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      Structural identification of the lipo-chitin oligosaccharide nodulation signals of Rhizobium loti.

      Molecular Microbiology
      Bacterial Proteins, metabolism, Carbohydrate Sequence, Chitin, analogs & derivatives, chemistry, physiology, Chromatography, High Pressure Liquid, Chromatography, Thin Layer, Fabaceae, microbiology, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Plants, Medicinal, Rhizobium, Spectrometry, Mass, Fast Atom Bombardment, Symbiosis

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          Abstract

          Rhizobium loti is a fast-growing Rhizobium species that has been described as a microsymbiont of plants of the genus Lotus. Nodulation studies show that Lotus plants are nodulated by R. loti, but not by most other Rhizobium strains, indicating that R. loti produces specific lipo-chitin oligosaccharides (LCOs) which are necessary for the nodulation of Lotus plants. The LCOs produced by five different Rhizobium loti strains have been purified and were shown to be N-acetylglucosamine pentasaccharides of which the non-reducing residue is N-methylated and N-acylated with cis-vaccenic acid (C18:1) or stearic acid (C18:O) and carries a carbamoyl group. In one R. loti strain, NZP2037, an additional carbamoyl group is present on the non-reducing terminal residue. The major class of LCO molecules is substituted on the reducing terminal residue with 4-O-acetylfucose. Addition of LCOs to the roots of Lotus plants results in abundant distortion, swelling and branching of the root hairs, whereas spot inoculation leads to the formation of nodule primordia.

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          Natural abundance nitrogen-15 NMR by enhanced heteronuclear spectroscopy

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            Application of phase sensitive two-dimensional correlated spectroscopy (COSY) for measurements of 1H-1H spin-spin coupling constants in proteins.

            Two-dimensional correlated spectroscopy (COSY) is used for measurements of proton-proton spin-spin coupling constants in protein 1H NMR spectra. High digital resolution along the frequency axis omega 2 is achieved by placing the carrier frequency in the center of the spectrum, using quadrature detection in both dimensions and presenting the spectrum in the phase sensitive mode. Compared to other techniques for studies of spin-spin coupling constants, COSY provides greatly improved spectral resolution. This is illustrated by experiments with H2O solutions of the small globular protein BUSI IIA (bull seminal inhibitor IIA).
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              Lotus japonicus, an autogamous, diploid legume species for classical and molecular genetics

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