RESUMEN
Interactions between legumes and rhizobia are controlled by the sequential exchange of symbiotic signals. Two different techniques, 2D-PAGE electrophoresis and differential display were used to study the effects of rhizobial signals on legume development. Application of variously substituted lipo-oligo-saccharidic Nod-factors to roots of Vigna unguiculata resulted in changes in the phosphorylation patterns of microsomal proteins. Reliable amino-acid sequences were obtained for one Nod-factor enhanced protein which was highly homologous to the 57-kDa subunit from Arabidopsis thaliana vacuolar membrane H(+)-ATPase. Immuno-blotting techniques demonstrated that Nod-factors cause rapid and massive increases of this enzyme in treated roots, suggesting that H(+)-ATPases play symbiotic roles. Concomitantly, we used differential display (DD) techniques on mRNA isolated from root-hairs to analyse early root responses to NGR234. Significant matches of several DD clones to known sequences were found. Clone D2.62 was homologous to a multitude of receptor kinases including S receptor-like kinases of A. thaliana and clone D4.1 showed similarities to Lotus japonicus phosphatidylinositol transfer-like protein III and late nodulin 16. Independent confirmatory analyses of these differentially expressed clones indicated expression at very low levels.
Asunto(s)
Fabaceae/microbiología , Rhizobium/fisiología , Fraccionamiento Celular/métodos , Membrana Celular/metabolismo , Clonación Molecular , Electroforesis en Gel Bidimensional , Fabaceae/genética , Fabaceae/metabolismo , Perfilación de la Expresión Génica , Immunoblotting , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Proteómica , ARN Mensajero/metabolismo , Análisis de Secuencia de Proteína , Análisis de Secuencia de ARN , Transducción de Señal , SimbiosisRESUMEN
A modified method for isolation of toxicologically significant substances after acid hydrolysis of biological material is proposed. Drugs and narcotics of different groups were identified, which are not disintegrated after acid hydrolysis of biological material and are isolated in high amounts.