[Antimicrobial properties of Raphanus sativus. Antimicrobial activity of extracts and ethereal oils from wild and garden varieties of Raphanus].
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Anti-Infective Agents↗
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[Antimicrobial properties of Raphanus sativus. II. Antimicrobial properties of a preparation from acid substances from Raphanus sativus var. niger].
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Anti-Bacterial Agents↗
Genetic analysis and genome mapping in Raphanus.
The first genetic map of the Raphanus genome was developed based on meiosis in a hybrid between Raphanus sativus (cultivated radish) and Raphanus raphanistrum (wild radish). This hybrid was used to produce a BC1 population of 54 individuals and an F2 population of 85 individuals. A total of 236 marker loci were assayed in these populations using a set of 144 informative Brassica RFLP probes previously used for genetic mapping in other crucifer species. The genetic maps derived from the BC1 and F2 populations were perfectly collinear and were integrated to produce a robust Raphanus map. Cytological observations demonstrated strict bivalent pairing in the R. sativus x R. raphanistrum hybrids. Productive pairing along the length of each chromosome was confirmed by the identification of nine extensive linkage groups and the lack of clustering of marker loci. Indeed, the distributions of both marker loci and crossovers was more random than those reported for other crop species. The genetic markers and the reference map of Raphanus will be of considerable value for future trait mapping and marker-assisted breeding in this crop, as well as in the intergenomic transfer of Raphanus genes into Brassica crops. The future benefits of comparative mapping with Arabidopsis and Brassica species are also discussed.
Chromosome Mapping↗
Mutations in Turnip mosaic virus genomes that have adapted to Raphanus sativus.
The genetic basis for virulence in potyviruses is largely unknown. Earlier studies showed that there are two host types of Turnip mosaic virus (TuMV); the Brassica/Raphanus (BR)-host type infects both Brassica and Raphanus systemically, whereas the Brassica (B)-host type infects Brassica fully and systemically, but not Raphanus. The genetic basis of this difference has been explored by using the progeny of an infectious clone, p35Tunos; this clone is derived from the UK1 isolate, which is of the B-host type, but rarely infects Raphanus systemically and then only asymptomatically. Two inocula from one such infection were adapted to Raphanus by passaging, during which the infectivity and concentration of the virions of successive infections increased. The variant genomes in the samples, 16 in total, were sequenced fully. Four of the 39 nucleotide substitutions that were detected among the Raphanus sativus-adapted variant genomes were probably crucial for adaptation, as they were found in several variants with independent passage histories. These four were found in the protein 1 (P1), protein 3 (P3), cylindrical inclusion protein (CI) and genome-liked viral protein (VPg) genes. One of four 'parallel evolution' substitutions, 3430G-->A, resulted in a 1100Met-->Ile amino acid change in the C terminus of P3. It seems likely that this site is important in the initial stages of adaptation to R. sativus. Other independent substitutions were mostly found in the P3, CI and VPg genes.
Adaptation, Physiological↗
Identification and classification of S haplotypes in Raphanus sativus by PCR-RFLP of the S locus glycoprotein (SLG) gene and the S locus receptor kinase (SRK) gene.
Polymorphism of the S-locus glycoprotein (SLG) and S-locus receptor kinase (SRK) genes in Raphanus sativus was analyzed by PCR-RFLP using SLG- and SRK-specific primers. Twenty four inbred lines of R. sativus could be grouped into nine S haplotypes. DNA fragments of SLG alleles specifically amplified from five S haplotypes by PCR with Class-I SLG-specific primers showed different profiles upon polyacrylamide-gel electrophoresis after digestion with restriction endonucleases. The five R. sativus SLG alleles were determined for their nucleotide sequences of DNA fragments. Comparison of the amino-acid sequences with a reported Brassica SLG (S(6)) showed 77-84% homology. Deduced amino-acid sequences showed 12-conserved cystein residues and three hypervariable regions which are characteristic of Brassicsa SLG. A DNA fragment was also amplified by PCR from two of each S haplotype with Class-II SLG-specific primers, and showed polymorphism when cleaved with restriction endonucleases. The nucleotide sequences of amplified DNA fragments of the Class-II SLG revealed about 60% similarity with those of the Class-I SLG. It is concluded that there exist both Class I and Class II S alleles in R. sativus, as in Brassica campestris and Brassica oleracea. PCR using SRK-specific primers amplified a DNA fragment of about 1.0 kb from seven of each S haplotype out of 24 tested. These DNA fragments showed high polymorphism in polyacrylamide-gel electrophoresis after digestion with restriction endonucleases. Nucleotide sequences of the DNA fragments amplified from the seven S haplotypes showed that the fourth and the fifth exons of SRK are highly conserved, and that there is high variation in the fifth intron, the sixth intron and seventh exon of the SRK which may be responsible for the polymorphic band patterns in PCR-RFLP analysis. The PCR-RFLP method has proven useful for the identification of S alleles in inbred lines and for listing S haplotypes in R. sativus. Phylogenic analysis of the SLG and SRK sequences from Raphanus and Brassica revealed that the Raphanus SLGs and SRKs did not form an independent cluster, but were dispersed in the tree, clustering together with Brassica SLGs and SRKs. Furthermore, SLGs and SRKs from Raphanus were both grouped into Class-I or Class-II S haplotypes. Therefore, these results suggest that the diversification of the SLG and SRK alleles occurred prior to the differentiation of the two genera Brassica and Raphanus.
Journal Article↗
Polymorphism of the S-locus glycoprotein gene (SLG) and the S-locus related gene (SLR1) in Raphanus sativus L. and self-incompatible ornamental plants in the Brassicaceae.
The S-locus glycoprotein gene, SLG, which participates in the pollen-stigma interaction of self-incompatibility, and its unlinked homologue, SLR1, were analyzed in Raphanus sativus and three self-incompatible ornamental plants in the Brassicaceae. Among twenty-nine inbred lines of R. sativus, eighteen S haplotypes were identified on the basis of DNA polymorphisms detected by genomic Southern analysis using Brassica SLG probes. DNA fragments of SLG alleles specifically amplified from eight S haplotypes by PCR with class I SLG-specific primers showed different profiles following polyacrylamide gel electrophoresis, after digestion with a restriction endonuclease. The nucleotide sequences of the DNA fragments of these eight R. sativus SLG alleles were determined. Degrees of similarity of the nucleotide sequences to a Brassica SLG (S6SLG) ranged from 85.6% to 91.9%. Amino acid sequences deduced from these had the twelve conserved cysteine residues and the three hypervariable regions characteristic of Brassica SLGs. Phylogenetic analysis of the SLG sequences from Raphanus and Brassica revealed that the Raphanus SLGs did not form an independent cluster, but were dispersed in the tree, clustering together with Brassica SLGs. These results suggest that diversification of the SLG alleles of Raphanus and Brassica occurred before differentiation of these genera. Although SLR1 sequences from Orychophragmus violaceus were shown to be relatively closely related to Brassica and Raphanus SLR1 sequences, DNA fragments that are highly homologous to the Brassica SLG were not detected in this species. Two other ornamental plants in the Brassicaceae, which are related more distantly to Brassica than Orychophragmus, also lacked sequences highly homologous to Brassica SLG genes. The evolution of self-incompatibility in the Brassicaceae is discussed.
Alleles↗
Toxic effects of Al-based coagulants on Brassica chinensis and Raphanus sativus growing in acid and neutral conditions.
The ecotoxicological effects of aluminum (Al)-based coagulants are of concern because of their wide-ranging applications in wastewater treatment and water purification. As important Al-based coagulants, AlCl(3) and PAC (polyaluminum-chloride) were selected as examples to examine the toxic effects on representative vegetables including the cabbage Brassica chinensis and the radish Raphanus sativus over a range of exposure concentrations in neutral (pH 7.00) and acidic (pH 4.00) conditions, using seed germination and root elongation in the early-growth stage as indicators of toxicity. The results showed that root elongation of the two vegetables was a more sensitive indicator than was seed germination for evaluating the toxicity of Al. As a single influencing factor, H(+) had no significant direct effects on root elongation of Brassica chinensis and Raphanus sativus under the experimental conditions. The toxicity of Al played the main role in inhibiting root elongation and seed germination and was strongly related to changes in pH. There was a markedly positive relationship between the inhibitory rate of root elongation, seed germination, and the concentration of Al at pH 4.00 (p < 0.01). The toxic effect of AlCl(3) on Brassica chinensis was less with a neutral pH than at pH 4.00, but Raphanus sativus was more susceptible to AlCl(3) toxicity at a neutral pH than at pH 4.00. Both Raphanus sativus and Brassica chinensis had a more toxic response to a low concentration (<64 mg . L(-1)) of PAC in a neutral condition than in an acidic condition. Undoubtedly, the Al toxicity caused by Al-based coagulants at a neutral pH is relevant when treatment solids are used in agriculture.
Aluminum↗
Transfer of resistance against the beet cyst nematode from radish (Raphanus sativus) to rape (Brassica napus) by monosomic chromosome addition.
In rape ( Brassica napus), no resistance to the beet cyst nematode (BCN) Heterodera schachtii is available. This study was carried out to determine the specific chromosome(s) of resistant radish ( Raphanus sativus) carrying the gene(s) for nematode resistance as a prequisite to convert rape from a host into a trap crop for this pest. A Raphanobrassica progeny of 25 plants was analyzed which segregated for all nine chromosomes of the Raphanus genome in a genetic background of synthetic rape. The number of radish chromosomes was determined by fluorescence in situ hybridization, using the Raphanus-specific DNA probe pURsN; and their type was identified by chromosome-specific randomly amplified polymorphic DNA markers. Five different multiple rape-radish chromosome additions (comprising the whole set of nine radish chromosomes, a-i) were selected and crossed to rape. For each cross-progeny, the number of cysts on plant roots was counted 42 days after inoculation with a L2 larvae suspension. Simultaneously, the plants were characterized for the presence or absence of individual radish chromosomes, using sets of chromosome-specific markers. Thus, the effect of each radish chromosome on cyst number was tested. Chromosome d had a major resistance effect, whereas the presence/absence of the other radish chromosomes had nearly no influence on cyst number. Plants with added chromosome d showed a resistance level comparable with that of the radish donor parent. The analysis in the cross to rape of a plant monosomic only for chromosome d confirmed the strong effect of this chromosome on nematode resistance. A further experiment comprising seven crosses using winter rape breeding lines and monosomic addition line d as pollen parent provided the same results on a broader genetic basis. In each case, the added chromosome d in a single dosage caused nearly the full resistance of the radish donor. Resistance was independent of the glucosinolate content in the roots. The possibilities for stabilizing BCN resistance in rape and its use for other crops and nematodes are discussed.
Animals↗
Hybridization between oilseed rape (Brassica napus) and different populations and species of Raphanus.
When cultivating genetically modified varieties, the spontaneous gene flow between crop and wild relatives could be of concern. We analyzed spontaneous hybridization between a transgenic male-sterile line of oilseed rape (Brassica napus, 2n = 38, AACC) and, as pollen donors, three European populations of wild radish (Raphanus raphanistrum, 2n = 18, Rr,Rr) and a variety of cultivated radish (Raphanus sativus, 2n = 18, RR). Seeds showed size and shape dimorphism that correlated to the frequency of hybrids. The offspring were scored morphologically and analyzed using DNA markers (inter-simple sequence repeats) to quantify hybrid frequencies. Seed set ranged from 0.4-1.2 seeds per pod, and 0.02-0.6 seeds per pod were confirmed as hybrids. The frequency of confirmed hybrids differed significantly among populations of R. raphanistrum. In the cross with a French population, all offspring were hybrids; in the cross with a Swiss population, 53% of the offspring were hybrids; and in the cross with a Danish population, only 2% of the offspring were found to be hybrids. The remaining offspring apparently belonged to two groups: the majority was B. napus-like plants, possibly of matromorphic origin, and a minority from the Danish cross seemed to carry fragments of the Raphanus genome. In the cross with a cultivated R. sativus, all offspring were found to be hybrids. This is the first report on spontaneous hybridization between B. napus and R. sativus. Hybrids from all cross-combinations had low pollen fertility (0-15%). If R. raphanistrum occurs where male-sterile B. napus is cultivated, large regional differences in hybridization frequencies between the species could complicate environmental risk assessment of transgenic oilseed rape.
Brassica napus↗
Determination of the three-dimensional solution structure of Raphanus sativus antifungal protein 1 by 1H NMR.
Raphanus sativus Antifungal Protein 1 (Rs-AFP1) is a 51 amino acid residue plant defensin isolated from radish (Raphanus sativus L.) seeds. The three-dimensional structure in aqueous solution has been determined from two-dimensional 1H NMR data recorded at 500 MHz using the DIANA/REDAC calculation protocols. Experimental constraints consisted of 787 interproton distances extracted from NOE cross-peaks, 89 torsional constraints from 106 vicinal interproton coupling constants and 32 stereospecific assignments of prochiral protons. Further refinement by simulated annealing resulted in a set of 20 structures having pairwise root-mean-square differences of 1.35(+/- 0.35) A over the backbone heavy atoms and 2.11(+/- 0.46) A over all heavy atoms. The molecule adopts a compact globular fold comprising an alpha-helix from Asn18 till Leu28 and a triple-stranded beta-sheet (beta 1 = Lys2-Arg6, beta 2 = His33-Tyr38 and beta 3 = His43-Pro50). The central strand of this beta-sheet is connected by two disulfide bridges (Cys21-Cys45 and Cys25-Cys47) to the alpha-helix. The connection between beta-strand 2 and 3 is formed by a type VIa beta-turn. Even the loop (Pro7 to Asn17) between beta-strand 1 and the alpha-helix is relatively well defined. The structure of Raphanus sativus Antifungal Protein 1 features all the characteristics of the "cysteine stabilized alpha beta motif". A comparison of the complete structure and of the regions important for interaction with the fungal receptor according to a mutational study, is made with the structure of gamma-thionin, a plant defensin that has no antifungal activity. It is concluded that this interaction is both electrostatic and specific, and some possible scenarios for the mode of action are given.
Amino Acid Sequence↗
[Production and cytogenetics of intergeneric hybrids between Ogura CMS Brassica campestris var. purpuraria and Raphanus sativus].
Crosses between Ogura CMS Brassica campestris var. purpuraria (AA, 2n = 20) and Raphanus sativus (RR, 2n = 18) were made and many intergeneric hybrids were produced. The F1 seedlings did not show chlorosis at low temperature. When red Raphanus sativus varieties were used as male parent, the leaf petiole and leaf vein of F1 plants were purple, and when white Raphanus sativus varieties were used as male parent, the leaf petiole and leaf vein of F1 plants were not purple. All the F1 plants had white flowers and normal honey glands. Male gametes of the F1 were highly sterile and female gametes of the F1 were partly fertile. Cytological studies indicated that chromosome number of the F1 was 2n = 19 as expected, the mean chromosome pairing pattern was 15.53 I + 1.34 II + 0.25 III + 0.01 IV. Most chromosomes exsistet as univalents, but there also exsisted some bivalents, trivalents and even tetravalents, suggesting that chromosome set A was partly homologous with chromosome set R.
Brassica↗
Multiple origins of cultivated radishes as evidenced by a comparison of the structural variations in mitochondrial DNA of Raphanus.
Configurations of mitochondrial coxI and orfB gene regions were analysed by polymerase chain reaction (PCR) in three wild and one cultivated species of Raphanus. A total of 207 individual plants from 60 accessions were used. PCR with five combinations of primers identified five different amplification patterns both in wild and cultivated radishes. While the mitochondrial DNA (mtDNA) type of Ogura male-sterile cytoplasm was distinguishable from the normal type, the mtDNAs of normal radishes were further classified into four types. The variations were common to wild and cultivated radishes, although contrasting features were found depending on the region of cultivation. These results provide evidence that cultivated radishes have multiple origins from various wild plants of Raphanus.
Biological Evolution↗
The effect of Trifolium, Raphanus, and Cistus pollen grains on some blood parameters and mesentery mast cells.
Three kinds of pollen taxa belonging to 3 families (Fabaceae--Trifolium spp., Brassicaceae--Raphanus spp. and Cistaceae--Cistus spp.) and commonly collected by honeybees were fed to mature male rats separately, in the form of 60 mg/animal/day for a 30-day period. The objective of this study was to investigate any positive effects or possible side effects of the use of pollen on the immune system. This was achieved through blood analysis and cell count on blood, hemoglobin, erythrocyte and immune system cells. The cell concentration of mast cells, degranulization and cell localization were investigated in prepared mesentery tissue samples. Histological investigations of the stomach and duedenum sections of pollen-fed rats were carried out to learn the reason for eosinophil gastroenteritis in the alimentary canal. The eosinophil and lymphocyte levels of rats fed with pollen of Trifolium spp., Raphanus spp., and Cistus spp. were observed to have increased blood cell counts, while neutrophil and monocyte levels decreased; different values were found in basophil leucocytes between the pollen groups. Differing reductions in mesentery mast cell concentration, degranulization and cell localization were found. Within the three separate pollens, the rats having been fed with Cistus spp. pollen were observed to have higher blood lymphocyte, eosinophil, hemoglobin and hematocrit values than those fed with the others, as well as low mesentery mast cell concentration. Hemoglobin values were determined to increase at a proportion of between 10.0-11.3%. No difference was found in other blood parameters. The fat proportion of the male rats fed with the three taxa was between 4.03-8.75%, while that for protein proportion was between 16.11-24.25%. Male rats receiving these taxa did not experience allergic reactions and it is possible to argue that the low protein and fat content of these pollens have a strengthening effect on the immune systems by the increase in lymphocyte content and the amount of hemoglobin leads to an increase of oxygen transport capacity in the tissues.
Animals↗
Homeobox gene diversification in the calcareous sponge, Sycon raphanus.
Knowledge of the developmental mechanisms in living basal metazoan phyla is crucial for understanding the genetic bases of morphological evolution in early animal history. We looked for homeobox genes in the calcareous sponge, Sycon raphanus, using the polymerase chain reaction. Partial sequences of eight homeoboxes were recovered, five of which are assignable to the NK-2 class of homeoboxes. The three remaining sequences are related members of a new class of homeoboxes, the Sycox class, showing limited similarity to bilaterian Lbx, Hlx, HEX, En, and Cad classes. Among the five NK-2 class homeoboxes are four closely related sequences occupying a divergent position within the class, the remaining one on the contrary showing high sequence similarity with members of the NK-2 family, a particular subgroup within the NK-2 class, previously known only from the Bilateria. This suggests that diversification of the NK-2 class occurred early in metazoan history. Altogether, the results reveal an unexpected diversification of homeobox genes in S. raphanus.
Amino Acid Sequence↗
The radish (Raphanus sativus L.) mitochondrial cox2 gene contains an ACG at the predicted translation initiation site.
The mitochondrial cox2 gene has been sequenced from radish (Raphanus sativus L.). The gene is interrupted by a 1346-bp group-II intron and contains an ACG codon as the predicted translation initiation site. Analysis of cox2 cDNAs indicates that the ACG codon is not converted to an AUG codon in the mRNA, although 15 other RNA editing sites were identified. The cox2 gene from Raphanus raphanistrum, and other varieties of R. sativus, also contain an ACG as the predicted start codon; plants in the closely related genus, Brassica, do not. Western-blot analyses indicate that cox2 proteins in radish mitochondria are the same size as those found in Brassica mitochondria and different from cox2 proteins in plants where cox2 is nuclear-encoded. This finding, along with the observation that cox2 sequences are not present in the nuclear genome of radish, suggests that ACG is utilized as the radish cox2 initiation codon.
Amino Acid Sequence↗
Characterization and phylogenetic analysis of a cDNA encoding the Fes/FER related, non-receptor protein-tyrosine kinase in the marine sponge sycon raphanus.
In search of ancient versions of phylogenetically conserved genes/proteins, which are typical for multicellular animals, we have decided to analyse marine sponges (Porifera), the most ancient and most primitive metazoan organisms. We report here the complete nucleotide sequence of Sycon raphanus cDNA coding for a 879 aa long protein, which displays high overall similarity in primary structure and organization of domains with non-receptor tyrosine kinases (TKs) from the Fes/FER family. The encoded protein, which we named Fes/FER_SR, has a highly conserved, 260 aa long tyrosine kinase domain at the C-terminus. Amino-terminal to the catalytic domain is an 85 aa long SH2 domain. The N-terminus is over 500 aa long and displays homology only with N-terminal domains of protein-tyrosine kinases (PTKs) from the Fes/FER family. Mammalian Fes/FER proteins show around 58% overall homology with Fes/FER_SR (identity and similarity) and lower homology was found with Drosophila melanogaster Fps (FER) protein (49%). Homologies in TK, SH2 and N-terminal domains are on average 78%, 65% and 49%, respectively. Fes/FER_SR shows next to best homology with the Abl family of non-receptor PTKs, while Src-related PTKs from the fresh-water sponge Spongilla lacustris are related only distantly to Fes/FER_SR. Phylogenetic analysis shows that the S. raphanus TK is indeed the most ancient known member of the Fes/FER family of non-receptor PTKs. The role of these PTKs in signal transduction in higher animals is still enigmatic; they are present in the nucleus as well as in the cytoplasm and FER is found in all cell types examined. The function of Fes/FER_SR in sponge, the most primitive multicellular animal which lacks specialized organ systems, remains to be elucidated.
Amino Acid Sequence↗
A study of the phylogeny of Brassica rapa, B. nigra, Raphanus sativus, and their related genera using noncoding regions of chloroplast DNA.
There are two evolutionary lineages in the genus Brassica: the rapa/oleracea lineage and the nigra lineage. Using nuclear DNA sequences such as the intergenic spacer between 5S rRNA genes and the internal transcribed spacer between 18S and 25S rRNA genes, we and others had previously demonstrated that Raphanus sativus is closely related to the nigra lineage. In the present study, we sequenced the chloroplast noncoding region between trnT and trnF and that between trnD and trnT in seven species and showed that R. sativus is more closely related to the rapa/oleracea lineage than to the nigra lineage. The conflicting results from nuclear DNA and chloroplast DNA support the hypothesis that Raphanus was derived from a hybridization between the rapa/oleracea and the nigra lineages. We estimated the date of this hybridization event to be 60% of the divergence time between the two Brassica lineages. In addition, the pattern and rate of nucleotide substitution were studied. There were more transversions than transitions in these noncoding regions, which have a high AT content. Furthermore, the proportion of transversions among the substitutions at a site increases with increasing A + T content of its two adjacent nucleotides. An influence of immediate 5(') pyrimidine on substitution pattern is also observed when both adjacent bases in the two DNA strands are A or T. The rate of nucleotide substitution in the trnL group I intron is only about one third of the rate in the nearby intergenic spacers in the trnT-trnF fragment. The rate of nucleotide substitution in the rapa/oleracea lineage is at least 1.5 times that in the nigra lineage.
Brassica rapa↗
Preliminary Studies on the Amino-acid Residues at Active Center of the Lysozyme from Raphanus sativus Leaves.
A 1ysozyme from Raphanus sativus leaves was purified by the method of affinity chromatography on a deaminated regenerated crab chitin column. The purified enzyme was crystallized and showed a single band on polyacrylamide gel disc electrophoresis. The functional groups at the active center of the enzyme were studied by the method of pH dynamics and chemical modification. It was found that carboxyl groups(Glu/Asp), tryptophanyl and histidyl residues were probably essential groups for the catalytic activity. The enzymatic activity was not affected when the enzyme was modified by reagents which could specifically react with tyrosyl, cysteinyl, arginyl and seryl/threonyl residues. It was inhibited by histamine and GlcNAc. The difference among Raphanus sativus lysozyme, HEWI and papaya lysozyme was discussed.
Journal Article↗