Biological and chemical characterization in relation to the yield of radish (Raphanus sativus L.) nourished with humus from plant residues.

The increase in prices of fertilizers, energy and other materials necessary for the industry triggered a global economic crisis. Reason that was investigated on the biological and chemical characteristics in relation to the yield of radish nourished with humus from plant residue. The objective was to determine the appropriate dose of humus to obtain greater yield and its relationship with the chemical and biological characteristics of the radish. It is based on applied methodology with an experimental approach; Therefore, the Completely Random Block Design model was used, which consisted of 3 blocks and 5 treatments that were T1 with 0, T2 with 4, T3 with 6, T4 with 8 and T5 with 10 t/ha of humus and They applied 15 days after sowing. The physical characteristics of the radish were evaluated and processed using analysis of variance and Duncan. Concentration of elements in leaves and stomatal density were also analyzed. It was determined that T5 stood out in total plant length with 28.95 cm, plant weight with 76.87 g, equatorial diameter with 4,404 cm and commercial yield with 20,296 t/ha. Nitrogen consumption in relation to yield with 247.44 kg/ha. Stomatal density 459 stomata/mm2 and profitability with 150% and nutrient concentration in leaves highlighted T4 with N, K, Ca, Mg, Mo and Zn. It concludes that T5 stood out with 20,296 t/ha, which differed by 26.04% in relation to the control (T1) with 15,011 t/ha. Therefore, this dose added nutrients to the soil that improved the availability for plant absorption and this influenced the concentration of nutrients in leaves such as N, P and Fe and stomatal density with 459 stomata/mm2, which had a response in good development, strengthening against environmental stress and therefore greater performance.

Oxytetracycline Increases the Residual Risk of Imidacloprid in Radish (Raphanus sativus) and Disturbs the Plant-Rhizosphere Microbiome Holobiont Homeostasis.

Antibiotics can be accidentally introduced into farmland by wastewater irrigation, and the environmental effects are still unclear. In this study, the effects of oxytetracycline on the residue of imidacloprid in soil and radishes were investigated. Besides, the rhizosphere microbiome and radish metabolome were analyzed. It showed that the persistence of imidacloprid in soil was unchanged, but the content of olefin-imidacloprid was increased by oxytetracycline. The residue of imidacloprid in radishes was increased by nearly 1.5 times, and the hazard index of imidacloprid was significantly raised by 1.5-4 times. Oxytetracycline remodeled the rhizosphere microbiome, including Actinobe, Elusimic, and Firmicutes, and influenced the metabolome of radishes. Especially, some amino acid metabolic pathways in radish were downregulated, which might be involved in imidacloprid degradation. It can be assumed that oxytetracycline increased the imidacloprid residue in radish through disturbing the plant-rhizosphere microbiome holobiont and, thus, increased the pesticide dietary risk.

Insights into the effect of manganese-based nanomaterials on the distribution trait and nutrition of radish (Raphanus sativus L.).

Manganese (Mn) is one of the essential elements for plant growth and is involved in the process of photosynthesis and seed germination. Herein, we applied two Mn-based nanoparticles, MnO2 and Mn3O4, to the soil to investigate their effects on radish growth, antioxidant system, and nutrients. The radish plant height after treatment with 10 mg/kg of MnO2 and Mn3O4 NPs were increased, compare to the control. In radish's shoot, MnO2 NPs at high concentrations (100 mg/kg) increased MDA activity by 58 % compared to the control group, while exposure to Mn3O4 NPs at the same concentration decreased MDA activity by 14 %. The nutrient content of radishes, such as soluble sugar and vitamin C, was improved. Moreover, single particle inductively coupled plasma mass spectrometry (SP ICP-MS) was used to understand the patterns of migration of Mn-based NPs in radish and subsequent impact on nutrients. We found that Mn-based NPs accumulated mainly in the roots of radish. Interestingly, the accumulation characteristics of MnO2 NPs and Mn3O4 NPs were different. MnO2 NPs accumulated more in radish leaves than in fruits, while the accumulation of Mn3O4 NPs gradually decreased from roots to leaves. Finally, we determined the mineral element content of the leaves, fruits, and roots of radish, and found that the uptake of main metallic mineral elements (e.g. Cu, Fe, Mg, Zn, Na, K) was inhibited by the application of Mn-based NPs. These findings underscore the importance of considering species and multifaceted impacts of Mn-based NPs as nanofertilizers for their wide application in agriculture.

Combinatorial preparation and structural characterization of anthocyanins and aglycones from Purple-heart Radish for evaluation of physicochemical stability and pancreatic lipase inhibitory activity.

In this study, an efficient approach to preparation of different anthocyanins from Purple-heart Radish was developed by combining microwave-assisted extraction (MAE), macroporous resin purification (MRP) and ultrasound-assisted acid hydrolysis (UAAH) for evaluation of physicochemical stability and pancreatic lipase (PL) inhibitory activity. By optimization of MAE, MRP and UAAH processes, the anthocyanins reached the yield of 6.081 ± 0.106 mg/g, the purity of 78.54 ± 0.62 % (w/w) and the content of 76.29 ± 1.31 % (w/w), respectively. With high-resolution UHPLC-Q-Orbitrap/MS, 15 anthocyanins were identified as pelargonins with diverse glucosides and confirmed by pelargonidin standard. By glycosylation, pelargonins exhibited higher stability in different pH, temperature, light, metal ions environments than that of pelargonidin. However, PL inhibitory assay, kinetic analysis and molecular docking demonstrated that pelargonidin had higher PL inhibitory activity than pelargonins even though with similar binding sites and a dose-effect relationship. The above results revealed that the effect of glycosylation and deglycosylation on PL inhibitory activity and physicochemical stability.

Phytochemical Profile, Bioactive Properties, and Se Speciation of Se-Biofortified Red Radish (Raphanus sativus), Green Pea (Pisum sativum), and Alfalfa (Medicago sativa) Microgreens.

The impact of selenium (Se) enrichment on bioactive compounds and sugars and Se speciation was assessed on different microgreens (green pea, red radish, and alfalfa). Sodium selenite and sodium selenate at a total concentration of 20 µM (1:1) lead to a noticeable Se biofortification (40-90 mg Se kg-1 DW). In green pea and alfalfa, Se did not negatively impact phenolics and antioxidant capacity, while in red radish, a significant decrease was found. Regarding photosynthetic parameters, Se notably increased the level of chlorophylls and carotenoids in green pea, decreased chlorophyll levels in alfalfa, and had no effect on red radish. Se treatment significantly increased sugar levels in green pea and alfalfa but not in red radish. Red radish had the highest Se amino acid content (59%), followed by alfalfa (34%) and green pea (28%). These findings suggest that Se-biofortified microgreens have the potential as functional foods to improve Se intake in humans.

Novel insights into the anti-asthmatic effect of Raphanus sativus L. (Raphani Semen): Targeting immune cells, inflammatory pathways and oxidative stress markers.

ETHNOPHARMACOLOGICAL RELEVANCE: Raphanus sativus L. is a well-known medicinal plant with traditional therapeutic applications in various common ailments including inflammation and asthma. AIMS OF THE STUDY: This study aimed to evaluate the chemical composition and anti-asthmatic potential of the hydro-methanolic extract of the leaves of R. sativus L. (Rs.Cr) using various in vitro and in vivo investigations. MATERIALS AND METHODS: The Rs.Cr was subjected to preliminary phytochemical analysis and HPLC profiling. The safety was assessed through oral acute toxicity tests in mice. The antiasthmatic effect of the extract was studied using milk-induced leukocytosis and ovalbumin (OVA)-induced allergic asthma models established in mice. While mast cell degranulation and passive paw anaphylaxis models were established in rats. Moreover, effect of the extract was studied on various oxidative and inflammatory makers. The antioxidant effect of the extract was also studied by in vitro DPPH method. RESULTS: The HPLC profiling of Rs.Cr showed the presence of important polyphenols in a considerable quantity. In toxicity evaluation, Rs.Cr showed no sign of morbidity or mortality with LD50 < 2000 mg/kg. The extract revealed significant mast cell disruption in a dose-dependent manner compared to the intoxicated group. Similarly, treatment with Rs.Cr and dexamethasone significantly (p < 0.001) reduced paw edema volume. Subcutaneous injection of milk at a dose of 4 mL/kg, after 24 h of its administration, showed an increase in the leukocyte count in the intoxicated group. Similarly, mice treated with dexamethasone and Rs.Cr respectively showed a significant decrease in leukocytes and eosinophils count in the ovalbumin-induced allergic asthma model. The extract presented a significant (p˂0.001) alleviative effect on the levels of SOD and GSH, MDA, IL-4, IL-5, and IL-13 in a dose-dependent manner as compared to the intoxicated group. Furthermore, the histological evaluation also revealed a notable decrease in inflammatory and goblet cell count with reduced mucus production. CONCLUSION: The current study highlights mechanism-based novel insights into the anti-asthmatic potential of R. sativus that also strongly supports its traditional use in asthma.

Broccoli, Kale, and Radish Sprouts: Key Phytochemical Constituents and DPPH Free Radical Scavenging Activity.

Our research group previously found that broccoli sprouts possess neuroprotective effects during pregnancy. The active compound has been identified as sulforaphane (SFA), obtained from glucosinolate and glucoraphanin, which are also present in other crucifers, including kale. Sulforaphene (SFE), obtained from glucoraphenin in radish, also has numerous biological benefits, some of which supersede those of sulforaphane. It is likely that other components, such as phenolics, contribute to the biological activity of cruciferous vegetables. Notwithstanding their beneficial phytochemicals, crucifers are known to contain erucic acid, an antinutritional fatty acid. The aim of this research was to phytochemically examine broccoli, kale, and radish sprouts to determine good sources of SFA and SFE to inform future studies of the neuroprotective activity of cruciferous sprouts on the fetal brain, as well as product development. Three broccoli: Johnny's Sprouting Broccoli (JSB), Gypsy F1 (GYP), and Mumm's Sprouting Broccoli (MUM), one kale: Johnny's Toscano Kale (JTK), and three radish cultivars: Black Spanish Round (BSR), Miyashige (MIY), and Nero Tunda (NT), were analyzed. We first quantified the glucosinolate, isothiocyanate, phenolics, and DPPH free radical scavenging activity (AOC) of one-day-old dark- and light-grown sprouts by HPLC. Radish cultivars generally had the highest glucosinolate and isothiocyanate contents, and kale had higher glucoraphanin and significantly higher sulforaphane content than the broccoli cultivars. Lighting conditions did not significantly affect the phytochemistry of the one-day-old sprouts. Based on phytochemistry and economic factors, JSB, JTK, and BSR were chosen for further sprouting for three, five, and seven days and subsequently analyzed. The three-day-old JTK and radish cultivars were identified to be the best sources of SFA and SFE, respectively, both yielding the highest levels of the respective compound while retaining high levels of phenolics and AOC and markedly lower erucic acid levels compared to one-day-old sprouts.

Physiological and biochemical markers of gamma irradiated white radish (Raphanus sativus).

PURPOSE: A field experiment was performed to investigate the impact of low-dose gamma rays on growth parameters and bioactive compounds of white radish. MATERIALS AND METHODS: White radish seeds were irradiated by gamma rays dose levels (10, 20, 40 and 80 Gy) beside control. Physiological and biochemical markers were done to follow the effect of gamma rays on white radish. RESULTS: The results revealed that gamma rays increased growth parameters with increasing irradiation to a dose of 40 Gy. The maximum increments were found at 14.64 (cm), 48.30 (cm), 20.84 (cm) and 5.51 (cm) for leaves number, leaves length, roots length and roots diameter, respectively, with a dose of 40 Gy. By increasing the irradiation dose to 80 Gy, the results showed reduction in all parameters studied. Ascorbic acid gave the maximum increase with the dose of 40 Gy, while phenols, flavonoids, antioxidant activity, peroxidase, and polyphenol oxidase showed the highest increase with the dose 80 of Gy in radish leaves. Similar trend was observed for the radish roots. Furthermore, the protein and isoenzyme profiles of peroxidase and polyphenol oxidase changed and induced alteration by different irradiation dose levels. CONCLUSION: Gamma rays can be a useful tool for increasing the growth and biochemical content of white radish plants and perhaps other food crops.

Metabolomic approach of azole fungicides in radish (Raphanus sativus): Perspective of functional metabolites.

Azole fungicides is one of the major fungicides in agricultural field. In this study, toxic effects of diniconazole (DIN), metconazole (MET), and tebuconazole (TEB) to radish leaves and roots were investigated using targeted metabolomics with gas chromatography-mass spectrometry (GC-MS/MS). Especially, the changes of functional chemicals, including phytosterols and glucosinolates evaluated. Radish leaves and roots were harvested after 7 days and 14 days from last exposure. In multivariate analysis, the experimental groups showed clear separation in PCA and PLS-DA score plots. Phytosterols and glucosinolates were significantly changed by azole fungicide. Six metabolic pathways which are affected by fungicides were selected and showed similar patterns regardless of the type of azole fungicide used. As a result, azole fungicide induces the defense mechanisms of plants and affects both primary and secondary metabolism.

Improvement of physicochemical characteristics, flavor profiles and functional properties in Chinese radishes via spontaneous fermentation after drying.

Spontaneously dried-fermented radishes have been consumed in China for hundreds of years and are usually fermented for a long time to acquire high quality. In this study, the spontaneously dried-fermented radishes with short-term manufacturing periods were made from five different varieties of radishes that grew in the same environment. In addition, the physicochemical characteristics (i.e., moisture content, soluble solid, and pH value), flavor profiles (i.e., free amino acids, organic acids, and volatile compounds), and functional properties (i.e., total phenolics content, total flavonoids content, sulforaphane content, and γ-aminobutyric acid [GABA] content) of these five raw radishes and spontaneously dried-fermented radishes were analyzed and compared. In detail, the content of volatile and nonvolatile compounds increased, especially in oxalic acid, succinic acid, and umami free amino acids. Furthermore, functional components, such as sulforaphane and GABA, were also enriched via spontaneous fermentation after drying. In addition, the results of principal component analysis, hierarchical clustering analysis, and redundancy analysis showed that there were significant discrepancies appeared when raw radishes were processed via spontaneous fermentation or not. These results suggested that the process of spontaneous fermentation after drying may contribute to improving the quality of fresh radishes. Notably, radishes with red skin and flesh were regarded as exceptional varieties for processing, because of the preferable flavor profiles and affluent functional substances via spontaneous fermentation after drying. Therefore, these findings could deliver a systematical insight into developing processed radishes with high quality. PRACTICAL APPLICATION: The spontaneously dried-fermented radishes were manufactured through the process of spontaneous fermentation after drying, which acquired tasty and healthy characteristics by accumulating the volatile and nonvolatile compounds as well as the functional components, like total phenolics, total flavonoids, sulforaphane, and γ-aminobutyric acid. Importantly, because of the excellent processing properties, the radishes with red skin and flesh could be more appropriate to produce spontaneously dried-fermented radishes. Our findings may provide a practical strategy for developing vegetable relishes with superb flavor profiles and good functional properties in pickled vegetables.

Understanding the nutraceutical diversity through a comparative analysis of the taproot metabolomes of different edible radish types via UHPLC-Q-TOF-MS.

Radishes are root vegetables that are rich in bioactive compounds and provide numerous health benefits, but the overall metabolic profiles of radish taproots and the metabolic differences among different edible types are not fully understood. In this research, we used UHPLC-Q-TOF-MS to identify the metabolites in cooked, processed and fruit radishes of ten varieties. In total, 264 metabolites belonging to 18 categories were detected. A multivariate analysis revealed that the metabolite composition differed among the three radish groups, and a comparative analysis showed that the significantly differentially accumulated metabolites were mainly amino acids and derivatives, lipids, flavonoids, hydroxycinnamate derivatives and carbohydrates. The accumulation of metabolites, particularly flavonoids, was greater in fruit radishes than in cooked and processed radishes. This work provides novel insights into the radish metabolomic profiles for assessment of the nutritional value of different edible radish types for humans.

An analysis of the content changes in free and combinative forms of organic selenium in radish sprouts cultivated with solutions of selenoamino acids.

Selenium (Se) is critical for human health, but human intake of Se is often inadequate. Organic forms of dietary Se are considered safe and more bioavailable than inorganic forms. Along with a generally high nutritional value, sprouts are sensitive to Se treatment. This study used selenomethionine and methylselenocysteine solutions to cultivate Se-enriched sprouts under an optimized hydroponic condition. Content change and transformations of the selenoamino acids were analyzed by a developed HPLC-ESI-MS/MS method. Uptake of both selenomethionine and methylselenocysteine was dose-dependent and involved active transport and passive diffusion, as demonstrated by the respiratory and aquaporin inhibition assays. Passive diffusion played a dominant role. Free methylselenocysteine was the predominant form in samples. Selenomethionine and methylselenocysteine were capable of mutual transformation. Moreover, the selenoprotein generation was associated with the increasing Se concentration of the culture solutions. The results provided scientific references for the efficient utilization of organic Se in sprouts.

Isolation and characterization of the anthocyanins derived from red radishes (Raphanus sativus L.) and the protective ability of ß-lactoglobulin against heat-induced oxidation.

This study employed the "two-step dialysis" method and AB-8 or D101 macroporous resin chromatography to isolate the anthocyanins in red radishes (ARR). The red radish juice was dialyzed twice at 3000 and 500 Da, respectively. UHPLC-QqQ-MS/MS revealed 24 types of ARRs, of which pelargonidin (Pg)-3-diglucoside-5-(malonyl)glucoside (P3D5MG), Pg-3-diglucoside-5-glucoside (P3D5G), Pg-3-(feruloyl)diglucoside-5-(malonyl)glucoside (P3FD5MG), Pg, and malvidin (Mv) represented the main compounds. The total anthocyanin content in the ARR prepared via the "two-step dialysis" method was 29.69% and 18.44% higher than that obtained using AB-8 and D101 macroporous resins, respectively. The ARRs inhibited heat-induced ß-lactoglobulin (ß-Lg) oxidation. The amino acid residue microenvironment and secondary ß-Lg structure were modified via ARR binding. The energy involved in P3D5MG and ß-Lg binding was -392 kJ/mol, which was significantly lower than that during the binding process of P3D5M, P3FD5MG, Pg, and Mv to ß-Lg (-338 to -168 kJ/mol). These results indicated that "two-step dialysis" was a promising method for deriving natural pigment with strong antioxidant activity from red radishes. PRACTICAL APPLICATION: As a natural food colorant, anthocyanins have attracted increasing attention in the food industry in recent years. This study used "two-step dialysis" to effectively separate ARRs. Moreover, the anthocyanins in ARR can bind to ß-Lg to protect against heating-induced oxidation. Therefore, ARRs may not only act as a food pigment but also as antioxidants.

Denitrifying halophilic archaea derived from salt dominate the degradation of nitrite in salted radish during pickling.

Salted radish is a popular high-salinity table food in China, and nitrite is always generated during the associated pickling process. However, this nitrite can be naturally degraded, and the underlying mechanism is unknown. Here, we identified the microbial groups that dominate the natural degradation of nitrite in salted radish and clarified the related metabolic mechanism. Based on dynamic monitoring of pH and the concentrations of nitrogen compounds as well as high-throughput sequencing analysis of the structural succession of microbial communities in the tested salted radish, we determined that the halophilic archaea derived from pickling salt dominate the natural degradation of nitrite via denitrification. Based on isolation, identification, nitrite reduction assays, and genome annotation, we further determined that Haloarcula, Halolamina, and Halobacterium were the key genera. These halophilic archaea might cope with high salt stress through the "salt-in" mechanism with the assistance of the accumulation of potassium ions, obtain electrons necessary for "truncated denitrification" from the metabolism of extracellular glucose absorbed from salted radish, and efficiently reduce nitrite to nitrogen, bypassing nitrite generation from nitrate reduction. The present study provides important information for the prevention and control of nitrite hazards in salted vegetables with high salinity, such as salted radish.

Raphani Semen (Raphanus sativus L.) Ameliorates Alcoholic Fatty Liver Disease by Regulating De Novo Lipogenesis.

In this study, we investigated the pharmacological effect of a water extract of Raphani Semen (RSWE) on alcoholic fatty liver disease (AFLD) using ethanol-induced AFLD mice (the NIAAA model) and palmitic acid (PA)-induced steatosis HepG2 cells. An RSWE supplement improved serum and hepatic triglyceride (TG) levels of AFLD mice, as well as their liver histological structure. To explore the molecular action of RSWE in the improvement of AFLD, we investigated the effect of RSWE on four major pathways for lipid homeostasis in the liver: free fatty acid transport, lipogenesis, lipolysis, and ß-oxidation. Importantly, RSWE decreased the mRNA expression of de novo lipogenesis-related genes, such as Srebf1, Cebpa, Pparg, and Lpin1, as well as the protein levels of these factors, in the liver of AFLD mice. That these actions of RSWE affect lipogenesis was confirmed using PA-induced steatosis HepG2 cells. Overall, our findings suggest that RSWE has the potential for improvement of AFLD by inhibiting de novo lipogenesis.

Radish sprout alleviates DSS-induced colitis via regulation of NF-kB signaling pathway and modifying gut microbiota.

In this study, we investigated the effects of radish sprout ethanol extract (RSE) on inflammatory responses in the macrophages and a mouse model of colitis. RSE administration was found to effectively inhibit the phosphorylation of IκB and, in turn, the production of pro-inflammatory enzymes and cytokines in lipopolysaccharide-stimulated macrophages. In dextran sulfate sodium (DSS)-colitis mice, RSE administration prevented body weight and colon length reduction, while decreasing inflammation and mucosal necrosis. The diversity of the fecal microbiota was significantly increased in the group treated with RSE. In addition, RSE administration decreased the relative abundance of the phylum Proteobacteria, which includes many pathogens, and increased the abundance of the genus Akkermansia. Beta diversity analyses showed that RSE administration restored the gut microbiota composition close to that of healthy mice. For the first time, we identified glycosides of sinapic acid as part of hydroxycinnamic acids in RSE with colitis-alleviating effects. Notably, 1,2-O-disinapoyl glucoside substantially decreased nitric oxide generation in LPS-stimulated macrophages.

[Effects of alcohol extract of radish seed on blood lipid, glucose and liver steatosis in ApoE-/- mice].

Objective: To investigate the effects and mechanisms of alcohol extract of Raphani seed (AERS) on blood lipid, blood glucose and hepatic steatosis in ApoE-/- mice. Methods: Sixty ApoE-/- mice were randomly divided into control group (CG), normal saline group (NG), rosiglitazone group (RG) and AERS treatment groups (AERS-L / M / H). Except CG, other groups were fed with high-fat, high-sugar and high-salt diet for 9 weeks. The mice in RG were treated with rosiglitazone (1.33 mg·kg-1, 0.2 ml ·10 g-1) by gavage daily. The mice in CG and NG were treated with equal volume of NS by intragastric administration daily. The mice in AERS groups were treated with AERS at the doses of 100, 300 and 900 mg·kg-1 for 9 weeks, respectively. FPG and Fins were detected. Insulin resistance index (IRI) and liver coefficient (LC) were calculated. The levels of ALT, AST, Leptin (LEP), TNF - α and blood lipid (TC, FFA, etc.) were tested. The expressions of proteins related to liver lipid metabolism (HMG-R、LDL-R、LEP-R) were detected by Western blot. Results: Compared with NG or RG, CG showed significant changes in liver appearance (color, swelling, etc.) and pathology (steatosis, hepatocyte necrosis, etc.), while AERS-M/H groups were similar to CG. Compared with CG, the serum levels of FPG, Fins, IRI, ALT, AST, TNF-α, LC, TG, LDL-C, FFA and LEP were increased significantly (P＜0.05) . Compared with NG, AERS could decrease the above mentioned indicators in a dose-dependent manner (P＜0.05). Compared with RG, the levels of FPG and Fins of AERS-H were increased significantly, while the level of IRI was decreased (P＜0.05). Compared with NG and RG, the protein expressions of HMG-R and LEP-R in AERS groups were decreased, while the protein expression of LDL-R was increased in a dose-dependent manner (P＜0.05). Conclusion: AERS can prevent the increase of blood lipid, glucose and hepatic steatosis induced by high-fat and high-sugar diet in ApoE-/- mice. The mechanism is related to the decrease of FFA and LEP, the inhibition of TNF-α, HMG-R, LEP-R protein expressions and the promotion of LDL-R protein expression.

Loss of the R2R3 MYB Transcription Factor RsMYB1 Shapes Anthocyanin Biosynthesis and Accumulation in Raphanus sativus.

The red or purple color of radish (Raphanus sativus L.) taproots is due to anthocyanins, which have nutritional and aesthetic value, as well as antioxidant properties. Moreover, the varied patterns and levels of anthocyanin accumulation in radish roots make them an interesting system for studying the transcriptional regulation of anthocyanin biosynthesis. The R2R3 MYB transcription factor RsMYB1 is a key positive regulator of anthocyanin biosynthesis in radish. Here, we isolated an allele of RsMYB1, named RsMYB1Short, in radish cultivars with white taproots. The RsMYB1Short allele carried a 4 bp insertion in the first exon causing a frame-shift mutation of RsMYB1, generating a truncated protein with only a partial R2 domain at the N-terminus. Unlike RsMYB1Full, RsMYB1Short was localized to the nucleus and the cytoplasm and failed to interact with their cognate partner RsTT8. Transient expression of genomic or cDNA sequences for RsMYB1Short in radish cotyledons failed to induce anthocyanin accumulation, but that for RsMYB1Full activated it. Additionally, RsMYB1Short showed the lost ability to induce pigment accumulation and to enhance the transcript level of anthocyanin biosynthetic genes, while RsMYB1Full promoted both processes when co-expressed with RsTT8 in tobacco leaves. As the result of the transient assay, co-expressing RsTT8 and RsMYB1Full, but not RsMYB1Short, also enhanced the promoter activity of RsCHS and RsDFR. We designed a molecular marker for RsMYB1 genotyping, and revealed that the RsMYB1Short allele is common in white radish cultivars, underscoring the importance of variation at the RsMYB1 locus in anthocyanin biosynthesis in the radish taproot. Together, these results indicate that the nonsense mutation of RsMYB1 generated the truncated protein, RsMYB1Short, that had the loss of ability to regulate anthocyanin biosynthesis. Our findings highlight that the frame shift mutation of RsMYB1 plays a key role in anthocyanin biosynthesis in the radish taproot.

An intrinsically disordered radish vacuolar calcium-binding protein (RVCaB) showed cryoprotective activity for lactate dehydrogenase with its hydrophobic region.

A soluble protein fraction from radish (Raphanus sativus L.) taproot had cryoprotective activity for lactate dehydrogenase (LDH). The activity was found mainly in the heat-stable fractions of soluble proteins. The cryoprotective protein, whose molecular mass was 43 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis, was obtained by successive chromatographies on TOYOPEARL SuperQ and TOYOPEARL DEAE. MALDI-TOF MS/MS analysis indicated that the purified protein was a radish vacuolar calcium-binding protein (RVCaB), which is reportedly related to calcium storage in the vacuoles of radish taproot. The purified RVCaB inhibited the cryoinactivation, cryodenaturation, and cryoaggregation of LDH. RVCaB had greater cryoprotective activity than general cryoprotectants. When RVCaB was divided into 15 segments (Seg01 to Seg15, 15 amino acids each), Seg03, which had a high hydrophobicity scale, showed remarkable cryoprotective activity. This indicated that RVCaB protected LDH from freezing and thawing damage presumably through a specific hydrophobic area (i.e., Seg03).

Toxic metal phytoextraction potential and health-risk parameters of some cultivated plants when grown in metal-contaminated river sediment of Danube, near an industrial town.

Toxic metal phytoextraction potential of some higher plants, the white mustard (Sinapis alba L.), perennial rye grass (Lolium perenne L.) and also two cultivated plants, as green pea (Pisum sativum L. var. Rajnai törpe), radish (Raphanus sativus L. var. Szentesi óriás vaj), was studied in a field experiment, along the river Danube in close vicinity of an industrial town, Dunaújváros, Hungary. Soil/sediment and the various plant organs (leaves, stems and roots) were assessed for the contamination with some potentially toxic elements (PTE), such as the cadmium (Cd), nickel (Ni), copper (Cu), and zinc (Zn). It was found that Cd and Ni concentration was below, while the Cu and Zn elements were above the Hungarian permissible limits in each of the studied soil/sediment samples. Bioconcentration factor (BAF) was less than 1 in the shoot biomass of test plant samples and followed the order of Cu > Zn > Cd and Ni. Phytoremediation potential of selected test plants was found to be rather limited. The translocation factor (TF) was more than 1 for Cu and Zn elements, at each test plants. Cadmium was translocated into the leaves in case of the radish, only. Considering of the potential human daily intake of metals (DIM), it was less than 1 both for the adults and for the children. Health risk index (HRI) values of children, however, were higher than 1 for the Cd in case of radish, and for Zn and Cu in case of the pea. Results suggest that consumption of these plants grown in gardens of contaminated sediments can result in some risks for citizens in the industrial town of Dunaújváros. Further studies are required to identify appropriate plants with greater toxic metal phytoextraction potential.