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PubMed2002

Cross-resistance to imidazolinone herbicides in chlorsulfuron-resistant Raphanus raphanistrum.

Raphanus raphanistrum L has evolved widespread resistance to sulfonylureas in the Western Australia (WA) wheat belt. With the introduction of imidazolinone-tolerant (IT) wheat (Tritcum aestivum L) and IT canola (Brassica napus L) in the WA wheat belt, it is important to understand the status of cross-resistance in this weed to sulfonylurea and imidazolinone (Imi) herbicides. A study was conducted to examine cross-resistance between chlorsulfuron and Imi herbicides (a mixture of imazapic and imazapyr) in 46 R raphanistrum populations collected from across the WA wheat belt. Plants were treated with herbicides and assessed for phytotoxicity under glasshouse conditions. Of the 46 R raphanistrum populations, 32 were resistant to chlorsulfuron and four were resistant to imazapic + imazapyr. Of the 70% chlorsulfuron-resistant populations, 13% showed cross-resistance to imazapic + imazapyr. However, the cross-resistant populations treated with imazapic + imazapyr showed a lower resistance level than the chlorsulfuron-treated populations. These results suggest that weed populations with such cross-resistance will not be controlled effectively by Imi herbicides. Although the resistance levels of the cross-resistant populations to Imi herbicides were low, the cross-resistance levels of R raphanistrum should be determined before growing IT crops, particularly IT canola.

Dose-Response Relationship, Drug↗

PubMed2005

Antioxidant effect of squeezed juice from black radish (Raphanus sativus L. var niger) in alimentary hyperlipidaemia in rats.

Black radish (Raphanus sativus L. var. niger) root has been used in folk medicine since antiquity as a natural drug for the stimulation of bile function. According to in vitro studies the squeezed juice from black radish root exhibited significant antioxidant properties. In the present study, the beneficial effect of the black radish juice on some free radical reactions in rats fed with a diet rich in lipids (20% sunflower oil, 2% cholesterol, 0.5% cholic acid in normal chow) was examined. Thiobarbituric acid reactive substances and conjugated diene concentrations were significantly higher, while the antioxidant enzyme activities and the free radical scavenging capacity were lower in hyperlipidaemic rats compared with normal controls. Supplementation of the lipid-rich diet with black radish juice resulted in a significant improvement of the parameters mentioned above. Although the exact mechanism of the biologically active compounds in black radish on the lipid metabolism and lipid peroxidation is not clear yet, a beneficial effect of the drug was evident in alimentary hyperlipidaemia.

Animals↗

PubMed2002

Effects of black radish root (Raphanus sativus L. var niger) on the colon mucosa in rats fed a fat rich diet.

The effect of black radish root (Raphanus sativus L. var niger) was studied on the structure and redox state of the colon mucosa in fat-rich diet fed rats. The epithelial lining disrupted, the number of enterocytes and the goblet cells reduced and inflammatory cells were observed in rats fed with a fat-rich diet. After treatment with granules from black radish root all of the histopathological changes and parameters of the redox state caused by the fat-rich diet were improved. The structure of the epithelial cells was similar to the controls, the number of goblet cells increased and no inflammation was observed.

Animals↗

PubMed2003

Somatic hybrids between Arabidopsis thaliana and cytoplasmic male-sterile radish (Raphanus sativus).

Somatic hybrids were produced by protoplast fusion between Arabidopsis thaliana ecotype Columbia and a male-sterile radish line MS-Gensuke ( Raphanus sativus) with the Ogura cytoplasm. Forty-one shoots were differentiated from the regenerated calli and established as shoot cultures in vitro. About 20 of these shoots were judged to be hybrids based on growth characteristics and morphology. Molecular analyses of 11 shoots were performed, confirming the hybrid features. Of these 11 shoots, eight were established as rooted plants in the greenhouse. Polymerase chain reaction and randomly amplified polymorphic DNA analyses of the nuclear genomes of all analyzed shoots and plants confirmed that they contained hybrid DNA patterns. Their chromosome numbers also supported the hybrid nature of the plants. Investigations of the organelles in the hybrids revealed that the chloroplast (cp) genome was exclusively represented by radish cpDNA, while the mitochondrial DNA configuration showed a combination of both parental genomes as well as fragments unique to the hybrids. Hybrid plants that flowered were male-sterile independent of the presence of the Ogura CMS-gene orf138.

Arabidopsis↗

PubMed2005

Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B. napus.

A protocol for producing transgenic radish (Raphanus sativus) was obtained by using both ultrasonic and vacuum infiltration assisted, Agrobacterium-mediated transformation. The Agrobacterium strain LBA4404 contained the binary vector pBI121-LEA (late embyogenesis abundant), which carried a Group 3 LEA gene, from Brassica napus. Among six combinations, Agrobacterium-mediated transformation assisted by a combination of 5-min sonication with 5-min vacuum infiltration resulted in the highest transformation frequency. The existence, integration and expression of transferred LEA gene in transgenic T(1) plants were confirmed by PCR, genomic Southern and Western blot analysis. Transgenic radish demonstrated better growth performance than non-transformed control plants under osmotic and salt stress conditions. Accumulation of Group 3 LEA protein in the vegetative tissue of transgenic radish conferred increased tolerance to water deficit and salt stress.

Base Sequence↗

PubMed2003

In vitro biosynthesis of galactans by membrane-bound galactosyltransferase from radish ( Raphanus sativus L.) seedlings.

We investigated a galactosyltransferase (GalT) involved in the synthesis of the carbohydrate portion of arabinogalactan-proteins (AGPs), which consist of a beta-(1-->3)-galactan backbone from which consecutive (1-->6)-linked beta-Gal p residues branch off. A membrane preparation from 6-day-old primary roots of radish ( Raphanus sativus L.) transferred [(14)C]Gal from UDP-[(14)C]Gal onto a beta-(1-->3)-galactan exogenous acceptor. The reaction occurred maximally at pH 5.9-6.3 and 30 degrees C in the presence of 15 mM Mn(2+) and 0.75% Triton X-100. The apparent K(m) and V(max) values for UDP-Gal were 0.41 mM and 1,000 pmol min(-1) (mg protein)(-1), respectively. The reaction with beta-(1-->3)-galactan showed a bi-phasic kinetic character with K(m) values of 0.43 and 2.8 mg ml(-1). beta-(1-->3)-Galactooligomers were good acceptors and enzyme activity increased with increasing polymerization of Gal residues. In contrast, the enzyme was less efficient on beta-(1-->6)-oligomers. The transfer reaction for an AGP from radish mature roots was negligible but could be increased by prior enzymatic or chemical removal of alpha- l-arabinofuranose (alpha- l-Ara f) residues or both alpha- l-Ara f residues and (1-->6)-linked beta-Gal side chains. Digestion of radiolabeled products formed from beta-(1-->3)-galactan and the modified AGP with exo-beta-(1-->3)-galactanase released mainly radioactive beta-(1-->6)-galactobiose, indicating that the transfer of [(14)C]Gal occurred preferentially onto consecutive (1-->3)-linked beta-Gal chains through beta-(1-->6)-linkages, resulting in the formation of single branching points. The enzyme produced mainly a branched tetrasaccharide, Galbeta(1-->3)[Galbeta(1-->6)] Galbeta(1-->3)Gal, from beta-(1-->3)-galactotriose by incubation with UDP-Gal, confirming the preferential formation of the branching linkage. Localization of the GalT in the Golgi apparatus was revealed on a sucrose density gradient. The membrane preparation also incorporated [(14)C]Gal into beta-(1-->4)-galactan, indicating that the membranes contained different types of GalT isoform catalyzing the synthesis of different types of galactosidic linkage.

Galactans↗

PubMed2005

Complete nucleotide sequences and genome characterization of double-stranded RNA 1 and RNA 2 in the Raphanus sativus-root cv. Yidianhong [corrected].

Four distinct double-stranded (ds) RNA bands were extracted from leaves of Raphanus sativus-root cv. Yidianhong [corrected] with yellowing at the leaf edge in China. Purified viral particles of 28-30 nm in diameter contained dsRNA segments with the same number and mobility as these extracted directly from radish leaves. The two major dsRNA segments, namely RasR 1 and RasR 2, were 1866 and 1791 bp in length, respectively. Computer analysis predicted that they both contained a single open reading frame (ORF) on their plus-stranded RNA, putatively encoding a RNA dependent RNA polymerase and a capsid protein similar to that encoded by members of the family Partitiviridae. In addition, both RasR 1 and RasR 2 were highly conserved at the 5' untranslated regions (UTR) and had an adenosine-uracil rich stretch at the 3' UTR, with an identical terminal motif (5'-AAAAUAAAACC-3'). Taken together, these results suggest that the two major dsRNA segments constitute the genome of a partitivirus infecting radish.

3' Untranslated Regions↗

PubMed2006

Complete nucleotide sequences of three dsRNA segments from Raphanus sativus-root cv. Yidianhong [corrected] with leaf yellow edge symptoms.

The two minor dsRNA bands, previously detected in symptomatic leaves of Raphanus sativus-root cv. Yidianhong [corrected] were subjected to further analysis. cDNA cloning and sequencing revealed that the smaller of the two dsRNA bands is actually a doublet consisting of two co-migrating dsRNA segments and the resulting three segments were designated as RasR 3, RasR 4, and RasR 5. RasR 3 was 1717 bp in length and potentially encoded a protein of about 55.3 kDa, containing all of the six conserved motifs shared by the RNA dependent RNA polymerases of members of the family Partitiviridae. RasR 4 and RasR 5, which co-migrated in the 5% polyacrylamide gel, were 1521 and 1485 bp in length and each encoded a putative protein of unknown function. Their molecular masses, as calculated from the deduced amino acid, were 38.2 and 38.8 kDa, respectively. The 5' UTRs of all three segments shared regions of high sequence similarities, but were distinct from those of the RasR 1 and RasR 2. Taken together, these results along with those described in the previous report, suggest that more than one partitivirus was co-infecting radish leaves.

5' Untranslated Regions↗

PubMed2006

Phosphorylation of H2AX histones in response to double-strand breaks and induction of premature chromatin condensation in hydroxyurea-treated root meristem cells of Raphanus sativus, Vicia faba, and Allium porrum.

Histone H2A variant H2AX is rapidly phosphorylated on the induction of DNA double-strand breaks by ionizing radiation and hydroxyurea-mediated replication arrest, resulting in the formation of gamma-H2AX foci along megabase chromatin domains nearby the sites of incurred DNA damage. In an attempt to establish a relationship between species-specific nuclear architecture and H2AX phosphorylation in S/G(2) phase-arrested root meristem cells, immunocytochemical comparisons using an antibody raised against human gamma-H2AX were made among three plants differing with respect to DNA contents: Allium porrum, representing a reticulate type of DNA package, Vicia faba, having semireticulate cell nuclei, and Raphanus sativus, characterised by a chromocentric type of chromatin. Another approach was aimed at determining possible correlations between the extent of hydroxyurea-induced phosphorylation of H2AX histones and the quantities of root meristem cells induced by caffeine to enter aberrant mitotic division (premature chromosome condensation). It was concluded that the higher-order structure of chromatin may contribute to the accessibility of molecular factors engaged in the recognition and repair of genetic lesions. Consequently, in contrast to A. porrum and V. faba, a diffuse chromatin in chromocentric cell nuclei of R. sativus may become more vulnerable both to generate DNA double-strand breaks and to recruit molecular elements needed to arrange the cell cycle checkpoint functions, and thus, more resistant to factors which allow the cells to enter premature chromosome condensation spontaneously. On the other hand, however, caffeine-mediated overriding of the S-M checkpoint control system resulted in the typical appearance of premature chromosome condensation, irrespective of the genomic content of DNA.

Antineoplastic Agents↗

PubMed2006

Osmotic water permeability of plasma and vacuolar membranes in protoplasts I: high osmotic water permeability in radish (Raphanus sativus) root cells as measured by a new method.

Intra- and transcellular water movements in plants are regulated by the water permeability of the plasma membrane (PM) and vacuolar membrane (VM) in plant cells. In the present study, we investigated the osmotic water permeability of both PM (P ( f1)) and VM (P ( f2)), as well as the bulk osmotic water permeability of a protoplast (P ( f(bulk))) isolated from radish (Raphanus sativus) roots. The values of P ( f(bulk)) and P ( f2) were determined from the swelling/shrinking rate of protoplasts and isolated vacuoles under hypo- or hypertonic conditions. In order to minimize the effect of unstirred layer, we monitored dropping or rising protoplasts (vacuoles) in sorbitol solutions as they swelled or shrunk. P ( f1) was calculated from P ( f(bulk)) and P ( f2) by using the 'three-compartment model', which describes the theoretical relationship between P ( f1), P ( f2) and P ( f(bulk)) (Kuwagata and Murai-Hatano in J Plant Res, 2007). The time-dependent changes in the volume of protoplasts and isolated vacuoles fitted well to the theoretical curves, and solute permeation of PM and VM was able to be neglected for measuring the osmotic water permeability. High osmotic water permeability of more than 500 mum s(-1), indicating high activity of aquaporins (water channels), was observed in both PM and VM in radish root cells. This method has the advantage that P ( f1) and P ( f2) can be measured accurately in individual higher plant cells.

Cell Membrane Permeability↗

PubMed2005

Within-plant variation in glucosinolate concentrations of Raphanus sativus across multiple scales.

Variation in chemical defenses remains underexplored. In particular, little is known about patterns of variation at small scales within leaves and spatial variation of induction. I examined variation in glucosinolate concentrations in the leaves of Raphanus sativus at several different spatial scales in two related experiments. I used samples equivalent in area to the amount an intermediate-sized caterpillar might eat in 1 d, a smaller scale than used in most previous studies. I examined variation due to induction and leaf age and small-scale spatial variation within leaves. The mean and variance of glucosinolate concentrations were higher in induced plants, young leaves, and the proximal half of leaves. Higher glucosinolate concentrations in the proximal half of leaves are previously unreported. Small-scale variation was extreme, accounting for 57% of the total random variation, and spatially random. There was no spatial autocorrelation found at scales as small as 1-2 cm. The high degree of small-scale, spatially random variation in glucosinolate concentrations in leaves is previously unreported. This small-scale variation and the variation caused by induction may have significant effects on herbivores and could be an important component of plant defense.

Glucosinolates↗

PubMed2005

Isolation and characterization of a novel PHGPx gene in Raphanus sativus.

A full-length cDNA encoding putative phospholipid hydroperoxide glutathione peroxidase (PHGPx) was cloned from Raphanus sativus. The cDNA, designated RsPHGPx, includes an open reading frame which encodes 197 amino acid residues. The alignment of amino acid sequences showed that RsPHGPx had the highest sequence homology to plant PHGPx and contained an N-terminal extension characteristic of a mitochondrial targeting peptide. Northern blot analysis indicated that RsPHGPx was constitutively and ubiquitously expressed during radish development, and its expression was differently regulated by various stress conditions. The expression of RsPHGPx in a yeast PHGPx-deletion mutant significantly rescued the mutant sensitivity to oxidation-sensitive linolenic acid, just as the yeast PHGPx3 gene did. This suggested that RsPHGPx encodes a functional PHGPx protein.

Amino Acid Sequence↗

PubMed2004

Phytoextraction of heavy metals by canola (Brassica napus) and radish (Raphanus sativus) grown on multicontaminated soil.

Phytoextraction can provide an effective in situ technique for removing heavy metals from polluted soils. The experiment reported in this paper was undertaken to study the basic potential of phytoextraction of Brassica napus (canola) and Raphanus sativus (radish) grown on a multi-metal contaminated soil in the framework of a pot-experiment. Chlorophyll contents and gas exchanges were measured during the experiment; the heavy metal phytoextraction efficiency of canola and radish were also determined and the phytoextraction coefficient for each metal calculated. Data indicated that both species are moderately tolerant to heavy metals and that radish is more so than canola. These species showed relatively low phytoremediation potential of multicontaminated soils. They could possibly be used with success in marginally polluted soils where their growth would not be impaired and the extraction of heavy metals could be maintained at satisfying levels.

Brassica napus↗

PubMed2005

Raphanus sativus and its isothiocyanates inhibit vascular smooth muscle cells proliferation and induce G(1) cell cycle arrest.

Mu (Raphanus sativus, Korean White Radish) crude extract (Mu-CE) has been studied for its anti-proliferative activity on mouse aortic smooth muscle cells. The abnormal growth of vascular smooth muscle cells (VSMC) is a prominent feature of vascular disease, including atherosclerosis, restenosis after angioplasty. We examined the mechanisms of the action of Mu-CE on VSMC proliferation. The viability of VSMC decreased to 35% at 24 h of treatment with Mu-CE. Treatment of Mu-CE showed potent inhibitory effects on the DNA synthesis of cultured VSMC. In addition, Mu-CE induced apoptosis using cell death ELISA assay. These inhibitory effects were associated with G1 cell cycle arrest. Treatment of Mu-CE, which induced a cell-cycle arrest in G1-phase, induced down-regulation of cyclins and CDKs and up-regulation of the CDK inhibitor p21 expression, whereas up-regulation of p27 by Mu-CE was not observed. Then, total isothiocyanates (ITC) including four different 4-(Methylthio)-3-butenyl isothiocyanate (MTBITC), allyl isothiocayanate (AITC), benzyl isothiocyanate (BITC), and phenethyl isothiocyanate (PEITC) was isolated from n-hexane extracts of Mu. When the VSMC were treated with ITC, the cell viability was significantly decreased. These findings indicate the efficacy of Mu-CE in inhibiting cell proliferation, G1- to S-phase cell-cycle progress on VSMC.

Animals↗

PubMed2004

Pharmacological basis for the gut stimulatory activity of Raphanus sativus leaves.

The crude extract of Raphanus sativus leaves (Rl.Cr) showed a dose-dependent (0.03-5.0 mg/ml) spasmogenicity in guinea-pig ileum and colon. The effect was insensitive to atropine pre-treatment but was completely abolished by pyrilamine indicating involvement of histaminergic (H(1)) receptors. The contractile effect at high doses (3.0-5.0mg/ml) was followed by relaxation. Rl.Cr also enhanced the transit of charcoal meal in mice at 30-100 mg/kg. The petroleum spirit, chloroform and aqueous fractions all showed histaminergic activity in ileum; aqueous fraction being more potent. The study shows the presence of a histaminergic component(s) along with a weak spasmolytic factor thus providing sound mechanistic basis for the traditional use of the plant in constipation.

Animals↗

PubMed2005

Purification and characterization of a cationic peroxidase Cs in Raphanus sativus.

A short distance migrating cationic peroxidase from Korean radish seeds (Raphanus sativus) was detected. Cationic peroxidase Cs was purified to apparent homogeneity and characterized. The molecular mass of the purified cationic peroxidase Cs was estimated to be about 44 kDa on SDS-PAGE. After reconstitution of apoperoxidase Cs with protohemin, the absorption spectra revealed a new peak in the Soret region around 400 nm, which is typical in a classical type III peroxidase family. The optimum pH of peroxidase activity for o-dianisidine oxidation was observed at pH 7.0. Kinetic studies revealed that the reconstituted cationic peroxidase Cs has Km values of 1.18 mM and of 1.27 mM for o-dianisidine and H2O2, respectively. The cationic peroxidase Cs showed the peroxidase activities for native substrates, such as coumaric acid, ferulic acid, and scopoletin. This result suggested that cationic peroxidase Cs plays an important role in plant cell wall formation during seed germination.

Gene Expression Regulation, Developmental↗

PubMed2006

Evaluation of dissipation mechanisms by Lolium perenne L, and Raphanus sativus for pentachlorophenol (PCP) in copper co-contaminated soil.

Though phytoremediation is widely studied in remediation of metal contaminated soils or organic contaminated soils, little information is available regarding the effectiveness and processes of phytoremediation of sites co-contaminated with organic and metal pollutants. Sites co-contaminated with organic and metal pollutants are common and considered to be a more complex problem as the two components often cause a synergistic effect on cytotoxicity as measured both by growth inhibition and colony-forming ability. In this paper, the dissipation mechanisms for pentachlorophenol (PCP) in copper co-contaminated soil by Lolium perenne L, and Raphanus sativus was investigated in a greenhouse experiment by monitoring the growth response of plants, evaluating the removal efficiency of extractable PCP, differentiating PCP residuals in strongly and loosely adhering rhizosphere soils, and analyzing the microbial activity in the rhizosphere. In copper co-contaminated soil with the initial PCP concentration of 50 mg/kg, plants grew better with the increment of soil Cu level (0, 150, 300 mg/kg), which implied that combinations of inorganic and organic pollutants sometimes exerted antagonistic effects on plant cytotoxicity. The observed higher PCP dissipation in soil spiked with 50 mg/kg PCP in the presence of Cu and the less difference of PCP residual between strongly and loosely adhering soils further suggests the occurrence of Cu-PCP interaction and the enhanced degradation and mass flow are two possible explanations. In copper co-contaminated soil with the initial PCP concentration of 100 mg/kg, however, both plant growth and microbial activity were inhibited with the increment of soil Cu level. The lowered degrading activity of microorganisms and the reduced mass flow were probably responsible for the significantly lower levels of PCP dissipation in copper co-contaminated soil. These results showed that remediation of sites co-contaminated with organic and metal pollutants is a complex problem and a more thorough understanding of the extent and mechanisms by which metals inhibit organic degradation is needed to develop phytoremediation of co-contaminated sites.

Biodegradation, Environmental↗

PubMed2005

Isolation of 4-methylthio-3-butenyl glucosinolate from Raphanus sativus sprouts (kaiware daikon) and its redox properties.

The most promising among glucosinolates (GLs) are those bearing in their aglycon an extra sulfur function, such as glucoraphasatin (4-methylthio-3-butenyl GL; GRH) and glucoraphenin (4-methylsulfinyl-3-butenyl GL; GRE). The GRE/GRH redox couple is typically met among secondary metabolites of Raphanus sativus L. and, whereas GRE prevails in seeds, GRH is the major GL in full-grown roots. During the 10 days of sprouting of R. sativus seeds, the GRE and GRH contents were determined according to the Eurpean Union official method (ISO 9167-1). In comparison to the seeds, the GRE content in sprouts decreased from about 90 to about 12 micromol g(-1) of dry weight (dw), whereas a 25-fold increase--from about 3 to 76 micromol g(-1) of dw--of the GRH content was measured. An efficient pure GRH gram-scale production process from R. sativus (kaiware daikon) sprouts resulted in significant yield improvement of up to 2.2% (dw basis). The reaction of GRH with both H2O2 and ABTS*+ radical cation was investigated. Whereas H2O2 oxidation of GRH readily resulted in complete transformation into GRE, ABTS*+ caused complete decay of the GL. Even though not directly related to its radical scavenging activity, the assessed reducing capacity of GRH suggests that R. sativus sprouts might possess potential for health benefits.

Algorithms↗