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Arabinogalactan-Proteins from Primary and Mature Roots of Radish (Raphanus sativus L.).

Organ-specific variations in blood group H-like activity were observed in developing radish plants. A temporary increase in serological activity was found to occur in the roots at the earlier stages of development. Arabinogalactan-proteins (AGPs) were isolated from primary and mature roots, and investigated for changes in their physicochemical properties, structure, and serological activities. These root AGPs were composed mainly of l-arabinose and d-galactose but were distinguishable from each other in their contents of l-fucose as well as of protein and hydroxyproline. The structures of the carbohydrate moieties of the root AGPs were essentially similar to those of AGPs isolated from seeds and mature leaves in that they consisted of consecutive (1-->3)-linked beta-d-galactosyl backbone chains having side chains of (1-->6)-linked beta-d-galactosyl residues, to which alpha-l-arabinofuranosyl residues were attached in the outer regions. One prominent feature of the primary root AGPs was that they contained appreciable amounts of l-fucose, which was presumably responsible for expression of the serological activity. In their immunological reactions with rabbit anti-radish leaf AGP antibody, the root AGPs were shown to share common antigenic determinant(s) with those of seed and leaf AGPs.

Journal Article↗

Tissue-specific expression and developmental regulation of cytochrome b561 genes in Arabidopsis thaliana and Raphanus sativus.

Ascorbate (Asc) is an essential molecule in many aspects of development and stress responses in plants and animals. Cytochromes b561 (cyts b561) are tightly coupled to Asc homeostasis. These proteins are found in mammalian tissues, where they are involved in the regeneration of Asc, serving the synthesis of catecholamine neurotransmitters, and in intestinal iron reduction. Plant genomes encode homologous membrane-associated, Asc-reducible cyts b561. The expression of these proteins in plants, however, has so far not been studied. We have now examined the expression of two Arabidopsis thaliana cyt b561-encoding genes-Artb561-1 and Artb561-2-using relative-quantitative RT-PCR and in situ hybridization (ISH) techniques. The genes show overlapping and distinct tissue- and organ-specific expression patterns. Transcripts of both genes are found in leaf epidermal cells, and expression seems to correlate with leaf maturation and cessation of cell elongation. Both genes are also expressed in the epidermal cell layer of stems and roots in the L1 layer of the shoot apex, in the vascular system of leaves, stems and roots, and in the root pericycle. In addition, Artb561-1 is expressed in the root cap, whereas Artb561-2 mRNA is found in the epidermis of lateral roots, in the root meristem, and in unfertilized ovules. These observations provide important information for the elucidation of the physiological function of cyts b561 in plants.

Journal Article↗

Analysis of plant genomes. IV. Isolation and characterization of satellite DNA components from two dicotyledons cucumber (Cucumis sativus) and radish (Raphanus sativus).

Satellite DNA fractions from cucumber and radish, two plants having low DNA contents and relatively small chromosomes, were isolated and characterized. Reassociation studies of satellite and total nuclear DNA showed that the satellite fractions in these two plants contain most of the rapidly reassociating DNA. Cucumber satellite I was found to contain one major component (70% of the total satellite) having a density of 1.706 g/cm3 and a Tm of 90.5 degrees C and a minor component with a density of 1.712 g/cm3 and a Tm of 93.5 degrees C. The complexity of the major component was estimated to be 3.8 X 10(5) daltons while that of the minor one was 12.9 X 10(7) daltons. Although cucumber satellite II banded as a single peak at a density of 1.700 g/cm3 in neutral CsCl gradients, it was observed to have a rather broad denaturation profile with a Tm of 86.5 degrees C. Its Cot curve was also broader than that of satellite I and one of its components (40% of the total) had a complexity of 5.8 X 10(5) daltons. Two satellite fractions were also observed in the case of radish DNA but only satellite I was isolated in a pure form and characterized. This radish satellite formed a sharp, symmetrical peak at a density of 1.698 g/cm3 in neutral CsCl gradients and underwent denaturation in a narrow temperature range of 6 to 7 degrees C. An analysis of the optical reassociation kinetics showed that this satellite contained a major and a minor component. The major component, which comprised 80% of the satellite, had a complexity of 12.9 X 10(5) daltons. Hybridization experiments revealed that the ribosomal DNA was present in satellite II.

Centrifugation, Density Gradient↗

Effects of semi-purified dietary fibers isolated from Lagenaria siceraria, Raphanus sativus and Lentinus edodes on fecal steroid excretions in rats.

Rats were fed diets, each of which contained 5% of the semi-purified dietary fiber (DF) preparations from three kinds of foods, continuously for a total of 7 weeks. The fat level was varied, and cholesterol (Chol) was added toward the end of the experimental period. After 7 weeks, decreases of serum and liver Chol concentrations were observed in the Shiitake (Lentinus edodes) group when compared to the cellulose (CP) group. In the absence of Chol, fecal excretions of several secondary bile acids and total bile acids were affected by the type of DF, showing an increase particularly in the Shiitake group. Total neutral steroid excretions were decreased in the test DF groups compared to the CP group at both fat levels, and were increased by Chol supplementation in all of the test DF groups. It was demonstrated that the effects of the three DF preparations on fecal steroid excretions somewhat differed depending on the dietary factors. In the absence of Chol, the lithocholic acid (LCA)/deoxycholic acid (DCA) ratio was significantly lowered in the test DF groups compared to the CP group by the fat level elevation, and was affected by the interaction of fiber with fat. This ratio responded more intensely to the dietary factors than the two other indices (composition ratios of fecal bile acids). However, this response was lost in the presence of Chol. These results suggest that further studies are warranted to examine if the intense response of the LCA/DCA ratio to the tested DF preparations and fat in the present study will also be shown to other DF and what significance it has in evaluating the function of DF.

Animals↗

[Effect of azacytidine on expression of traits concomitant with tumor formation in the radish (Raphanus sativus) in vitro].

The effect of azacytidine, a demethylating agent, on the expression of traits concomitant with tumor formation was studied in inbred radish (Rhaphanus sativus) lines carrying genomic sequences homologous to the tmr/tml genes of Agrobacteriium tumefaciens. AzaC was found to have no effect on the traits studied, which provided evidence that the capacity for tumor formation in radish lines does not depend on the level of methylation of these sequences.

Agrobacterium tumefaciens↗

Mechanisms of differential pollen donor performance in wild radish, Raphanus sativus (Brassicaceae).

In order to understand the characters on which sexual selection might operate in plants, it is critical to assess the mechanisms by which pollen competition and mate choice occur. To address this issue we measured a number of postpollination characters, ranging from pollen germination and pollen tube growth to final seed paternity, in wild radish. Crosses were performed using four pollen donors on a total of 16 maternal plants (four each from four families). Maternal plants were grown under two watering treatments to evaluate the effects of maternal tissue on the process of mating. The four pollen donors differed significantly in number of seeds sired and differed overall in the mating characters measured. However, it was difficult to associate particular mechanistic characters with ability to sire seeds, perhaps because of interactions among pollen donors within styles or among pollen donors and maternal plants. The process of pollen tube growth and fertilization differed substantially among maternal watering treatments, with many early events occurring more quickly in stressed plants. Seed paternity, however, was somewhat more even among pollen donors used on stressed maternal plants, suggesting that when maternal tissue is more competent, mating is slowed and is more selective.

Journal Article↗

Analysis of two novel classes of plant antifungal proteins from radish (Raphanus sativus L.) seeds.

Two novel classes of antifungal proteins were isolated from radish seeds. The first class consists of two homologous proteins (Rs-AFP1 and Rs-AFP2) that were purified to homogeneity. They are highly basic oligomeric proteins composed of small (5-kDa) polypeptides that are rich in cysteine. Both Rs-AFPs have a broad antifungal spectrum and are among the most potent antifungal proteins hitherto characterized. In comparison with many other plant antifungal proteins, the activity of the Rs-AFPs is less sensitive to the presence of cations. Moreover, their antibiotic activity shows a high degree of specificity to filamentous fungi. The amino-terminal regions of the Rs-AFPs show homology with the derived amino acid sequences of two pea genes specifically induced upon fungal attack, to gamma-thionins and to sorghum alpha-amylase inhibitors. The radish 2S storage albumins were identified as the second novel class of antifungal proteins. All isoforms inhibit growth of different plant pathogenic fungi and some bacteria. However, their antimicrobial activities are strongly antagonized by cations.

Amino Acid Sequence↗