Immunological Evidence of Thaumatin-Like Proteins during Tobacco Floral Differentiation.

Tobacco proteins that share homology with thaumatin, a sweet protein of Thaumatococcus daniellii Benth., are produced in various physiological situations such as pathogenesis-related stress or water deficit stress. Using purified polyclonal anti-thaumatin antibodies, we have detected other thaumatin-like proteins in tobacco (Nicotiana tabacum var Samsun) that have been related with floral differentiation. Thaumatin-like proteins with apparent molecular masses of 42.6, 31.6, and 26.3 kilodaltons were found in immature and mature flower organs in vivo, and others of 46.7, 41.7, and 27.5 kilodaltons were exclusively detected in thin cell layer explants forming flowers. In situ immunolocalization revealed their synthesis in newly differentiated floral meristems, in tracheids, and in parenchyma cells.

An Examination of the Plastid DNA of Hypohaploid Nicotiana plumbaginifolia Plants.

DNA was extracted from different morphological types of hypohaploid Nicotiana plumbaginifolia plants. The cellular levels of chloroplast DNA (expressed as percent of total DNA) were found to be approximately two- to threefold higher in two albino hypohaploids than in a green hypohaploid. The level of chloroplast DNA in the green hypohaploid was not significantly different from either in vitro or in vivo grown haploid N. plumbaginifolia plants. Molecular hybridization with DNA probes for the large subunit of ribulose bisphosphate carboxylase from spinach and with Pvull fragments representing the entire Nicotiana tabacum chloroplast genome revealed no gross qualitative differences in the chloroplast DNAs of hypohaploid plants. Based on these observations we have concluded that the lack of chloroplast function observed in the albino forms of hypohaploid N. plumbaginifolia plants is not due to changes in the chloroplast genome.

Light- and sugar-mediated control of direct de novo flower differentiation from tobacco thin cell layers.

The nature of organs neoformed from tobacco (Nicotiana tabacum cv Samsun) thin cell layers is influenced by the quantity of light supplied and on the sequence of this supply. It is observed that glucose exhibits similar effects. In the presence of glucose at 167 millimolar, continuous light of 50 watts per square meter is required for optimal flower differentiation in vitro. However, 50 watts per square meter irradiance limited to 6 days is sufficient to trigger flower formation in 80% of the explants provided that light is applied from day 6 to day 11 of culture. This critical phase may correspond to the initiation phase during which soluble sugars are mainly needed as carbon energy source rather than as osmoregulators. Under continuous or precise sequential sugar deprivations, either no organogenesis occurs, or abnormal structures or buds are formed. Therefore, light per se is not sufficient to induce flower differentiation. Conversely, a specific quantitative combination of glucose and sucrose almost substitutes for the light requirement for differentiation of anthers and pistils. These observations suggest that, during the sequence of events leading to flower differentiation, light acts on energy-dependent sugar uptake and metabolism and on the increase of reducing potential of chloroplasts.

Distribution of Lectins in Tissues, Derived Callus, and Roots of Psophocarpus tetragonolobus (Winged Bean).

The distribution of lectin in parental tissues, roots formed de novo from parental stem tissue, and derived callus cells of Psophocarpus tetragonolobus has been measured by hemagglutinating activity and radioimmunoassay. The antisera used for the radioimmunoassay was raised in rabbits to lectin isolated from seeds by affinity chromatography using insolubilized hog gastric mucin. The distribution of lectin in buffer extracts of the tissues (or cells) and the extracellular medium favors the tissues for in vitro grown roots, regardless of the culture conditions used. The lectin content of the extracellular medium is more significant for callus, regardless of its conditions of culture. The lectin activity of extracts of in vitro grown roots was higher than that of mature roots from whole plants. Differences in relative levels of lectin activity measured by hemagglutination and by radioimmunoassay, and differences in saccharide inhibition of hemagglutination, suggest the presence of multiple lectins in extracts of different tissues.