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M Schultze

Publications and source records attributed to M Schultze.

At least 19 recordsLinked to original sources
PubMed2006

Control of electron localization in molecular dissociation.

We demonstrated how the subcycle evolution of the electric field of light can be used to control the motion of bound electrons. Results are presented for the dissociative ionization of deuterium molecules (D2 --> D+ + D), where asymmetric ejection of the ionic fragment reveals that light-driven intramolecular electronic motion before dissociation localizes the electron on one of the two D+ ions in a controlled way. The results extend subfemtosecond electron control to molecules and provide evidence of its usefulness in controlling reaction dynamics.

Journal Article↗
PubMed2005

Modification of a continuous flow method for analysis of trace amounts of nitrate in iron-rich sediment pore-waters of mine pit lakes.

Nitrate was analysed in pore-waters with high ionic strength. Extremely high concentrations of dissolved ferrous iron interfere common analytical methods. The automatic photometrical procedure based on the cadmium reduction method is often used for analysis of nitrate in water samples (continuous flow analysis CFA). An integrated dialysis usually serves for sample dilution and (or) sample purification (from particles). Iron was precipitated as iron hydroxide due to the imidazole buffer system (pH 7.5). The dialysis membrane is an effective barrier for iron hydroxide particles to prevent interferences within the cadmium column or the flow-cell. However, dialysis membrane is blocked successively after analysis of several iron-rich pore-water samples by agglomeration of precipitated iron. The blocking of nitrate diffusion through the dialysis membrane is tantamount to a decrease of analytical sensitivity to recognise by decreasing photometrical extinction. Minimising the iron deposition within the dialyser solved the problem. A simple modification of the CFA apparatus was found to keep the analytical sensitivity nearly constant: The mixing coil for the imidazole buffer was considerably elongated. Nearly all iron hydroxide was deposited at the glass coil surface installed before the dialyser. At least 50 iron-rich samples could be analysed within one sample queue with a loss of sensitivity <10%. The recovery of nitrate was about 95%, demonstrated by spiking experiments.

Chemical Precipitation↗
PubMed2000

N-deacetylation of Sinorhizobium meliloti Nod factors increases their stability in the Medicago sativa rhizosphere and decreases their biological activity.

Nod factors excreted by rhizobia are signal molecules that consist of a chitin oligomer backbone linked with a fatty acid at the nonreducing end. Modifications of the Nod factor structures influence their stability in the rhizosphere and their biological activity. To test the function of N-acetyl groups in Nod factors, NodSm-IV(C16:2,S) from Sinorhizobium meliloti was enzymatically N-deacetylated in vitro with purified chitin deacetylase from Colletotrichum lindemuthianum. A family of partially and completely deacetylated derivatives was produced and purified. The most abundant chemical structures identified by mass spectrometry were GlcN(C16:2)-GlcNAc-GlcNH2-GlcNAc(OH)(S), GlcN(C16,2)-GlcNAc-GlcNH2-GlcNH2(OH)(S), and GlcN(C16:2)-GlcNH2-GlcNH2-GlcNH2(OH)(S). In contrast to NodSm-IV(C16:2,S), the purified N-deacetylated derivatives were stable in the rhizosphere of Medicago sativa, indicating that the N-acetyl groups make the carbohydrate moiety of Nod factors accessible for glycosyl hydrolases of the host plant. The N-deacetylated derivatives displayed only a low level of activity in inducing root hair deformation. Furthermore, the N-deacetylated molecules were not able to stimulate Nod factor degradation by M. sativa roots, a response elicited by active Nod factors. These data show that N-acetyl groups of Nod factors are required for biological activity.

Amidohydrolases↗
PubMed2000

Nod factors of Rhizobium leguminosarum bv. viciae and their fucosylated derivatives stimulate a nod factor cleaving activity in pea roots and are hydrolyzed in vitro by plant chitinases at different rates.

Nod factors (NFs) are rhizobial lipo-chitooligosaccharide signals that trigger root nodule development in legumes. Modifications of NF structures influence their biological activity and affect their degradation by plant chitinases. Nodulation of certain pea cultivars by Rhizobium leguminosarum bv. viciae requires modification of NFs at the reducing end by either an O-acetyl or a fucosyl group. Fucosylated NFs were produced by an in vitro reaction with NodZ fucosyltransferase and purified. Their biological activity on pea was tested by measuring their capacity to stimulate the activity of a hydrolase that cleaves NFs. Nonmodified and fucosylated NFs displayed this activity at nano- to picomolar concentrations, while a sulfated NF from Sinorhizobium meliloti was inactive. In an additional series of experiments, the stability of non-modified and fucosylated NFs in the presence of purified tobacco chitinases was compared. The presence of the fucosyl group affected the degradation rates and the accessibility of specific cleavage sites on the chitooligosaccharide backbone. These results suggest that the fucosyl group in NFs also weakens the interaction of NFs with certain chitinases or chitinase-related proteins in pea roots.

Chitinases↗
PubMed2000

How alfalfa root hairs discriminate between Nod factors and oligochitin elicitors.

Using ion-selective microelectrodes, the problem of how signals coming from symbiotic partners or from potential microbial intruders are distinguished was investigated on root hairs of alfalfa (Medicago sativa). The Nod factor, NodRm-IV(C16:2,S), was used to trigger the symbiotic signal and (GlcNAc)(8) was selected from (GlcNAc)(4-8), to elicit defense-related reactions. To both compounds, root hairs responded with initial transient depolarizations and alkalinizations, which were followed by a hyperpolarization and external acidification in the presence of (GlcNAc)(8). We propose that alfalfa recognizes tetrameric Nod factors and N-acetylchitooligosaccharides (n = 4-8) with separate perception sites: (a) (GlcNAc)(4) and (GlcNAc)(6) reduced the depolarization response to (GlcNAc)(8), but not to NodRm-IV(C16:2, S); and (b) depolarization and external alkalization were enhanced when NodRm-IV(C16:2,S) and (GlcNAc)(8) were added jointly without preincubation. We suggest further that changes in cytosolic pH and Ca(2+) are key events in the transduction, as well as in the discrimination of signals leading to symbiotic responses or defense-related reactions. To (GlcNAc)(8), cells responded with a cytosolic acidification, and they responded to NodRm-IV(C16:2,S) with a sustained alkalinization. When both agents were added jointly, the cytosol first alkalized and then acidified. (GlcNAc)(8) and NodRm-IV(C16:2,S) transiently increased cytosolic Ca(2+) activity, whereby the response to (GlcNAc)(8) exceeded the one to NodRm-IV(C16:2,S) by at least a factor of two.

Calcium↗
PubMed1999

Nod factors modulate the concentration of cytosolic free calcium differently in growing and non-growing root hairs of Medicago sativa L.

Using Ca(2+)-selective microelectrodes, the concentration of free calcium ([Ca(2+)]) in the cytosol has been measured in root hair cells of Medicago sativa L. in the presence of nodulation (Nod) factors. Growing root hairs of M. sativa displayed a steep apical [Ca(2+)] gradient, i.e. 604-967 nM in the tip compared with 95-235 nM in the basal region. When tested within the first 5 to 10 µm of the tip, addition of NodRm-IV(C16:2,S) decreased the cytosolic [Ca(2+)], whereas an increase was observed when tested behind the tip. Overall, this led to a partial dissipation of the [Ca(2+)] gradient. The Ca(2+) response was specific: it was equally well observed in the presence of NodRm-IV(Ac,C16:2,S), reduced with NodRm-IV(C16:0,S), but not with chitotetraose, the nonactive glucosamine backbone. In contrast to growing root hairs, non-growing root hairs without a tip-to-base cytosolic [Ca(2+)] gradient responded to NodRm-IV(C16:2,S) with an increase in cytosolic [Ca(2+)] at the tip as well as at the root hair base. We suggest that the response to Nod factors depends on the stage of development of the root hairs, and that changes in cytosolic [Ca(2+)] may play different roles in Nod-factor signaling: changes of cytosolic [Ca(2+)] in the apical part of the root hair may be related to root hair deformation, while the increase in [Ca(2+)] behind the tip may be essential for the amplification of the Nod signal, for its propagation and transduction to trigger downstream events.

Journal Article↗
PubMed1999

Elevation of the cytosolic free [Ca2+] is indispensable for the transduction of the Nod factor signal in alfalfa.

In root hairs of alfalfa (Medicago sativa), the requirement of Ca(2+) for Nod factor signaling has been investigated by means of ion-selective microelectrodes. Measured 50 to 100 microm behind the growing tip, 0.1 microM NodRm-IV(C16:2,S) increased the cytosolic free [Ca2+] by about 0.2 pCa, while the same concentration of chitotetraose, the nonactive glucosamine backbone, had no effect. We demonstrate that NodRm-IV(C16:2,S) still depolarized the plasma membrane at external Ca(2+) concentrations below cytosolic values if the free EGTA concentration remained low (</=0.01 mM). Externally added Sr(2+) was able to replace Ca(2+), and to some extent even enhanced the Nod-factor-induced depolarization, whereas with Mg(2+) it was decreased. This suggests that the Nod factor response is triggered by Ca(2+) from external stores. The addition of the endomembrane Ca(2+)-ATPase inhibitor 2,5-di(t-butyl)-1, 4-benzohydroquinone, which presumably mobilizes Ca(2+) from Ins(1,4, 5)P(3)-sensitive stores, mimicked the Nod factor response, i.e. increased the cytosolic free [Ca2+], triggered Cl(-)-efflux, depolarized the plasma membrane, and alkalized the root hair space. In all cases a refractory state toward Nod factor perception was produced, indicating a shortcut of Nod factor signal transduction by releasing Ca(2+) from internal stores. These latter results strongly support the idea that an elevation of cytosolic free [Ca2+] is indispensable for the transduction of the Nod factor signal, which is consistent with the role of Ca(2+) as a second messenger.

Calcium↗
PubMed1998

Purification and characterization of a novel chitinase-lysozyme, of another chitinase, both hydrolysing Rhizobium meliloti Nod factors, and of a pathogenesis-related protein from Medicago sativa roots.

The symbiosis between Rhizobium meliloti and Medicago sativa (Leguminosae) involves the interaction of lipochito-oligosaccharides (Nod factors) excreted by bacteria with specific proteins of the host plant. The cleavage of Nod factors can be used as an enzymic assay to identify novel hydrolytic enzymes. Here a soluble extract of 3-day-old roots was fractionated by anion exchange, affinity chromatography, gel filtration and native electrophoresis. Two acidic chitinases (pI 4.6-5.4), CHIT24 and CHIT36, designated in accordance with their molecular mass in kDa, were separated. CHIT24 cleaves all tested Nod factors to produce lipotrisaccharides with the preference NodRm-V(S)>NodRm-IV >NodRm-IV(S)>=NodRm-IV(Ac,S); it also hydrolyses colloidal 3H-chitin and has lysozyme activity. The kinetics of Nod factor degradation by CHIT24 depends on substrate structural parameters, namely the length of the oligosaccharide chain and sulphation (S) at the reducing end, but not much on acetylation (Ac) at the non-reducing end. The 25-residue N-terminal sequence of CHIT24 has no similarity with known chitinases or lysozymes, indicating that it is a novel type of hydrolase. CHIT36 also hydrolyses NodRm-V(S) into NodRm-III, but it is inactive towards NodRm-IV(S) and NodRm-IV(Ac,S) formed by R. meliloti. Finally, a 17 kDa protein, P17, was co-purified with CHIT24. It neither degrades Nod factors nor exhibits lysozyme activity and shows complete identity, at the 15-residue N-terminal sequence, with a class 10 pathogenesis-related protein, PR-10.

Amino Acid Sequence↗
PubMed1998

Regulation of symbiotic root nodule development.

Symbiosis between rhizobia and leguminous plants leads to the formation of N2-fixing root nodules. The interaction of rhizobia and plants shows a high degree of host specificity based on the exchange of chemical signals between the symbiotic partners. The plant signals, flavonoids exuded by the roots, activate the expression of nodulation genes, resulting in the production of the rhizobial lipochitooligosaccharide signals (Nod factors). Nod factors act as morphogens that, under conditions of nitrogen limitation, induce cells within the root cortex to divide and to develop into nodule primordia. This review focuses on how the production of Nod factors is regulated, how these signals are perceived and transduced by the plant root, and the physiological conditions and plant factors that control the early events leading to root nodule development.

Fabaceae↗
PubMed1996

The role of lipochitooligosaccharides in root nodule organogenesis and plant cell growth.

Lipochitooligosaccharides (Nod signals) excreted by rhizobia induce the formation of symbiotic root nodules in leguminous plants. This process is host plant specific, depending on the structural modifications of Nod signals. Rapid responses of plant roots in single cell assays have provided powerful tools in dissecting Nod signal transduction pathways and in elucidating the molecular basis of host specificity. Recent findings indicate that lipochitooligosaccharides, as well as symbiosis-related genes, also function in non legumes, pointing to a general role for these elements in plant morphogenesis.

Antigens, Bacterial↗
PubMed1995

In vitro sulfotransferase activity of Rhizobium meliloti NodH protein: lipochitooligosaccharide nodulation signals are sulfated after synthesis of the core structure.

The Rhizobium common nod gene products NodABC are involved in the synthesis of the core lipochitooligosaccharide (Nod factor) structure, whereas the products of the host-specific nod genes are necessary for diverse structural modifications, which vary in different Rhizobium species. The sulfate group attached to the Rhizobium meliloti Nod signal is necessary for activity on the host plant alfalfa, while its absence renders the Nod factor active on the non-host plant vetch. This substituent is therefore a major determinant of host specificity. The exact biosynthetic pathway of Nod factors has not been fully elucidated. In particular, it is not known why some chemical modifications are introduced with high fidelity whereas others are inaccurate, giving rise to a family of different Nod factor structures produced by a single Rhizobium strain. Using protein extracts and partially purified recombinant NodH protein obtained from Escherichia coli expressing the R. meliloti nodH gene, we demonstrate here NodH-dependent in vitro sulfotransferase activity. Kinetic analyses with Nod factors, chitooligosaccharides, and their deacetylated derivatives revealed that Nod factors are the preferred substrate for the sulfate transfer. Moreover, the tetrameric Nod factor, NodRm-IV, was a better substrate than the trimer, NodRm-III, or the pentamer, NodRm-V. These data suggest that the core lipochitooligosaccharide structure must be synthesized prior to its host-specific modification with a sulfate group. Since in R. meliloti tetrameric Nod factors are the most abundant and the most active ones, high affinity of NodH for the appropriate tetrameric substrate guarantees its modification and thus contributes to the fidelity of host-specific behavior.

Bacterial Proteins↗
PubMed1994

Activation of the cell cycle machinery and the isoflavonoid biosynthesis pathway by active Rhizobium meliloti Nod signal molecules in Medicago microcallus suspensions.

We have shown that treatment of Medicago microcallus suspensions with the cognate Rhizobium meliloti Nod signal molecule NodRm-IV(C16:2,S) can modify gene expression both qualitatively and quantitatively. At concentrations of 10(-6) - 10(-9) M, this host specific plant morphogen but not the inactive non-sulfated molecule stimulated cell cycle progression as indicated by the significantly enhanced thymidine incorporation, elevated number of S phase cells, increase in kinase activity of the p34cdc2-related complexes and enhancement of the level of expression of several cell cycle marker genes, the histone H3-1, the cdc2Ms and the cyclin cycMs2. The presented data suggest that at least part of the physiological role of the Nod factor may be linked to molecular events involved in the control of the plant cell division cycle. In situ hybridization experiments with antisense H3-1 RNA probe indicated that only certain cells of the calli were able to respond to the Nod factor. High (10(-6) M) but not low (10(-9) M) concentrations of the active Nod factors induced the expression of the isoflavone reductase gene (IFR), a marker gene of the isoflavonoid biosynthesis pathway in most callus cells. Our results indicate that Medicago cell responses to the Nod signal molecules can be investigated in suspension cultures.

CDC2 Protein Kinase↗
PubMed1994

Alfalfa Enod12 genes are differentially regulated during nodule development by Nod factors and Rhizobium invasion.

MsEnod12A and MsEnod12B are two early nodulin genes from alfalfa (Medicago sativa). Differential expression of these genes was demonstrated using a reverse transcription-polymerase chain reaction approach. MsEnod12A RNA was detected only in nodules and not in other plant tissues. In contrast, MsEnod12B transcripts were found in nodules and also at low levels in roots, flowers, stems, and leaves. MsEnod12B expression was enhanced in the root early after inoculation with the microsymbiont Rhizobium meliloti and after treatment with purified Nod factors, whereas MsEnod12A induction was detected only when developing nodules were visible. In situ hybridization showed that in nodules, MsEnod12 expression occurred in the infection zone. In empty Fix- nodules the MsEnod12A transcript level was much reduced, and in spontaneous nodules it was not detectable. These data indicate that MsEnod12B expression in roots is related to the action of Nod factors, whereas MsEnod12A expression is associated with the invasion process in nodules. Therefore, alfalfa possesses different mechanisms regulating MsEnod12A and MsEnod12B expression.

Amino Acid Sequence↗
PubMed1993

[References for after-care of tumor patients].

Recent publications have questioned the benefit of extensive routine follow-up measures in all patients that have undergone treatment for head and neck cancer. Both the poor detection rate of asymptomatic early tumor lesions and the severe limitations that are encountered with regard to further treatment options require an individual follow-up protocol mainly depending on site, size, and treatment of the respective tumor. The objective is to increase the efficacy of the follow-up in head and neck cancer patients carried out in both clinics and private practice.

Aftercare↗
PubMed1992

Rhizobium meliloti produces a family of sulfated lipooligosaccharides exhibiting different degrees of plant host specificity.

We have shown that a Rhizobium meliloti strain overexpressing nodulation genes excreted high amounts of a family of N-acylated and 6-O-sulfated N-acetyl-beta-1,4-D-glucosamine penta-, tetra-, and trisaccharide Nod factors. Either a C(16:2) or a C(16:3) acyl chain is attached to the nonreducing end subunit, whereas the sulfate group is bound to the reducing glucosamine. One of the tetrasaccharides is identical to the previously described NodRm-1 factor. The two pentasaccharides as well as NodRm-1 were purified and tested for biological activity. In the root hair deformation assay the pentasaccharides show similar activities on the host plants Medicago sativa and Melilotus albus and on the non-host plant Vicia sativa at a dilution of up to 0.01-0.001 microM, in contrast to NodRm-1, which displays a much higher specific activity for Medicago and Melilotus than for Vicia. The active concentration range of the pentasaccharides is more narrow on Medicago than on Melilotus and Vicia. In addition to root hair deformation, the different Nod factors were shown to induce nodule formation on M. sativa. We suggest that the production of a series of active signal molecules with different degrees of specificity might be important in controlling the symbiosis of R. meliloti with several different host plants or under different environmental conditions.

Bacterial Adhesion↗
PubMed1992

Rhizobium nodM and nodN genes are common nod genes: nodM encodes functions for efficiency of nod signal production and bacteroid maturation.

Earlier, we showed that Rhizobium meliloti nodM codes for glucosamine synthase and that nodM and nodN mutants produce strongly reduced root hair deformation activity and display delayed nodulation of Medicago sativa (Baev et al., Mol. Gen. Genet. 228:113-124, 1991). Here, we demonstrate that nodM and nodN genes from Rhizobium leguminosarum biovar viciae restore the root hair deformation activity of exudates of the corresponding R. meliloti mutant strains. Partial restoration of the nodulation phenotypes of these two strains was also observed. In nodulation assays, galactosamine and N-acetylglucosamine could substitute for glucosamine in the suppression of the R. meliloti nodM mutation, although N-acetylglucosamine was less efficient. We observed that in nodules induced by nodM mutants, the bacteroids did not show complete development or were deteriorated, resulting in decreased nitrogen fixation and, consequently, lower dry weights of the plants. This mutant phenotype could also be suppressed by exogenously supplied glucosamine, N-acetylglucosamine, and galactosamine and to a lesser extent by glucosamine-6-phosphate, indicating that the nodM mutant bacteroids are limited for glucosamine. In addition, by using derivatives of the wild type and a nodM mutant in which the nod genes are expressed at a high constitutive level, it was shown that the nodM mutant produces significantly fewer Nod factors than the wild-type strain but that their chemical structures are unchanged. However, the relative amounts of analogs of the cognate Nod signals were elevated, and this may explain the observed host range effects of the nodM mutation. Our data indicate that both the nodM and nodN genes of the two species have common functions and confirm that NodM is a glucosamine synthase with the biochemical role of providing sufficient amounts of the sugar moiety for the synthesis of the glucosamine oligosaccharide signal molecules.

Acetylglucosamine↗
PubMed1990

The reverse transcriptase gene of cauliflower mosaic virus is translated separately from the capsid gene.

Cauliflower mosaic virus (CaMV) possesses start codons at the beginning of its reverse transcriptase (RT) gene (ORF V) suggesting that, unlike in retroviruses and retrotransposons, it is translated independently from the capsid gene (ORF IV). To test this hypothesis a mutational analysis of the CaMV ORF IV/V overlapping region was performed. Mutants in which both ORFs are separated by stop codons in all three reading frames are viable and stable, while mutations affecting the first two AUG codons of ORF V are either lethal or unstable, giving rise to true and second site reversions. Mutants in which the AUG codons were replaced by ACG or AAG reverted only slowly and ACG could direct the synthesis of small amounts of reporter enzyme in transfected plant protoplasts, showing that this codon can act in plant cells as a weak start codon. CaMV has apparently developed a strategy for translation of the RT gene different from that in retroviruses and retrotransposons, but similar to that of hepadnaviruses, another group of pararetroviruses. The separate translation of the RT gene as a common feature of pararetroviruses might reflect the difference in their life cycle in comparison with retroviruses.

Base Sequence↗