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PubMed1991

New root growth of Douglas-fir seedlings at low carbon dioxide concentration.

New root growth of conifer nursery seedlings is dependent on light, but whether this is necessary only for photosynthesis, or also has some other root growth promoting effect is unknown. This question was investigated using one-year-old Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings grown at two nurseries from the same seed lot and taken from cold storage in March and April. New root production was examined in two 10-day root growth capacity (RGC) experiments. Seedlings were subjected to one of four treatments: (1) control, (2) exclusion of light, (3) low CO(2) concentration, and (4) girdling to separate the phloem connection between shoot and root. Shoots were enclosed in Plexiglas cuvettes and supplied with scrubbed air to reduce the CO(2) concentration to 11 to 22 microl l(-1) in the light. The closed system used in Experiment 1 was opened to the atmosphere briefly each day to restore O(2) to ambient, but this was unnecessary for the open system used in Experiment 2. In Experiment 1, new root production was affected by treatments in the order: control > low CO(2) concentration > dark > girdling. Greater new root production in the low CO(2) treatment compared with the dark treatment was attributed to brief increases in CO(2) to ambient concentrations when O(2) was restored each day. In Experiment 2, new root production at the low CO(2) concentration in the light was essentially the same as in darkness, but only 17% of the control value. Thus light appeared to play no part in new root production other than permitting photosynthesis. Limited production of new roots occurred in the absence of photosynthesis, which was further reduced by girdling, presumably because, after girdling, only root system reserves could be used.

Journal Article↗
PubMed1998

Effects of lidocaine on nucleus pulposus-induced nerve root injury. A neurophysiologic and histologic study of the pig cauda equina.

STUDY DESIGN: Application of autologous nucleus pulposus on nerve roots and treatment with local application of lidocaine in the pig. OBJECTIVES: Studies of the effects of lidocaine on nucleus pulposus-exposed nerve roots. SUMMARY OF BACKGROUND DATA: Nerve root infiltration may improve radicular symptoms beyond the pharmacologic duration of local anesthetics, but the mechanisms for this effect are not known. METHODS: Nucleus pulposus was harvested from a lumbar disc and placed onto the sacrococcygeal cauda equina in pigs. In Series 1, early lidocaine treatment of nucleus pulposus-induced nerve root injury, pigs received 2% lidocaine (n = 5) or saline (n = 5) before and after surgery. Nerve conduction velocity and histologic appearance were studied after 3 days. In Series 2, delayed lidocaine treatment of nucleus pulposus-induced nerve root injury, after 7 days 2% lidocaine was administered epidurally to nucleus pulposus-exposed (n = 4) and -nonexposed (n = 4) nerve roots. Nerve conduction velocity, muscle action potentials, and histologic appearance were assessed. RESULTS: In Series 1, early treatment with lidocaine limited the reduction in nerve conduction velocity. The epidural inflammation was less in lidocaine treated animals. In Series 2, nerve conduction velocity was lower in nucleus pulposus-exposed animals than in nonexposed animals. The initial reduction of nerve conduction velocity and muscle action potential was similar between the groups, but the recovery of muscle action potential was slower and less complete in nucleus pulposus-exposed nerve roots. There was minimal histologic nerve injury in both series and in both protocols. CONCLUSIONS: Early treatment with lidocaine may reduce nucleus pulposus-induced nerve root injury. Lidocaine induced a delayed recovery in nerve roots exposed to nucleus pulposus. Further studies are needed to clarify the therapeutic effects of nerve root infiltration and the pathophysiology of nucleus pulposus-induced nerve root injury.

Anesthetics, Local↗
PubMed2001

Clinical features of extraforaminal lumbar disc herniation based on the radiographic location of the dorsal root ganglion.

STUDY DESIGN: The relations between the location of the dorsal root ganglion and pre- and postoperative symptoms were reviewed retrospectively in 27 patients who underwent radiculography and posterior discectomy. OBJECTIVES: To evaluate the clinical features and surgical outcome of extraforaminal lumbar disc herniation based on the location of dorsal root ganglion. SUMMARY OF BACKGROUND DATA: The location of dorsal root ganglia has been reported to be correlated with a variety of radicular symptoms. Extraforaminal lumbar disc herniation has several specific clinical features, one of which is severe radicular pain. However, there is no report in the literature on the association between the location of the dorsal root ganglia and the severity of the symptoms of extraforaminal lumbar disc herniation. METHODS: The radiographic location of the dorsal root ganglion of each compressed nerve root was determined by preoperative direct radiculograms. All patients were classified into the following three groups according to the location of dorsal root ganglion: intraspinal, intraforaminal, and extraforaminal. The incidences of these locations were 5 of 27 (18.5%), 15 of 27 (55.5%), and 7 of 27 (25.9%), respectively. The relation between the location of the dorsal root ganglion and clinical parameters such as the level of the compressed nerve root, the degree of limitation on straight leg raising test, the severity of the pre- and postoperative subjective symptoms (leg pain, low back pain, and walking capacity), clinical signs (sensory and motor disturbance), and the recovery rate were investigated. RESULTS: The degree of limitation on the straight leg raising test in the extraforaminal group tended to be low, compared with that in the intraspinal and intraforaminal groups. Low back pain in the extraforaminal group was more severe than that in the intraspinal and intraforaminal groups. Preoperative leg pain in the extraforaminal group was significantly more severe that that in the intraspinal group, and the walking capacity in the extraforaminal group tended to be lower than that in the intraspinal and intraforaminal groups. No significant differences were found between the location of dorsal root ganglion and the preoperative sensory or motor disturbance and surgical outcomes. CONCLUSION: The location of the dorsal root ganglion might influence the severity of radicular symptoms (pain and walking distance tolerance) in patients with extraforaminal lumbar disc herniation.

Adult↗
PubMed1995

Differential Expression of Proteins and mRNAs from Border Cells and Root Tips of Pea.

Many plants release large numbers of metabolically active root border cells into the rhizosphere. We have proposed that border cells, cells produced by the root cap meristem that separate from the rest of the root upon reaching the periphery of the cap, are a singularly differentiated part of the root system that modulates the environment of the plant root by producing specific substances to be released into the rhizosphere. Proteins synthesized in border cells exhibit profiles that are very distinct from those of the root tip (root cap, root meristem, and adjacent cells). In vivo-labeling experiments demonstrate that 13% of the proteins that are abundant in preparations from border cells are undetectable in root tip preparations. Twenty-five percent of the proteins synthesized by border cells in a 1-h period are rapidly excreted into the incubation medium. Quantitative variation in levels of specific marker proteins, including glutamine synthetase, heat-shock protein 70, and isoflavone reductase, also occurs between border cells and cells in the root tip. mRNA differential-display assays demonstrate that these large qualitative and quantitative differences in protein expression are correlated with similarly distinct patterns of gene expression. These observations are consistent with the hypothesis that a major switch in gene expression accompanies differentiation into root border cells, as expected for cells with specialized functions in plant development.

Journal Article↗
PubMed1995

Sugar-Binding Activity of Pea Lectin Expressed in White Clover Hairy Roots.

Introduction of the pea (Pisum sativum L.) lectin (PSL) gene into white clover (Trifolium repens L.) hairy roots facilitates nodulation by the nitrogen-fixing bacterium Rhizobium leguminosarum biovar viciae, which normally nodulates pea and not white clover (C.L. Diaz, L.S. Melchers, P.J.J. Hooykaas, B.J.J. Lugtenberg, and J.W. Kijne [1989] Nature 338: 579-581). Here, we show that PSL is functionally expressed in transgenic white clover hairy roots transformed with the PSL gene. PSL could be isolated from these roots by affinity chromatography. Immunoanalysis of PSL showed the presence of polypeptides corresponding to the PSL precursor and its [beta] subunits. In addition, we developed a highly sensitive localization technique based on specific binding of a glycan moiety of rat IgE to PSL. Similar to the situation in pea roots, PSL appeared to be localized on the external cell surface of elongated epidermal cells and on the tips of emerging and growing root hairs of transgenic white clover hairy roots. PSL was not observed on normal white clover roots and on hairy roots without the PSL gene. These results show that (a) in transgenic white clover hairy roots, PSL is correctly processed and targeted to root cells susceptible to rhizobial infection, and (b) like in pea roots, PSL is surface bound with at least one of its two sugar-binding sites available for (rhizobial) ligands.

Journal Article↗
PubMed1996

An Ethylene-Mediated Increase in Sensitivity to Auxin Induces Adventitious Root Formation in Flooded Rumex palustris Sm.

The hormonal regulation of adventitious root formation induced by flooding of the root system was investigated in the wetland species Rumex palustris Sm. Adventitious root development at the base of the shoot is an important adaptation to flooded conditions and takes place soon after the onset of flooding. Decreases in either endogenous auxin or ethylene concentrations induced by application of inhibitors of either auxin transport or ethylene biosynthesis reduced the number of adventitious roots formed by flooded plants, suggesting an involvement of these hormones in the rooting process. The rooting response during flooding was preceded by increased endogenous ethylene concentrations in the root system. The endogenous auxin concentration did not change during flooding-induced rooting, but a continuous basipetal transport of auxin from the shoot to the rooting zone appeared to be essential in maintaining stable auxin concentrations. These results suggest that the higher ethylene concentration in soil-flooded plants increases the sensitivity of the root-forming tissues to endogenous indoleacetic acid, thus initiating the formation of adventitious roots.

Journal Article↗
PubMed1998

Expression of a Soybean Hydroxyproline-Rich Glycoprotein Gene Is Correlated with Maturation of Roots

A novel extensin gene has been identified in soybean (Glycine max L.) that encodes a hydroxyproline-rich glycoprotein (SbHRGP3) with two different domains. In this study expression of SbHRGP3 was investigated during soybean root development. SbHRGP was expressed in roots of mature plants, as well as seedlings, and showed a distinct pattern of expression during root development. The expression of SbHRGP3 increased gradually during root development of seedlings and reached a maximum while the secondary roots were maturing. The maximum expression level was contributed mainly by the secondary roots rather than by the primary root. Furthermore, expression of SbHRGP3 was preferentially detected in the regions undergoing maturation of the primary and secondary roots. These results imply that the expression of SbHRGP3 is regulated in an organ- and development-specific manner and that in soybean SbHRGP3 expression may be required for root maturation, presumably for the cessation of root elongation. Wounding and sucrose in combination enhanced expression of SbHRGP3 in roots, whereas both wounding and sucrose were required for the expression of SbHRGP3 in leaves.

Journal Article↗
PubMed1976

Influence of Helminthosporium maydis, Race T, Toxin on Potassium Uptake in Maize Roots.

The effect of a toxin extract of Helminthosporium maydis, race T on K(+) ((86)Rb) uptake by excised root segments of normal (N) and Texas cytoplasmic male-sterile (T) versions of corn inbred W64A was investigated. The uptake of K(+) was inhibited in both N and T roots by the toxin. This was true for both basal (freshly excised) and augmented (pretreated with aeration) K(+) uptake. Augmented uptake was more toxin-sensitive than basal uptake (irrespective of cytoplasm type), and the augmented uptake in T roots was seven to eight times more toxin-sensitive than in N roots.Specific zones of roots differed in their basal and augmented K(+) uptake rates as well as their toxin sensitivities. The root apex of T was more sensitive to toxin than the apex of N roots when basal K(+) uptake was measured. In mature zones of the root, T was more sensitive than N when augmented rates were measured. During the development of the augmented K(+) uptake capacity in either N or T roots, the sensitivity to the toxin did not change; uptake in N roots was inhibited by 10 to 25% and uptake in T roots was inhibited by 70 to 80%.The difference in toxin sensitivity of K(+) uptake between N and T roots may be due to N possessing a protective mechanism which is deficient in T.

Journal Article↗
PubMed1978

Sites of absorption and translocation of iron in barley roots: tracer and microautoradiographic studies.

Absorption and translocation of labeled Fe were measured at various locations along the length of intact seminal axes and lateral roots of iron-sufficient (+Fe) and iron-stressed (-Fe) barley (Hordeum vulgare) plants. In seminal axes of +Fe plants, rates of translocation were very much higher in a zone 1 to 4 cm from the root tip than elsewhere in the root. Lateral roots of high rates of translocation were also restricted to a narrow band of maturing or recently matured cells. In -Fe plants the patterns of uptake and translocation were essentially the same as in +Fe plants but the rates were 7- to 10-fold higher. The amount of labeled Fe bound to the root itself was not increased by Fe stress and its distribution along the root seemed inversely related to the ability to translocate Fe.Microautoradiographic studies showed that most of the iron bound to roots was held in an extracellular peripheral band in which iron seemed to be precipitated. This process may be assisted by microbial colonies but did not depend on them since it was seen, although to a lesser extent, in sterile roots. In zones from which iron was translocated there was evidence that internal root tissues became labeled readily, but as translocation declined with distance from the root tip, radial penetration of Fe appeared to become progressively less. The results are discussed in relation to possible changes in the pH or redox potential of the surface of the root.

Journal Article↗
PubMed1978

Effect of l-Canavanine on Nitrate Reductase in Corn Roots.

l-Canavanine inhibits the appearance of nitrate reductase (NADH-nitrate oxidoreductase, EC 1.6.6.1) in both root tips and mature root sections of corn (Zea mays L.). Ten-fold more canavanine was required to cause a 50% reduction in the level of nitrate reductase activity (NRA) in root tips than in mature root sections. For example with one particular batch of seeds 500 mum canavanine was effective in root tips whereas only 50 mum was required in mature root sections. In root tips arginine (1 mm) completely reversed the effect of 1 mm canavanine. In mature root sections higher concentrations of arginine (approximately 5 mm) were required for a complete reversal of the canavanine effect. Additions of canavanine to roots after a period of 3 hours with 5 mm KNO(3) resulted in a loss of NRA. NO(3) (-) protected nitrate reductase from this inactivation in both root tip and mature root sections.

Journal Article↗
PubMed1982

Effect of changes in shoot carbon-exchange rate on soybean root nodule activity.

The effect of short- and long-term changes in shoot carbon-exchange rate (CER) on soybean (Glycine max [L.] Merr.) root nodule activity was assessed to determine whether increases in photosynthate production produce a direct enhancement of symbiotic N(2) fixation. Shoot CER, root + nodule respiration, and apparent N(2) fixation (acetylene reduction) were measured on intact soybean plants grown at 700 microeinsteins per meter per second, with constant root temperature and a 14/10-hour light/dark cycle. There was no diurnal variation of root + nodule respiration or apparent N(2) fixation in plants assayed weekly from 14 to 43 days after planting. However, if plants remained in darkness following their normal dark period, a significant decline in apparent N(2) fixation was measured within 4 hours, and decreasing CO(2) concentration from 320 to 90 microliters CO(2) per liter produced diurnal changes in root nodule activity. Increasing shoot CER by 87, 84, and 76% in 2-, 3-, and 4-week-old plants, respectively, by raising the CO(2) concentration around the shoot from 320 to 1,000 microliters CO(2) per liter, had no effect on root + nodule respiration or acetylene-reduction rates during the first 10 hours of the increased CER treatment. When the CO(2)-enrichment treatment was extended in 3-week-old plants, the only measured parameter that differed significantly after 3 days was shoot CER. After 5 days of continuous CO(2) enrichment, root + nodule respiration and acetylene reduction increased, but such changes reflected an increase in root nodule mass rather than greater specific root nodule activity. The results show that on a 24-hour basis the process of symbiotic N(2) fixation in soybean plants grown under controlled environmental conditions functioned at maximum capacity and was not limited by shoot CER. Whether N(2)-fixation capacity was limited by photosynthate movement to root nodules or by saturation of metabolic processes in root nodules is not known.

Journal Article↗
PubMed1983

Interaction of Combined Nitrogen with the Expression of Root-Associated Nitrogenase Activity in Grasses and with the Development of N(2) Fixation in Soybean (Glycine max L. Merr.).

Soluble root N concentrations of corn, sorghum, pearl millet, rice, wild rice, and soybeans were determined and related to measurements of nitrogenase activity and changes in availability of combined N to plants. In corn, sorghum, and pearl millet, applications of fertilizer N increased soluble root N concentrations, but root-associated nitrogenase activity was negligible in control and treated plants. Applications of NH(4)NO(3) to rice increased the water soluble root N concentrations and inhibited root-associated nitrogenase activity. In wild rice, root-associated nitrogenase activity was absent during vegetative growth and developed at the reproductive growth stage. The soluble root N concentration decreased progressively as wild rice grew indicating that the availability of combined N in the root environment declined. Therefore, development of nitrogenase activity in wild rice is associated with the change in availability of combined N in the root environment. The development of nitrogenase activity in wild rice was probably not due to colonization of roots by N(2)-fixing bacteria because most probable numbers of recovery did not significantly vary throughout the plants' growth cycle. In field-grown soybeans with or without fertilizer N application, we also observed a relationship between a decrease in soluble root N concentration and the development of nitrogenase activity.

Journal Article↗
PubMed1983

Gravity-Induced Polar Transport of Calcium across Root Tips of Maize.

Calcium movement across primary roots of maize (Zea mays, L.) was determined by application of (45)Ca(2+) to one side of the root and collection of radioactivity in an agar receiver block on the opposite side. Ca movement across the root tip was found to be at least 20 times greater than movement across the elongation zone. The rapid movement of Ca across the tip was severely inhibited in roots from which the root cap had been removed. Ca movement across the tip was also strongly retarded in roots pretreated with 2,4-dinitrophenol or potassium cyanide. Orientation of roots horizontally had no effect on Ca movement across the elongation zone but caused a strong asymmetry in the pattern of Ca movement across the tip. In gravistimulated roots, the movement of Ca from top to bottom increased while movement from bottom to top decreased. The data indicate that gravistimulation induces polar movement of Ca toward the lower side of the root cap. An earlier report (Lee, Mulkey, Evans 1983 Science 220: 1375-1376) from this laboratory showed that artificial establishment of calcium gradients at the root tip can cause gravitropic-like curvature. Together, the two studies indicate that Ca plays a key role in linking gravistimulation to the gravitropic growth response in roots.

Journal Article↗
PubMed1990

Soybean Roots Retain the Seed Urease Isozyme Synthesized during Embryo Development.

Roots of young soybean (Glycine max [L.] Merr.) plants (up to 25 days old) contain two distinct urease isozymes, which are separable by hydroxyapatite chromatography. These two urease species (URE1 and URE2) differ in: (a) electrophoretic mobility in native gels, (b) pH dependence, and (c) recognition by a monoclonal antibody specific for the seed ("embryo-specific") urease. By these parameters root URE1 urease is similar to the abundant embryo-specific urease isozyme, while root URE2 resembles the "ubiquitous" urease which has previously been found in all soybean tissues examined (leaf, embryo, seed coat, and cultured cells). The embryo-specific and ubiquitous urease isozymes are products of the Eu1 and Eu4 structural genes, respectively. Roots of the eu1-sun/eu1-sun genotype, which lacks the embryo-specific urease (i.e. ;seed urease-null'), contain no URE1 urease activity. Roots of eu4/eu4, which lacks ubiquitous urease, lack the URE2 (leaflike) urease activity. From these genetic and biochemical criteria, then, we conclude that URE1 and URE2 are the embryo-specific and ubiquitous ureases, respectively. Adventitious roots generated from cuttings of any urease genotype lack URE1 activity. In seedling roots the seedlike (URE1) activity declines during development. Roots of 3-week-old plants contain 5% of the total URE1 activity of the radicle of 4-day-old seedlings, which, in turn, has approximately the same urease activity level as the dormant embryonic axis. The embryo-specific urease incorporates label from [(35)S]methionine during embryo development but not during germination, indicating that there is no de novo synthesis of the embryo-specific (URE1) urease in the germinating root. We conclude that the seedlike urease (URE1) found in roots of young soybean plants is a remnant of the Eu1-encoded, abundant, embryo-specific urease which accumulates in the embryonic root axis during seed development.

Journal Article↗
PubMed1992

Effect of inhibition of abscisic Acid accumulation on the spatial distribution of elongation in the primary root and mesocotyl of maize at low water potentials.

Previous work showed that accumulation of endogenous abscisic acid (ABA) acts both to maintain primary root growth and inhibit shoot growth in maize seedlings at low water potentials (psi(w)) (IN Saab, RE Sharp, J Pritchard, GS Voetberg [1990] Plant Physiol 93: 1329-1336). In this study, we have characterized the growth responses of the primary root and mesocotyl of maize (Zea mays L. cv FR27 x FRMo 17) to manipulation of ABA levels at low psi(w) with a high degree of spatial resolution to provide the basis for studies of the mechanism(s) of ABA action. In seedlings growing at low psi(w) and treated with fluridone to inhibit carotenoid (and ABA) biosynthesis, ABA levels were decreased in all locations of the root and mesocotyl growing zones compared with untreated seedlings growing at the same psi(w). In the root, low psi(w) (-1.6 megapascals) caused a shortening of the growing zone, as reported previously. The fluridone treatment was associated with severe inhibition of root elongation rate, which resulted from further shortening of the growing zone. In the mesocotyl, low psi(w) (-0.3 megapascal) also resulted in a shortened growing zone. In contrast with the primary root, however, fluridone treatment prevented most of the inhibition of elongation and the shortening of the growing zone. Final cell length measurements indicated that the responses of both root and mesocotyl elongation to ABA manipulation at low psi(w) involve large effects on cell expansion. Measurements of the relative changes in root and shoot water contents and dry weights after transplanting to a psi(w) of -0.3 megapascal showed that the maintenance of shoot elongation in fluridone-treated seedlings was not attributable to increased water or seed-reserve availability resulting from inhibition of root growth. The results suggest a developmental gradient in tissue responsiveness to endogenous ABA in both the root and mesocotyl growing zones. In the root, the capacity for ABA to protect cell expansion at low psi(w) appears to decrease with increasing distance from the apex. In the mesocotyl, in contrast, the accumulation of ABA at low psi(w) appears to become increasingly inhibitory to expansion as cells are displaced away from the meristematic region.

Journal Article↗
PubMed1983

Enumeration and Localization of N(2)-Fixing Bacteria Associated with Roots of Spartina alterniflora Loisel.

Numbers and possible locations of N(2)-fixing bacteria were investigated in roots of Spartina alterniflora Loisel, which support nitrogenase activity in the undisturbed native habitat. N(2)-fixing bacteria were recovered in cultures both from S. alterniflora roots and from the surrounding sediment, and they formed a greater proportion of the bacteria recovered from root homogenates than from salt-marsh sediment. N(2)-fixing bacteria were recovered in high numbers from the rhizoplane of S. alterniflora after roots were treated with 1 or 5% chloramine-T for 1 h or with 1% NaOCl for 1 or 2 h. Immersing S. alterniflora roots in 5% NaOCl for 1 h was more effective in distinguishing bacteria inside the roots since this treatment nearly eliminated N(2)-fixing bacteria recoverable from the rhizoplane, although high numbers of N(2)-fixing bacteria were recovered from homogenates of roots treated with 5% NaOCl for 1 h. However, this treatment was less effective with roots of Zea mays L. (Funks G4646) and Sorghum bicolor (L.) Moench (CK-60 A), indicating that techniques to surface sterilize roots should be evaluated for different plants. Bacteria were observed by light and electron microscopy inter- and intracellularly in the cortex and in the aerenchyma of S. alterniflora roots. This study clearly shows that bacteria, including N(2) fixers, colonize the interior of roots of S. alterniflora growing in a Chesapeake Bay, Maryland, salt marsh.

Journal Article↗
PubMed1999

Mapping of sugar and amino acid availability in soil around roots with bacterial sensors of sucrose and tryptophan

We developed a technique to map the availability of sugars and amino acids along live roots in an intact soil-root matrix with native microbial soil flora and fauna present. It will allow us to study interactions between root exudates and soil microorganisms at the fine spatial scale necessary to evaluate mechanisms of nitrogen cycling in the rhizosphere. Erwinia herbicola 299R harboring a promoterless ice nucleation reporter gene, driven by either of two nutrient-responsive promoters, was used as a biosensor. Strain 299RTice exhibits tryptophan-dependent ice nucleation activity, while strain 299R(p61RYice) expresses ice nucleation activity proportional to sucrose concentration in its environment. Both biosensors exhibited up to 100-fold differences in ice nucleation activity in response to varying substrate abundance in culture. The biosensors were introduced into the rhizosphere of the annual grass Avena barbata and, as a control, into bulk soil. Neither strain exhibited significant ice nucleation activity in the bulk soil. Both tryptophan and sucrose were detected in the rhizosphere, but they showed different spatial patterns. Tryptophan was apparently most abundant in soil around roots 12 to 16 cm from the tip, while sucrose was most abundant in soil near the root tip. The largest numbers of bacteria (determined by acridine orange staining and direct microscopy) occurred near root sections with the highest apparent sucrose or tryptophan exudation. High sucrose availability at the root tip is consistent with leakage of photosynthate from immature, rapidly growing root tissues, while tryptophan loss from older root sections may result from lateral root perforation of the root epidermis.

Journal Article↗
PubMed1991

Rhizobium lipopolysaccharide modulates infection thread development in white clover root hairs.

The interaction between Rhizobium lipopolysaccharide (LPS) and white clover roots was examined. The Limulus lysate assay indicated that Rhizobium leguminosarum bv. trifolii (hereafter called R. trifolii) released LPS into the external root environment of slide cultures. Immunofluorescence and immunoelectron microscopy showed that purified LPS from R. trifolii 0403 bound rapidly to root hair tips and infiltrated across the root hair wall. Infection thread formation in root hairs was promoted by preinoculation treatment of roots with R. trifolii LPS at a low dose (up to 5 micrograms per plant) but inhibited at a higher dose. This biological activity of LPS was restricted to the region of the root present at the time of exposure to LPS, higher with LPS from cells in the early stationary phase than in the mid-exponential phase, incubation time dependent, incapable of reversing inhibition of infection by NO3- or NH4+, and conserved among serologically distinct LPSs from several wild-type R. trifolii strains (0403, 2S-2, and ANU843). In contrast, infections were not increased by preinoculation treatment of roots with LPSs from R. leguminosarum bv. viciae strain 300, R. meliloti 102F28, or members of the family Enterobacteriaceae. Most infection threads developed successfully in root hairs pretreated with R. trifolii LPS, whereas many infections aborted near their origins and accumulated brown deposits if pretreated with LPS from R. meliloti 102F28. LPS from R. leguminosarum 300 also caused most infection threads to abort. Other specific responses of root hairs to infection-stimulating LPS from R. trifolii included acceleration of cytoplasmic streaming and production of novel proteins. Combined gas chromatography-mass spectroscopy and proton nuclear magnetic resonance analyses indicated that biologically active LPS from R. trifolii 0403 in the early stationary phase had less fucose but more 2-O-methylfucose, quinovosamine, 3,6-dideoxy-3-(methylamino)galactose, and noncarbohydrate substituents (O-methyl, N-methyl, and acetyl groups) on glycosyl components than did inactive LPS in the mid-exponential phase. We conclude that LPS-root hair interactions trigger metabolic events that have a significant impact on successful development of infection threads in this Rhizobium-legume symbiosis.

Blotting, Western↗