Plasmodiophora brassicae (Woronin, 1877), a biotrophic, obligate parasite, is the causal agent of clubroot disease in brassicas. The clubroot pathogen has been reported in more than 80 countries worldwide, causing economic losses of hundreds of millions every year. Despite its widespread impact, very little is known about the molecular strategies it employs to induce the characteristic clubs in the roots of susceptible hosts during infection, nor about the mechanisms it uses to overcome genetic resistance. Here, we provide the first telomere-to-telomere complete genome of P. brassicae. We generated ∼27 Gb of Illumina, Oxford Nanopore, and PacBio HiFi data from resting spores of strain Pb3A and produced a 25.3 Mb assembly comprising 20 chromosomes, with an N50 of 1.37 Mb. The BUSCO score, the highest reported for any member of the group Rhizaria (Eukaryota: 88.2%), highlights the limitations within the Eukaryota database for members of this lineage. Using available transcriptomic data and protein evidence, we annotated the Pb3A genome, identifying 10,521 protein-coding gene models. This high-quality, complete genome of P. brassicae will serve as a crucial resource for the plant pathology community to advance the much-needed understanding of the evolution of the clubroot pathogen.

Fodder radish is widely used as a livestock supplement, however, the nutritional value of fodder radish under different water conditions remains insufficiently understood. This study aimed to assess the chemical components and in vitro, ruminal dry matter degradability of two fodder radish genotypes (Endurance and Line 2) subjected to three irrigation regimes: well-watered (W1), moderate water stress (W2), and severe water stress (W3). The analysis revealed statistically significant effects of the main factors on the chemical composition and estimates of fodder radish leaves and tubers, particularly in terms of Crude Protein (CP) and Ether Extract (EE) across genotypes. Both Endurance and Line 2 leaves exhibited interaction effects on N, P, Ca, Mg, K, Na, Fe, Zn, Cu, Mn and Al. Meanwhile only Na, K, Zn, and Cu were affected in tubers. Endurance tubers, specifically, displayed significantly higher (p < 0.05) CP content, with Line 2 tubers showing the highest CP content under W1. Furthermore, Endurance leaves had higher levels of Neutral Detergent Fibre, EE, and Non-Structural Carbohydrate (NSC) compared to Line 2 leaves under W1. Notable differences in tuber fibres were found, specifically in Acid Detergent Fibre for Endurance, with W3 exhibiting a higher concentration level. Both genotypes displayed higher NSC under W3. Significant variations in macro and mmicro minerals were observed between water levels in both genotypes. In terms of in vitro degradability during the 24 h and 48 h incubation periods, all treatments met the acceptable level of 60-80 %. Regardless of water regimes, both Endurance and Line 2 showed nutrient concentrations meeting the minimum requirements for optimal animal production. Though, Line 2 exhibits significantly higher nutritional value and in vitro ruminal dry matter degradability than Endurance, evident in both leaves and tubers. Notably, moderate water stress conditions yielded better nutritional quality and in vitro ruminal dry matter degradability compared to both well-watered and severe water stress treatments. This suggests that applying 180-220 mm of water per season can also yield better nutritive value of these genotypes.

Most nonoccupational human exposure to thallium (Tl) occurs via consumption of contaminated food crops. Brassica cultivars are common crops that can accumulate more than 500 μg Tl g-1. Knowledge of Tl uptake and translocation mechanisms in Brassica cultivars is fundamental to developing methods to inhibit Tl uptake or conversely for potential use in phytoremediation of polluted soils. Brassica cultivars (25 in total) were subjected to Tl dosing to screen for Tl accumulation. Seven high Tl-accumulating varieties were selected for follow-up Tl dosing experiments. The highest Tl accumulating Brassica cultivars were analyzed by synchrotron-based micro-X-ray fluorescence to investigate the Tl distribution and synchrotron-based X-ray absorption near-edge structure spectroscopy (XANES) to unravel Tl chemical speciation. The cultivars exhibited different Tl tolerance and accumulation patterns with some reaching up to 8300 μg Tl g-1. The translocation factors for all the cultivars were >1 with Brassica oleracea var. acephala (kale) having the highest translocation factor of 167. In this cultivar, Tl is preferentially localized in the venules toward the apex and along the foliar margins and in minute hot spots in the leaf blade. This study revealed through scanning electron microscopy and X-ray fluorescence analysis that highly Tl-enriched crystals occur in the stoma openings of the leaves. The finding is further validated by XANES spectra that show that Tl(I) dominates in the aqueous as well as in the solid form. The high accumulation of Tl in these Brassica crops has important implications for food safety and results of this study help to understand the mechanisms of Tl uptake and translocation in these crops.

Colletotrichum higginsianum causes anthracnose disease in brassicas. The availability of the C. higginsianum genome has paved the way for the genome-wide exploration of genes associated with virulence/pathogenicity. However, delimiting the biological functions of these genes remains an arduous task due to the recalcitrance of C. higginsianum to genetic manipulations. Here, we report a CRISPR/Cas9-based system that can knock out the genes in C. higginsianum with a staggering 100% homologous recombination frequency (HRF). The system comprises two vectors: pCas9-Ch_tRp-sgRNA, in which a C. higginsianum glutaminyl-tRNA drives the expression of sgRNA, and pCE-Zero-HPT carrying a donor DNA cassette containing the marker gene HPT flanked by homology arms. Upon co-transformation of the C. higginsianum protoplasts, pCas9-Ch_tRp-sgRNA causes a DNA double-strand break in the targeted gene, followed by homology-directed replacement of the gene with HPT by pCE-Zero-HPT, thereby generating loss-of-function mutants. Using the system, we generated the knockout mutants of two effector candidates (ChBas3 and OBR06881) with a 100% HRF. Interestingly, the ΔChBas3 and ΔOBR06881 mutants did not seem to affect the C. higginsianum infection of Arabidopsis thaliana. Altogether, the CRISPR/Cas9 system developed in the study enables the targeted deletion of genes, including effectors, in C. higginsianum, thus determining their biological functions.

Green tissues and seeds from cruciferous vegetables growing in conventional and ecological conditions (Brassica carinata; Brassica rapa; Eruca vesicaria and Sinapis alba) were analyzed to determine their contents of glucosinolates, isotihiocyanates (ITCs) and inorganic micronutrients (Ca, Cr, Cu, Fe, Mn, Ni, Se and Zn), and the bioaccessibility of these compounds. Regarding total contents and bioaccessibility values of these compounds, no clear difference was found between the organic and conventional systems. Glucosinolates bioaccessibility present in green tissues were high, with values around 60-78%. In additon, it was quantified in bioaccessible fraction ITCs concentrations such as Allyl - ITC; 3 - Buten - 1 - yl - ITC and 4 - Penten - 1 - yl - ITC. Trace elements bioaccessibility in green tissues was also high for Ca (2.26-7.66 mg/g), Cu (0.60-2.78 µg/g), Se (9.93-74.71 µg/Kg) and Zn (12.98-20.15 µg/g). By contrast, the bioaccessibility of glucosinolates and trace elements in cruciferous seeds was extremely low. With the exception of Cu, these bioaccessibility percentages did not exceed 1% in most cases.

Alternaria leaf blight and head rot is an important disease of broccoli and other cole crops. With no resistant host varieties, fungicides are utilized to manage this disease. However, anecdotal evidence suggests that, in southeastern U.S. broccoli-producing states, there is a loss of disease control through the use of quinone outside inhibitor (QoI) fungicides. To understand why there is a reduced sensitivity to QoI fungicides in these states, we isolated Alternaria spp. from symptomatic lesions on cole crops from Georgia and Virginia (two states with observations of loss of fungicide sensitivity) as well as New York (a state with no observations of loss of fungicide sensitivity). Using multilocus sequencing and phylogenetic analysis, we identified two species, Alternaria brassicicola and A. japonica. Whereas A. brassicicola was isolated in all states, A. japonica was only isolated in Georgia. Next, we wanted to determine the sensitivity of these isolates to azoxystrobin-an active ingredient in some QoI fungicides-by estimating the effective concentration at which only 50% of spores germinate (EC50). The EC50 of A. brassicicola ranged from 0.01 to 0.17 ppm, whereas that of A. japonica was 8.1 to 28.1 ppm. None of the known target-site mutations that confer resistance to QoI fungicides were identified during screening of either species. A. japonica was first reported on the east coast of the United States in 2020 in South Carolina. The substantially higher EC50 value suggests that its emergence in the southeastern United States may play at least a part in the observed loss of disease control. However, further in planta and field studies are needed to thoroughly test this hypothesis.

In this study, the effects of red (R), blue (B) and far-red (FR) LED lights and their combination (R + B, R + FR, B + FR, R + B + FR) together with white (W) LED light as control, on the growth, nutritional quality and the glucosinolates of brassica microgreens were determined. Fresh and dry weights were increased with W, R, R + FR lights in broccoli and cabbage and with the R + B + FR and B + FR in radish microgreens. Soluble solids content (SSC) (%) was highest with W, R and B lights in broccoli and cabbage. The highest titratable acidity (TA) (%) was determined with B, FR, R + FR, B + FR in broccoli and W, R + FR, R + B in cabbage. In radish, lower TA was determined. In broccoli microgreens, glucoraphanin content and total GSLs were increased with B light whereas in cabbage, the combination of R + B revealed the highest aliphatics, In radish, glucoraphenin was highest in B light and the glucoraphasatin in R, FR, R + FR and B lights.

Brassica plants play an important role in common agricultural practices, such as livestock feed or biofumigation, due to the bioactivity of the natural degradation products of glucosinolate metabolites. Therefore, the ability to survey comprehensive glucosinolate profiles for individual brassicas is essential for informing proper species selection for the intended application. Current methods for glucosinolate identification and quantification involve complex or unconventional procedures, and proper reference materials are not readily available. Therefore, researchers with limited resources that require glucosinolate profiles are at an extreme disadvantage. In this work, a simple and accurate HPLC-MS method was developed and validated to build preliminary glucosinolate profiles for three agriculturally relevant forage brassica varieties [turnip (B. rapa L.), canola (B. napus L.), and rapeseed (B. napus L.)]. The average glucosinolate content across three herbage collection dates for canola, rapeseed and turnip were 2.9 ± 0.9 mg g-1, 6.4 ± 1.3 mg g-1, and 14 ± 3.4 mg g-1, respectively. GLS concentrations are reported in milligrams of glucosinolate, calculated as sinigrin equivalents, per gram of dry plant material. This semi-quantitative approach for reporting total GLS content in brassicas is accurate within 15%. Several minor individual glucosinolates were identified that have not been previously reported in canola, rapeseed and turnip species, including glucotropaeolin and 4-hydroxyglucobrassicin (canola), glucoraphanin and glucoberteroin (rapeseed), and glucosinalbin and glucobarbarin (turnip). This non-targeted screen of several forage brassica varieties demonstrates the inherent variation in both the individual glucosinolate content and the total glucosinolate profile among brassicas, and highlights the importance of such glucosinolate characterization in agricultural practices. Additionally, the method developed in this study can be used as a tool for researchers with limited resources to build accurate glucosinolate profiles of brassica plants.

UNASSIGNED: The development of nanocarriers of plant origin, such as plant cell membranes, has recently been investigated. Also, plant bioactive compounds as sulforaphane (SFN) from broccoli have recognized antioxidant or anticancer properties.
UNASSIGNED: To investigate the capacity of membrane vesicles from broccoli (BM-vesicles) to encapsulate SFN and their application in the cancer cell line.
UNASSIGNED: Physicochemical analysis was carried out to characterize BM-vesicles through different approaches: dynamic light scattering, transmission electron microscopy, stopped-flow analysis, and proteomic analysis. They were applied at different concentrations (BM-vesicles at 0.04-0.00315% of protein and SFN at 5, 25, and 100 µM) in SK-MEL-28 cells during 24 h for studying cytotoxicity and gene expression.
UNASSIGNED: The entrapment efficiency was 41%. The anticancer activity tested in cells showed a decrease in proliferation when SFN in BM-vesicles was utilized. Expression patterns when SFN was applied in an encapsulated form showed a reduction of cancer markers and an increase of AQP3. Also, the metabolism of SFN occurred inside of cells, and higher SFN penetrated when it was encapsulated.
UNASSIGNED: The results showed that encapsulated SFN was better absorbed by melanoma cells providing metabolism products and a reduction of cancer molecular markers. Also aquaporin, AQP3 was pointed to as an important marker since it appeared to play a key role in homeostasis due to the importance of water transport in biological processes.
UNASSIGNED: These results indicate that SFN and SFN encapsulated in BM-vesicles have a high activity for the inhibition of melanocyte development. Therefore, BM-vesicles could serve as nanocarriers for drugs.

Animal manure application to soils is considered to be one of the main cause of antibiotic and bacterial pathogen spread in the environment. Pig livestock, which is the source of one of the most used fertilizer for cultivated land, is also a hotspot for antibiotics and antibiotic-resistant bacteria. Besides harsh chemical and physical sanitization treatments for the abatement of antibiotics and bacterial load in livestock waste, more sustainable and environmentally friendly strategies need to be considered. In this context, the use of natural substances which are proved useful for pest and disease control is currently under exploration for their role in the reduction of bacterial pathogen population. Among these, plants and derived products from the Brassicaceae family, characterized by the presence of a defensive glucosinolate-myrosinase enzymatic system, have been successfully exploited for years in agriculture using the so-called biofumigation technique against crop diseases. Although the application of biofumigation to suppress a range of soil borne pests has been well documented, no studies have been examined to reduce bacterial population in animal waste. In the present study, the release and the antibacterial activity of bioactive compounds deriving from different Brassicaceae defatted seed meals against pathogens and bacterial population in pig manure is addressed. Rapistrum rugosum and Brassica nigra defatted seed meals were found to be the most active products against tested pathogens and able to significantly reduce the bacterial load in the manure.