Brassica Species Research Articles

Comprehensive Quality Assessment of Brassica napus L. Seeds via HPTLC, LC-QToF, and Anatomical Investigation.

The Brassicaceae family, commonly referred to as cruciferous plants, is globally cultivated and consumed, with the Brassica genus being particularly renowned for its functional components. These vegetables are rich sources of nutrients and health-promoting phytochemicals, garnering increased attention in recent years. This study presents a comprehensive microscopic, chromatographic, and spectroscopic characterization of Brassica napus L. seeds from Kazakhstan aimed at elucidating their morphological features and chemical composition. Microscopic analysis revealed distinct localization of flavonoids, total lipids, and alkaloids. High-performance thin-layer chromatography (HPTLC) analysis of seed extracts demonstrated a complex chemical profile with significant quantities of non-polar compounds in the hexane extracts. Additionally, methanolic extracts revealed the presence of diverse chemical compounds, including alkaloids, flavonoids, and glucosinolates. The chemical composition exhibited varietal differences across different Brassica species, with B. napus L. seeds showing higher concentrations of bioactive compounds. Furthermore, liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QToF-MS) analysis provided insights into the chemical composition, with sinapine isomers, feruloyl, and sinapoyl choline derivatives as major compounds in the seeds. This study contributes to a better understanding of the chemical diversity and quality control methods' approximations of B. napus L. seeds, highlighting their importance in functional food and nutraceutical applications.

Genome-wide characterization of Related to ABI3/VP1 transcription factors among U's triangle Brassica species reveals a negative role for BnaA06.RAV3L in seed size

The transcription factors Related to ABI3/VP1 (RAV) are crucial for various plant processes and stress responses. Although the U's triangle Brassica species genomes have been released, the knowledge regarding the RAV family is still limited. In this study, we identified 123 putative RAV genes across the six U's triangle Brassica species (Brassica rapa, 14; Brassica oleracea, 14; Brassica nigra, 13; Brassica carinata, 27; Brassica juncea, 28; Brassica napus, 27). Phylogenetic analysis categorized them into three groups. The RAV genes exhibited diversity in both functional and structural aspects, particularly in gene structure and cis-acting elements within their promoters. The expression analysis revealed that BnaRAV genes in Group 1/2 exhibited diverse expression patterns across various tissues, while those in Group 3 did not show expression except for BnaRAV3L-2 and BnaRAV3L-6, which were exclusively expressed in seeds. Furthermore, the seed-specific expression of BnaA06. RAV3L (BnaRAV3L-2) was confirmed through promoter-GUS staining. Subcellular localization studies demonstrated that BnaA06.RAV3L is localized to the nucleus. The overexpression of BnaA06. RAV3L in Arabidopsis led to a remarkable inhibition of seed-specific traits such as seed width, seed length, seed area, and seed weight. This study provides insights into the functional evolution of the RAV gene family in U triangle Brassica species. It establishes a foundation for uncovering the molecular mechanisms underlying the negative role of RAV3L in seed development.

Assessment of Drought Tolerance of Two Cultivars of Indian Mustard (Brassica juncea) Using Morphological and Physiological Parameters

Aims: Drought stress may affect the Morphology, biochemistry, growth, and physiology of Brassica juncea (Indian mustard), a low-cost, oil-yielding, and economically significant plant. In this paper assessing various growth, morphological, and physiological parameters under different irrigation regimes could offer some significant information for the selection of a suitable and appropriate genotype for breeding. Study Design: Two commonly grown cultivars (RH-725 and RH-749) of mustard in Haryana under four irrigation regimes were evaluated. The applied Irrigation regimes were: control (double irrigation: once at the 50% flowering and another at the 50% fruiting stages), early irrigation (at 50% flowering only), late irrigation (at 50% fruiting only), and stress (no irrigation). The plants were exposed to short-term water shortages during the vegetative and reproductive growth stages. Drought stress in both stages had a negative effect on the morphological and physiological parameters of mustard. Place and Duration of Study: An open field experiment was performed at the Nursery of Kurukshetra University, Kurukshetra, Haryana, India, located at the latitude of 290–95° north and longitude of 760–82° east, with a height of 258.4 m above sea level. Crop sown under field conditions from October to March for two consecutive years (2018-2019, 2019-2020) at a temperature of 30-32 °C (days) and 15-25 °C (nights). Methodology: Measured adaptability using 15 morphological, 4 physiological, and 7 different growth parameters. PCA data revealed that physiological and growth parameters are more sensitive than morphological parameters in distinguishing the control and drought treatments. Alterations in physiological parameters occurred at all three levels of water stress. Growth analysis and yield parameters decreased with the severity of stress. Drought-stress treatments gradually reduced the Morphological and physiological traits of mustard. Results: The study revealed a relationship between Brassica species' adaptation to drought stress and the rapidity, severity, and duration of the drought event. This highlights the importance of considering these factors while selecting genotypes for breeding programmes aimed at improving drought tolerance in B. juncea. Overall, the research provides valuable information for selecting suitable and appropriate genotypes for breeding drought-tolerant varieties of B. juncea. By understanding the specific physiological (relative water content, leaf osmotic potential, membrane stability index, electrolytic index) and growth parameters (plant height, root length, fresh and dry weight of leaves and roots, leaf area etc.) that are more sensitive to drought stress. Furthermore, this study revealed a relationship between Brassica species' adaptation to drought stress and the rapidity, severity, and duration of the drought event. Based on PCA values, the cultivar RH-749 performed better both morphologically and physiologically under all stress conditions compared to RH-725. This suggests that RH-749 is more adapted and tolerant to drought stress, making it a potential candidate for breeding and cultivation in water-limited environments.

Assembly and comparative analysis of the complete mitochondrial genome of Brassica rapa var. Purpuraria

BackgroundPurple flowering stalk (Brassica rapa var. purpuraria) is a widely cultivated plant with high nutritional and medicinal value and exhibiting strong adaptability during growing. Mitochondrial (mt) play important role in plant cells for energy production, developing with an independent genetic system. Therefore, it is meaningful to assemble and annotate the functions for the mt genome of plants independently. Though there have been several reports referring the mt genome of in Brassica species, the genome of mt in B. rapa var. purpuraria and its functional gene variations when compared to its closely related species has not yet been addressed.ResultsThe mt genome of B. rapa var. purpuraria was assembled through the Illumina and Nanopore sequencing platforms, which revealed a length of 219,775 bp with a typical circular structure. The base composition of the whole B. rapa var. purpuraria mt genome revealed A (27.45%), T (27.31%), C (22.91%), and G (22.32%). 59 functional genes, composing of 33 protein-coding genes (PCGs), 23 tRNA genes, and 3 rRNA genes, were annotated. The sequence repeats, codon usage, RNA editing, nucleotide diversity and gene transfer between the cp genome and mt genome were examined in the B. rapa var. purpuraria mt genome. Phylogenetic analysis show that B. rapa var. Purpuraria was closely related to B. rapa subsp. Oleifera and B. juncea. Ka/Ks analysis reflected that most of the PCGs in the B. rapa var. Purpuraria were negatively selected, illustrating that those mt genes were conserved during evolution.ConclusionsThe results of our findings provide valuable information on the B.rapa var. Purpuraria genome, which might facilitate molecular breeding, genetic variation and evolutionary researches for Brassica species in the future.

Cytotoxicity of allyl isothiocyanate on insulin-producing INS-1E cells.

Allyl isothiocyanate (AITC) is the pungent ingredient of brassica species, used as a food additive and flavoring agent, including condiments such as wasabi, horseradish, and mustard. Currently, there is much evidence that AITC modulates glucose and lipids metabolism. Interestingly, AITC has been shown to improve glycaemia, and insulin action along with the induction of a deepened decline in blood insulin levels in T2DM rats. Therefore, in the present study, we characterized the role of AITC at a wide concentration range (5, 10, 25, 50, 100 μM) in controlling viability, proliferation, apoptosis, mitochondrial condition, mRNA expression of encoding pancreatic and duodenal homeobox 1 (Pdx1), and Ins1, Ins2 genes, and insulin content in INS-1E cells. The INS-1E cell line is a suitable, and well-characterized model to study beta cell functions. We demonstrate that AITC reduced the viability (p≤0.001) (also in the presence of transient receptor potential cation subfamily A member 1 (TRPA1) selective antagonist; HC-030031; p≤0.05), and proliferation of INS-1E cells (p≤0.001). AITC evoked a significant reduction of mitochondrial membrane potential (p≤0.01) and decreased the intracellular level of adenosine triphosphate (ATP) (p≤0.001) without influence on reactive oxygen species (ROS) level. Additionally, AITC inhibited the insulin mRNA expression (p≤0.001) in INS-1E cells along with insulin content (p≤0.05). Mitochondrial dysfunction is proposed to be a significant disruption mechanism of AITC in INS-1E cells, and it was independent of ROS, and the influx of external calcium.

Genome-wide association studies of root system architecture traits in a broad collection of Brassica genotypes.

The root systems of Brassica species are complex. Eight root system architecture (RSA) traits, including total root length, total root surface area, root average diameter, number of tips, total primary root length, total lateral root length, total tertiary root length, and basal link length, were phenotyped across 379 accessions representing six Brassica species (B. napus, B. juncea, B. carinata, B. oleracea, B. nigra, and B. rapa) using a semi-hydroponic system and image analysis software. The results suggest that, among the assessed species, B. napus and B. oleracea had the most intricate and largest root systems, while B. nigra exhibited the smallest roots. The two species B. juncea and B. carinata shared comparable root system complexity and had root systems with larger root diameters. In addition, 313 of the Brassica accessions were genotyped using a 19K Brassica single nucleotide polymorphism (SNP) array. After filtering by TASSEL 5.0, 6,213 SNP markers, comprising 5,103 markers on the A-genome (covering 302,504 kb) and 1,110 markers on the C-genome (covering 452,764 kb), were selected for genome-wide association studies (GWAS). Two general linear models were tested to identify the genomic regions and SNPs associated with the RSA traits. GWAS identified 79 significant SNP markers associated with the eight RSA traits investigated. These markers were distributed across the 18 chromosomes of B. napus, except for chromosome C06. Sixty-five markers were located on the A-genome, and 14 on the C-genome. Furthermore, the major marker-trait associations (MTAs)/quantitative trait loci (QTLs) associated with root traits were located on chromosomes A02, A03, and A06. Brassica accessions with distinct RSA traits were identified, which could hold functional, adaptive, evolutionary, environmental, pathological, and breeding significance.

Biocontrol potential of cell-free supernatant of Paenibacillus chitinolyticus against Plasmodiophora brassicae in two important Brassica species

Plasmodiophora brassicae is a serious threat to Brassica crops worldwide, resulting in substantial economic losses for growers. Biocontrol with chitinolytic bacteria producing chitinase is gaining attention as a natural alternative to chemicals. This approach is favored due to the essential role chitinases play in protecting against chitin-containing pathogens. Given that chitin is a major component in the resting spores of P. brassicae and plays a crucial role during pathogenesis, it is probable that Paenibacillus chitinolyticus, producing a high level of chitinase, could suppress P. brassicae by targeting chitin in a critical stage of this pathogen’s life cycle. Our research aimed to evaluate the effect of various applications of P. chitinolyticus on clubroot suppression in two economically important Brassica species: Chinese cabbage and rapeseed. The effectiveness of the cell-free supernatant (CFS) of an endemic strain of P. chitinolyticus from the Czech Republic at five different time points was studied in the greenhouse by measuring the disease severity index. Early application of P. chitinolyticus decreased the disease index significantly within both plant species resulting in increased shoot dry weight. In conclusion, the CFS of P.chitinolyticus has significant antagonistic activity against clubroot in Chinese cabbage and rapeseed in the early developmental stages of clubroot occurrence and holds the potential as a biofertilizer as well as bioprotectant agent in clubroot management of P. brassicae.

The ilv2 gene, encoding acetolactate synthase for branched chain amino acid biosynthesis, is required for plant pathogenicity by Leptosphaeria maculans

BackgroundControl of blackleg disease of canola caused by the fungus Leptosphaeria maculans relies on strategies such as the inhibition of growth with fungicides. However, other chemicals are used during canola cultivation, including fertilizers and herbicides. There is widespread use of herbicides that target the acetolactate synthase (ALS) enzyme involved in branched chain amino acid synthesis and low levels of these amino acids within leaves of Brassica species. In L. maculans the ilv2 gene encodes ALS and thus ALS-inhibiting herbicides may inadvertently impact the fungus.Methods and resultsHere, the impact of a commercial herbicide targeting ALS and mutation of the homologous ilv2 gene in L. maculans was explored. Exposure to herbicide had limited impact on growth in vitro but reduced lesion sizes in plant disease experiments. Furthermore, the mutation of the ilv2 gene via CRISPR-Cas9 gene editing rendered the fungus non-pathogenic.ConclusionHerbicide applications can influence disease outcome, but likely to a minor extent.

Multiomics analysis of a resistant European turnip ECD04 during clubroot infection reveals key hub genes underlying resistance mechanism

The clubroot disease has become a worldwide threat for crucifer crop production, due to its soil-borne nature and difficulty to eradicate completely from contaminated field. In this study we used an elite resistant European fodder turnip ECD04 and investigated its resistance mechanism using transcriptome, sRNA-seq, degradome and gene editing. A total of 1751 DEGs were identified from three time points after infection, among which 7 hub genes including XTH23 for cell wall assembly and two CPK28 genes in PTI pathways. On microRNA, we identified 17 DEMs and predicted 15 miRNA-target pairs (DEM-DEG). We validated two pairs (miR395-APS4 and miR160-ARF) by degradome sequencing. We investigated the miR395-APS4 pair by CRISPR-Cas9 mediated gene editing, the result showed that knocking-out APS4 could lead to elevated clubroot resistance in B. napus. In summary, the data acquired on transcriptional response and microRNA as well as target genes provide future direction especially gene candidates for genetic improvement of clubroot resistance on Brassica species.

A chromosome-scale assembly of Brassica carinata (BBCC) accession HC20 containing resistance to multiple pathogens and an early generation assessment of introgressions into B.juncea (AABB).

Brassica carinata (BBCC) commonly referred to as Ethiopian mustard is a natural allotetraploid containing the genomes of Brassica nigra (BB) and Brassica oleracea (CC). It is an oilseed crop endemic to the northeastern regions of Africa. Although it is under limited cultivation, B. carinata is valuable as it is resistant/highly tolerant to most of the pathogens affecting widely cultivated Brassica species of the U's triangle. We report a chromosome-scale genome assembly of B. carinata accession HC20 using long-read Oxford Nanopore sequencing and Bionano optical maps. The assembly has a scaffold N50 of ~39.8 Mb and covers ~1.11 Gb of the genome. We compared the long-read genome assemblies of the U's triangle species and found extensive gene collinearity between the diploids and allopolyploids with no evidence of major gene losses. Therefore, B. juncea (AABB), B. napus (AACC), and B. carinata can be regarded as strict allopolyploids. We cataloged the nucleotide-binding and leucine-rich repeat immune receptor (NLR) repertoire of B.carinata and, identified 465 NLRs, and compared these with the NLRs in the other Brassica species. We investigated the extent and nature of early-generation genomic interactions between the constituent genomes of B. carinata and B. juncea in interspecific crosses between the two species. Besides the expected recombination between the constituent B genomes, extensive homoeologous exchanges were observed between the A and C genomes. Interspecific crosses, therefore, can be used for transferring disease resistance from B. carinata to B. juncea and broadening the genetic base of the two allotetraploid species.

Laser capture microdissection transcriptome (LCM RNA-seq) reveals BcDFR is a key gene in anthocyanin synthesis of non-heading Chinese cabbage

BackgroundPurple non-heading Chinese cabbage [Brassica campestris (syn. Brassica rapa) ssp. chinensis] has become popular because of its richness in anthocyanin. However, anthocyanin only accumulates in the upper epidermis of leaves. Further studies are needed to investigate the molecular mechanisms underlying the specific accumulation of it.ResultsIn this study, we used the laser capture frozen section method (LCM) to divide purple (ZBC) and green (LBC) non-heading Chinese cabbage leaves into upper and lower epidermis parts (Pup represents the purple upper epidermis, Plow represents the purple lower epidermis, Gup represents the green upper epidermis, Glow represents the green lower epidermis). Through transcriptome sequencing, we found that the DIHYDROFLAVONOL 4-REDUCTASE-encoding gene BcDFR, is strongly expressed in Pup but hardly in others (Plow, Gup, Glow). Further, a deletion and insertion in the promoter of BcDFR in LBC were found, which may interfere with BcDFR expression. Subsequent analysis of gene structure and conserved structural domains showed that BcDFR is highly conserved in Brassica species. The predicted protein-protein interaction network of BcDFR suggests that it interacts with almost all functional proteins in the anthocyanin biosynthesis pathway. Finally, the results of the tobacco transient expression also demonstrated that BcDFR promotes the synthesis and accumulation of anthocyanin.ConclusionsBcDFR is specifically highly expressed on the upper epidermis of purple non-heading Chinese cabbage leaves and regulates anthocyanin biosynthesis and accumulation. Our study provides new insights into the functional analysis and transcriptional regulatory network of anthocyanin-related genes in purple non-heading Chinese cabbage.

Genome-wide identification and evolutionary analysis of the NRAMP gene family in the AC genomes of Brassica species

BackgroundBrassica napus, a hybrid resulting from the crossing of Brassica rapa and Brassica oleracea, is one of the most important oil crops. Despite its significance, B. napus productivity faces substantial challenges due to heavy metal stress, especially in response to cadmium (Cd), which poses a significant threat among heavy metals. Natural resistance-associated macrophage proteins (NRAMPs) play pivotal roles in Cd uptake and transport within plants. However, our understanding of the role of BnNRAMPs in B. napus is limited. Thus, this study aimed to conduct genome-wide identification and bioinformatics analysis of three Brassica species: B. napus, B. rapa, and B. oleracea.ResultsA total of 37 NRAMPs were identified across the three Brassica species and classified into two distinct subfamilies based on evolutionary relationships. Conservative motif analysis revealed that motif 6 and motif 8 might significantly contribute to the differentiation between subfamily I and subfamily II within Brassica species. Evolutionary analyses and chromosome mapping revealed a reduction in the NRAMP gene family during B. napus evolutionary history, resulting in the loss of an orthologous gene derived from BoNRAMP3.2. Cis-acting element analysis suggested potential regulation of the NRAMP gene family by specific plant hormones, such as abscisic acid (ABA) and methyl jasmonate (MeJA). However, gene expression pattern analyses under hormonal or stress treatments indicated limited responsiveness of the NRAMP gene family to these treatments, warranting further experimental validation. Under Cd stress in B. napus, expression pattern analysis of the NRAMP gene family revealed a decrease in the expression levels of most BnNRAMP genes with increasing Cd concentrations. Notably, BnNRAMP5.1/5.2 exhibited a unique response pattern, being stimulated at low Cd concentrations and inhibited at high Cd concentrations, suggesting potential response mechanisms distinct from those of other NRAMP genes.ConclusionsIn summary, this study indicates complex molecular dynamics within the NRAMP gene family under Cd stress, suggesting potential applications in enhancing plant resilience, particularly against Cd. The findings also offer valuable insights for further understanding the functionality and regulatory mechanisms of the NRAMP gene family.

Shotgun approach reveals distinctive lipid profiles in brassica oilseeds: A high-resolution ESI-ToF-MS study

Members of Brassica seed oil are important sources of nutritionally superior edible oil. There are no comprehensive reports on complete lipidomic profile of these oilseed crops. In this study, the lipidomic profiling of edible oil from 7 different cultivated species of Brassica seed oils was performed by shotgun approach using electrospray High Resolution Time of Flight-Mass Spectrometry (ESI-ToF-MS). The mass spectrum under positive polarity revealed 1098 lipids under different lipid classes including sphingolipids, phospholipids and different storage lipids. Under negative polarity, 70 lipids including free fatty acids (FFA), cardiolipins and phospholipids were detected. Erucic acid in FFA form was found to be most abundant in both Yellow and Brown sarson. Brassica napus contains almost all forms of cardiolipins (CL). Out of 26 different species of cardiolipins detected in negative ion mode, CL 56:1 (FA 18:1) and CL 56:1 (FA 22:1) were present only in brown sarson. Similarly, CL 56:2 (FA 18:2) and CL 56:1 (FA 22:1) were present only in Yellow and Brown sarson. These findings enhance our understanding of the nutritional diversity in Brassica seed oils, emphasizing the significance of lipidomic analysis for elucidating the molecular composition of edible oils.

Alternaria brassicicola‐induced ROS accumulation during black spot disease differentially affects antioxidant efficiency, phenolic content and susceptibility of Brassica species

AbstractEarly‐stage responses of Brassica juncea, B. napus and B. oleracea during black spot disease and the development of Alternaria brassicicola were evaluated. Infection of plant cells by the fungus occurred mainly through direct penetration or by appressoria and, rarely, stomata in B. juncea and B. oleracea; in B. napus, penetration was mostly direct. The process of conidial germination, germ tubes and appressoria formation on a leaf surface was correlated with the post‐inoculation time and host species (p < 0.05). Changes in the leaf surface were observed at successful infection sites as bright, wax‐deprived areas. Significant differences in progression of infection between Brassica species were observed, with B. oleracea being the most susceptible and B. napus the least. Accumulation of superoxide anion radicals and hydrogen peroxide was not only observed in plant cells, but also in A. brassicicola germ tubes and appressoria at 8 and 12 h post‐inoculation (hpi). Enhanced production of reactive oxygen species (ROS) and lipid peroxidation during infection triggered diverse patterns of enzymatic and non‐enzymatic antioxidant activities and phenolic compound contents in response to A. brassicicola. In each Brassica species, a different antioxidant was the most active. With their antioxidant properties, phenolic compounds also played an essential role in these interactions. This phenomenon could be related to the disparate levels of susceptibility of the Brassica species to A. brassicicola; for example, the most susceptible, B. oleracea, showed high ROS accumulation with decreasing tendency during disease progression, unchanged phenolics content, and the highest catalase activity, differentiating it from B. juncea and B. napus.

Pathotyping Systems and Pathotypes of Plasmodiophora brassicae-Navigating toward the Optimal Classification.

Plasmodiophora brassicae Woronin, an obligate biotrophic soil-borne pathogen, poses a significant threat to cruciferous crops worldwide by causing the devastating disease known as clubroot. Pathogenic variability in P. brassicae populations has been recognized since the 1930s based on its interactions with Brassica species. Over time, numerous sets of differential hosts have been developed and used worldwide to explore the pathogenic variability within P. brassicae populations. These sets encompass a range of systems, including the Williams system, the European Clubroot Differential set (ECD), the Brassica napus set, the Japanese Clubroot Differential Set, the Canadian Clubroot Differential Set (CCS), the Korean Clubroot Differential Set, and the Chinese Sinitic Clubroot Differential set (SCD). However, all existing systems possess both advantages as well as limitations regarding the detection of pathotypes from various Brassica species and their corresponding virulence pattern on Brassica genotypes. This comprehensive review aims to compare the main differential systems utilized in classifying P. brassicae pathotypes worldwide. Their strengths, limitations, and implications are evaluated, thereby enhancing our understanding of pathogenic variability.

Heavy metal tolerance and accumulation in the Brassica species (Brassica chinensis var. parachinensis and Brassica rapa L.): A pot experiment

This study delves into the heavy metal tolerance and accumulation capabilities of Brassica chinensis var. parachinensis (B. chinensis) and Brassica rapa L. (B. rapa) in a pot experiment, specifically focusing on cadmium (Cd), chromium (Cr) and lead (Pb). Agricultural topsoils were spiked with varying concentrations of these heavy metals (0 mg/kg, 75 mg/kg, 150 mg/kg, 225 mg/kg and 300 mg/kg) for each element. The experiment involved cultivating 15 pots each of B. chinensis and B. rapa over 60 days. Results indicated that both Brassica species experienced delayed germination, with B. chinensis exhibiting a significant drop in germination percentage to 53 % at the highest concentration (300 mg/kg), while B. rapa showed a tendency for an increased germination percentage of up to 80 % at elevated metal concentrations; however, these differences were not statistically significant. Both B. chinensis and B. rapa demonstrated a stable decline in growth rate from 0.05 cm/day to 0.04 cm/day with increasing heavy metal concentrations, and the he reduction in relative growth rate was significant at the highest concentration compared to the control. The stress tolerance index revealed a significant decrease in plant heights for B. chinensis, in contrast to the stable performance of B. rapa, showcasing the tolerance of B. rapa to toxic conditions. Despite insignificant differences in fresh biomass due to metal treatments, B. chinensis consistently yielded higher biomass, yet it had a lower edible index due to its higher root biomass. Leaf areas increased significantly in both species at higher soil treatments, while root lengths remained unchanged, suggesting their resilience to elevated heavy metal concentrations. Analysis of plant tissues (leaves, stems and roots) using ICP-OES revealed that B. rapa accumulated the highest Cd concentration (864 mg/kg), whereas B. chinensis accumulated the highest Pb concentration (953 mg/kg) in root parts. Both species significantly accumulated Cr in roots, demonstrating a sequestration mechanism. These findings suggest that both species, particularly, B. rapa possess strong tolerance and accumulation capabilities for non-essential heavy metals, making them potential hyperaccumulators for green remediation techniques in toxic soil environments. Understanding the molecular mechanisms driving these responses and validating phytoremediation potential in real-world scenarios is essential for developing sustainable soil management practices.

Evolutionary dynamics of the cytoskeletal profilin gene family in Brassica juncea L. reveal its roles in silique development and stress resilience

Essential to plant adaptation, cell wall (CW) integrity is maintained by CW-biosynthesis genes. Cytoskeletal actin-(de)polymerizing, phospholipid-binding profilin (PRF) proteins play important roles in maintaining cellular homeostasis across kingdoms. However, evolutionary selection of PRF genes and their systematic characterization in family Brassicaceae, especially in Brassica juncea remain unexplored. Here, a comprehensive analysis of genome-wide identification of BjPRFs, their phylogenetic association, genomic localization, gene structure, and transcriptional profiling were performed in an evolutionary framework. Identification of 23 BjPRFs in B. juncea indicated an evolutionary conservation within Brassicaceae. The BjPRFs evolved through paralogous and orthologous gene formation in Brassica genomes. Evolutionary divergence of BjPRFs indicated purifying selection, with nonsynonymous (dN)/synonymous (dS) value of 0.090 for orthologous gene-pairs. Hybrid homology-modeling identified evolutionary distinct and conserved domains in BjPRFs which suggested that these proteins evolved following the divergence of monocot and eudicot plants. RNA-seq profiles of BjPRFs revealed their functional evolution in spatiotemporal manner during plant-development and stress-conditions in diploid/amphidiploid Brassica species. Real-Time PCR experiments in seedling, vegetative, floral and silique tissues of B. juncea suggested their essential roles in systematic plant development. These observations underscore the expansion of BjPRFs in B. juncea, and offer valuable evolutionary insights for exploring cellular mechanisms, and stress resilience.

Low Nitrogen Input Mitigates Quantitative but Not Qualitative Reconfiguration of Leaf Primary Metabolism in Brassica napus L. Subjected to Drought and Rehydration.

In the context of climate change and the reduction of mineral nitrogen (N) inputs applied to the field, winter oilseed rape (WOSR) will have to cope with low-N conditions combined with water limitation periods. Since these stresses can significantly reduce seed yield and seed quality, maintaining WOSR productivity under a wide range of growth conditions represents a major goal for crop improvement. N metabolism plays a pivotal role during the metabolic acclimation to drought in Brassica species by supporting the accumulation of osmoprotective compounds and the source-to-sink remobilization of nutrients. Thus, N deficiency could have detrimental effects on the acclimation of WOSR to drought. Here, we took advantage of a previously established experiment to evaluate the metabolic acclimation of WOSR during 14 days of drought, followed by 8 days of rehydration under high- or low-N fertilization regimes. For this purpose, we selected three leaf ranks exhibiting contrasted sink/source status to perform absolute quantification of plant central metabolites. Besides the well-described accumulation of proline, we observed contrasted accumulations of some "respiratory" amino acids (branched-chain amino acids, lysineand tyrosine) in response to drought under high- and low-N conditions. Drought also induced an increase in sucrose content in sink leaves combined with a decrease in source leaves. N deficiency strongly decreased the levels of major amino acids and subsequently the metabolic response to drought. The drought-rehydration sequence identified proline, phenylalanine, and tryptophan as valuable metabolic indicators of WOSR water status for sink leaves. The results were discussed with respect to the metabolic origin of sucrose and some amino acids in sink leaves and the impact of drought on source-to-sink remobilization processes depending on N nutrition status. Overall, this study identified major metabolic signatures reflecting a similar response of oilseed rape to drought under low- and high-N conditions.

Identification, Evolutionary Dynamics, and Gene Expression Patterns of the ACP Gene Family in Responding to Salt Stress in Brassica Genus.

Acyl carrier proteins (ACPs) have been reported to play a crucial role in responding to biotic and abiotic stresses, regulating growth and development. However, the biological function of the ACP gene family in the Brassica genus has been limited until now. In this study, we conducted a comprehensive analysis and identified a total of 120 ACP genes across six species in the Brassica genus. Among these, there were 27, 26, and 30 ACP genes in the allotetraploid B. napus, B. juncea, and B. carinata, respectively, and 14, 13, and 10 ACP genes in the diploid B. rapa, B. oleracea, and B. nigra, respectively. These ACP genes were further classified into six subclades, each containing conserved motifs and domains. Interestingly, the majority of ACP genes exhibited high conservation among the six species, suggesting that the genome evolution and polyploidization processes had relatively minor effects on the ACP gene family. The duplication modes of the six Brassica species were diverse, and the expansion of most ACPs in Brassica occurred primarily through dispersed duplication (DSD) events. Furthermore, most of the ACP genes were under purifying selection during the process of evolution. Subcellular localization experiments demonstrated that ACP genes in Brassica species are localized in chloroplasts and mitochondria. Cis-acting element analysis revealed that most of the ACP genes were associated with various abiotic stresses. Additionally, RNA-seq data revealed differential expression levels of BnaACP genes across various tissues in B. napus, with particularly high expression in seeds and buds. qRT-PCR analysis further indicated that BnaACP genes play a significant role in salt stress tolerance. These findings provide a comprehensive understanding of ACP genes in Brassica plants and will facilitate further functional analysis of these genes.

Impact of biotic stresses on the Brassicaceae family and opportunities for crop improvement by exploiting genotyping traits.

Utilizing RNAi, miRNA, siRNA, lncRNA and exploiting genotyping traits can help safeguard the food supply from illnesses and pest damage to Brassicas, as well as reduce yield losses caused by plant pathogens and insect pests. In the natural environment, plants face significant challenges in the form of biotic stress, due to various living organisms, leading to biological stress and a sharp decline in crop yields. To cope with these effects, plants have evolved specialized mechanisms to mitigate these challenges. Plant stress tolerance and resistance are influenced by genes associated with stress-responsive pathogens that interact with various stress-related signaling pathway components. Plants employ diverse strategies and mechanisms to combat biological stress, involving a complex network of transcription factors that interact with specific cis-elements to regulate gene expression. Understanding both plant developmental and pathogenic disease resistance mechanisms can allow us to develop stress-tolerant and -resistant crops. Brassica genus includes commercially important crops, e.g., broccoli, cabbage, cauliflower, kale, and rapeseed, cultivated worldwide, with several applications, e.g., oil production, consumption, condiments, fodder, as well as medicinal ones. Indeed, in 2020, global production of vegetable Brassica reached 96.4 million tones, a 10.6% rise from the previous decade. Taking into account their commercial importance, coupled to the impact that pathogens can have in Brassica productivity, yield losses up to 60%, this work complies the major diseases caused due to fungal, bacterial, viral, and insects in Brassica species. The review is structured into three parts. In the first part, an overview is provided of the various pathogens affecting Brassica species, including fungi, bacteria, viruses, and insects. The second part delves into the exploration of defense mechanisms that Brassica plants encounter against these pathogens including secondary metabolites, duplicated genes, RNA interference (RNAi), miRNA (micro-RNA), siRNA (small interfering RNA), and lncRNA (long non-coding RNA). The final part comprehensively outlines the current applications of CRISPR/Cas9 technology aimed at enhancing crop quality. Taken collectively, this review will contribute to our enhanced understanding of these mechanisms and their role in the development of resistance in Brassica plants, thus supporting strategies to protect this crucial crop.