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Brassinosteroids (BRs) and polyamines (PAs) are well-established growth regulators playing key roles in stress management among plants. In the present study, we evaluated the effects of epibrassinolide (EBL, an active BR) and spermidine (Spd, an active PA) on the tolerance of radish to oxidative stress induced by Cr (VI) metal. Our investigation aimed to study the impacts of EBL (10(-9) M) and/or Spd (1 mM) on the biochemical and physiological responses of radish (Raphanus sativus L.) under Cr-stress. Applications of EBL and/or Spd were found to improve growth of Cr-stressed seedlings in terms of root length, shoot length and fresh weight. Our data also indicated that applications of EBL and Spd have significant impacts, particularly when applied together, on the endogenous titers of PAs, free and bound forms of IAA and ABA in seedlings treated with Cr-stress. Additionally, co-applications of EBL and Spd modulated more remarkably the titers of antioxidants (glutathione, ascorbic acid, proline, glycine betaine and total phenol) and activities of antioxidant enzymes (guaicol peroxidase, catalase, superoxide dismutase and glutathione reductase) in Cr-stressed plants than their individual applications. Attenuation of Cr-stress by EBL and/or Spd (more efficient with EBL and Spd combination) was also supported by enhanced values of stress indices, such as phytochelatins, photosynthetic pigments and total soluble sugars, and reduction in malondialdehyde and H(2)O(2) levels in Cr-treated seedlings. Diminution of ROS production and enhanced ROS scavenging capacities were also noted for EBL and/or Spd under Cr-stress. However, no significant reduction in Cr uptake was observed for co-application of EBL and Spd when compared to their individual treatments in Cr-stressed seedlings. Taken together, our results demonstrate that co-applications of EBL and Spd are more effective than their independent treatments in lowering the Cr-induced oxidative stress in radish, leading to improved growth of radish seedlings under Cr-stress.

Red-fleshed radish (Raphanus sativus L.) is a unique cultivar whose taproot is rich in anthocyanins beneficial to human health. However, the frequent occurrence of white-fleshed mutants affects the purity of commercially produced radish and the underlying mechanism has puzzled breeders for many years. In this study, we combined quantitative trait location by genome resequencing and transcriptome analyses to identify a candidate gene (RsMYB1) responsible for anthocyanin accumulation in red-fleshed radish. However, no sequence variation was found in the coding and regulatory regions of the RsMYB1 genes of red-fleshed (MTH01) and white-fleshed (JC01) lines, and a 7372 bp CACTA transposon in the RsMYB1 promoter region occurred in both lines. A subsequent analysis suggested that the white-fleshed mutant was the result of altered DNA methylation in the RsMYB1 promoter. This heritable epigenetic change was due to the hypermethylated CACTA transposon, which induced the spreading of DNA methylation to the promoter region of RsMYB1. Thus, RsMYB1 expression was considerably down-regulated, which inhibited anthocyanin biosynthesis in the white-fleshed mutant. An examination of transgenic radish calli and the results of a virus-induced gene silencing experiment confirmed that RsMYB1 is responsible for anthocyanin accumulation. Moreover, the mutant phenotype was partially eliminated by treatment with a demethylating agent. This study explains the molecular mechanism regulating the appearance of white-fleshed mutants of red-fleshed radish.

Due to the high solubility of Cd in water, it is considered a potential toxin which can cause cancer in humans. In plants, it is associated with the development of oxidative stress due to the generation of reactive oxygen species. To overcome this issue, the roles of different plant hormones are vital. Strigolactones, one of such natural plant hormones, show promise in alleviating cadmium toxicity by mitigating its harmful effects. Acidified biochar (AB) can also effectively mitigate cadmium toxicity via ion adsorption and pH buffering. However, the combined effects of strigolactone and AB still need in-depth investigations in the context of existing literature. This study aimed to assess the individual and combined impacts of SLs (0 and 25 µM) and AB (0 and 0.75% w/w) on radish growth under Cd toxicity, i.e., 0 and 20 mg Cd/kg soil. Using a fully randomized design (CRD), each treatment was administered in four replicates. In comparison to the control under 20 mg Cd/kg soil contamination, the results showed that 25 µM strigolactone + 0.75% AB significantly improved the following: radish shoot length (~ 17%), root length (~ 47%), plant fresh weight (~ 28%), plant dry weight (~ 96%), chlorophyll a (~ 43%), chlorophyll b (~ 31%), and total chlorophyll (~ 37%). It was also noted that 0.75% AB was more pronounced in decreasing antioxidant activities than 25 µM strigolactone under 20 mg Cd/ kg soil toxicity. However, performing 25 µM strigolactone + 0.75% AB was far better than the sole application of 25 µM strigolactone and 0.75% AB in decreasing antioxidant activities in radish plants. In conclusion, by regulating antioxidant activities, 25 µM strigolactone + 0.75% AB can increase radish growth in cadmium-contaminated soils.

Carmine radish ( Raphanus sativus L.) is cultivated in Fuling, Chongqing, for its red color. Dof -TFs are critical in regulating plant growth, development, stress responses, and signal transduction.This work comprehensively examined the structure, evolution, and expression of the carmine radish Dof gene and its behavior under cadmium (Cd) stress. The radish genome has 59 RsDofs , which are divided into nine clusters (A: 8, B1: 10, B2: 10, C1: 3, C2.1: 5, C2.2: 4, C3: 11, D1: 4, and D2: 4). Phylogenetic tree analysis revealed significant Dof gene family resemblance between Arabidopsis thaliana and Brassica napus . Perhaps segment duplication resulted in RsDof gene family expansion. Cd stress-induced RsDof expression patterns were studied using an RNA-seq atlas and qRT-PCR. The majority of RsDofs were tissue-specific and Cd-sensitive. The involvement of RsDof genes in Cd stress response and anthocyanin synthesis was verified using qRT-PCR. RsDof33 is involved in Cd stress response and anthocyanin synthesis. A. thaliana overexpressed the recombinant fusion protein RsDof33-GFP, which was localized to the nucleus, resulting in fewer rosette leaves, delayed flowering, and higher anthocyanin concentration. RsDof33 -expressing plants had significantly higher transcript levels of the auxin biosynthetic genes YUCCA ( AtYUC2 ), auxin efflux carrier ( AtPIN4 ), and AtKNAT2 , which are involved in leaf shape development, as well as AtPAL , AtCHS , AtCHI , AtDFR , AtLDOX , and AtUF3GT . These findings indicate that RsDofs are critical to plant development and stress responses.

Lead (Pb), one of the most toxic heavy metals, can be absorbed and accumulated by plant roots and then enter the food chain resulting in potential health risks for human beings. The radish (Raphanus sativus L.) is an important root vegetable crop with fleshy taproots as the edible parts. Little is known about the mechanism by which radishes respond to Pb stress at the molecular level. In this study, Next Generation Sequencing (NGS)-based RNA-seq technology was employed to characterize the de novo transcriptome of radish roots and identify differentially expressed genes (DEGs) during Pb stress. A total of 68,940 assembled unique transcripts including 33,337 unigenes were obtained from radish root cDNA samples. Based on the assembled de novo transcriptome, 4,614 DEGs were detected between the two libraries of untreated (CK) and Pb-treated (Pb1000) roots. Gene Ontology (GO) and pathway enrichment analysis revealed that upregulated DEGs under Pb stress are predominately involved in defense responses in cell walls and glutathione metabolism-related processes, while downregulated DEGs were mainly involved in carbohydrate metabolism-related pathways. The expression patterns of 22 selected genes were validated by quantitative real-time PCR, and the results were highly accordant with the Solexa analysis. Furthermore, many candidate genes, which were involved in defense and detoxification mechanisms including signaling protein kinases, transcription factors, metal transporters and chelate compound biosynthesis related enzymes, were successfully identified in response to heavy metal Pb. Identification of potential DEGs involved in responses to Pb stress significantly reflected alterations in major biological processes and metabolic pathways. The molecular basis of the response to Pb stress in radishes was comprehensively characterized. Useful information and new insights were provided for investigating the molecular regulation mechanism of heavy metal Pb accumulation and tolerance in root vegetable crops.

Water scarcity is a foremost environmental concern and is expected to hasten in the forthcoming years due to severe fluctuations in weather patterns worldwide. The present work was designed to explore the potential role of alpha-tocopherol (α-Toc), a form of vitamin E, on the morphological, physio-biochemical, and cellular antioxidant responses of two radish genotypes grown under drought conditions (38 ± 3% of field capacity). The individual and combined applications of α-Toc (100 ppm) were used as T0- Control, T1- Control + TF (TF-alpha-tocopherol), T2- Drought (D), and T3- D + TF with three replications. In general, drought conditions cause a marked reduction in, growth traits such as root length (RL), shoot dry weight (SDW), and shoot fresh weight (SFW). However, the sole and combined applications of α-Toc significantly enhanced the SDW, SFW, and RL in both radish genotypes. Drought stress causes a significant upsurge in hydrogen peroxide (H 2 O 2 ) and lipid peroxidation (LPX) in leaves. At the same time, exogenous fertigation of α-Toc protects the membranes by reducing the level of LPX, enhancing antioxidants such as catalase (CAT) and peroxidase (POX) to scavenge the reactive oxygen species (ROS), and enhancing the osmolyte as total soluble proteins to maintain cell internal osmotic potential. Also, the α-Toc enhanced the photosynthetic pigments and significantly increased photosynthetic activity in the Early Milo (G2) as compared to Laal Pari (G1) genotype under drought, enhancing water use efficiency by maintaining transpiration rate and stomatal conductance. The α-Toc also regulates the beneficial inorganic ions (K + , Ca 2+ , and PO₄³⁻) in the shoots of both genotypes. Our present findings demonstrate the potential role of α-Toc in mitigating drought stress and infer that it can enhance plant growth under drought conditions.

Glucosinolates (GSLs) are secondary metabolites whose degradation products confer intrinsic flavors and aromas to Brassicaceae vegetables. Several structures of GSLs are known in the Brassicaceae, and the biosynthetic pathway and regulatory networks have been elucidated in Arabidopsis (Arabidopsis thaliana). GSLs are precursors of chemical defense substances against herbivorous pests. Specific GSLs can act as feeding blockers or stimulants, depending on the pest species. Natural selection has led to diversity in the GSL composition even within individual species. However, in radish (Raphanus sativus), glucoraphasatin (4-methylthio-3-butenyl glucosinolate) accounts for more than 90% of the total GSLs, and little compositional variation is observed. Because glucoraphasatin is not contained in other members of the Brassicaceae, like Arabidopsis and cabbage (Brassica oleracea), the biosynthetic pathways for glucoraphasatin remain unclear. In this report, we identified and characterized a gene encoding GLUCORAPHASATIN SYNTHASE 1 (GRS1) by genetic mapping using a mutant that genetically lacks glucoraphasatin. Transgenic Arabidopsis, which overexpressed GRS1 cDNA, accumulated glucoraphasatin in the leaves. GRS1 encodes a 2-oxoglutarate-dependent dioxygenase, and it is abundantly expressed in the leaf. To further investigate the biosynthesis and transportation of GSLs in radish, we grafted a grs1 plant onto a wild-type plant. The grafting experiment revealed a leaf-to-root long-distance glucoraphasatin transport system in radish and showed that the composition of GSLs differed among the organs. Based on these observations, we propose a characteristic biosynthesis pathway for glucoraphasatin in radish. Our results should be useful in metabolite engineering for breeding of high-value vegetables.

The radish ( Raphanus sativus L.) is an important root vegetable crop. In this study, the metabolite profiling analysis of radish roots exposed to lead (Pb) and cadmium (Cd) stresses has been performed using gas chromatography-mass spectrometry (GC-MS). The score plots of principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA) showed clear discrimination between control and Pb- or Cd-treated samples. The metabolic profiling indicated Pb or Cd stress could cause large metabolite alteration mainly on sugars, amino acids and organic acids. Furthermore, an integrated analysis of the effects of Pb or Cd stress was performed on the levels of metabolites and gene transcripts from our previous transcriptome work in radish roots. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of integration data demonstrated that exposure of radish to Pb stress resulted in profound biochemical changes including carbohydrate metabolism, energy metabolism and glutathione metabolism, while the treatment of Cd stress caused significant variations in energy production, amino acid metabolism and oxidative phosphorylation-related pathways. These results would facilitate further dissection of the mechanisms of heavy metal (HM) accumulation/tolerance in plants and the effective management of HM contamination in vegetable crops by genetic manipulation.

Resistance to auxinic herbicides is increasing in a range of dicotyledonous weed species, but in most cases the biochemical mechanism of resistance is unknown. Using 14C-labelled herbicide, the mechanism of resistance to 2,4-dichlorophenoxyacetic acid (2,4-D) in two wild radish (Raphanus raphanistrum L.) populations was identified as an inability to translocate 2,4-D out of the treated leaf. Although 2,4-D was metabolized in wild radish, and in a different manner to the well-characterized crop species wheat and bean, there was no difference in metabolism between the susceptible and resistant populations. Reduced translocation of 2,4-D in the latter was also not due to sequestration of the herbicide, or to reduced uptake by the leaf epidermis or mesophyll cells. Application of auxin efflux or ABCB transporter inhibitors to 2,4-D-susceptible plants caused a mimicking of the reduced-translocation resistance phenotype, suggesting that 2,4-D resistance in the populations under investigation could be due to an alteration in the activity of a plasma membrane ABCB-type auxin transporter responsible for facilitating long-distance transport of 2,4-D.

This article analyzes the effects of different rates of nitrogen (0, 30, 60, 90, and 120 kg N ha –1 ) and sulfur (0, 15, 30 kg S ha –1 ) fertilizers on energy balance in oilseed radish biomass production. Energy inputs (EI) were determined at 6.8–7.1 GJ ha –1 . Nitrogen application increased EI by 34% (30 kg ha –1 ) and 135% (120 kg ha –1 ). The energy output of seeds and total biomass peaked after the application of 60 kg N ha –1 + 15 kg S ha –1 and 30 kg N ha –1 + 15 kg S ha –1 , respectively. The energy gain from seeds and total biomass peaked in response to 30 kg N ha –1 + 15 kg S ha –1 . The energy efficiency ratio (EER) of seeds peaked in response to 15 kg S ha –1 (without N fertilization). The EER of total biomass was highest in the absence of N and S fertilization. Nitrogen decreased the EER of seeds and total biomass by 47% and 56%, respectively. The N-induced decrease in the EER was reduced by 4–8% (seeds) and 4–10% (total biomass) when S was applied.