to be acceptable alternatives to the in vivo method in evaluating the water absorption of ointments. It is a simple and useful method for preliminary formulation studies of ointments for the treatment of skin ulcers with exudate.

Animals↗

Epithelial cell kinetics with atelocollagen membranes: a study in rats.

A recent development in guided tissue regeneration procedures is the use of resorbable membranes, which eliminate the need for subsequent surgical removal. In this study we performed flap surgery in rats with (experimental) or without (control) implantation of one of the newer materials, atelocollagen. We observed the gingival epithelial cell kinetics using 3H-thymidine and examined the extent of gingival epithelium migration. Histological observations at day 1 on the experimental side demonstrated regenerated epithelium apposed to the collagen membrane with an intervening layer of necrotic tissues and/or fibrinous exudate. There was no observable proliferation of regenerated epithelium toward the root apex. On day 14, the regenerated epithelium migrated apically along the treated root surface in the control group. By contrast, on day 14 in the experimental group, the regenerated epithelium contacted the root surface at the cemento-enamel junction (CEJ). Apical to the CEJ, there was new cementum formation with inserting connective tissue fibers. Autoradiographs from day 1 experimental sides demonstrated labeled cells in the basal cell layers from oral, crevicular, and junctional epithelium. From day 1 to day 5, labeling indices of oral epithelium and regenerating crevicular epithelium on experimental sides were lower than on control sides. These histological and autoradiographic findings suggest that atelocollagen membrane inhibits apical migration of regenerating epithelium and accelerates connective tissue reattachment in part by inhibiting the mitotic function of basal epithelial cells in early stages of wound healing.

Animals↗

Physiological basis of reduced AL tolerance in ditelosomic lines of Chinese Spring wheat.

Aluminum tolerance was assessed in the moderately Al-tolerant wheat (Triticum aestivum L.) cultivar Chinese Spring and a set of ditelosomic lines derived from Chinese Spring. Three ditelosomic lines lacking chromosome arms 4DL, 5AS and 7AS, respectively, exhibited decreased Al tolerance relative to the euploid parent Chinese Spring based on reduced root growth in Al-containing solutions. The physiological basis of the reduced Al tolerance was investigated. Measurements by inductively coupled argon plasma mass spectroscopy of root apical Al accumulation demonstrated that two of these three lines had a decreased ability to exclude Al from the root apex, the site of Al phytotoxicity. As Al-induced malate exudation has been suggested to be an important physiological mechanism of Al tolerance in wheat, this parameter was quantified and malate exudation was shown to be smaller in all three deletion lines compared with Chinese Spring. These results suggest that the decreased Al tolerance in at least two of the three ditelosomic lines is due to the loss of different genes independently influencing a single Al-tolerance mechanism, rather than to the loss of genes encoding alternative Al-tolerance mechanisms.

Adaptation, Physiological↗

Quantification of endotoxins in necrotic root canals from symptomatic and asymptomatic teeth.

The purpose of this investigation was to quantify the concentration of endotoxin in necrotic root canals and investigate the possible relationship between the concentration of endotoxin and endodontic signs and symptoms. Samples were collected from root canals of 50 patients requiring endodontic treatment due to necrosis of the pulpal tissue. Anaerobic techniques were used to determine the number of c.f.u. in each sample. A quantitative chromogenic Limulus amoebocyte lysate assay was used to measure the concentration of endotoxin in each sample. The presence of c.f.u. was detected by culture in all samples (range 10(2)-5x10(6)). In samples from cases of patients with spontaneous pain, the mean c.f.u. was 1.43x10(6) while in asymptomatic cases it was 9.1x10(4). Endotoxin was present in all the samples studied [range 2390.0-22100.0 endotoxin units (EU) ml-1]. The mean concentration of endotoxin in samples from patients with spontaneous pain was 18540.0 EU ml-1 while in asymptomatic cases it was 12030.0 EU ml-1. Asymptomatic cases generally had lower levels of endotoxin (i.e. a negative association). A positive association was found between endotoxin and symptomatic cases (e.g. spontaneous pain, tenderness to percussion, pain on palpation, swelling and purulent exudates). This study showed that endotoxin is present in high concentrations in root canals of symptomatic teeth. There was a positive correlation between the concentration of endotoxin in the root canal and the presence of endodontic signs and symptoms.

Bacteria, Anaerobic↗

Radial transport of ions in roots.

Measurements were made of the relative amounts of (86)Rb, (36)Cl, and (32)P accumulated in the cortex and stele of intact roots of corn (Zea mays), either detached or attached to their shoots. Both 4- and 7-day-old roots accumulated as much or more (86)Rb in the stele as in the cortex. In experiments with (36)Cl, cortex and stele accumulated the same amount, except for 4-day-old and 7-day-old attached roots, in which the cortex contained more (36)Cl than the stele after 23 hr. An additional study of (32)P uptake showed greater accumulation in the cortex than the stele for a short period of time, but as much in the stele as in the cortex after 8 to 24 hr. Transport of (86)Rb, (36)Cl, and (32)P into the xylem exudate increased with increasing accumulation of these ions in stele and cortex of the root. These experiments show no consistent difference between cortex and stele of intact corn roots with respect to their ability to accumulate several kinds of ions.

Journal Article↗

Expression of inflammatory cytokine genes in vivo by human alveolar bone-derived polymorphonuclear leukocytes isolated from chronically inflamed sites of bone resorption.

Alveolar bone-derived polymorphonuclear leukocytes (PMNs) were characterized for their ability to produce inflammatory cytokines such as interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF alpha), and IL-6 in vivo. Periapical exudates (PE) were collected from periapical lesions with chronic periapical periodontitis through root canals. Cells and noncellular supernatants were then isolated by centrifugation. The concentration of cytokines present in the noncellular supernatants were determined by ELISA. High concentrations of IL-1 alpha, IL-1 beta, and IL-6 were detected in PE, however, TNF alpha was not. PE contains predominantly PMNs ( > 95% of residing cells) with a few percent of lymphocytes and/or macrophages. These alveolar bone-derived PMNs were purified by the Ficoll-Hypaque gradient method and were analyzed for cytokine mRNA expression using the cytokine-specific reverse-transcription polymerase chain reaction. Highly purified PMNs ( > 99.5%) isolated from PE expressed significant levels of mRNA for IL-alpha, IL-1 beta, and TNF alpha. IL-6 mRNA was not detected, although a high concentration of IL-6 was detected in supernatants of PE by ELISA. The IL-6 secretion in PE could be derived from macrophages, T lymphocytes, osteoblasts, or fibroblasts around periapical lesions. These data strongly suggest that human PMNs derived from alveolar bone can spontaneously produce IL-1 alpha, IL-1 beta, and TNF alpha at sites of inflammation, and probably initiate inflammation and regulate augmentation of bone resorption in vivo.

Adolescent↗

Lateral Transport of Ions into the Xylem of Corn Roots: II. Evaluation of a Stelar Pump.

When an excised corn (Zea mays) root pretreated with chloride was exposed for 10 minutes to pulse labeling with (30)Cl and then transferred to unlabeled chloride, the activity in the xylem exudate reached a maximum about 4 minutes after pulse labeling was discontinued and then declined sharply. The rate at which labeled chloride was transported across the root into the xylem and basipetally therein was on the order of 75 to 250 centimeters per hour. Consequently, symplasmic movement of chloride in corn roots is fast and may not be rate-limiting in transfer from the root surface to the xylem. Experiments on pulse labeling with (22)Na gave similar results. A large fraction of the absorbed (22)Na was not translocated into the exudate but was tightly sequestered in a cell compartment, probably the vacuole.Electron probe analysis was used to reveal the pattern of potassium distribution in cross sections taken 10 to 11 millimeters from the tip. The cytoplasm and vacuoles of the xylem parenchyma cells accumulated potassium to a much greater extent than cortical and other stelar cells. Ultrastructural studies showed that the cytoplasm of the xylem parenchyma cells contains numerous membrane systems. It was concluded that the xylem parenchyma cells secrete ions from the symplasm into the conducting vessels, and it was suggested that this secretion is driven across the plasmalemma by a carrier-mediated transport.

Journal Article↗