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Effects of pollen load size and composition on pollen donor performance in wild radish, Raphanus sativus (Brassicaceae).

A critical concern in the debate over the importance of sexual selection in plants is whether the nonrandom mating demonstrable in greenhouse crosses can occur in the field. Field populations likely experience smaller and more variable pollen load sizes than those that have been used in many greenhouse experiments. Therefore, we performed a greenhouse experiment in which we varied both pollen load size and composition in wild radish, Raphanus sativus, and examined the paternity of seeds. We used five maternal plants and four pairs of pollen donors. We were able to produce pollen loads of 40, 118, and 258 grains per stigma. The smallest of the pollen loads was scant enough to result in a slight, but significant reduction in seed number per fruit. While variation in pollen load composition significantly affected the proportions of seeds fathered by different donors, variation in pollen load size did not. The relative performance of different donors was constant across pollen load sizes, suggesting that, for this species, differential performance of pollen donors can occur at pollen load sizes that are likely to occur in field populations.

Journal Article↗

[Production and cytogenetics of hybrids of Ogura CMS Brassica campestris var. purpuraria x Raphanus sativus x Brassica napus].

Crosses of Ogura CMS Brassica campestris var. purpuraria x Raphanus sativus x Brassica napus were made and four hybrids were produced. One plant (PRN-1) was mosaic with yellow and milk white flowers and some flowers had both yellow and white petals. The others (PRN-2, -3, -4) had white flowers. PRN-4 had degenerated anthers, the other three had three to six anthers and could produce some pollens, but the pollens of PRN-2 were unstainable by I2-KI solution. PRN-2 had four normal honey glands, PRN-1 and PRN-3 had two, and PRN-4 had none. PRN-2 had normal leaf color and the other three showed different degrees of chlorophyll deficiency at low temperature. The chromosome number of PRN-1 was 2n = 38 and had the mean chromosome paring configuration of 14.67 I + 10.07 II + 1.06 III, and its chromosome set constitution might be AACR. This chromosome constitution may be due to the fertilization of female gamete of n = 19 (AR) with male gamete of n = 19 (AC) from B. napus. The occurrence of mosaic flower color in this plant may be attributed to the chromosome abnormalities caused by wide hybridization, such as chromosome deficiency and the formation of chromosome fragments and chromosome bridges. The chromosome number of PRN-2 was 2n = 35 and the mean chromosome paring configuration was 13.89 I + 8.33 II + 1.33 III + 0.11 IV. The chromosome number of PRN-3 was 2n = 33 and the mean chromosome paring configuration was 14.00 I + 7.82 II + 1.00 III + 0.09 IV. The chromosome number of PRN-4 was not determined as there was no pollen mother cell formation. Chromosome bridges and laggards were observed in PRN-1-3. Some seeds were harvested from PRN-1-3 but none was harvested from PRN-4 when backcrossed with B. napus. It seems possible for us to overcome the chlorophyll deficiency and honey gland abnormality and restore the male fertility in Ogura CMS by introduction of the nucleus of R. sativus into this cytoplasmic male sterile line.

Aneuploidy↗

[The effects of aqueous extracts of Raphanus niger on an experimental influenza infection in mice and on the enzyme polymorphism in lung tissue extracts].

Prophylactic effects of Raphanus niger water extract against A/PR 8/34 influenza virus experimental infection was tested in mice, by means of the following parameters: mortality mean survival time and hemagglutinating (HA) titre of lung extract. Variations of the isoenzymatic structures of peroxidase and acid and alkaline phosphatases were studied too. The results revealed the significant reduction of mortality and of hemagglutinating titre and the augmentation of mean survival time. Modifications of the isoenzymatic structures, especially of the phosphatases, are characteristic for the illness state as well as for the one induced by R. niger extract treatment.

Acid Phosphatase↗

[Immunomodulation with natural products. I. Effect of an aqueous extract of Raphanus sativus niger on experimental influenza infection in mice].

A Raphanus sativus niger water extract was administered by intranasal instillations to mice before inoculation of the influenza virus A/PR 8/34 (H1N1) strain by the same route. The extract ensured some protection against the experimental influenza infection. A significant decrease of the hemagglutinin titre of the mouse lung homogenate was noted, as well as a decrease of the mortality rate and a significant increase of the rate of survival as compared to the untreated controls.

Animals↗

Changes in microtubule arrays during the differentiation of cortical root cells of Raphanus sativus.

Cortical microtubule arrays in meristematic and differentiated cortical cells from root tips of Raphanus sativus were studied using both immunofluorescence and dry cleaving. Length, density and orientation of the cortical microtubules were measured. Between individual, non-dividing cells of the meristematic zone the mean microtubule length varied from 0.9 micron to 1.3 micron and the density varied from 1.7 micron to 3.2 micron microtubule/micron 2 membrane. The direction of the cortical tubules, running parallel to each other in individual cells, appeared to be more or less perpendicular to the root axis, at angles of 85 degrees to 95 degrees. In elongated cortical cells, the mean length had increased to values between 2.6 and 6.7 micron, while the density had decreased to 0.9 to 1.9 micron/micron 2. Microtubules remained parallel to each other within one cell, although their angle with the root axis changed to highly variable values: between 10 degrees to 80 degrees. The results clearly show that important changes occur in the cytoskeleton during the differentiation of cortical cells. It is argued that these changes might be related directly to the morphogenesis of these cells.

Cell Differentiation↗

[Effect of insulin on germination and ionic exchange in Raphanus sativus (author's transl)].

Hypoglycemic sulfamide BZ-55 activates or inhibits germination of Raphanus sativus, depending upon the dosis. Since this drug acts upon the glycemia by increasing the secretion and action of insulin, the influence of this hormone on germination and ionic changes (Na+-K+) between seeds and culture medium, were studied. Seeds were incubated during 72 h with different concentrations of insulin in 10 ml deionized water or in 10 ml 18 mM K+ (KCl) solutions at 37 degrees C in vapor saturated atmosphere. A solution of 0.125 IU insulin/ml in water increases the germination to 110% whereas 0.175 IU insulin/ml inhibits it to 40% against controls. Further increases in insulin concentration always inhibit germination. Similar results have been obtained with K+ containing media. Germination rate changes in a small concentration range suggest that insulin might affect an enzymatic activity in the seed.

Blood Glucose↗

Identification by 2D-page analysis of salt-stress induced proteins in radish (Raphanus sativus).

The response of Raphanus sativus to salinity has been investigated. Salt-responsive proteins were detected by comparative 2D-PAGE analysis. Three polypeptides of 22, 28 and 28.5 kDa were isolated and subjected to microsequencing. The P22 possesses homology with members of the Kunitz family of trypsin inhibitor; the P28.5 is homologous to ascorbate peroxidase of several plant species while P28 did not exhibit significant homology with sequences archived in available databases.

Amino Acid Sequence↗

Effect of Japanese radish (Raphanus sativus) sprout (Kaiware-daikon) on carbohydrate and lipid metabolisms in normal and streptozotocin-induced diabetic rats.

No information is available about the effects of Japanese radish sprout (JRS) on diabetes. To clarify the effects, the influence of JRS on carbohydrate and lipid metabolisms was investigated in normal and streptozotocin-induced diabetic rats. The rats were fed a diet containing 0%, 2.5% or 5% of JRS ad libitum for 21 days. Compared with the corresponding control groups, the JRS-fed normal rats showed lower plasma levels of total cholesterol (TC), triglycerides (TG), phospholipids (PL), fructosamine, glucose and insulin and higher plasma levels of low-density lipoprotein-cholesterol, whereas the JRS-fed diabetic rats showed lower plasma levels of fructosamine, glucose and insulin without changes in the plasma lipid parameters. JRS also decreased the hepatic TC, TG and PL levels in the normal rats and the TG level in the diabetic rats. These results showed that JRS had a hypoglycemic activity in both the normal and diabetic rats and partly improved lipid metabolism in the normal rats. JRS has the potential to alleviate hyperglycemia in cases where diabetes is present and to serve in the primary prevention of diabetes mellitus.

Animals↗

The effectiveness of insect-pollination test to evaluate the level of self-incompatibility and their genetic analysis in radish (Raphanus sativus L.).

We have tried an insect-pollination self-incompatibility (SI) test to strictly evaluate the level of SI as a model for the actual F(1) seed production field using radish as experimental material. Twelve inbred lines, homozygous for the S-alleles, were used in the artificial self-pollination and the insect-pollination SI test. There was a positive correlation ( r = 0.606) between the results by these two methods. Some lines showed a low level of SI in the insect-pollination test despite showing a high level of SI in the artificial self-pollination test. On the other hand, no lines showing a low level of SI in artificial self-pollination had a high level of SI in insect-pollination. These results show that the insect-pollination SI test can be considered to be a more reliable and stricter method than the artificial self-pollination test with respect to an evaluation of SI levels. We have raised and analyzed an F(2) population and F(3) lines derived from an F(1) cross between a line showing a high level of SI (R00-04) and one showing a low level. The rate of self-seed settings of the F(2) population showed a binomial distribution. There were 39 high-level SI plants to 15 low-level SI plants. This result and F(3) progeny tests suggested that the high level of SI which R00-04 showed is controlled by a dominant gene.

Animals↗

Development of a molecular marker specific to a novel CMS line in radish (Raphanus sativus L.).

In this study, we have investigated the cytoplasmic male sterility (CMS) of a novel male sterile radish line, designated NWB CMS. The NWB CMS was crossed with 16 fertile breeding lines, and all the progenies were completely male sterile. The degree of male sterility exhibited by NWB CMS is more than Ogura CMS from the Cruciferae family. The NWB CMS was found to induce 100% male sterility when crossed with all the tested breeding lines, whereas the Ogura CMS did not induce male sterility with any of the breeding lines. PCR analysis revealed that the molecular factor that influenced Ogura CMS, the orf138 gene, was absent in the NWB CMS line, and that the orf138 gene was not also expressed in this CMS line. In order to identify the cytoplasmic factors that confer male sterility in the NWB CMS line, we carried out RFLP analyses with 32 mitochondrial genes, all of which were used as probes. Fourteen genes exhibited polymorphisms between the NWB CMS line and other radish cultivars. Based on these RFLP data, intergenic primers were developed in order to amplify the intergenic regions between the polymorphic genes. Among these, a primer pair at the 3' region of the atp6 gene (5'-cgcttggactatgctatgtatga-3') and the 5' region of the nad3 gene (5'-tcatagagaaatccaatcgtcaa-3') produced a 2 kbp DNA fragment as a result of PCR. This DNA fragment was found to be specific to NWB CMS and was not present in other CMS types. It appears that this fragment could be used as a DNA marker to select NWB CMS line in a radish-breeding program.

Blotting, Northern↗

Delimitation of the fertility restorer locus Rfk1 to a 43-kb contig in Kosena radish (Raphanus sativus L.).

We are pursuing a positional cloning strategy to isolate the fertility restoration gene Rfk1 from radish. Random polymorphic DNA-sequence-tagged site (RAPD-STS) markers tightly linked to the gene in radish were isolated, and a RAPD map surrounding the Rfk1 locus was constructed. We surveyed 948 F2 plants with adjacent RAPD-STS markers to isolate recombinants for bulk segregant analysis. This analysis was effective in isolating tightly linked amplification fragment length polymorphism (AFLP) markers surrounding the gene of interest. Ten tightly linked AFLP markers were obtained and used to construct a high-resolution map of the region. The closest AFLP-STS markers flanking Rfk1 were 0.1 cM and 0.2 cM away. Using the four adjacent AFLP markers, we screened lambda and cosmid libraries. The lambda and cosmid clones were aligned by examination of end sequences and restriction fragment length polymorphism (RFLP) patterns for each clone, and by hybridization to the DNA isolated from recombinants. Finally, we constructed a 198-kb contig encompassing the Rfk1 gene and comprising 20 lambda and two cosmid clones. By analysis of the breakpoints in recombinants with the rfk1/rfk1 or Rfk1/- genotype, the Rfk1 locus could be assigned to a 43-kb region comprising four lambda clones and one cosmid clone. This pinpoint localization in the radish genome has made it possible for us to identify the gene by sequence analysis and genetic transformation of cytoplasmic male-sterile Brassica napus plants.

Blotting, Southern↗

Antioxidant constituents of radish sprout (Kaiware-daikon), Raphanus sativus L.

Methanol extracts of 11 kinds of commonly available vegetables were examined for hydroxyl radical scavenging potency using the bleomycin-Fe method. In this method, the iron ion and bleomycin in water form hydroxyl radicals, and the scavenging activity is monitored by the modified thiobarbituric acid method. All extracts showed scavenging capacity, even though the activity of some of them was lower than that of l-ascorbic acid. Those vegetables were classified into three groups according to their activity, groups showing strong activity, moderate activity, and weak activity, as compared to the activity of l-ascorbic acid at the same concentration. Among them, the methanol extract of radish sprout (Japanese name "kaiware-daikon") exhibited the highest potency (1.8 times as l-ascorbic acid). Then, we investigated the constituents of the methanol extract of radish sprout and the contribution to the overall activity of each compound by examining their activity. As the result, several kinds of sinapinic acid esters and flavonoids were isolated with high radical scavenging potency, which must contribute substantially to the activity.

Antioxidants↗

Selenium species bioaccessibility in enriched radish (Raphanus sativus): a potential dietary source of selenium.

An in vitro gastrointestinal method was employed to predict the potential bioavailability of selenium and its species from radish, belonging to the Brassicaceae family, grown in hydroponics media in the presence of inorganic selenium, such as Na2SeO3 and Na2SeO4. A low transformation of Se into organic forms was observed in radish plants grown in Se(VI)-enriched culture media. On the contrary, in those plants exposed to selenite, >95% of the total selenium was found as selenocystine (SeCys2), selenomethionine (SeMet), and Se-methylselenocysteine (SeMetSeCys). The concentrations of these species in fresh samples remained almost unaltered after a simulated gastrointestinal digestion. Therefore, a high selenium content of Se-methylselenocysteine (65%), previously reported as a cancer chemopreventive species, remained in the potentially bioabsorbable fraction. As these plants usually undergo a short development cycle, these results suggest that radish enriched in selenite could be a good choice as an organoselenium supplement for the human diet and animal feed.

Biological Availability↗

Subcellular localization and possible functions of gamma-glutamyltransferase in the radish (Raphanus sativus L.) plant.

Previously we reported the purification of soluble gamma-glutamyltransferases (GGTs) from radish cotyledon. Subcellular fractionation of radish cells revealed that soluble GGT is a vacuolar enzyme. Acivicin, a GGT inhibitor, mediated the in vivo catabolism inhibition of the glutathione S-conjugate generated from endogenous glutathione and exogenously supplied monochlorobimane. Thus soluble GGT is possibly involved in the catabolism of glutathione S-conjugates.

Glutathione↗