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The article presents the results of a three-year field study that was conducted in Poland to evaluate the yield and quality of seeds and oil from traditional (SAM) and canola-quality white mustard (SAC) in response to different N fertilizer rates (0, 40, 80, 120, and 160 kg ha -1 ). Seed yields were 25% higher in SAM than SAC. The seeds of SAC contained more crude fat (by 3%) and crude fiber (by 6%) than the seeds of SAM. In turn, the seeds of SAM were a richer source of total protein (by 7%). The content of glucosinolates (GSLs) was 8-10 times lower in the seeds of SAC than SAM. The seeds of both mustard cultivars were most abundant in γ-tocopherol (γ-T) (90-94%). The seeds of SAC were characterized by a higher content of γ-T and a lower α-T/γ-T ratio than SAM seeds. White mustard oil contained mostly MUFAs (69-75%). However, C22:1 accounted for more than 50% of MUFAs in the oil SAM. In the oil SAC, the proportion of C22:1 did not exceed 6%, whereas C18:1 accounted for nearly 85% of total MUFAs. Nitrogen fertilization induced a significant increase in seed yields (by 33%), a decrease in crude fat content (by 3-4%), and an increase in total protein content (by 4%), and crude fiber content (by 7%). Nitrogen decreased GSL levels by 31% in SAM seeds. In SAC, N fertilization induced differences in the qualitative composition of GSLs, but did not affect the total GSL content of seeds. Higher N rates increased the content of α-T and γ-T, the α-T/γ-T ratio, and total T content. Nitrogen fertilization decreased the content of C18:3 in the seeds of the SAM. In SAC, the application of N decreased the content of C18, C18:1, and C18:3, and increased the biosynthesis of C18:2, C20:1, and C22:1., (© 2024. The Author(s).)

A pot experiment at Punjab Agricultural University, Ludhiana, India, assessed rice residue biochar's effect on remediating lead and cadmium in soil and on Indian mustard growth. Soil spiked with Cd (0, 10, 25 mg kg⁻¹) and Pb (0, 25, 50 mg kg⁻¹) was amended with biochar (0, 0.5, 1, 2% w/w) in a completely randomized design with three replications. Adding 2% biochar reduced DTPA-Pb in soil and Pb in shoots by 61% and 56%, respectively. Biochar increased dry matter yield (DMY) by 18% at 1% but decreased by 19% at 2%. The 1% biochar increased shoot Fe, Mn, Zn, and Cu by 11.39%, 26.74%, 5.89%, and 17.72%, respectively. Rice residue biochar significantly improved the DMY and micronutrient content of Indian mustard by reducing lead contamination in co-contaminated soil., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Brassica juncea (L.) Czern is an important oilseed crop affected by various abiotic stresses like drought, heat, and salt. These stresses have detrimental effects on the crop's overall growth, development and yield. Various Transcription factors (TFs) are involved in regulation of plant stress response by modulating expression of stress-responsive genes. The myeloblastosis (MYB) TFs is one of the largest families of TFs associated with various developmental and biological processes such as plant growth, secondary metabolism, stress response etc. However, MYB TFs and their regulation by non-coding RNAs (ncRNAs) in response to stress have not been studied in B. juncea. Thus, we performed a detailed study on the MYB TF family and their interactions with miRNAs and Long non coding RNAs., Results: Computational investigation of genome and proteome data presented a comprehensive picture of the MYB genes and their protein architecture, including intron-exon organisation, conserved motif analysis, R2R3 MYB DNA-binding domains analysis, sub-cellular localization, protein-protein interaction and chromosomal locations. Phylogenetically, BjuMYBs were further classified into different subclades on the basis of topology and classification in Arabidopsis. A total of 751 MYBs were identified in B. juncea corresponding to 297 1R-BjuMYBs, 440 R2R3-BjuMYBs, 12 3R-BjuMYBs, and 2 4R-BjuMYBs types. We validated the transcriptional profiles of nine selected BjuMYBs under drought stress through RT-qPCR. Promoter analysis indicated the presence of drought-responsive cis-regulatory components. Additionally, the miRNA-MYB TF interactions was also studied, and most of the microRNAs (miRNAs) that target BjuMYBs were involved in abiotic stress response and developmental processes. Regulatory network analysis and expression patterns of lncRNA-miRNA-MYB indicated that selected long non-coding RNAs (lncRNAs) acted as strong endogenous target mimics (eTMs) of the miRNAs regulated expression of BjuMYBs under drought stress., Conclusions: The present study has established preliminary groundwork of MYB TFs and their interaction with ncRNAs in B. juncea and it will help in developing drought- tolerant Brassica crops., (© 2024. The Author(s).)

Background: India's oilseed economy falls short of self-sufficiency and is supplemented by huge imports every year. Increasing national productivity of the major oilseeds is confronted with yield losses due to diverse biotic and abiotic stresses. The productivity of Indian mustard (Brassica juncea Linnaeus), belonging to the family Brassicaceae, is significantly reduced due to damage caused by mustard aphids (Lipaphis erysimi Kaltenbach, Hemiptera: Aphididae). Rapid colonization by the nymphs makes it difficult to protect the crop through agrochemicals. Aphids release effector molecules to modulate the host-defence responses. Glucosinolates (GSLs) extensively found in Brassicaceae family, are hydrolysed by myrosinase into toxic compounds that deter herbivore insects., Methods: Here, we investigated the differential activation of the glucosinolate-myrosinase pathway in mustard manifesting susceptibility and resistance to different aphid species. Mustard plants were challenged by two different aphid species mustard aphid and cowpea aphid (Aphis craccivora Koch, Hemiptera: Aphididae) leading to complete host-susceptibility in one case and resistance in the other, respectively. Differential regulation of the GSL biosynthetic pathway and myrosinase activity was assessed by gene expression study and ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC- QToF-ESL-MS)., Results: Gene expression study identified selective transcriptional attenuation of the key GSL biosynthetic and myrosinase gene in mustard when challenged with mustard aphid. In contrary, the activation of GSL biosynthetic genes in conjunction with myrosinase at the transcriptional level was profound in mustard, when challenged with cowpea aphid. UPLC-MS analysis showed higher turnover in the hydrolysis of glucosinolates by myrosinase which led to concomitant generation of glucose as byproduct in response to cowpea aphid in mustard plants., Conclusion: GSL-myrosinase pathway is specifically attenuated by the successful aphid species in mustard and thus plays a pivotal role in determining the outcome of the B. juncea-aphid interaction. The results open up a new genetic modification strategy for developing resistance against aphids., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Brassica juncea is a crucial oilseed crop, and its seeds possess high economic value as they are a source of edible oil. In order to understand the role of long non coding RNAs (lncRNAs) in the regulation of seed development, we carried out computational analysis using transcriptome data of developing seeds of two contrasting genotypes of B. juncea, Pusajaikisan (PJK) and Early Heera 2 (EH2). The seeds were sampled at three stages, 15, 30, and 45 days after pollination. We identified 1,539 lncRNAs, of which 809 were differentially expressed. We also carried out extensive characterization and functional analysis of seed lncRNAome. The expression patterns were analysed using k-means clustering, and the targets were analysed using pathway, transcription factor, and GO enrichment, as well as ortholog information. We shortlisted a total of 25 robust lncRNA candidates for seed size, oil content, and seed coat color. We also identified 4 lncRNAs as putative precursors of miRNAs regulating seed development. Moreover, a total of 28 miRNA-lncRNA-mRNA regulatory networks regulating seed traits were identified. We also developed a comprehensive database, (BrassIca juncea database or "BIJ" ( https://bij.cuh.ac.in/ ), which provides seed omics as well as other functional genomics and genetics data in an easily accessible form. These candidate lncRNAs are suitable for including in crop improvement programs through molecular breeding, as well as for future validations through genome editing. Together, the knowledge of these candidate lncRNAs and availability of BIJ database shall leverage the crop improvement efforts in B. juncea., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Understanding the stability of photosynthetic pigments is crucial for developing crop cultivars with high productivity and resilience to the environmental stresses. This study leveraged GGE biplot, WAASB, and MTSI indices to assess the stability of content and composition of photosynthetic pigments in leaves and siliques of 286 Brassica juncea (L.) Czern. genotypes across three environments. The GGE biplot analysis identified NRCQR-9901 as the best genotype in terms of chlorophyll 'a' under conditions of high irradiance and long days (E1). For chlorophyll 'b' and total chlorophyll, NC-533728 performed the best. AJ-2 and NPJ-208 had the maximum total carotenoids levels in leaves. RLC-2 was characterized by maximum values for chlorophyll a, chlorophyll b, and total chlorophyll in the siliques. The low irradiance, short days, and moderate to high temperatures (E2) seemed perfect for the synthesis of photosynthetic pigments. NPJ-182 shows the maximum concentrations of chlorophyll 'a', total chlorophyll, and total carotenoids in leaves. Conversely, IC-597869, RE-389, and IC-597894 exhibited the highest concentrations of chlorophyll 'b' under an environment characterized by low light intensity, shorter daylights, and low temperatures (E3) during flowering and siliqua formation stages. The combined analysis found NPJ-182, NC-533728, CN-105233, RLC-2, CN-101846, JA-96, PBR-357, JM-3, and DTM-34 as top performers with high stability. Comparative transcriptome analysis with two stable and high-performing genotypes (PBR-357 and DTM-34) and two average performers revealed upregulation of critical photosynthesis-related genes (ELIP1, CAB3.1, ELIP1.5, and LHCB5) in top performers. This study identified promising trait donors for use in breeding programs aimed at improving the mustard crop's photosynthetic efficiency, productivity, and stability., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Soil acidity is a divesting factor that restricts crop growth and productivity. Conversely, silicon nanomaterials (Si-NMs) have been praised as a blessing of modern agricultural intensification by overcoming the ecological barrier. Here, we performed a sequential study from seed germination to the yield performance of mustard ( Brassica campestris ) crops under acid-stressed conditions. The results showed that Si-NMs significantly improved seed germination and seedling growth under acid stress situations. These might be associated with increased antioxidant activity and the preserve ratio of GSH/GSSG and AsA/DHA, which is restricted by soil acidity. Moreover, Si-NMs in field regimes significantly diminished the acid-stress-induced growth inhibitions, as evidenced by increased net photosynthesis and biomass accumulations. Again, Si-NMs triggered all the critical metrics of crop productivity, including the seed oil content. Additionally, Si-NMs, upon dolomite supplementation, further triggered all the metrics of yields related to farming resilience. Therefore, the present study highlighted the crucial roles of Si-NMs in sustainable agricultural expansion and cropping intensification, especially in areas affected by soil acidity.

Cd is highly mobile, non-essential trace element, that has become serious environmental issue due to its elevated concentration in soil. The present study was taken up to work out salutary effect of melatonin (Mlt) and PGPR ((Pseudomonas putida (Pp), Pseudomonas fluorescens (Pf) in 10 days old Cd stressed (0.3 mM) Brassica juncea L. seedlings. The present work investigated growth characteristics, photosynthetic pigments, secondary metabolites in melatonin-PGPR inoculated B. juncea seedlings. It was backed by molecular studies entailing RT-PCR and transcriptomic analyses. Our results revealed, substantial increase in photosynthetic pigments and secondary metabolites, after treatment with melatonin, P.putida, P. fluorescens in Cd stressed B. juncea seedlings, further validated with transcriptome analysis. Comparative transcriptome analyses identified 455, 5953, 3368, 2238 upregulated and 4921, 430, 137, 27 down regulated DEGs, Cn-vs-Cd, Cd-vs-Mlt, Cd-vs-Mlt-Pp-Pf, Cd-vs-Mlt-Pp-Pf-Cd comparative groups respectively. In depth exploration of genome analyses (Gene ontology, Kyoto encyclopaedia of genes), revealed that Cd modifies the expression patterns of most DEGs mainly associated to photosystem and chlorophyll synthesis. Also, gene expression studies for key photosynthetic genes (psb A, psb B, CHS, PAL, and PSY) suggested enhanced expression in melatonin-rhizobacteria treated Cd stressed B. juncea seedlings. Overall, results provide new insights into probable mechanism of Mlt-PGPR induced protection to photosynthesis in Cd stressed B. juncea plants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Trichomes are specialized structures derived from epidermal cells. Non-glandular trichomes primarily protect plants from herbivores and intense ultraviolet radiation by acting as a physical barrier. Recent research has highlighted the importance of homeodomain leucine zipper (HD-Zip) IV transcription factors (TFs) in promoting trichome development. In this study, an HD-Zip III TF called PHAVOLUTA (BjPHVa) was identified as a negative regulator of non-glandular trichome initiation in Brassica juncea. Genome editing of BjPHVa resulted in a significant increase in trichome number in B. juncea. Co-expression networks revealed a strong association between trichome development and the HD-Zip family, which was supported by transcriptomic analysis findings. An R2R3-MYB TF, BjGL1a, a key regulator of trichome development, was found to be associated with BjPHVa-regulated trichome development. Knockdown of BjGL1a expression resulted in reduced trichome number in B. juncea. BjPHVa was observed to interact directly with BjGL1a while binding to the BjGL1a promoter, resulting in the inhibition of BjGL1a transcription. These results provide new insights into the identification of regulators involved in trichome development and offer new opportunities to enhance resistance to predicted stresses through genome editing targeting PHVa within Brassicaceae., (© 2024 Scandinavian Plant Physiology Society.)

Salt stress is a prevalent environmental issue that disrupts the redox balance and metabolic processes in plants, leading to reduced crop growth and productivity. Currently, over 6.74 million hectares in India are salt-affected, and about 75% of this land lies in states that are the major cultivators of edible oilseed crops (rapeseed-mustard). Therefore, this study focused on the efficacy of glycine betaine (GB) supplementation in mitigating the detrimental effects of salt stress in Brassica juncea L. (Indian mustard) plants. Indian mustard plants were subjected to salt stress [0, 50, 100, and 150 mM sodium chloride] 20 days after sowing (DAS), while a foliar spray of 20 mM GB was applied to the foliage at 50 and 70 DAS. The data showed that salt stress substantially reduced growth, photosynthetic rate, membrane stability, and yield by significantly increasing lipid peroxidation, ion toxicity, cell death, electrolyte leakage, and reactive oxygen species accumulation that triggered oxidative stress. Supplementation with 20 mM GB provided tolerance to plants against salt-induced toxicity since it substantially increased growth, biomass, water content, nutrient uptake, and photosynthetic efficiency. Additionally, GB enhances the accumulation of osmolytes, enhances the antioxidant defence system, improves ionic balance, and enhances cell viability. Taken together, the obtained data provides deeper insights into the beneficial effect of the exogenous GB application that could have biotechnological uses to enhance crop stress tolerance in challenging environments., (© 2024 Scandinavian Plant Physiology Society.)

The present research primarily focuses on Brassica juncea's physiological and cytological responses to low and high temperature stress at 4 °C and 44 °C respectively, along with elucidating the protective role of 28-Homobrassinolide (28-homoBL). Cytological investigations performed in floral buds of Brassica juncea L. under temperature (24, 4, 44 °C) stress conditions depict the presence of some abnormalities associated with cytomixis such as chromosome stickiness or agglutination, pycnotic nature of chromatin, irregularities in spindle formation, disoriented chromatins, and non-synchronous chromatin material condensation in Brassicaceae family that subsisted at diploid level (2n = 36). Spindle abnormalities produce various size pollen grains such as sporads micronuclei at some stages of microsporogenesis, polyads, triads, dyads that irrupted the productiveness of pollen grains. Furthermore, sugars play an imperative role in protecting plants under stress besides being energy sources. Therefore, the present study revealed accumulation of total soluble sugars (TSS), with 28-homoBL treatment which pinpoints protective role of 28-homoBL under temperature stress. Sugar profiling was done by using high-performance liquid chromatography (HPLC) which helped in analyzing different sugars both quantitatively and qualitatively under 28-homoBL and temperature stress conditions. The results indicate that the 28-homoBL treatment substantially enhances plant tolerance to heat stress, as evident by higher mitotic indices, fewer chromosomal abnormalities, and significantly more sugar accumulation. The findings of the study acknowledge the potential of 28-homoBL in inducing temperature stress tolerance in B. juncea along with improving the metabolic stability thereby implying application of 28-homoBL in crop strengthening under variable temperature conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Heatwaves, expected to become more frequent, pose a significant threat to plant biomass production. This experiment was designed to estimate heatwave influence on Brassica juncea phytoremediation when superimposed on different CO 2 levels. A 7-day heatwave was generated during the species flowering stage. Heatwaves decreased all B. juncea dry weights. The lowest species dry weight was recorded when the heatwave was accompanied by 250 ppm CO 2 , in which the biomass significantly decreased by 40.0% relative to that of no heatwave under the same atmospheric CO 2 conditions. Heatwave superposition with 250 ppm CO 2 reduced the Cd content in B. juncea aerial parts by 28.1% relative to that of identical environmental conditions without heatwave, whereas the opposite result was observed under 550 ppm CO 2 conditions. The heatwave caused oxidative damage to B. juncea under all CO 2 conditions, as manifested by increased malondialdehyde levels in the plant shoots. With heatwave superposition, antioxidant enzyme activity was enhanced by exposure to 400 and 550 ppm CO 2 . Considering biomass yield generation and Cd uptake capacity, heatwave superposition decreased the B. juncea phytoremediation effects, and high atmospheric CO 2 conditions could alleviate detrimental effects to a certain extent. This study uniquely examines the combined effects of heatwaves and varying CO 2 levels on phytoremediation, providing microscopic insights into oxidative damage and enzyme activity, highlighting the potential for CO 2 enrichment to mitigate heatwave impacts, and offering comprehensive analysis for future agricultural practices and environmental management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Brassica juncea depends heavily on nitrogen (N) fertilizers for growth and accumulation of seed protein. However, it is an inefficient mobilizer of applied N which leads to accumulation of excess N in the soil, posing environmental risks. Hence, it is imperative to systematically examine spatial-temporal pattern of crop N to efficiently manage N application. The Kjeldahl method is commonly used to estimate N status of crops but it is a destructive method that entails the use of perilous and expensive chemicals. Near-infrared reflectance spectroscopy (NIRS) offers a safe, accurate, and non-destructive alternative for large-scale screening of seed metabolites. Currently, no NIRS model exists to quickly estimate N content in shoots and roots from large germplasm sets in any rapeseed-mustard crop. Developing such a model is essential to breed for enhanced nitrogen use efficiency (NUE). We used 738 shoot and 346 root samples from a B. juncea diversity set to construct the NIRS models. A diverse range of genetic variation in N content was recorded in the stem (0.21-6.61%) and root (0.15-3.04%) tissues of the crop raised on two different N levels (N0 and N100). Modified partial least squares (MPLS) method was employed to establish a regression equation linking reference N values with spectral changes. The developed models exhibited strong associations with reference values, with RSQ values of 0.884 for stem and 0.645 for roots. Furthermore, external validation confirms the reliability of the developed models. The developed models have strong predictive capabilities for rapid and reliable N estimation in various tissues of B. juncea plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Clubroot has become a major obstacle in rapeseed production. Breeding varieties resistant to clubroot is the most effective method for disease management. However, the clubroot-resistant germplasm of rapeseed remains limited. To tackle this challenge, we synthesized the clubroot-resistant mustard, CT19, via distant hybridization, and subsequently an F 2 segregating population was created by intercrossing CT19 with a clubroot-susceptible germplasm CS15. A major-effect clubroot resistance QTL qCRa3-1 on chromosome A03 was identified through QTL scanning. Transcriptome analyses of CT19 and CS15 revealed that the mechanisms conferring resistance to Plasmodiophora brassica likely involved the regulation of flavonoid metabolism, fatty acid metabolism, and sulfur metabolism. By combining the results from transcriptome, QTL mapping, and gene sequencing, a candidate gene BjuA03.BNT1, encoding NLR (nucleotide-binding domain leucine-rich repeat-containing receptors) protein, was obtained. Intriguingly, comparing with CT19, a base T insertion was discovered in the BjuA03.BNT1 gene's coding sequence in CS15, resulting an alteration within the LRR conserved domain. Overexpression of BjuA03.BNT1 from CT19 notably enhanced the resistance to clubroot in Arabidopsis. Our investigations revealed that BjuA03.BNT1 regulated the resistance to clubroot by modulating fatty acid synthesis and the structure of cell wall. These results are highly relevant for molecular breeding to improve clubroot resistance in rapeseed., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Indian mustard (Brassica juncea; Brassicaceae) is an edible, oilseeds-yielding crop widely consumed as a food spice owing to its richness in nutrients with several health benefits. The current study aims to dissect the B. juncea metabolome heterogeneity among its different organs including leaf, stem, flower, and seed. Moreover, assessing the metabolome differences between two different varieties RH-725 and RH-761 grown at the same conditions. Gas chromatography-mass spectrometry (GC-MS) post-silylation was used to dissect the composition of nutrient metabolites coupled to multivariate data analysis. Variation in sulphur aglycones was measured using headspace-solid phase-microextraction HS-SPME coupled to GC-MS. A total of 101 nutrient metabolites were identified with the abundance of sugars represented by monosaccharides in all organs, except for seeds which were enriched in disaccharides (sucrose). α-Linolenic acid was detected as a marker fatty acid in leaf from RH-725 at 12.5 µg/mg. Malic acid was detected as a significant variant metabolite between the two varieties as detected in the leaf from the RH-725 variety at ca. 128.2 µg/mg compared to traces in RH-761. 7 Volatile sulphur compounds were detected at comparable levels in RH-725 and RH-761, with 3-butenyl isothiocyanate was the most abundant at 0.8-2 ng/mg., (© 2024. The Author(s).)

Background: Zha cai, a pickled vegetable with unique flavors, is produced by fermenting fresh mustard tubers. In this study, the main physicochemical indices and volatile flavor compounds were determined in three fermentation periods. The bacterial and fungal communities in the three fermentation periods of zha cai were also monitored using high-throughput sequencing. Key microbial communities were identified based on significant correlations with flavor substances., Results: Firmicutes and Proteobacteria were the main bacterial phyla found within the three fermentation periods. Lactic acid bacteria, namely Lactobacillus, was the predominant bacteria found at the genus level. Ascomycetes and Stenotrophomonas were the major fungal phyla found in the three fermentation periods. Yeast, namely Debaryomyces, was the predominant fungus found at the genus level. A total of 42 bacterial genera were negatively correlated with volatile flavor substances of zha cai, and 37 bacterial genera were positively correlated. Meanwhile, a total of 47 genera of fungi were negatively correlated with the volatile flavor substances of zha cai, while 50 genera were positively correlated. Several microbial genera were significantly correlated with volatile flavor compounds, including Lactobacillus, Halomonas, Rhodococcus, and Debaryomyces., Conclusion: This study identified the microbial classes that positively regulate the flavor of zha cai which could provide valuable help for flavor modulation in zha cai production. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The purple leaves of Brassica napus are abundant in anthocyanins, which are renowned for their role in conferring distinct colors, stress tolerance, and health benefits, however the genetic basis of this trait in B. napus remains largely unelucidated. Herein, the purple leaf B. napus (PL) exhibited purple pigments in the upper epidermis and a substantial increase in anthocyanin accumulation, particularly of cyanidin, compared to green leaf B. napus (GL). The genetic control of the purple leaf trait was attributed to a semi-dominant gene, pl, which was mapped to the end of chromosome A03. However, sequencing of the fragments amplified by the markers linked to pl indicated that they were all mapped to chromosome B05 from B. juncea. Within this B05 chromosomal segment, the BjMYB113 gene-specific marker showed perfect co-segregation with the purple leaf trait in the F 2 population, suggesting that the BjMYB113 introgression from B. juncea was the candidate gene for the purple leaf trait in B. napus. To further verify the function of candidate gene, CRISPR/Cas9 was performed to knock out the BjMYB113 gene in PL. The three myb113 mutants exhibited evident green leaf phenotype, absence of purple pigments in the adaxial epidermis, and a significantly reduced accumulation of anthocyanin compared to PL. Additionally, the genes involved in positive regulatory (TT8), late anthocyanin biosynthesis (DFR, ANS, UFGT), as well as transport genes (TT19) were significantly suppressed in the myb113 mutants, further confirming that BjMYB113 was response for the anthocyanin accumulation in purple leaf B. napus. This study contributes to an advanced understanding of the regulation mechanism of anthocyanin accumulation in B. napus., (© 2024. The Author(s).)

Understanding the heavy metals (HMs) tolerance mechanism is crucial for improving plant growth in metal-contaminated soil. In order to evaluate the lead (Pb) tolerance mechanism in Brassica species, a comparative proteomic study was used. Thirteen-day-old seedlings of B. juncea and B. napus were treated with different Pb(NO 3 ) 2 concentrations at 0, 3, 30, and 300 mg/L. Under 300 mg/L Pb(NO 3 ) 2 concentration, B. napus growth was significantly decreased, while B. juncea maintained normal growth similar to the control. The Pb accumulation was also higher in B. napus root and shoot compared to B. juncea. Gel-free proteomic analysis of roots revealed a total of 68 and 37 differentially abundant proteins (DAPs) in B. juncea and B. napus-specifically, after 300 mg/L Pb exposure. The majority of these proteins are associated with protein degradation, cellular respiration, and enzyme classification. The upregulated RPT2 and tetrapyrrole biosynthesis pathway-associated proteins maintain the cellular homeostasis and photosynthetic rate in B. juncea. Among the 55 common DAPs, S-adenosyl methionine and TCA cycle proteins were upregulated in B. juncea and down-regulated in B. napus after Pb exposure. Furthermore, higher oxidative stress also reduced the antioxidant enzyme activity in B. napus. The current finding suggests that B. juncea is more Pb tolerant than B. napus, possibly due to the upregulation of proteins involved in protein recycling, degradation, and tetrapyrrole biosynthesis pathway., Competing Interests: Declaration of competing interest All the authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Leaf mustard (Brassica juncea L.) is explored for its biofumigant properties, derived from its secondary metabolites, particularly allyl isothiocyanate (AITC), produced during the enzymatic breakdown of glucosinolates like sinigrin. The research examines eight leaf mustard cultivars developed in Yeosu city, South Korea, focusing on their genetic characteristics, AITC concentration and nitriles formation rates from glucosinolates. Results indicate that the allelopathic effects, largely dependent on AITC concentration and enzymatic activity, vary across cultivar. Sinigrin and AITC constitute 79% and 36%, respectively, of glucosinolate and its hydrolysis products. The cultivar 'Nuttongii' demonstrates significant potential for inhibiting weeds, exhibiting the highest AITC concentration at 27.47 ± 6.46 µmole g -1 These outcomes highlight the importance of selecting mustard cultivars for biofumigation based on their glucosinolate profiles and hydrolysis product yields. The study also identifies a significant genetic influence on AITC and nitrile formation, suggesting that epithiospecifier protein modulation could enhance both allelopathic and other beneficial effects. Collectively, the research underscores the promise of mustard as a sustainable, environmentally friendly alternative to traditional herbicides., (© 2024. The Author(s).)

Background: An attempt has been made to explore the nutritional profile of pink oyster mushrooms that have been grown in various agricultural residues, including sugarcane bagasse, rice straw, coconut coir and sawdust, along with other nutrient supplements such as defatted mustard and chickpea powder, for appropriate growth and fruiting body formation in a short span of time. The spawn production was experimented with five different grain varieties. The study became interesting when the observations differed slightly from the traditional practices, with the addition of defatted mustard supplements resulting in a positive correlation with respect to reducing the fruiting time, as well as improving yield and the nutritional profile of Pleurotus djamor., Results: An elevated yield of 651.93 g kg -1 was recorded in the medium where the RS and DM were used in the ratio of 1:0.01 (rice straw +1% w/w defatted mustard) bag, whereas, in terms of protein content, a maximum yield of 32.57 ± 0.79 mg g -1 was observed when SB:DM was in the same ratio (sugarcane bagasse +1% w/w defatted mustard) bag., Conclusion: To confer the best outcomes from the screened substrates, a series of experiments were performed by varying the concentration of RS and SB, with 1% w/w DM. It is worth noting that the highest protein content of 32.76 ± 0.38 mg g -1 was obtained along with the total yield of 702.56 ± 2.9 g kg -1 of mushroom when the ratio of RS:SB was 0.7:0.3. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Using natural clinoptilolite (NCP) as a carrier and alginate (Alg)-calcium as an active species, the porous silicon calcium alginate nanocomposite (Alg-Ca-NCP) was successfully fabricated via adsorption-covalence-hydrogen bond. Its structural features and physicochemical properties were detailed investigated by various characterizations. The results indicated that Alg-Ca-NCP presented the disordered lamellar structures with approximately uniform particles in size of 300-500 nm. Specially, their surface fractal evolutions between the irregular roughness and dense structures were demonstrated via the SAXS patterns. The results elucidated that the abundant micropores of NCP were beneficial for unrestricted diffusing of Alg-Ca, which was conducive to facilitate a higher loading and sustainable releasing. The Ca content of leaf mustard treated with Alg-Ca-NCP-0.5 was 484.5 mg/100g on the 21st day, higher than that by water (CK) and CaCl 2 solution treatments, respectively. Meanwhile, the prepared Alg-Ca-NCPs presented the obvious anti-aging effects on peroxidase drought stress of mustard leaves. These demonstrations provided a simple and effective method to synthesize Alg-Ca-NCPs as delivery nanocomposites, which is useful to improve the weak absorption and low utilization of calcium alginate by plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Heavy metals are one of the most damaging environmental toxins that hamper growth of plants. These noxious chemicals include lead (Pb), arsenic (As), nickel (Ni), cadmium (Cd) and chromium (Cr). Chromium is one of the toxic metal which induces various oxidative processes in plants. The emerging role of nanoparticles as pesticides, fertilizers and growth regulators have attracted the attention of various scientists. Current study was conducted to explore the potential of zinc oxide nanoparticles (ZnONPs) alone and in combination with plant growth promoting rhizobacteria (PGPR) Klebsiella sp. SBP-8 in Cr stress alleviation in Brassica juncea (L.). Chromium stress reduced shoot fresh weight (40%), root fresh weight (28%), shoot dry weight (28%) and root dry weight (34%) in B. juncea seedlings. Chromium stressed B. juncea plants showed enhanced levels of malondialdehyde (MDA), electrolyte leakage (EL), hydrogen peroxide (H 2 O 2 ) and superoxide ion (O 2 • - ). However, co-supplementation of ZnONPs and Klebsiella sp. SBP-8 escalated the activity of antioxidant enzymes i.e., superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) in B. juncea grown in normal and Cr-toxic soil. It is further proposed that combined treatment of ZnONPs and Klebsiella sp. SBP-8 may be useful for alleviation of other abiotic stresses in plants., Competing Interests: Declaration of competing interest All the authors declare no conflict of interest., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Soil monitoring in abandoned mine areas is important from the perspective of ecological and human health risk. Arsenic (As) is a predominant metalloid contaminant in abandoned mine area and its behavior has been influenced by various soil characteristics. Bioindicator can be a useful tool in terms of testing the extent to which they are uptaken by plants bioavailability. Eighteen soils near the mine tailings dam were collected to investigate the effect of As contamination on As absorption by Brassica juncea. The pH range of the experimental soils was between 4.90 and 8.55, and the total As concentrations were between 34 mg kg -1 and 3017 mg kg -1 . The bioavailability of As was evaluated by Olsen method, and B. juncea was cultivated in eighteen soils for 3 weeks. Principal component analysis, correlation, and multiple regression analysis were performed to estimate a significant factor affecting As uptake by B. juncea. All statistical results indicated that As bioavailability in soil is the main factor affecting As uptake in root and shoot of B. juncea. Although translocation process, the amount of As in shoot was exponentially explained by As bioavailability in soil. This result suggests that the contamination and bioavailability of As can be confirmed only by analyzing the shoot of B. juncea, which is be easily found in environmental ecosystem, and implies the applicability of B. juncea as a bioindicator for the monitoring of As contamination and its behavior in soil ecosystem., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Jeong-Gyu Kim reports financial support was provided by Korea University. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Brassica juncea (mustard) is a vegetable crop of Brassica , which is widely planted in China. The yield and quality of stem mustard are greatly influenced by the transition from vegetative growth to reproductive growth, i.e., flowering. The WRKY transcription factor family is ubiquitous in higher plants, and its members are involved in the regulation of many growth and development processes, including biological/abiotic stress responses and flowering regulation. WRKY71 is an important member of the WRKY family. However, its function and mechanism in mustard have not been reported. In this study, the BjuWRKY71-1 gene was cloned from B . juncea . Bioinformatics analysis and phylogenetic tree analysis showed that the protein encoded by BjuWRKY71-1 has a conserved WRKY domain, belonging to class Ⅱ WRKY protein, which is closely related to BraWRKY71-1 in Brassica rapa . The expression abundance of BjuWRKY71-1 in leaves and flowers was significantly higher than that in roots and stems, and the expression level increased gradually along with plant development. The result of subcellular localization showed that BjuWRKY71-1 protein was located in nucleus. The flowering time of overexpressing BjuWRKY71-1 Arabidopsis plants was significantly earlier than that of the wild type. Yeast two-hybrid assay and dual-luciferase reporter assay showed that BjuWRKY71-1 interacted with the promoter of the flowering integrator BjuSOC1 and promoted the expression of its downstream genes. In conclusion, BjuWRKY71-1 protein can directly target BjuSOC1 to promote plant flowering. This discovery may facilitate further clarifying the molecular mechanism of BjuWRKY71-1 in flowering time control, and creating new germplasm with bolting and flowering tolerance in mustard.

Obesity is a global health concern. Recent research has suggested that the development of anti-obesity ingredients and functional foods should focus on natural products without side effects. We examined the effectiveness and underlying mechanisms of Brassica juncea extract (BJE) in combating obesity via experiments conducted in both in vitro and in vivo obesity models. In in vitro experiments conducted in a controlled environment, the application of BJE demonstrated the ability to suppress the accumulation of lipids induced by MDI in 3T3-L1 adipocytes. Additionally, it downregulated adipogenic-related proteins peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer-binding protein-α (C/EBP-α), adipocyte protein 2 (aP2), and lipid synthesis-related protein acetyl-CoA carboxylase (ACC). It also upregulated the heat generation protein peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) and fatty acid oxidation protein carnitine palmitoyltransferase-1 (CPT-1). The oral administration of BJE decreased body weight, alleviated liver damage, and inhibited the accumulation of lipids in mice with diet-induced obesity resulting from a high-fat diet. The inhibition of lipid accumulation by BJE in vivo was associated with a decreased expression of adipogenic and lipid synthesis proteins and an increased expression of heat generation and fatty acid oxidation proteins. BJE administration improved obesity by decreasing adipogenesis and activating heat generation and fatty acid oxidation in 3T3-L1 cells and in HFD-induced obese C57BL/6J mice. These results suggest that BJE shows potential as a natural method for preventing metabolic diseases associated with obesity.

S-nitrosoglutathione reductase (GSNOR). a master regulator of NO homeostasis, is a single-copy gene in most plants. In Lotus japonicus, two GSNOR isoforms were identified exhibiting similar kinetic properties but differential tissue-specific expressions. Previously, a genome-wide identification in Brassica juncea revealed four copies of GSNOR, each encoding proteins that vary in subunit molecular weights and pI. Here, we report multiple forms of GSNOR using 2D immunoblot which showed 4 immunopositive spots of 41.5 kDa (pl 5.79 and 6.78) and 43 kDa (pl 6.16 and 6.23). To confirm, purification of GSNOR using anion-exchange chromatography yielded 2 distinct pools (GSNOR-A & GSNOR-B) with GSNOR activities. Subsequently, affinity-based purification resulted in 1 polypeptide from GSNOR-A and 2 polypeptides from GSNOR-B. Size exclusion-HPLC confirmed 3 GSNORs with molecular weight of 87.48 ± 2.74 KDa (GSNOR-A); 87.36 ± 3.25 and 82.74 ± 2.75 kDa (GSNOR-B). Kinetic analysis showed K m of 118 ± 11 μM and V max of 287 ± 22 nkat/mg for GSNOR-A, whereas K m of 96.4 ± 8 μM and V max of 349 ± 15 nkat/mg for GSNOR-B. S-nitrosylation and inhibition by NO showed redox regulation of all BjGSNORs. Both purified GSNORs exhibited variable denitrosylation efficiency as depicted by Biotin Switch assay. To the best of our knowledge, this is the first report confirming multiple isoforms of GSNOR in B. juncea., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Myrosinases constitute an important component of the glucosinolate-myrosinase system responsible for interaction of plants with microorganisms, insects, pest, and herbivores. It is a distinctive feature of Brassicales. Multiple isozymes of myrosinases are present in the vacuoles. Active myrosinases are also present in the apoplast and the nucleus however, the similarity or difference in the biochemical properties with the vacuolar myrosinases are not known. Here, we have attempted to isolate, characterize, and identify myrosinases from seeds, seedlings, apoplast, and nucleus to understand these forms. 2D-CN/SDS-PAGE coupled with western blotting and MS have shown low abundant myrosinases (65/70/72/75 kDa) in seeds and seedlings and apoplast & nucleus of seedlings to exist as dimers, oligomers, and as protein complex. Nuclear membrane associated form of myrosinase was also identified. The present study for the first time has shown enzymatically active myrosinase-alpha-mannosidase complex in seedlings. Both 65 and 70 kDa myrosinase in seedlings were S-nitrosated. Nitric oxide donor treatment (GSNO) led to 25% reduction in myrosinase activity which was reversed by DTT suggesting redox regulation of myrosinase. These S-nitrosated myrosinases might be a component of NO signalling in B. juncea., Competing Interests: Declaration of Competing Interest The authors have no relevant financial or non-financial interests to disclose., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Both microplastics (MPs) and cadmium (Cd) are common contaminants in farmland systems, is crucial for assessing their risks for human health and environment, and little research has focused on stress responses mechanisms of crops exposed to the combined pollution. The present study investigated the impact of polyethylene (PE) and polypropylene (PP) microplastics (MPs), in combination with Cd, on the physiological and metabolomic changes as well as rhizosphere soil of potherb mustard. Elevated levels of PEMPs and PPMPs were found to impede nutrient uptake in plants while promoting premature flowering, and the concomitant effect is lower crop yields. The substantial improvement in Cd bioavailability facilitated by MPs in rhizosphere soil, especially in high concentrations of MPs, then elevated bioavailability of Cd contributed to promoted Cd accumulation in plants, with distinct effects depending on the type and concentration of MPs. The presence of MPs Combined exposure to high concentrations of MPs and Cd resulted in alterations in plant physiology and metabolomics, including decreased biomass and photosynthetic parameters, elevated levels of reactive oxygen species primarily H 2 O 2 , increased antioxidant enzyme activities, and modifications in metabolite profiles. Overall, our study assessed the potential impact on food security (the availability of cadmium to plant) and crops stress responses regarding the contamination of MPs and Cd, providing new insights for future risk assessment in agriculture., Competing Interests: Declaration of competing interest All authors declare that they have no known competing financial interests or personal relationships that could appear to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Drought, a widespread abiotic stressor, exerts a profound impact on agriculture, impeding germination and plant growth, and reducing crop yields. In the present investigation, the osmotolerant rhizobacteria Bacillus casamancensis strain MKS-6 and Bacillus sp. strain MRD-17 were assessed for their effects on molecular processes involved in mustard germination under osmotic stress conditions. Enhancement in germination was evidenced by improved germination percentages, plumule and radicle lengths, and seedling vigor upon rhizobacterial inoculation under no stress and osmotic stress conditions. Under osmotic stress, rhizobacteria stimulated the production of gibberellins and reserve hydrolytic enzymes (lipases, isocitrate lyase, and malate synthase), bolstering germination. Furthermore, these rhizobacteria influenced the plant hormones such as gibberellins and abscisic acid (ABA), as well as signalling pathways, thereby promoting germination under osmotic stress. Reduced proline and glycine betaine accumulation, and down-regulation of transcription factors BjDREB1_2 and BjDREB2 (linked to ABA-independent signalling) in rhizobacteria-inoculated seedlings indicated that bacterial treatment mitigated water deficit stress during germination, independently of these pathways. Hence, the advantageous attributes exhibited by these rhizobacterial strains can be effectively harnessed to alleviate drought-induced stress in mustard crops, potentially through the development of targeted bio-formulations., (© 2024 Scandinavian Plant Physiology Society.)

In this study, the morphological and physiological responses of Brassica juncea to the stresses of Cadmium (Cd) and trichlorfon (TCF), and the phytoremediation potential of B. juncea to Cd and TCF were investigated under hydroponics. Results showed that Cd exhibited strong inhibition on biomass and root morphology of B. juncea as Cd concentration increased. The chlorophyll a fluorescence intensity and chlorophyll content of B. juncea decreased with the increased Cd concentration, whereas the malondialdehyde and soluble protein contents and superoxide dismutase activity increased. TCF with different concentrations showed no significant influence on these morphological and physiological features of B. juncea . The biomass and physiological status of B. juncea were predominantly regulated by Cd level under the co-exposure of Cd and TCF. B. juncea could accumulate Cd in different plant parts, as well as showed efficient TCF degradation performance. A mutual inhibitory removal of Cd and TCF was observed under their co-system. The present study clearly signified the physiological responses and phytoremediation potential of B. juncea toward Cd and TCF, and these results suggest that B. juncea can be used as an effective phytoremediation agent for the Cd-TCF co-contamination in water.

Salinity stress is a well-known abiotic stress that has been shown to have a negative impact on crop growth, production, and soil richness. The current study was intended to ameliorate salt stress in Indian mustard ( Brassica juncea L.), keeping in mind the detrimental influence of salt stress. A pot experimentation was executed on B. juncea to examine the efficacy of exogenous application of triacontanol (TRIA) and hydrogen sulfide (H 2 S) (NaHS donor), either alone or in combination, on growth attributes, metabolites, and antioxidant defense system exposed to salt stress at three distinct concentrations (50, 100 and 150 mM NaCl). Increase in the concentration of oxidative markers (malondialdehyde and hydrogen peroxide) was found which results in inhibited growth of B. juncea . The growth characteristics of plant, such as root and shoot length, fresh and dry weight under salt stress, were improved by foliar application of TRIA (150 µM) and H 2 S (25 µM) alone as well as in combination. Additionally, salt stress reduced the levels of protein, metabolites (flavonoids, phenolic and anthocyanin), antioxidant enzyme activity including that of ascorbate peroxidase, catalase, polyphenol oxidase and guaiacol peroxidase as well as the level of ascorbic acid and glutathione (non-enzymatic antioxidants). However, application of TRIA and H 2 S alone or in grouping substantially raised the content of protein, metabolites and antioxidant defense system in plants of B. juncea .

Repeated exposure to pathogens leads to evolutionary selection of adaptive traits. Many species transfer immunological memory to their offspring to counteract future immune challenges. Transfer factors such as those found in the colostrum are among the many mechanisms where transfer of immunologic memory from one generation to the next can be achieved for an enhanced immune response. Here, a library of 100 plants with high protein contents was screened to find plant-based proteins that behave like a transfer factor moiety to boost human immunity. Aqueous extracts from candidate plants were tested in a human peripheral blood mononuclear cell (PBMC) cytotoxicity assay using human cancerous lymphoblast cells-with K562 cells as a target and natural killer cells as an effector. Plant extracts that caused PBMCs to exhibit enhanced killing beyond the capability of the colostrum-based transfer factor were considered hits. Primary screening yielded an 11% hit rate. The protein contents of these hits were tested via a Bradford assay and Coomassie-stained SDS-PAGE, where three extracts were confirmed to have high protein contents. Plants with high protein contents underwent C18 column fractionation using methanol gradients followed by membrane ultrafiltration to isolate protein fractions with molecular weights of <3 kDa, 3-30 kDa, and >30 kDa. It was found that the 3-30 kDa and >30 kDa fractions had high activity in the PBMC cytotoxicity assay. The 3-30 kDa ultrafiltrates from the top two hits, seeds from Raphanus sativus and Brassica juncea , were then selected for protein identification by mass spectrometry. The majority of the proteins in the fractions were found to be seed storage proteins, with a low abundance of proteins involved in plant defense and stress response. These findings suggest that Raphanus sativus or Brassica juncea extracts could be considered for further characterization and immune functional exploration with a possibility of supplemental use to bolster recipients' immune response.

Wild Brassica juncea is a troublesome weed that infests wheat fields in China. Two suspected wild B. juncea populations (19-5 and 19-6) resistant to acetolactate synthase (ALS) inhibitors were collected from wheat fields in China. To clarify their resistance profiles and resistance mechanism, the resistance levels of populations 19-5 and 19-6 to ALS-inhibiting herbicides and their underlying target-site resistance mechanism were investigated. The results showed that the 19-5 population exhibited resistance to tribenuron-methyl, pyrithiobac‑sodium and florasulam, while the 19-6 population was resistant to tribenuron-methyl, pyrithiobac‑sodium, imazethapyr and florasulam. Using the homologous cloning method, two ALS genes were identified in wild B. juncea, with one gene (ALS1) encoding 652 amino acids and the other (ALS2) encoding 655 amino acids. Pro-197-Arg mutation on ALS2 and Trp-574-Leu mutation on ALS1, together with the combination of these two mutations in a single plant, were observed in both 19-5 and 19-6 populations. ALS2 enzymes carrying the Pro-197-Arg mutation were cross-resistant to tribenuron-methyl, pyrithiobac‑sodium, imazerthapyr and florasulam, with resistance index (RI) values of 6.23, 32.81, 7.97 and 1162.50, respectively. Similarly, ALS1 enzymes with Trp-574-leu substitutions also displayed high resistance to these four herbicides (RI values ranging from 132.61 to 3375.00). In addition, the combination of Pro-197-Arg (ALS2) and Trp-574-Leu (ALS1) mutations increased the resistance level of the ALS enzyme to ALS inhibitors, with its RI values 3.83-214.19, 6.88-37.34, 1.91-31.82 and 2.03-5.90-fold higher than a single mutation for tribenuron-methyl, pyrithiobac‑sodium, imazerthapyr and florasulam, respectively. Collectively, Pro-197-Arg mutation on ALS2, Trp-574-Leu mutation on ALS1 and the combination of Pro-197-Arg (ALS2) and Trp-574-Leu (ALS1) mutations in wild B. juncea could endow broad-spectrum resistance to ALS inhibitors, which might provide guides for establishing effective strategies to prevent or delay such resistance evolution in this weed., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Cruciferous plants manufacture glucosinolates (GSLs) as special and important defense compounds against insects. However, how insect feeding induces glucosinolates in Brassica to mediate insect resistance, and how plants regulate the strength of anti-insect defense response during insect feeding, remains unclear. Here, mustard ( Brassica juncea ), a widely cultivated Brassica plant, and beet armyworm ( Spodoptera exigua ), an economically important polyphagous pest of many crops, were used to analyze the changes in GSLs and transcriptome of Brassica during insect feeding, thereby revealing the plant-insect interaction in Brassica plants. The results showed that the content of GSLs began to significantly increase after 48 h of herbivory by S. exigua , with sinigrin as the main component. Transcriptome analysis showed that a total of 8940 DEGs were identified in mustard challenged with beet armyworm larvae. The functional enrichment results revealed that the pathways related to the biosynthesis of glucosinolate and jasmonic acid were significantly enriched by upregulated DEGs, suggesting that mustard might provide a defense against herbivory by inducing JA biosynthesis and then promoting GSL accumulation. Surprisingly, genes regulating JA catabolism and inactivation were also activated, and both JA signaling repressors (JAZs and JAMs) and activators (MYCs and NACs) were upregulated during herbivory. Taken together, our results indicate that the accumulation of GSLs regulated by JA signaling, and the regulation of active and inactive JA compound conversion, as well as the activation of JA signaling repressors and activators, collectively control the anti-insect defense response and avoid over-stunted growth in mustard during insect feeding.

Salt stress is an alarming abiotic stress that reduces mustard growth and yield. To attenuate salt toxicity effects, plant growth-promoting rhizobacteria (PGPR) offers a sustainable approach. Among the various PGPR, Pseudomonas fluorescens (P. fluorescens NAIMCC-B-00340) was chosen for its salt tolerance (at 100 mM NaCl) and for exhibiting various growth-promoting activities. Notably, P. fluorescens can produce auxin, which plays a role in melatonin (MT) synthesis. Melatonin is a pleiotropic molecule that acts as an antioxidant to scavenge reactive oxygen species (ROS), resulting in stress reduction. Owing to the individual role of PGPR and MT in salt tolerance, and their casual nexus, their domino effect was investigated in Indian mustard under salt stress. The synergistic action of P. fluorescens and MT under salt stress conditions was found to enhance the activity of antioxidative enzymes and proline content as well as  promote the production of secondary metabolites. This led to reduced oxidative stress following effective ROS scavenging, maintained photosynthesis, and improved growth. In mustard plants treated with MT and P. fluorescens under salt stress, eight flavonoids showed significant increase. Kaempferol and cyanidin showed the highest concentrations and are reported to act as antioxidants with protective functions under stress. Thus, we can anticipate that strategies involved in their enhancement could provide a better adaptive solution to salt toxicity in mustard plants. In conclusion, the combination of P. fluorescens and MT affected antioxidant metabolism and flavonoid profile that could be used to mitigate salt-induced stress and bolster plant resilience., (© 2023 Scandinavian Plant Physiology Society.)

Hyperaccumulation of heavy metal in agricultural land has hampered yield of important crops globally. It has consequently deepened concerns regarding the burning issue of food security in the world. Among heavy metals, Chromium (Cr) is not needed for plant growth and found to pose detrimental effects on plants. Present study highlights the role of exogenous application of sodium nitroprusside (SNP, exogenous donor of NO) and silicon (Si) in alleviating detrimental ramification of Cr toxicity in Brassica juncea. The exposure of B. juncea to Cr (100 μM) under hydroponic system hampered the morphological parameters of plant growth like length and biomass and physiological parameters like carotenoid and chlorophyll contents. It also resulted in oxidative stress by disrupting the equilibrium between ROS production and antioxidant quenching leading to accumulation of ROS such as hydrogen peroxide (H 2 O 2 ) and superoxide (O 2 •‾ ) radicle which causes lipid peroxidation. However, application of Si and SNP both individually and in combination counteracted oxidative stress due to Cr by regulating ROS accumulation and enhancing antioxidant metabolism by upregulation of antioxidant genes of DHAR, MDHAR, APX and GR. As the alleviatory effects were more pronounced in plants treated with combined application of Si and SNP; therefore, our findings suggest that dual application of these two alleviators can be used to mitigate Cr stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Biosynthesis, stabilization, and storage of carotenoids are vital processes in plants that collectively contribute to the vibrant colors observed in flowers and fruits. Despite its importance, the carotenoid storage pathway remains poorly understood and lacks thorough characterization. We identified two homologous genes, BjA02.PC1 and BjB04.PC2, belonging to the esterase/lipase/thioesterase (ELT) family of acyltransferases. We showed that BjPCs in association with fibrillin gene BjFBN1b control the stable storage of carotenoids in yellow flowers of Brassica juncea. Through genetic, high-resolution mass spectrometry and transmission electron microscopy analyses, we demonstrated that both BjA02.PC1 and BjB04.PC2 can promote the accumulation of esterified xanthophylls, facilitating the formation of carotenoid-enriched plastoglobules (PGs) and ultimately producing yellow pigments in flowers. The elimination of BjPCs led to the redirection of metabolic flux from xanthophyll ester biosynthesis to lipid biosynthesis, resulting in white flowers for B. juncea. Moreover, we genetically verified the function of two fibrillin genes, BjA01.FBN1b and BjB05.FBN1b, in mediating PG formation and demonstrated that xanthophyll esters must be deposited in PGs for stable storage. These findings identified a previously unknown carotenoid storage pathway that is regulated by BjPCs and BjFBN1b, while offering unique opportunities for improving the stability, deposition, and bioavailability of carotenoids., (© 2023 The Authors New Phytologist © 2023 New Phytologist Foundation.)

Nano-phytoremediation is anticipated as a potential technology for the remediation of heavy metals from soil sites. This study evaluated the feasibility of using titanium dioxide nanoparticles (TiO 2 NPs) at various concentrations (0, 100, 250, 500 mg/kg) along with a hyperaccumulator, Brassica juncea L., for effective removal of Cadmium (Cd) from the soil. Plants were grown for a whole life cycle in soil containing 10 mg/kg of Cd and spiked TiO2 NPs. We analyzed the plants for Cd tolerance, phytotoxicity, Cd removal, and translocation. Brassica plants displayed high Cd tolerance with a significant increase in plant growth, biomass, and photosynthetic activity in a concentration-dependent manner. Cd removal from the soil at TiO 2 NPs concentrations of 0, 100, 250, and 500 mg/kg treatment was 32.46%, 11.62%, 17.55%, and 55.11%, respectively. The translocation factor for Cd was found to be 1.35, 0.96, 3.73, and 1.27 for 0, 100, 250, and 500 mg/kg concentrations. The results of this study indicate that TiO 2 NPs applications in the soil can minimize Cd stress in plants and lead to its efficient removal from soil. Thus, the association of nanoparticles with the phytoremediation process can lead to good application prospects for the remediation of contaminated soil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Mustard (Brassica campestris L.) is a major oilseed crop that plays a crucial role in agriculture. Nevertheless, a number of abiotic factors, drought in particular, significantly reduce its production. Phenylalanine (PA) is a significant and efficacious amino acid in alleviating the adverse impacts of abiotic stressors, such as drought. Thus, the current experiment aimed to evaluate the effects of PA application (0 and 100 mg/L) on brassica varieties i.e., Faisal (V1) and Rachna (V2) under drought stress (50% field capacity). Drought stress reduced the shoot length (18 and 17%), root length (12.1 and 12.3%), total chlorophyll contents (47 and 45%), and biological yield (21 and 26%) of both varieties (V1 and V2), respectively. Foliar application of PA helped overcome drought-induced losses and enhanced shoot length (20 and 21%), total chlorophyll contents (46 and 58%), and biological yield (19 and 22%), whereas reducing the oxidative activities of H 2 O 2 (18 and 19%), MDA concentration (21 and 24%), and electrolyte leakage (19 and 21%) in both varieties (V1 and V2). Antioxidant activities, i.e., CAT, SOD, and POD, were further enhanced under PA treatment by 25, 11, and 14% in V1 and 31, 17, and 24% in V2. Overall findings suggest that exogenous PA treatment reduced the drought-induced oxidative damage and improved the yield, and ionic contents of mustard plants grown in pots. It should be emphasized, however, that studies examining the impacts of PA on open-field-grown brassica crops are still in their early stages, thus more work is needed in this area., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

A field study was conducted to investigate the influence of MgO-NPs priming on growth and development of mustard. Priming of mustard seeds before sowing with MgO-NPs at concentration 10, 50, 100, and 150 μg/ml enhanced the vegetative parameters of plants, with considerable increase in leaf area. MgO-NPs exposure increased the photosynthetic pigment accumulation in mustard that led to increase in biomass, carbohydrate content, and the yield in terms of total grain yield. Increased chlorophyll has simultaneously increased the oxidative stress in plants, and hence stimulated their antioxidant potential. A consistent increase was observed in the content of mustard polyphenols and activity of SOD, CAT, and APX on MgO-NPs exposure. MgO-NPs induced oxidative stress further reduced the protein content and bioavailability in mustard. We further, evaluated the influence of MgO-NPs on the quality of mustard harvested seeds. The seeds harvested from nanoprimed mustard possessed increased antioxidant potential and reduced oxidative stress. The carbohydrate and protein accumulation was significantly enhanced in response to nanopriming. Reduced chlorophyll content in seeds obtained from nanoprimed mustard indicated their potential for disease resistance and stability on long term storage. Therefore, the seeds harvested from MgO-NPs primed mustard were biochemically rich and more stable. Therefore, MgO-NPs priming can be potentially used as a novel strategy for growth promotion in plants where leaves are economically important and a strategy to enhance the seed quality under long term storage conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Indian mustard (Brassica juncea L. Czern and Coss) is an important oil and vegetable crop frequently affected by seasonal drought stress during seed germination, which retards plant growth and causes yield loss considerably. However, the gene networks regulating responses to drought stress in leafy Indian mustard remain elusive. Here, we elucidated the underlying gene networks and pathways of drought response in leafy Indian mustard using next-generation transcriptomic techniques. Phenotypic analysis showed that the drought-tolerant leafy Indian mustard cv. 'WeiLiang' (WL) had a higher germination rate, antioxidant capacity, and better growth performance than the drought-sensitive cv. 'ShuiDong' (SD). Transcriptome analysis identified differentially expressed genes (DEGs) in both cultivars under drought stress during four germination time points (i.e., 0, 12, 24, and 36 h); most of which were classified as drought-responsive, seed germination, and dormancy-related genes. In the Kyoto Encyclopedia of Genes and Genome (KEGG) analyses, three main pathways (i.e., starch and sucrose metabolism, phenylpropanoid biosynthesis, and plant hormone signal transduction) were unveiled involved in response to drought stress during seed germination. Furthermore, Weighted Gene Co-expression Network Analysis (WGCNA) identified several hub genes (novel.12726, novel.1856, BjuB027900, BjuA003402, BjuA021578, BjuA005565, BjuB006596, novel.12977, and BjuA033308) associated with seed germination and drought stress in leafy Indian mustard. Taken together, these findings deepen our understanding of the gene networks for drought responses during seed germination in leafy Indian mustard and provide potential target genes for the genetic improvement of drought tolerance in this crop., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Novel pesticides broflanilide (BFI), afidopyropen (ADP), and flupyradifurone (FPO) have been widely used and become the new organic pollutants. However, uptake, translocation and residual distribution of BFI, ADP, and FPO in plants remain unclear. Therefore, residues distribution, uptake, and translocation of BFI, ADP, and FPO were investigated in mustard field trials and hydroponic experiments. The field results indicated that the residues of BFI, ADP, and FPO were 0.001-1.87 mg/kg at 0-21 d and dissipated fast in mustard (half-lives=5.2-11.3 d). More than 66.5 % of FPO residues were distributed in the cell-soluble fractions because of their high hydrophilicity, while hydrophobic BFI and ADP were primarily stored in the cell walls and organelles. The hydroponic data showed that the foliar uptake rates of BFI, ADP, and FPO were weak (bioconcentration factors<1), but the root uptake rate was strong (bioconcentration factors>1). The upward and downward translations of BFI, ADP, and FPO were limited (translation factor<1). BFI and ADP are uptake by roots via apoplast pathway, and FPO is uptake via symplastic pathway. This study contributes to the understanding of the formation of pesticide residues in plants and provides a reference for safe application and risk assessment of BFI, ADP, and FPO., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Two significant soil degradation processes that pose a hazard to our ecosystems are soil salinization and sodification. The information on potential of salicylic acid (SA) and trehalose (Tre) to induce abiotic stress signaling and triggers physio-biochemical responses in crop plants is limited. Therefore, the present study was aimed to investigate the efficacy of 5 μM SA and/or 10 mM Tre in improving the growth, photosynthesis, ion homeostasis, nutrient acquisition, antioxidant defense system and yield of mustard plants growing under sodium chloride (NaCl) stress (0, 50, 100 and 150 mM NaCl). The data showed that increasing NaCl stress concentration decreased growth, photosynthesis, membrane permeability, ion homeostasis and yield in a dose-dependent manner while increasing considerably enzymatic antioxidant enzyme activities, compatible solute accumulation, sodium ion and oxidative stress biomarkers linearly with increasing NaCl stress concentration. The spray of SA, Tre, and SA + Tre played diversified roles in enhancing NaCl stress tolerance in mustard at morpho-physiological and biochemical levels. The combined SA + Tre application proved best and completely neutralized the NaCl stress-induced suppression in growth, photosynthesis, ion homeostasis, nutrient acquisition and yield by significantly enhancing the activities of enzymatic antioxidants, compatible solutes accumulation, water status and membrane permeability, while reducing considerably osmotic stress, reactive oxygen species generation, lipid peroxidation, cell death and sodium uptake in mustard. The SA + Tre application enhanced relative water content by 23%, net photosynthetic rate by 48%, superoxide dismutase activity by 51% and seed yield per plant by 64%, while decreased superoxide anion content by 26%, sodium ion content by 36% and malondialdehyde content by 25% over 0 mM NaCl treatment. Our findings indicate that the co-application of SA + Tre can be a suitable approach to palliate the ill effect of NaCl stress in mustard plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Environmental stresses affect the yield and productivity of Brassica crops. Catalases are important antioxidant enzymes involved in reducing excess hydrogen peroxide produced by environmental stresses. In the present study, nine and seven CAT family members in two oilseed Brassica species (B. juncea and B. rapa) were identified with complete characterization based on gene and protein structure. Phylogenetic classification categorized CAT proteins into three classes and differentiated the monocot and dicot-specific CAT proteins. Further, the gene and protein characterizations revealed a high degree of conservation across the CAT family members. Differences were observed in the CAT-HEME binding affinity in CAT1, CAT2, and CAT3 isozymes, which could suggest their differential enzyme activities in different conditions. Furthermore, protein-protein interaction with other antioxidant proteins suggested their coordinated role in ROS scavenging mechanisms. Notably, the differential gene expression of BjuCATs and BraCATs and CAT enzyme activities suggested their crucial roles in major abiotic stresses faced by Brassica species. Promoter analysis in BjuCATs and BraCATs suggested the presence of abiotic-stress responsive cis-regulatory elements. Gene regulatory network analysis suggested miRNA and TF mediated stress response in BjuCATs and BraCATs. CAT family screening and characterization in Brassica sp. has established a basic ground for further functional validation in abiotic and heavy-metal stresses which can help in developing stress tolerant crops., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Due to global warming, increasing incidences of higher-than-normal temperatures have been observed, which adversely affect seed germination, crop growth, and productivity. Several reports are available on the effect of inoculation with rhizobacteria on plant growth and biochemical attributes; however, information on their influence on seed germination and plant stress levels is lacking. In the present study, under heat stress, we studied the effect of three thermotolerant rhizobacterial strains on mustard seed germination, seedling vigor, and plant growth. Effect of inoculation with the rhizobacterial strains on the plant stress levels, biochemical attributes and antioxidant activity was also determined. Under heat stress, inoculation with the rhizobacterial strains improved seed germination and seedling fresh weight and plumule length; while only Bacillus licheniformis SSA 61 inoculated plants showed better radicle length. There was a concomitant decrease in the plant ethylene levels in the inoculated treatments. Inoculated plants showed higher shoot fresh weight, however, Bacillus sp. MRD-17 inoculated plants only improved root growth. There was significant increase in most of the plant biochemical parameters and activities of antioxidant enzymes superoxide dismutase, catalase, and ascorbate peroxidase. Significant reduction in proline and total sugar content was noted in the inoculated treatments; while increase in the amino acid and phenolics content was observed. A further increase in the antioxidant enzyme activity was recorded in most of the inoculated treatments compared with no stress. Thus, our study indicated that thermotolerant rhizobacterial strains reduced plant stress levels; enhanced seed germination, seedling vigor, plant biomass, and thermotolerance of mustard., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Glucoraphanin (GRA) is a precursor of sulforaphane (SFN), which can be synthesized by the enzyme myrosinase. In this study, we developed and validated HPLC analytical methods for the determination of GRA and SFN in mustard seed powder (MSP), broccoli sprout powder (BSP), and the MSP-BSP mixture powder (MBP), and evaluated their anti-adipogenic effects in 3T3-L1 adipocytes. We found that the analysis methods were suitable for the determination of GRA and SFN in MSP, BSP, and MBP. The content of GRA in BSP was 131.11 ± 1.84 µmol/g, and the content of SFN in MBP was 162.29 ± 1.24 µmol/g. In addition, BSP and MBP effectively decreased lipid accumulation content without any cytotoxicity. Both BSP and MBP significantly inhibited the expression of adipogenic proteins and increased the expression of proteins related to lipolysis and lipid metabolism. BSP and MBP inhibited the expression of adipocyte protein 2 (aP2), CCAAT/enhancer-binding protein- α (C/EBP- α ), and peroxisome proliferator-activated receptor- γ (PPAR- γ ) in 3T3-L1 adipocytes, and inhibited the expression of fatty acid synthase (FAS) through AMP-activated protein kinase (AMPK). Meanwhile, BSP and MBP also increased the expression of the lipolysis-related proteins, uncoupling protein-1 (UCP-1) and carnitine palmitoyltransferase-1 (CPT-1). Moreover, MBP exerted anti-adipogenic to a greater extent than BSP in 3T3-L1 preadipocytes., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

Plant growth-promoting rhizobacteria (PGPR) have a positive effect on plant development and being a promising way to enhance crop productivity and as substitution of chemical fertilizers. Selenium (Se) is an important trace element and its intake is usually lower than the daily minimum amount required for humans; hence, there is a demand on the design of Se biofortification strategies. Here, the genetic traits known to be associated with Plant-Growth Promotion (PGP) and Se biotransformation of Exiguobacterium sp. S17 were evaluated through genome analysis. Its growth-promoting capacity was tested through plant-growth promotion assays in laboratory and field conditions, using Brassica juncea (indian mustard), Beta vulgaris (chard), and Lactuca sativa (lettuce). Additionally, the Se biotransformation ability of Exiguobacterium sp. S17 was evaluated and the obtained selenized bacteria were tested in mustard plants. The sequenced bacteria genome revealed the presence of multiple genes involved in important functions regarding soil and plant colonization, PGP and Se biotransformation. Moreover, it was demonstrated that Exiguobacterium sp. S17 enhanced plant growth and could be useful to produce Se accumulation and biofortification in accumulator plants such as mustard. Thereby, Exiguobacterium sp. S17 might be used for developing new, sustainable, and environmentally friendly agro-technological strategies., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Metabolic profiles in xylem sap are considered a fundamental mechanism for Cadmium (Cd) detoxification in plants. However, the metabolic mechanism of Brassica juncea xylem sap in response to Cd is still unclear. Here, we investigated the effects on the metabolomics of B. juncea xylem sap treated with Cd at different times by utilizing a nontargeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomics method for further elucidating the response mechanism of Cd exposure. The findings indicated that 48 h and 7 days Cd exposure caused significant differences in metabolic profiles of the B. juncea xylem sap. Those differential metabolites are primarily involved in amino acids, organic acids, lipids, and carbohydrates, and most of them were downregulated, which played essential roles in response to Cd stress. Furthermore, B. juncea xylem sap resisted 48-h Cd exposure via regulation of glycerophospholipid metabolism, carbon metabolism, aminoacyl-tRNA biosynthesis, glyoxylate and dicarboxylate metabolism, linoleic acid metabolism, C5-branched dibasic acid metabolism, alpha-linolenic acid metabolism, cyanoamino acid metabolism, ABC transporters, biosynthesis of amino acids, and pyrimidine metabolism; whereas alpha-linolenic acid metabolism, glycerophospholipid metabolism, photosynthesis, and oxidative phosphorylation were regulated for resisting 7-day Cd exposure., (© 2023 Scandinavian Plant Physiology Society.)

Chlorpyrifos (CP) is a commonly used organophosphorous pesticide that is frequently utilised in the agricultural industry because of its great efficiency and inexpensive cost. The focus of the present study was to assess the impact of CP toxicity on Brassica juncea L. and to unravel the ameliorative potential of phytohormone, 24-epibrassinolide (EBL) mediated plant-microbe (Pseudomonas aeruginosa (B1), Burkholderia gladioli (B2)) interaction in B. juncea L. The maximum significant increment in the total chlorophyll, carotenoids, xanthophyll, anthocyanin and flavonoid content with EBL and B2 treatment in CP stressed B. juncea seedlings on spectrophotometric analysis were observed. Autofluorescence imaging of photosynthetic pigments i.e. chlorophyll, carotenoids, and total phenols with confocal microscopy showed maximum fluorescence with EBL and B2. Furthermore, when compared to CP stressed seedlings, scanning electron microscopy (SEM) study of the abaxial surface of leaves revealed a recovery in stomatal opening. The supplementation of EBL and PGPR (plant growth promoting rhizobacteria) improved the level of psb A (D1 subunit PSII) and psb B (CP 47 subunit of PSII) genes expression. The expression analysis of chalcone synthase (CHS), Phenylalanine ammonialyase (PAL), Phyotene synthase (PSY) with RT-PCR system showed up-regulation in the expression when supplemented with EBL and PGPR. As a result, the current study suggests that EBL and PGPR together, can reduce CP-induced toxicity in B. juncea seedlings and recovering the seedling biomass., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Unrestricted disposal of tannery solid waste (TSW) into agricultural soils has resulted in the contamination of heavy metals (HMs) such as chromium (Cr) cadmium (Cd), Copper (Cu), and Zinc (Zn) along with the severe potential to degrade the environmental quality around the world. In the present study, a combined phyto- and myco-remediation strategy was evaluated to enhance the growth, ionic contents, and phytoextraction potential of Brassica juncea and Vigna radiata for HMs from TSW-contaminated soil. A pot experiment was conducted in the greenhouse using single or combined inoculation of Trichoderma pseudokoningii (Tp) and Aspergillus niger (An) in B. juncea and V. radiata under TSW-contaminated soil at different doses (0, 50, and 100%). The results showed that the growth parameters of both B. juncea and V. radiata were severely affected under 50 and 100% TSW treatment. The combined inoculation of both the fungal species ameliorated the positive impacts of 50 and 100% TSW application on growth and ionic contents accumulation in B. juncea and V. radiata . The combined application of An + Tp at 100% TSW enhanced the shoot length (87.8, 157.2%), root length (123.9, 120.6%), number of leaves (184.2, 175.0%), number of roots (104.7, 438.9%), and dry weight (179.4, 144.8%) of B. juncea and V. radiata , respectively as compared to control with any fungal treatment at 100% TSW. A single application of An at different doses of TSW enhanced the metal concentration in B. juncea , whereas Tp increased the concentration of the metals in V. radiata . The concentration of Cr in roots (196.2, 263.8%), shoots (342.4, 182.2%), Cu in roots (187.6, 137.0%), shoots (26.6, 76.0%), Cd in roots (245.2, 184.6%), shoots (142.1, 73.4%), Zn in roots (73.4, 57.5%), shoots (62.9, 57.6%), in B. juncea were increased by the application of An at 50 and 100% treatment levels of TSW, respectively compared to control (C). Moreover, the HMs (Cr, Cu, Cd, and Zn) uptake was also improved under 50 and 100% TSW with the combined inoculation of Tp + An in both B. juncea and V. radiata . In conclusion, the combined inoculation of Tp + An was more effective in metal removal from TSW-treated soil.NOVELTY STATEMENTLimited studies have been conducted on filamentous fungi systematically under metal-contaminated sites for their diversity, metal tolerance, and their potential in enhancing the phytoremediation potential of different crop plants.In the present study, single and/or combined inoculation of fungal strains was found effective in alleviating different metals stress in tannery solid waste contaminated soil by improving defense mechanisms and plant growth due to the association between fungal strains and plants.The combined application of both fungal strains had an additive effect in enhancing the bioaccumulation capacity of B. juncea and V. radiata compared to their single inoculation.