Search Results (11)

Glucosinolates and their degradation products have a wide range of actions and are important components of plant defense. NSP2 (nitrile-specific protein 2) is a key regulator in the breakdown process of glucosinolates. However, the precise function of NSP2 in plant disease resistance beyond its role in glucosinolate degradation is still unclear. In this study, we discovered that NSP2 which was induced by Pst DC3000, influenced PR genes expression and reactive oxygen burst. Additionally, omics analysis revealed that NSP2 was engaged in plant-pathogen interaction and several hormone signal transduction pathways. Furthermore, immunoprecipitation-tandem mass spectrometry analysis (IP-MS), bimolecular fluorescence complementation (BiFC), and co-immunoprecipitation demonstrated that NSP2 interacts with MPK3. Genetic analysis shows that NSP2 may be a function downstream of MPK3. Upon pathogen inoculation, NSP2 protein levels increase while MPK3 protein levels decrease. Moreover, the level of phosphorylated NSP2 decreases. Taken together, this study sheds light on a new mode of synergistic action between NSP2 and MPK3 in the disease resistance process. Full article

The glucosinolate transporters 1/2/3 (GTR1/2/3) from the Nitrate and Peptide transporter Family (NPF) play an essential role in the transport, accumulation, and distribution of the specialized plant metabolite glucosinolates. Due to representing both antinutritional and health-promoting compounds, there is increasing interest in characterizing GTRs from various plant species. We generated seven artificial glucosinolates (either aliphatic or benzenic) bearing different fluorophores (Fluorescein, BODIPY, Rhodamine, Dansylamide, and NBD) and investigated the ability of GTR1/2/3 from Arabidopsis thaliana to import the fluorescent glucosinolates (F-GSLs) into oocytes from Xenopus laevis. Five out of the seven F-GSLs synthesized were imported by at least one of the GTRs. GTR1 and GTR2 were able to import three F-GSLs actively above external concentration, while GTR3 imported only one actively. Competition assays indicate that the F-GSLs are transported by the same mechanism as non-tagged natural glucosinolates. The GTR-mediated F-GSL uptake is detected via a rapid and sensitive assay only requiring simple fluorescence measurements on a standard plate reader. This is highly useful in investigations of glucosinolate transport function and provides a critical prerequisite for elucidating the relationship between structure and function through high-throughput screening of GTR mutant libraries. The F-GSL themselves may also be suitable for future studies on glucosinolate transport in vivo. Full article

Heat stress is one of the most common agrometeorological risks in crop production in the middle and lower reaches of the Yangtze River in China. This study aimed to investigate whether glutamic acid (Glu) or poly-γ-glutamic acid (γ-PGA) biostimulants can improve the thermotolerance of a cool-season Chinese cabbage (Brassica rapa L. ssp. pekinensis) crop. Priming with Glu (2.0 mM) or γ-PGA (20 mg·L⁻¹) was conducted at the third leaf stage by applying as daily foliar sprays for 5 days before 5 days of heat stress (45 °C in 16-h light/35 °C in 8-h dark). Coupled with morpho-physiological and biochemical analyses, transcriptomes of Glu or γ-PGA-primed Chinese cabbage under heat stress were examined by RNA-seq analysis. The results showed that the thermotolerance conferred by Glu and γ-PGA priming was associated with the increased parameters of vegetative growth, gas exchange, and chlorophyll fluorescence. Compared with the control, the dry weights of plants treated with Glu and γ-PGA increased by 51.52% and 39.39%, respectively. Glu and γ-PGA application also significantly increased the contents of total chlorophyll by 42.21% and 23.12%, and carotenoid by 32.00% and 24.00%, respectively. In addition, Glu- and γ-PGA-primed plants markedly inhibited the levels of malondialdehyde, electrolyte leakage, and super-oxide anion radical, which was accompanied by enhanced activity levels of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD). Enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) categories within the differentially expressed genes (DEGs) functional clusters of RNA-seq data indicated that the expression levels of the genes for DNA replication, DNA repair system, linoleic acid metabolism, cysteine and methionine metabolism, glutathione metabolism, purine and pyrimidine metabolism, carotenoid biosynthesis, and plant–pathogen interaction were commonly up-regulated by both Glu and γ-PGA priming. Glu treatment enhanced the expression levels of the genes involved in aliphatic glucosinolate and 2-oxocarboxylic acid, while γ-PGA treatment activated carotenoid cleavage reaction to synthesize abscisic acid. Taken together, both Glu and γ-PGA have great potential for the preadaptation of Chinese cabbage seedlings to heat stress, with Glu being more effective than γ-PGA. Full article

The effects of different ratios of red:white light on the biomass, photosynthetic capacity, phytochemical accumulation, and antioxidant capacity of pak choi were investigated. There were three treatments: red:white = 1:2 (LR), red:white = 1:1 (MR), red:white = 2:1 (HR) with white light as control (CK) at photosynthetic photon flux density (PPFD) of 250 μmol·m⁻²·s⁻¹. In LR treatment, chlorophyll fluorescence and nitrate content were reduced, while the photosynthetic rate (Y(ll)), the contents of soluble protein, soluble sugar, vitamin C, flavonoids, polyphenols, glucosinolates, FRAP, and 2,2-diphenyl-1-bitter acyl radical scavenging increased. The chlorophyll content was enhanced in MR treatment, and the photosynthetic rate and glucosinolate content increased in HR treatment. The most suitable light for the cultivation of pak choi in the plant factory was red:white = 1:2. Full article

Ultrasounds (US) and LED illumination are being studied to optimize yield and quality. The objective was to evaluate the effect of a pre-sowing US treatment combined with a postharvest photoperiod including LEDs on rocket sprouts’ quality and phytochemicals during shelf life. A US treatment (35 kHz; 30 min) applied to seeds and a postharvest photoperiod of 14 h fluorescent light (FL) + 10 h White (W), Blue (B), Red (R) LEDs or Darkness (D) were assayed. Antioxidants as phenolics and sulfur compounds (glucosinolates and isothiocyanates) were periodically monitored over 14 days at 5 °C. The US treatment increased the sulforaphane content by ~4-fold compared to CTRL seeds and sprouts. The phenolic acids and the flavonoid biosynthesis were enhanced by ~25%, ~30%, and ~55% under photoperiods with W, B, and R, respectively, compared to darkness. The total glucosinolate content was increased by >25% (W) and >45% (B and R) compared to darkness, which also reported increases of ~2.7-fold (W), ~3.6-fold (B), and ~8-fold (R) of the sulforaphane content as a main isothiocyanate. Postharvest lighting is an interesting tool to stimulate the secondary metabolism, while a US treatment was able to increase the sulforaphane content in seeds and sprouts, although no synergistic effect was reported. Full article

In complex foods, bioactive secondary plant metabolites (SPM) can bind to food proteins. Especially when being covalently bound, such modifications can alter the structure and, thus, the functional and biological properties of the proteins. Additionally, the bioactivity of the SPM can be affected as well. Consequently, knowledge of the influence of chemical modifications on these properties is particularly important for food processing, food safety, and nutritional physiology. As a model, the molecular structure of conjugates between the bioactive metabolite benzyl isothiocyanate (BITC, a hydrolysis product of the glucosinolate glucotropaeolin) and the whey protein α-lactalbumin (α-LA) was investigated using circular dichroism spectroscopy, anilino-1-naphthalenesulfonic acid fluorescence, and dynamic light scattering. Free amino groups were determined before and after the BITC conjugation. Finally, mass spectrometric analysis of the BITC-α-LA protein hydrolysates was performed. As a result of the chemical modifications, a change in the secondary structure of α-LA and an increase in surface hydrophobicity and hydrodynamic radii were documented. BITC modification at the ε-amino group of certain lysine side chains inhibited tryptic hydrolysis. Furthermore, two BITC-modified amino acids were identified, located at two lysine side chains (K32 and K113) in the amino acid sequence of α-LA. Full article

The effect of salt treatment on Brassica carinata (BC) microgreens grown under different light wavelengths on glucosinolates (GLs) and phenolic compounds were evaluated. Quantifiable GLs were identified using ultra-high performance-quadrupole time of flight mass spectrometry. Extracts’ ability to activate antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT)) was evaluated on human colorectal carcinoma cells (HCT116). Furthermore, BC compounds’ ability to activate expression of nuclear transcription factor-erythroid 2 related factor (Nrf2) and heme-oxygenase-1 (HO-1) proteins was examined using specific antibodies on HCT116 cells. Sinigrin (SIN) was the abundant GLs of the six compounds identified and its content together with total aliphatic GLs increased in saline conditions. Fluorescent (FL) and blue plus red (B1R1) lights were identified as stable cultivation conditions for microgreens, promoting biomass and glucobrassicin contents, whereas other identified individual and total indole GLs behaved differently in saline and non-saline environments. Blue light-emitting diodes and FL light in saline treatments mostly enhanced SIN, phenolics and antioxidant activities. The increased SOD and CAT activities render the BC microgreens suitable for lowering oxidative stress. Additionally, activation of Nrf2, and HO-1 protein expression by the GLs rich extracts, demonstrate their potential to treat and prevent oxidative stress and inflammatory disorders. Therefore, effective salt treatments and light exposure to BC microgreens present an opportunity for targeted regulation of growth and accumulation of bioactive metabolites. Full article

Antiviral treatments inhibiting Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication may represent a strategy complementary to vaccination to fight the ongoing Coronavirus disease 19 (COVID-19) pandemic. Molecules or extracts inhibiting the SARS-CoV-2 chymotripsin-like protease (3CL^Pro) could contribute to reducing or suppressing SARS-CoV-2 replication. Using a targeted approach, we identified 17 plant products that are included in current and traditional cuisines as promising inhibitors of SARS-CoV-2 3CL^Pro activity. Methanolic extracts were evaluated in vitro for inhibition of SARS-CoV-2 3CL^Pro activity using a quenched fluorescence resonance energy transfer (FRET) assay. Extracts from turmeric (Curcuma longa) rhizomes, mustard (Brassica nigra) seeds, and wall rocket (Diplotaxis erucoides subsp. erucoides) at 500 µg mL⁻¹ displayed significant inhibition of the 3CL^Pro activity, resulting in residual protease activities of 0.0%, 9.4%, and 14.9%, respectively. Using different extract concentrations, an IC₅₀ value of 15.74 µg mL⁻¹ was calculated for turmeric extract. Commercial curcumin inhibited the 3CL^Pro activity, but did not fully account for the inhibitory effect of turmeric rhizomes extracts, suggesting that other components of the turmeric extract must also play a main role in inhibiting the 3CL^Pro activity. Sinigrin, a major glucosinolate present in mustard seeds and wall rocket, did not have relevant 3CL^Pro inhibitory activity; however, its hydrolysis product allyl isothiocyanate had an IC₅₀ value of 41.43 µg mL⁻¹. The current study identifies plant extracts and molecules that can be of interest in the search for treatments against COVID-19, acting as a basis for future chemical, in vivo, and clinical trials. Full article

Chinese cabbage (Brassica rapa) is a perennial crucifer vegetable that has long been used for forage. Crucifers are rich sources of glucosinolates (GSLs), which are anti-carcinogenic in humans and involved in plant defense responses. Myeloblastosis (MYB) proteins are a large family of transcription factors (TFs) in plants and play major regulatory roles in many biological processes. We identified 14 functional R2R3-MYB genes involved in glucosinolate biosynthesis in B. rapa ssp. pekinensis. Bioinformatic analysis of their phylogeny, protein motifs, gene interaction network, and molecular characteristics showed that Chinese cabbage MYB genes are comparable to those of Arabidopsis thaliana. The expression levels of the 14 BrMYB genes under fluorescent lamp, blue, and red light were quantitated using qRT-PCR analysis. Almost all of the R2R3-BrMYBs were upregulated and expressed more under red light than under fluorescent lamp or blue light, except BrMYB34s. We also calculated the total GSLs under each light condition. The total GSL content was higher under red light than under fluorescent lamp or blue light. Furthermore, the individual glucosinolates, comprised of four aliphatic GSLs (progoitrin, sinigrin, gluconapin, and glucobrassicanapin) and one indolic GSL (glucobrassicin), were higher under red light than the other light conditions. The relationships between light quality and glucosinolate biosynthesis require further investigation. Full article

Sisymbrium officinale is a wild Brassicaceae species that is known for its use in the alleviation of vocal inflammatory states. Since this species is particularly rich in bioactive compounds, there is an interest for developing cultivation protocols to use this plant as a leafy vegetable harvested at the 13th BBCH growth stage. Two wild populations of S. officinale (L.) Scop., denominate MI (Milan) and BG (Bergamo), have been used, and three different levels of nutrients (g/m²) have been provided such as 13 N, 7 P₂O₅, 8 K₂O (100%); 9 N, 5 P₂O₅, 5.5 K₂O (70%); and 6.5 N, 3.5 P₂O₅, 4.0 K₂O (50%). The effects of different fertilization levels were evaluated on the yield, leaf pigments (chlorophylls, carotenoids), nitrate concentration, sugars content, and on the antioxidant compounds such as anthocyanins, total phenols and glucosinolates (GLS). Plant stress monitoring was performed by measuring the chlorophyll a fluorescence. Results indicated that yield was not affected and ranged from 0.18 to 0.47 kg/m², and differences were not statistically significant for chlorophylls, carotenoids, and total sugars content. Nitrate concentrations were higher in the BG wild population (4388.65 mg/kg FW) during the second cultivation cycle and lower (1947.21 mg/kg FW) in the same wild population during the first cultivation cycle, both at the 100% fertilization level. Total phenols and anthocyanins were higher in the highest fertilization rate in the MI wild population during the second cycle. The GLS were higher in the lowest fertilization rate in the BG wild population (19 µmol/g FW) grown at the fertilization level of the 50%, and significant differences were observed in the second cycle. In conclusion, the 50% fertilization level can be considered the most suitable for our experimental conditions. Full article

Volatile allyl isothiocyanate (AITC) derives from the biodegradation of the glucosinolate sinigrin and has been associated with growth inhibition in several plants, including the model plant Arabidopsis thaliana. However, the underlying cellular mechanisms of this feature remain scarcely investigated in plants. In this study, we present evidence of an AITC-induced inhibition of actin-dependent intracellular transport in A. thaliana. A transgenic line of A. thaliana expressing yellow fluorescent protein (YFP)-tagged actin filaments was used to show attenuation of actin filament movement by AITC. This appeared gradually in a time- and dose-dependent manner and resulted in actin filaments appearing close to static. Further, we employed four transgenic lines with YFP-fusion proteins labeling the Golgi apparatus, endoplasmic reticulum (ER), vacuoles and peroxisomes to demonstrate an AITC-induced inhibition of actin-dependent intracellular transport of or, in these structures, consistent with the decline in actin filament movement. Furthermore, the morphologies of actin filaments, ER and vacuoles appeared aberrant following AITC-exposure. However, AITC-treated seedlings of all transgenic lines tested displayed morphologies and intracellular movements similar to that of the corresponding untreated and control-treated plants, following overnight incubation in an AITC-absent environment, indicating that AITC-induced decline in actin-related movements is a reversible process. These findings provide novel insights into the cellular events in plant cells following exposure to AITC, which may further expose clues to the physiological significance of the glucosinolate-myrosinase system. Full article