Effects of Supplementing Selenium-Enriched Cardamine violifolia to Laying Hens on Egg Quality and Yolk Antioxidant Capacity during Storage at 4 °C and 25 °C.

Oxidative stress occurs in the process of egg storage. Antioxidants as feed additives can enhance egg quality and extend the shelf life of eggs. Selenium-enriched Cardamine violifolia (SEC) has strongly antioxidant properties. The objective of this study was to assess the effects of dietary supplementation with SEC on egg quality and the yolk antioxidant capacity of eggs stored at 4 °C and 25 °C. Four hundred fifty 65-week-old, Roman hens that were similar in laying rate (90.79 ± 1.69%) and body weight (2.19 ± 0.23 kg) were divided into 5 groups. The birds were fed diets supplemented with 0 mg/kg selenium (Se) (CON), 0.3 mg/kg Se from sodium selenite (SS), 0.3 mg/kg Se from Se-enriched yeast (SEY), 0.3 mg/kg Se for selenium-enriched Cardamine violifolia (SEC) or 0.3 mg/kg Se from Se-enriched Cardamine violifolia and 0.3 mg/kg Se from Se-enriched yeast (SEC + SEY) for 8 weeks. The eggs were collected on the 8th week and were analyzed for egg quality and oxidative stability of yolk during storage at 4 °C or 25 °C for 0, 2, 4, or 6 weeks. Dietary SEC and SEC + SEY supplementation increased the Haugh unit (HU) and albumen foam stability in eggs stored at 4 °C and 25 °C (p < 0.05). SS and SEC supplementation increased the yolk index in eggs stored at 25 °C (p < 0.05). SEC or SEC + SEY slowed down an increase in albumen pH and gel firmness in eggs stored at 4 °C and 25 °C (p < 0.05). Moreover, SEC or SEC + SEY alleviated the increase in malonaldehyde (MDA), and the decrease in total antioxidant capacity (T-AOC) level and total superoxide dismutase (T-SOD) activity in yolks stored at 4 °C and 25 °C (p < 0.05). These results indicate that SEC mitigated egg quality loss and improved the antioxidant capacity of yolks during storage. SEC supplementation would be advantageous to extend the shelf life of eggs.

Investigation of selenium and selenium species in Cardamine violifolia using in vitro digestion coupled with a Caco-2 cell monolayer model.

Inadequate Se intake can enhance vulnerability to certain health risks, with supplementation lessening these risks. This study investigated the bioavailability of Se and Se species in five Se compounds and in Se-rich Cardamine violifolia using in vitro digestion coupled with a Caco-2 cell monolayer model, which enabled the study of Se transport and uptake. Translocation results showed that SeCys2 and MeSeCys had high translocation rates in C. violifolia leaves (CVLs). The uptake rate of organic Se increased with time, and MeSeCys exhibited a higher uptake rate than that for SeCys2 and SeMet. The translocation mechanisms of SeMet, Se(IV), and Se(VI) were passive transport, whereas those of SeCys2 and MeSeCys were active transport. The bioavailability of organic Se was higher than that of inorganic Se, with a total Se bioavailability in CVLs of 49.11 %. This study would provide a theoretical basis for the application of C. violifolia in the functional food.

Selenium content, chemical composition and volatile components of essential oil and hydrosol from flowers of Cardamine violifolia.

Cardamine violifolia is a unique selenium hyperaccumulating vegetable in China, but its flowers are commonly wasted in large-scale cultivation. To better utilize this resource, this study explored the selenium content, chemical composition, and volatile organic compounds (VOCs) of hydro-distilling essential oil (EO) and hydrosol from C. violifolia flowers. ICP-MS results indicated that the EO and hydrosol contained selenium reaching 13.66±2.82 mg/kg and 0.0084±0.0013 mg/kg, respectively. GC-MS analysis revealed that organic acids, hydrocarbons, and amines were the main components of EO. Additionally, benzyl nitrile, benzaldehyde, benzyl isothiocyanate, benzyl alcohol, megastigmatrienone, and 2-methoxy-4-vinylphenol also existed in considerable amounts. The hydrosol extract had fewer components, mainly amines. HS-SPME-GC-MS corresponded to the composition analysis and aromatic compounds were the prevalent VOCs, while HS-GC-IMS primarily identified C2-C10 molecular alcohols, aldehydes, ethers, and sulfur-containing compounds. This study first described the chemical composition and VOC profiles of EO and hydrosol from selenium hyperaccumulating plant.

Systemic subchronic toxicity and comparison of four selenium nutritional supplements by 90-day oral exposure in Sprague-Dawley rats.

To evaluate and compare the safety of four selenium supplements, namely Se-enriched peptides (SeP), yeast selenium (SeY), L-Se-methylselenocysteine (L-SeMc) and sodium selenite (Na2SeO3), the subchronic toxicity study was designed by 90-day gavage administration in Sprague-Dawley rats. The doses of SeP, SeY, L-SeMc and Na2SeO3 were 0.15, 0.30 and 0.60 mg/kg bw/day, with additional dose of 0.45 mg/kg L-SeMc (All dose calculated as Se). Symptoms like growling, hair loss and significant weight loss were found at 0.60 mg/kg of L-SeMc, but not in other groups. At the dose of 0.60 mg/kg, females in Na2SeO3, SeY and L-SeMc groups showed significant elevations in ALT and/or ALP. Pathologic manifestations such as bile duct hyperplasia and cholestasis were predominantly found in females at 0.6 mg/kg of L-SeMc and SeY groups, and in males at same dose of L-SeMc group showed marked testicular atrophy. 0.60 mg/kg of SeY and Na2SeO3, and 0.30, 0.45, 0.60 mg/kg of L-SeMc induced significant reductions in sperm motility rates, rapid movement and amount. In conclusion, the NOAEL of SeP, SeY, L-SeMc, Na2SeO3 was all 0.30 mg/kg for female, and 0.60, 0.30, 0.15 and 0.30 mg/kg for male respectively. Liver and reproductive organs are possible toxic target organs of hyper selenium.

Selenium speciation and volatile flavor compound profiles in the edible flowers, stems, and leaves of selenium-hyperaccumulating vegetable Cardamine violifolia.

Cardamine violifolia is a unique selenium (Se)-hyperaccumulating vegetable in China. The total Se content and Se speciation of three edible parts, including flowers, stems, and leaves were detected by HPLC-ICP-MS. Volatile organic compounds (VOCs) greatly impact food flavor. The VOCs of three samples were analyzed by E-nose, HS-GC-IMS, and HS-SPME-GC-MS. The results showed that the total Se content in flowers was significantly higher than that in leaves and was the lowest in stems. Organic Se accounts for more than 98% of the total Se content, primarily selenocystine, followed by methyl selenocysteine. A total of 102 VOCs were identified from C. violifolia, mainly esters, aldehydes, alcohols, and ketones. Flowers contained abundant VOCs, while stems and leaves contained fewer but similar profiles. Moreover, multivariate statistical analysis was applied to investigate the VOC variations and marker VOCs. This work can provide useful knowledge for understanding the Se characteristics and flavor of C. violifolia.

Se-Enriched Cardamine violifolia Improves Laying Performance and Regulates Ovarian Antioxidative Function in Aging Laying Hens.

As a selenium-enriched plant, Cardamine violifolia (SEC) has an excellent antioxidant function. The edibility of SEC is expected to develop new sources of organic Se supplementation for human and animal nutrition. This study was conducted to investigate the effects of SEC on laying performance and ovarian antioxidant capacity in aging laying hens. A total of 450 laying hens were assigned to five treatments. Dietary treatments included the following: a basal diet (diet without Se supplementation, CON) and basal diets supplemented with 0.3 mg/kg Se from sodium selenite (SS), 0.3 mg/kg Se from Se-enriched yeast (SEY), 0.3 mg/kg Se from SEC, or 0.3 mg/kg Se from SEC and 0.3 mg/kg Se from SEY (SEC + SEY). Results showed that supplementation with SEC tended to increase the laying rate, increased the Haugh unit of eggs, and reduced the FCR. SEC promoted ovarian cell proliferation, inhibited apoptosis, and ameliorated the maintenance of follicles. SEC, SEY, or SEC + SEY increased ovarian T-AOC and decreased MDA levels. SEC increased the mRNA abundance of ovarian selenoproteins. SEC and SEC + SEY increased the mRNA abundance of Nrf2, HO-1, and NQO1, and decreased the mRNA abundance of Keap1. These results indicate that SEC could potentially to improve laying performance and egg quality via the enhancement of ovarian antioxidant capacity. SEC exerts an antioxidant function through the modulation of the Nrf2/Keap1 signaling pathway.

Effects of Different Drying Methods on the Selenium Bioaccessibility and Antioxidant Activity of Cardamine violifolia.

Understanding the effects of drying on the selenium (Se) content and Se bioaccessibility of Se-rich plants is critical to dietary supplementation of Se. The effects of five common drying methods (far-infrared drying (FIRD), vacuum drying (VD), microwave vacuum drying (MVD), hot air drying (HD), and freeze vacuum drying (FD)) on the content and bioaccessibility of Se and Se species in Cardamine violifolia leaves (CVLs) were studied. The content of SeCys2 in fresh CVLs was the highest (5060.50 µg/g of dry weight (DW)); after FIRD, it had the lowest selenium loss, with a loss rate of less than 19%. Among all of the drying processes, FD and VD samples had the lowest Se retention and bioaccessibility. FIRD, VD, and FD samples have similar effects on antioxidant activity.

Spatial Metallomics Reveals Preferable Accumulation of Methylated Selenium in a Single Seed of the Hyperaccumulator Cardamine violifolia†.

Cardamine violifolia is a Se hyperaccumulator found in Enshi, China. In this study, spatial metallomics was applied to visualize the distribution and speciation of Se in a single seed of C. violifolia. It was found that Se reached 1729.89 ± 28.14 mg/kg and the main Se species were SeCys and SeMet in bulk seeds. Further in situ study on a single seed found that the methylated Se species located mostly in the episperm. This is the first visualized evidence of the in situ distribution of methylated Se species in the seeds of C. violifolia. In all, spatial metallomics finds a preferable accumulation of methylated Se species in the seed coat, which deepens the understanding of the tolerance of Se by C. violifolia. The protocol applied in this study may also be used for the understanding of the tolerance of heavy metals/metalloids in other hyperaccumulators.

Microbial functional communities and the antibiotic resistome profile in a high-selenium ecosystem.

Enshi City, in the Hubei Province of China, is known as the world capital of selenium with the most abundant selenium resource. An important selenium hyperaccumulator plant, Cardamine violifolia, was found to naturally grow in this high-selenium ecosystem. However, relatively little is known about the impact of the selenium levels on microbial community and functional shifts in C. violifolia rhizosphere. Here, we tested the hypothesis that underground microbial diversity and function vary along a selenium gradient, including antibiotic resistance genes (ARGs). Comprehensive metagenomic analyses, such as taxonomic investigation, functional detection, and ARG annotation, showed that selenium, mercury, cadmium, lead, arsenic, and available phosphorus and potassium were correlated with microbial diversity and function. Thaumarchaeota was exclusively dominant in the highest selenium concentration of mine outcrop, and Rhodanobacter and Nitrospira were predominant in the high-selenium ecosystem. The plant C. violifolia enriched a high concentration of selenium in the rhizosphere compared to those in the bulk soil, and it recruited Variovorax and Polaromonas in its rhizosphere. Microbial abundance showed a trend of increasing first and then decreasing from low to high selenium concentrations. Annotation of ARGs showed that the multidrug resistance genes adeF, mtrA, and poxtA, the aminoglycoside resistance gene rpsL, and the sulfonamide resistant gene sul2 were enriched in the high-selenium system. It was discovered that putative antibiotic resistant bacteria displayed obvious differences in the farmland and the soils with various selenium concentrations, indicating that a high-selenium ecosystem harbors the specific microbes with a higher capacity to enrich or resist selenium, toxic metals, or antibiotics. Taken together, these results reveal the effects of selenium concentration and the selenium hyperaccumulator plant C. violifolia on shaping the microbial functional community and ARGs. Metalloid selenium-inducible antibiotic resistance is worth paying attention to in future.

Cardamine violifolia as a potential Hg hyperaccumulator and the cellular responses.

Cardamine violifolia belongs to the Brassicaceae family and is a selenium (Se) hyperaccumulator found in Enshi, China. In this study, C. violifolia was found to accumulate mercury (Hg) in its roots and aboveground parts at concentrations up to 6000 µg/g. In the seedling and mature stages, the bioaccumulation factors (BAFS) of Hg reached 1.8-223, while the translocation factor (TF) for Hg reached 1.5. We observed a significant positive correlation between THg concentrations in plant tissues and those in the soil (r2 = 0.71-0.84). Synchrotron radiation X-ray fluorescence with focused X-ray (µ-SRXRF) showed that Hg was translocated from the roots to shoots through the vascular bundle and was transported through the leaf veins in leaves. Transmission electron microscopy showed that root cells were more tolerant to Hg than leaf cells. These findings provide insights into the mechanisms of Hg hyperaccumulation in C. violifolia. Overall, we demonstrated that C. violifolia is a promising Hg hyperaccumulator that may be used for phytoremediating Hg-contaminated farmlands.

Effect of Cardamine violifolia on Plasma Biochemical Parameters, Anti-Oxidative Capacity, Intestinal Morphology, and Meat Quality of Broilers Challenged with Lipopolysaccharide.

Cardamine violifolia is a newly discovered selenium (Se)-enriched plant rich in MeSeCys and SeCys and has a strong antioxidant capacity. This study aimed to investigate the effects of Cardamine violifolia on plasma biochemical indices, antioxidant levels, intestinal morphology, and meat quality of broilers under acute LPS-induced oxidative stress by comparing it with inorganic Se (sodaium selenite). A total of 240 one-day-old Ross 308 broilers were fed a basal diet and divided into four groups: (1) SeNa-SS, fed a diet supplied with 0.3 mg/kg Se from sodium selenite, and injected with 0.9% sterile saline, (2) SeCv-SS, fed a diet supplied with 0.3 mg/kg Se from Cardamine violifolia, and injected with 0.9% sterile saline, (3) SeNa-LPS, fed a diet supplied with 0.3 mg/kg Se from sodium selenite, and injected with 0.5 mg/kg LPS, (4) SeCv-LPS, fed a diet supplied with 0.3 mg/kg Se from Cardamine violifolia and injected with 0.5 mg/kg LPS. The experiment lasted for 42 days. Sterile saline or LPS was injected intraperitoneally two hours before slaughter, and blood and tissue samples were collected for testing. The results showed that compared with SeNa, SeCv significantly reduced the plasma levels of aspartate aminotransferase, alanine aminotransferase, and urea nitrogen after LPS challenge (p < 0.05), and increased the plasma levels of total antioxidant capacity and glutathione peroxidase, decreased malondialdehyde content in LPS-challenged broilers (p < 0.05). In addition, compared with SeNa, SeCv supplementation increased villus height and the ratio of villus height to crypt depth of jejunum and ileum after LPS challenge (p < 0.05). Additionally, SeCv could increase the redness of breast and thigh muscle, and decrease drip loss, cooking loss, and shear force (p < 0.05). In conclusion, our results indicated that supplementing with 0.3 mg/kg Se from Cardamine violifolia alleviated tissue injury after LPS challenge, increased antioxidant capacity, and improved meat quality of breast and thigh muscle after stress.

Selenium-enriched Cardamine violifolia improves growth performance with potential regulation of intestinal health and antioxidant function in weaned pigs.

This study was conducted to evaluate the effects of different Selenium (Se) sources on growth performance, intestinal function and antioxidant status of weaned piglets. A total of 300 weaned pigs were randomly allocated to 5 treatment groups with 5 replicates of 12 pigs/pen. The control group was corn-soybean basal diet without any additional Se supplement. The experimental diets were supplemented with 0.3 mg/kg of Se from sodium selenite (SS), Se-enriched yeast (SEY), Se-enriched Cardamine violifolia (SEC) and 0.3+0.3 mg/kg of Se from SEY and SEC, respectively. The trial lasted for 4 weeks. The results showed that diets supplementation with SEY, SEC or SEY+SEC could improve average daily gain and reduce feed/gain ratio during the entire study. Compared with the control group, SEC or SEY+SEC improved intestinal morphology, indicated by greater villus height and villus height/ crypt depth ratio. In addition, SEC or SEY+SEC also increased maltase and lactase activities as well as tight junction protein expression. Different Se sources decreased malondialdehyde (MDA) concentration and improved superoxide dismutase (SOD) activity in serum. In the jejunum, SEY or SEC reduced MDA concentration and increased total antioxidant capacity (T-AOC) compared with the control group. Moreover, SEY+SEC increased the antioxidant parameters including SOD and T-AOC in the jejunum. Dietary SEY or SEC supplementation significantly increased the mRNA expression of selenoproteins including thioredoxin reductase 1 (TXNRD1), selenoprotein I (SELENOI), selenoprotein S (SELENOS), and selenoprotein P (SELENOP) in the jejunum. In conclusion, organic Se sources, especially Cardamine violifolia, improve growth performance, potentially by regulating intestinal function, antioxidant capacity and selenoprotein expression in piglets.

Selenium-enriched peptides isolated from Cardamine violifolia are potent in suppressing proliferation and enhancing apoptosis of HepG2 cells.

Selenium (Se)-enriched peptides were isolated from Cardamine violifolia by enzymatic hydrolysis and ultrafiltration. S3 (molecular weight [MW] distribution of 3-5 kDa) exhibited the strongest inhibitory effect on HepG2 cells and was thus screened using the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide assay; it was found to have a high organic Se content. Its amino acid sequence was determined using HPLC-MS/MS. We then examined its ability to inhibit tumor cell proliferation and found that it arrested tumor cells in the S phase; moreover, it could induce cancer cell apoptosis. Following S3 treatment, we observed a decrease in mitochondrial membrane potential and an increase in cell calcium content. Upon S3 treatment at 60 µg/ml, the relative activities of caspase-3 and caspase-9 increased by 1.48 times and 2.17 times, and the contents of PI3K and AKT decreased from 2.05 ng/L and 1.95 ng/L to 0.71 ng/L and 0.50 ng/L, respectively, when compared with the control group. Transcriptomic analysis revealed significant changes in the PI3K-AKT pathway following S3 treatment. This study thus established a foundation for additional development of Se-enriched peptides from C. violifolia as a functional food. PRACTICAL APPLICATION: Cardamine violifolia is a Se-tolerant cruciferous plant that can metabolically transform inorganic Se into organic Se that exists in the form of a selenoprotein. Se-enriched peptide obtained by extraction and enzymolysis of selenoprotein, as an organic combination of organic Se and peptide, possess valuable biological activities. In this paper, the effect of Se-enriched peptides of C. violifolia on tumor cells was studied via cell experiments, and its mechanism was preliminarily discussed, which should provide a theoretical basis for developing functional foods containing C. violifolia.

Selenium-Enriched Cardamine violifolia Increases Selenium and Decreases Cholesterol Concentrations in Liver and Pectoral Muscle of Broilers.

BACKGROUND: Supernutrition of selenium (Se) in an effort to produce Se-enriched meat may inadvertently cause lipid accumulation. Se-enriched Cardamine violifolia (SeCv) contains >80% of Se in organic forms. OBJECTIVES: This study was to determine whether feeding chickens a high dose of SeCv could produce Se-biofortified muscle without altering their lipid metabolism. METHODS: Day-old male broilers were allocated to 4 groups (6 cages/group and 6 chicks/cage) and were fed either a corn-soy base diet (BD, 0.13-0.15 mg Se/kg), the BD plus 0.5 mg Se/kg as sodium selenite (SeNa) or as SeCv, or the BD plus a low-Se Cardamine violifolia (Cv, 0.20-0.21mg Se/kg). At week 6, concentrations of Se and lipid and expression of selenoprotein and lipid metabolism-related genes were determined in the pectoral muscle and liver. RESULTS: The 4 diets showed no effects on growth performance of broilers. Compared with the other 3 diets, SeCv elevated (P < 0.05) Se concentrations in the pectoral muscle and liver by 14.4-127% and decreased (P < 0.05) total cholesterol concentrations by 12.5-46.7% and/or triglyceride concentrations by 28.8-31.1% in the pectoral muscle and/or liver, respectively. Meanwhile, SeCv enhanced (P < 0.05) muscular α-linolenic acid (80.0%) and hepatic arachidonic acid (58.3%) concentrations compared with SeNa and BD, respectively. SeCv downregulated (P < 0.05) the cholesterol and triglyceride synthesis-related proteins (sterol regulatory element binding transcription factor 2 and diacylglycerol O-acyltransferase 2) and upregulated (P < 0.05) hydrolysis and ß-oxidation of fatty acid-related proteins (lipoprotein lipase, fatty acid binding protein 1, and carnitine palmitoyltransferase 1A), as well as selenoprotein P1 and thioredoxin reductase activity in the pectoral muscle and/or liver compared with SeNa. CONCLUSIONS: Compared with SeNa, SeCv effectively raised Se and reduced lipids in the liver and muscle of broilers. The effect was mediated through the regulation of the cholesterol and triglyceride biosynthesis and utilization-related genes.

Impact of selenium on rhizosphere microbiome of a hyperaccumulation plant Cardamine violifolia.

Cardamine violifolia is the only selenium hyperaccumulation plant found in China. It has been developed as a source of medicinal and edible products that we can consume as selenium supplements. Many planting approaches have been developed to increase the selenium content of C. violifolia for nutrient biofortification. However, the contribution of rhizosphere microbes of C. violifolia to selenium enrichment has not been investigated. In this study, four types of selenium, i.e., selenate, selenite, nanoparticles selenium from Bacillus subtilis (B. subtilis-Se), and organic selenium from yeast (yeast-Se), were added to the soil that C. violifolia was grown in, respectively. Selenate led to the greatest accumulation of selenium in C. violifolia, followed by selenite, B. subtilis-Se, and yeast-Se. Except for yeast-Se, the concentration of selenium in C. violifolia positively correlated with the amount of selenium added to the soil. Furthermore, the different types of exogenous selenium exhibited distinct effects on the rhizosphere microbiome of C. violifolia. Alpha and beta diversity analyses demonstrated that rhizosphere microbiome was more obviously affected by selenium from B. subtilis and yeast than from selenate and selenite. Different microbial species were enriched in the rhizosphere of C. violifolia under various exogenous selenium treatments. B. subtilis-Se application enhanced the abundance of Leucobacter, Sporosarcina, Patulibacter, and Denitrobacter, and yeast-Se application enriched the abundance of Singulishaera, Lactobacillus, Bdellovibrio, and Bosea. Bosea and the taxon belonging to the order Solirubrobacterales were enriched in the samples with selenate and selenite addition, respectively, and the abundances of these were linearly related to the concentrations of selenate and selenite applied in the rhizosphere of C. violifolia. In summary, this study revealed the response of the rhizosphere microbiome of C. violifolia to exogenous selenium. Our findings are useful for developing suitable selenium fertilizers to increase the selenium hyperaccumulation level of this plant.

Integration analysis of PacBio SMRT- and Illumina RNA-seq reveals candidate genes and pathway involved in selenium metabolism in hyperaccumulator Cardamine violifolia.

BACKGROUND: Cardamine violifolia, native to China, is one of the selenium (Se) hyperaccumulators. The mechanism of Se metabolism and tolerance remains unclear, and only limited genetic information is currently available. Therefore, we combined a PacBio single-molecule real-time (SMRT) transcriptome library and the Illumina RNA-seq data of sodium selenate (Na2SeO4)-treated C. violifolia to further reveal the molecular mechanism of Se metabolism. RESULTS: The concentrations of the total, inorganic, and organic Se in C. violifolia seedlings significantly increased as the Na2SeO4 treatment concentration increased. From SMRT full-length transcriptome of C. violifolia, we obtained 26,745 annotated nonredundant transcripts, 14,269 simple sequence repeats, 283 alternative splices, and 3407 transcription factors. Fifty-one genes from 134 transcripts were identified to be involved in Se metabolism, including transporter, assimilatory enzyme, and several specific genes. Analysis of Illumina RNA-Seq data showed that a total of 948 differentially expressed genes (DEGs) were filtered from the four groups with Na2SeO4 treatment, among which 11 DEGs were related to Se metabolism. The enrichment analysis of KEGG pathways of all the DEGs showed that they were significantly enriched in five pathways, such as hormone signal transduction and plant-pathogen interaction pathways. Four genes related to Se metabolism, adenosine triphosphate sulfurase 1, adenosine 5'-phosphosulfate reductase 3, cysteine (Cys) desulfurase 1, and serine acetyltransferase 2, were regulated by lncRNAs. Twenty potential hub genes (e.g., sulfate transporter 1;1, Cys synthase, methionine gamma-lyase, and Se-binding protein 1) were screened and identified to play important roles in Se accumulation and tolerance in C. violifolia as concluded by weighted gene correlation network analysis. Based on combinative analysis of expression profiling and annotation of genes as well as Se speciation and concentration in C. violifolia under the treatments with different Na2SeO4 concentrations, a putative Se metabolism and assimilation pathway in C. violifolia was proposed. CONCLUSION: Our data provide abundant information on putative gene transcriptions and pathway involved in Se metabolism of C. violifolia. The findings present a genetic resource and provide novel insights into the mechanism of Se hyperaccumulation in C. violifolia.

Effects of different exogenous selenium on Se accumulation, nutrition quality, elements uptake, and antioxidant response in the hyperaccumulation plant Cardamine violifolia.

Cardamine violifolia (Brassicaceae) is a novel selenium(Se) hyperaccumulation plant with rich nutrients, and serves as a good source of special vegetables in Enshi, China. The present study aimed to investigate the effects of the application of selenate, selenite, and Se yeast (50-800 mg/L) on the growth, Se accumulation, nutrient uptake, and antioxidant response of C. violifolia. The results showed that the Se accumulation efficiency was selenate > selenite > Se yeast, the maximum Se concentration could achieve over 7000 mg/kg, and about 90% was organic Se. The major Se speciation found was mainly SeCys2 and the proportion of various Se species were affected by the Se forms and concentrations. Besides, the plant growth, nutrition quality indexes, element uptakes, and antioxidant responses indicated that 200 mg/L selenate was optimum for C. violifolia to accumulate Se without much impacts, while to obtain more proportion of organic Se, 200 mg/L selenite might be a better choice.

Antioxidant Activity of Selenium-Enriched Peptides from the Protein Hydrolysate of Cardamine violifolia.

Cardamine violifolia is a selenium (Se)-enriched plant found in China. In this study, the Se-enriched peptides of C. violifolia (CP) were isolated using a 1 kDa ultrafiltration membrane after enzymatic hydrolysis by alkaline and neutral proteases. The peptides were separated by DEAE-Sepharose FF anion-exchange chromatography and purified using preparative high-performance liquid chromatography (prep-HPLC). The component with the highest antioxidant activity, CPR13, was identified by comparing the 1,1-diphenyl-2-picrylhydrazyl (DPPH•), hydroxyl (•OH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+ •), and superoxide ( O 2 - · ) radical scavenging ability of each fraction. At a concentration of 0.1 mg/mL, the DPPH•, •OH, O 2 - · , and ABTS+ • scavenging activities of CPR13 were 89.2%, 26.3%, 40.6%, and 42.9%, respectively. Amino acid sequences were obtained by liquid chromatography combined with mass spectrometry as follows: GRVGSSSC, GRAGGSYM, GHPNFKLNCSGG, GTKSCKA, ASSNARDMI, TAGGCYIPI, and KNCALQ. The seleno-amino acids were identified as selenomethionine, methylselenocysteine, and selenocysteine. Correlation analysis among organic Se content, peptide content, and antioxidant activity revealed that organic Se plays a greater role in free radical scavenging than peptides, and that the organic Se content of the Se-enriched peptides was positively correlated with their antioxidant ability (P < 0.05). It indicated that CP has a great potential as natural functional materials for dietary supplement.