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In Brassica plants, methionine-derived aliphatic glucosinolates are chemically diverse natural products that serve as plant defense compounds, as well as molecules with dietary healthpromoting effects. During their biosynthesis, methylthioalkylmalate synthase (MAMS) catalyzes the elongation reaction of the aliphatic chain. The MAMS-catalyzed condensation of 4-methylthio-2-oxobutanoic acid and acetyl-CoA generates a 2-malate derivative that either enters the pathway for the synthesis of C3-glucosinolates or undergoes additional extension reactions, which lead to C4- to C9-glucosinolates. Recent determination of the x-ray crystal structure of MAMS from Brassica juncea (Indian mustard) provided insight on the molecular evolution of MAMS, especially substrate specificity changes, from the leucine biosynthesis enzyme aisopropylmalate synthase but left details of the reaction mechanism unanswered. Here we use the B. juncea MAMS2A (BjMAMS2A) isoform to analyze the kinetic and catalytic mechanisms of this enzyme. Initial velocity studies indicate that MAMS follows an ordered bi bi kinetic mechanism, which based on the x-ray crystal structure, involves binding of 4- methylthio-2-oxobutanoic acid followed by acetyl-CoA. Examination of the pH-dependence of kcat and kcat/Km are consistent with acid/base catalysis. Site-directed mutagenesis of three residues originally proposed to function in the reaction mechanism--Arg89 (R89A, R89K, R89Q), Glu227 (E227A, E227D, E227Q), and His388 (H388A, H388N, H388Q, H388D, and H388E)--showed that only two mutants (E227Q and H388N) retained activity. Based on available structural and biochemical data, a revised reaction mechanism for MAMScatalyzed elongation of methionine-derived aliphatic glucosinolates is proposed, which is likely also conserved in a-isopropylmalate synthase from leucine biosynthesis in plants and microbes. [ABSTRACT FROM AUTHOR]

Here we report that non-thermal atmospheric-pressure plasma exposure can improve Brassica juncea (leaf mustard) seed germination rate from 50 % to 98 %. The commercially relevant germination rate was achieved by plasma exposure for only 10 minutes and the effect sustains at least for one month under an appropriate storage condition. Improved germination by plasma exposure was also observed for Brassica rapa subsp. pekinensis (Chinese cabbage) seeds. The plasma device used is simple. No pure gas flow system is necessary and it is easy to handle. A large number of seeds can be treated by simply scaling up the device. Plasma exposure can be a practical method for improving seed germination of crop plants important for agriculture. • Non-thermal atmospheric-pressure plasma exposure can improve Brassica seed germination rate. • The effect sustains at least for one month under an appropriate storage condition. • The plasma device used is simple and a large number of seeds can be treated. • Plasma exposure can be a practical method for improving seed germination in agriculture. [ABSTRACT FROM AUTHOR]

• Continuous effluents and wastewater discharge contaminants the soil in the vicinity of Lyari River, Karachi. • Helianthus annuus, Zea mays and Brassica juncea plants successfully grown in the heavy metals contaminated soil. • Z. mays exhibited the higher tolerance against HMCS (based on growth attributes). • H. annuus was the best hyper accumulator plant related to other tested crops. • The physiological and biochemical attributes of plant are summarized and discussed. • Perspectives are presented for the potentials of pytoremediation in sustainable agriculture. Industrial and domestic effluents are often disposed of untreated or undertreated in developing countries resulting in the deposition of heavy metals in soil and water bodies. Karachi is the largest commercial hub of the country with many industries and the densely developed urban area. Lyari is one of the two major rivers of Karachi which has become a dumping site of the city's effluent. In the present study three crops- Helianthus annuus, Zea mays and Brassica juncea were grown in the heavy metals contaminated soil (HMCS) of Lyari River to explore their tolerance and phytoremediation potential. All the tested plants exhibited improved germination and growth in HMCS. The shoot length and plant biomass of all three cultivated species were high in HMCS. However, the tolerance against HMCS (based on growth attributes) of Z. mays was higher than other crops. The photochemistry and relative chlorophyll content of all plants were unaffected with HMCS. Although, H 2 O 2 content was high (around 25 %) but no change in antioxidant enzyme was found in H. annuus and B. juncea species under HMCS. In contrast, the enhanced activity of peroxidase and catalase enzymes in the shoot of Z. mays along with no increase in H 2 O 2 content is related with better plant growth in HMCS. H. annuus and B. juncea accumulated 40 % more Cu in shoots that are associated with high BAF and TF. High Zn accumulation in root (20 % more than shoot) of three cultivated species is related with <1 TF. Higher accumulation (40 % more than control) of Fe in H. annuus and Z. mays is linked with their BAF values. Among tested heavy metals, Mn accumulation was least (< 19 mg Kg−1) in all species. This study concluded that, Zea mays could be used as a best hyper-tolerant plant while H. annuus as the best hyper accumulator. [ABSTRACT FROM AUTHOR]

• Heavy metal tolerant and PGPR rhizobacteria (Bacillus cereus, Bacillus safensis, Bacillus pumilus, Serratia marcescens, Lelliottia amnigena, Pseudomonas koreensis and Klebsiella variicola) isolated from Musa rhizosphere growing in coal mining area. • Inoculation of the PGPR consortia in the Brassica sp. and Helianthus annuus growing in Cd/Cu contaminated soil has improved its tolerance to these heavy metals over the control. • PGPR can be used for the bioremediation of the contaminated soil. In soils polluted with heavy metals, plant growth promoting rhizobacteria (PGPR) with heavy metal tolerance can aid plants to adapt to heavy metal contaminated conditions. Heavy metal tolerant rhizobacteria were isolated from Musa rhizosphere growing the Tsurang Riverbank, Nagaland, India, a coal mining area. A total of 26 bacterial isolates/strains were tested for heavy metals and salt tolerance and some growth-promoting traits like IAA, siderophore production and phosphate solubilisation. Of the 26 strains, 7 strains (TSU1, TSU11, TSU12, TSU3, TSU4, TSU7, and TSU8) supported better plant growth. Different isolates displayed variable heavy metal tolerance (30 to 8070 μg/ml). Strains TSU8 and TSU4 exhibited the highest salt tolerance (12 %). Based on the 16S rRNA sequencing, the isolates were identified as TSU3 (Bacillus cereus) (OQ547230), TSU4 (Bacillus safensis) (OQ547231), TSU8 (Bacillus pumilus) (OQ547232), TSU11 (Klebsiella variicola) (OQ947091), TSU12 (Lelliottia amnigena) (OQ947097), TSU7 (Pseudomonas koreensis) (OQ947088) and TSU1 (Serratia marcescens) (OQ726408). A successful consortium of bacterial isolates was formulated and inoculated on Brassica juncea and Helianthus annuus with heavy metal Cd/Cu (150 µgml−1) in the potted soil. This consortium improved growth performance significantly in both fresh and dry weight basis. In B. juncea, enhancement in root length and weight were recorded by 53 and 93 % respectively against control, while, fresh weight of shoot was improved by 72 % on soil adjunct with PGPR and 67 % with PGPR in heavy metal contaminated condition. Similar trend was also registered on dry weight basis. Whereas, PGPR consortia treated H. annuus registered increased shoot and root lengths both fresh and dry weight basis. This consortium with these seven bacterial isolates may contribute to microbe assisted phytoremediation under heavy metal polluted area. [ABSTRACT FROM AUTHOR]

Salinization has become a major concern for the growth and development of plants, resulting in significant crop losses. Beneficial microbial symbionts are utilized as promising tools in alleviation of adverse effects of salt stress. Piriformospora (Serendipita) indica is one of the cost-effective, eco-friendly, and stress relieving biostimulant that positively regulates the growth of a broad host range of plants. The present study was to investigate the impact of P. indica on Indian mustard (Brassica juncea L.) cultivars (Pusa Jaikisan and Pusa Bold) under different levels of salinity stress (0, 50, 100, 150, 200 and 300 mM NaCl) by analyzing morphological, physiological and biochemical attributes. Results revealed that the plant biomass, chlorophyll content, and relative water content were reduced and Na+ and Cl‒ ions uptake increased with increasing NaCl concentrations. The maximum reduction in the measured traits was observed at 300 mM NaCl in both cultivars [tolerant (Pusa Jaikisan) and sensitive (Pusa Bold)]. Additionally, the oxidative stress markers (TBARS, MG, H 2 O 2, and EL) accumulation were increased with increasing salt concentrations. However, P. indica colonization enhanced the plant performance by improving the biomass, chlorophyll, and water content, and reduced levels of oxidative stress markers (TBARS, MG, H 2 O 2, and EL) in both cultivars comparatively more in Pusa Jaikisan than Pusa Bold under salinity stress. Additionally, the maximum activities of methyl glyoxal detoxifying enzymes (Gly I & Gly II) along with the components of ROS scavenging machinery (SOD, APX, GPX, GR, AsA, and GSH) in the P. indica colonized plants conferred salt tolerance in comparison to non-colonized cultivars under salinity stress. The results indicate that P. indica promotes the growth and tolerance of mustard plants under salinity stress, thus providing opportunity for further assessment in terms of use in sustainable agriculture. [Display omitted] • Salinity stress can disrupt normal growth and development of plants thus pose serious threat to the global agriculture and cause significant yield loss. • Salinity induced production reactive oxygen species (ROS) and methyl glyoxal (MG) cause oxidative stress which disrupts the vital metabolic pathways. • Piriformospora (syn. Serendipita, Basidiomycota) indica colonized plants showed improved photosynthetic characteristics, physiological and morphological characteristics in mustard plants under salinity stress. • P. indica, acts as potential biostimulant thus confers salinity tolerance by regulating the ROS metabolism and MG detoxification in Brassica. [ABSTRACT FROM AUTHOR]

In the current study, we successfully synthesized the zinc oxide nanoparticles (ZnO-NPs) via an efficient, facile, and cost-effective method of the modified hydrothermal synthesis approach, and examined their effectiveness in alleviating arsenic (As) stress in Brassica juncea. The synthesized ZnO-NPs were analyzed using a variety of experimental techniques viz. XRDs, SEM micrographs, EDAX/Mapping pattern, Raman Spectroscopy Pattern, and X-ray photoemission spectroscopy analysis. All of these analyses revealed that ZnO-NPs were highly pure with no internal defects and could potentially be used in plant applications. Hence, we further determined the effect of these nanoparticles in modulation of As tolerance in B. juncea plants by examining the various growth attributes, photosynthesis parameters, antioxidant activities, and enzyme activities. Our results demonstrated that As-stress inhibited growth, photosynthesis-related parameters, reduced protein content, as well as carbonic anhydrase, nitrate reductase, and RuBisCO; however, these attributes were substantially up-regulated by the supply of ZnO-NPs to the leaves. Furthermore, As stress-induced increases in malondialdehyde and H 2 O 2 levels are partially reversed by ZnO-NP in the As-stressed plants. Overall, supplementation positively regulated As tolerance in B. juncea by altering the key enzymes involved in ROS detoxification, which in turn prevented As-induced oxidative damage to the plant tissues. Hence, our study clarified the potential involvement of the newly synthesized ZnO-NPs in alleviating As stress in plants. Furthermore, this is the first report to elucidate the effect of the ZnO-NPs synthesized by the hydrothermal reaction route on As-stress modulation in B. juncea. [Display omitted] • Arsenic (As) stress affects growth and biomass yield in mustard plants. • As toxicity decreased pigment system and alters biochemical attributes and antioxidants. • Supplementation of zinc oxide nanoparticles (ZnO-NPs) enhanced growth and protects pigments. • ZnO-NPs regulates antioxidant system and As uptake in mustard. [ABSTRACT FROM AUTHOR]

• The foliar feeding of epibrassinolide is an eco-friendly and sustainable approach to influence the productivity of vegetables crops, including mustard. • Epibrassinolide supplementation improve the biomass and enzymatic activity of crop plants. • Epibrassinolide foliage spray regulates the leaf gas exchange parameters and stomatal behavior. • Epibrassinolide application enhance the longevity of root cells, seed yield and oil content of mustard crop. Epibrassinolide (EBL) is an emerging plant growth regulator of the brassinosteroids group that has the potential to modulate various morpho-physiological processes in plants. Therefore, due to its key role in boosting plant functioning mechanisms, it is widely used in agriculture to enhance crop growth and yield. The present research work was planned to investigate the role of exogenously applied EBL on the mustard crop by assessing morphological, physiochemical, and yield features. This randomized pot experiment contains three mustard cultivars i.e., Rohini (Roh), Chutki (Chk), and Nath Sona (Nas) and various graded levels of EBL, 0 (Control), 10−8 M, 10−7 M, 10−6 M, 10−5 M, and 10−4 M. EBL was applied twice by giving first spray at 50 days after sowing (DAS) and the second at 70 DAS. Evaluation of plants was done at 90 DAS to study phenological traits, gas exchange attributes, non-enzymatic antioxidant activities, and microscopical parameters. The yield and quality features were studied at 120 DAS. The results revealed that treatment of 10−6 M EBL proved effective and increased fresh weight by 17.76 %, 19.11 %, and 19.34 %, chlorophyll content by 26.50 %, 26.59 %, and 25.86 %, net photosynthetic rate by 45.67 %, 57.88 % and 49.76 %, protein content by 66.98 %, 68.62 % and 69.47 %, proline content by 25.70 %, 24.05 % and 21.87 %, ascorbate content by 94.91 %, 89.99 % and 87.43 %, carbonic anhydrase activity by 32.43 %, 30.84 % and 29.93 %, seed yield by 44.85 %, 51.36 %, and 57 % and oil yield by 68.75 %, 72.71 % and 74.34 % in cultivars Nas, Roh, and Chk respectively over the control plants. Moreover, 10−6 M EBL supplementation significantly improved stomatal aperture width and root cell longevity. Cultivar Nas, followed by cultivars Roh and Chk, showed better responses on nearly all characters studied. The effect of EBL treatment in terms of enhancing the performance of the crop being in the order of 10−6 M > 10−7 M > 10−8 M > 10−5 M > 10−4 M. It may be concluded that among sprayed EBL levels, two sprays of 10−6 M EBL proved efficacious in improving the phenotypic, physiochemical, and yield traits of mustard cultivars. [ABSTRACT FROM AUTHOR]

Salinity is a crucial problem affecting the productivity of crops across the globe and poses a serious threat to agricultural food production and food security. Salinity stress adversely affects the growth and development of plants by reducing growth attributes, chlorophyll content, and relative water content and further, led to nutritional toxicities and high osmotic stress. The present investigation was employed to assess the response of ten mustard [ Brassica juncea (L.) Czern. & Coss.] genotypes under different regimes of salt concentrations (0, 50, 100, 150, 200, 300 mM NaCl) for tolerance potential using principal component analysis (PCA). The PCA analysis of studied attributes found that Pusa Jai Kisan was salt-tolerant and Pusa Bold was susceptible under different salt conditions compared to the rest of the mustard genotypes. The reduction in morphological attributes, the maximum quantum yield of PSII (Fv/Fm), total chlorophyll, and relative water content was lower in Pusa Jai Kisan and higher in Pusa Bold with increasing concentrations of salinity. In addition, the accumulation of Na+ and Cl‒ ions was increased with increasing salinity stress. Based on growth performance, ten genotypes of mustard were ranked depending on their tolerance to salinity stress: Pusa Jai Kisan > Pusa Mustard 30 > Pusa Jagannath > Pusa Mustard 28 > Pusa Vijay > Pusa Tarak > Pusa Mustard 25 > Pusa Mustard 27 > Pusa Mahak > Pusa Bold, respectively. Thus, PCA analysis can be used as a powerful tool for the identification of tolerant and susceptible genotypes under salinity stress. The understanding of mustard responses towards salinity stress could help in the selection of tolerant genotypes and this selection may play a crucial role in further improving the effectiveness of breeding. [Display omitted] • Soil salinity impacts the global agriculture negatively and limits the crop production globally. • Differential response of ten Indian mustard genotypes under different regimes of salinity (0, 50, 100, 200, 300 mM NaCl) was noted. • Increasing concentration of salinity reduced the morphological traits, total chlorophyll and relative water content in all the ten genotypes studied. • Principal component analysis revealed Pusa Jai Kisan as promising candidate for developing salinity tolerant variety. [ABSTRACT FROM AUTHOR]

Salt stress is one of the common environmental threats to crop growth, development, and productivity. Plant growth regulators (PGRs) are natural messengers and are known to play pivotal roles at different stages of the growth and development of plants under various environmental conditions. Keeping in mind the importance of PGRs in stress management, a factorial randomized pot experiment was conducted to evaluate the efficiency of three selected PGRs, namely gibberellic acid (GA 3), salicylic acid (SA) and triacontanol (Tria) for the amelioration of NaCl stress in mustard. Plants were subjected to four concentrations of NaCl (0, 50, 100 and 150 mM). Two foliar sprays of PGRs (GA 3 , SA and Tria), each at 5 μM were applied to the foliage of plants using a hand sprayer. The increasing levels of NaCl decreased growth, physio-biochemical, histochemical and yield parameters in a dose-dependent manner while increasing activities of antioxidant enzymes, contents of osmolytes and oxidative stress biomarkers linearly with increasing levels of NaCl. The spray of GA 3 , SA and Tria under stressed-free and stressed conditions improved the aforesaid attributes while decreasing the generation of stress biomarkers. Of sprayed PGRs, SA proved to be the best for alleviating the adverse effect of NaCl stress. Furthermore, it provides experimental data for its possible biotechnological applications in mustard crops exposed to high concentrations of salinity and possibly to other environmental stresses which have associated oxidative stress. • NaCl stress adversely affects the growth and productivity of mustard. • PGRs regulate diverse physio-biochemical processes in the oilseed crop. • Exogenous GA 3 , SA, and Tria mitigate NaCl-induced osmotic and oxidative damage in mustard. • Exogenous SA, GA 3 , and Tria mitigated NaCl-induced toxicities by improving the inherent defense system. • The PGR and SA proved to be the most effective to increase NaCl stress tolerance in mustard. [ABSTRACT FROM AUTHOR]

[Display omitted] • N quantification by NIRS in stem and root of B. juncea is achieved for first time. • Diverse germplasm collections allowed rapid and reliable NIRS calibration. • The developed NIRS equations displayed the best prediction performance. 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. [ABSTRACT FROM AUTHOR]

• Heat map and PCA biplot identifies two ideotypes with regard to N allocation. • Higher root biomass and volume helps to absorb nutrients more efficiently. • The activity of NR from pre-anthesis and post-anthesis is correlated with the final seed products, such as oil content. • Rise in NiR activity during post-anthesis correlated with N assimilation for seed component storage, particularly proteins, which aligns with the rising seed N content at this stage. • N -allocation follows the pattern: seed > siliquae husk > stover > root. The study explores the effect of varying levels of nitrogen (N) application on N allocation in Indian mustard (Brassica juncea) genotypes from vegetative to the generative phases. The use of a heat map and PCA biplot analysis identifies two distinct sets of ideotypes. The first set is represented by genotype IC212031 having "Stay-green" phenotype with effective N regulating enzymes, high N uptake (NUpt) and better root development. The second set of ideotype is represented by HB9902 having delayed flowering phenology phases. The study highlights the relevance of Nitrate reductase (NR) in the final oil buildup in seeds (r = 0.5, p = 0.09). The considerable rise in NR activity from pre-anthesis to post-anthesis stages translates to higher seed storage compounds such as oil (r =−0.65, p = 0.03). And the developed prediction models confirmed the significance of these stages (pre-anthesis and post-anthesis) in the accumulation of seed storage components with emphasis on oil content. Similarly, this prediction model helps us to study the cause and effect relationship between various factors and oil content. The notable rise in Nitrite reductase (NiR) activity during the post-anthesis phase suggests its role in assimilating N for the storage of seed components, most likely proteins. Further, the inverse relationship between seed N (−0.53, p = 0.09) and oil content (−0.65, p = 0.03) confirmed that at the post anthesis stage the allocation of N towards precursor carbon compounds happens, which in turn forms either proteins or lipids. This becomes a critical factor in determining the final composition of the seed. Irrespective of N treatment and genotype, the N allocation follows the pattern: seed > siliquae husk > stover > root. However, higher N application rates primarily result in increased NUpt, but not necessarily improve utilisation. Overall, the study highlights the roles of root characteristics, N metabolising enzymes, and flowering phenology in enhancing N allocation and seed production in Indian mustard offering valuable insights for breeding strategies to enhance seed yield, oil content and nutrient efficiency in oilseed crops. [Display omitted] [ABSTRACT FROM AUTHOR]

• Brassica juncea cultivated globally as oilseed used as spice and greens and have nutritional, medicinal, and economic benefits. • Brassica traits improve with the application of transformation, nanotechnology, and in vitro techniques. • Molecular markers measure and illustrate the variation in genes among different varieties and gene pools of Indian mustard. • This review focuses on advancing crop development for enhanced production to overcome global demand. Brassica juncea (L.) Czern & Coss. commonly known as Indian mustard is a versatile oilseed crop which is widely cultivated in India as well as in some parts of Canada, Russia, China, and Australia. While the plant's seeds serve as a spice, its vegetative parts are edible and used as green vegetable. Apart from its usage as vegetable oil, B. juncea has various other applications in biofertilizer, chemical and paint industries. The plant also shows medicinal properties due to its anti-cancer, antioxidant, and anti-inflammatory activities. Furthermore, mustard oil is a valuable reservoir of unsaturated fatty acids, resulting in a significant surge in demand relative to supply during the past decade. Also, the production of this crop is severely hindered by a number of biotic and abiotic stresses which cause significant yield losses. Therefore, it is important to create mustard varieties that can withstand a wide range of agroclimatic conditions and provide high yields while being resistant to biotic and abiotic challenges. Plant tissue culture, ovary culture, and Agrobacterium -mediated genetic transformation are the important methods of propagation and regeneration in Brassica species. In addition, molecular marker studies have shown new directions for crop improvement, with different combinations of promoters and markers influencing traits such as seed germination, enhanced oil content, disease resistance, and abiotic stress handling. Marker-assisted selection (MAS) and SSR markers are crucial in molecular investigations due to their ability to reveal genetic diversity and resistance features. In recent years nanotechnological interventions have also provided novel avenues for crop improvement. Various metal and metal oxide nanoparticles have been used as nanofertilizers for crop improvement. Future research should focus on developing stress-resistant varieties using advanced breeding techniques, genetic engineering and nanotechnological interventions. [Display omitted] [ABSTRACT FROM AUTHOR]

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. • Identified a major clubroot resistance QTL qCRa3–1 in B. juncea. • Candidate clubroot-risistance gene BjuA03.BNT1 encodes NLR protein. • Overexpression of BjuA03.BNT1 enhances clubroot resistance. • T insertion in BjuA03.BNT1 CDS crucial for clubroot resistance. [ABSTRACT FROM AUTHOR]

● With the rapid development of new breeding techniques, the ecological impacts of transgenic plants receive wide concern again, particularly for potential gene flow from transgenic crops to their relatives. The transgene insertion position, number of gene copies, and flanking sequence of exogenous genes integrated into the recipient genome affect the genetic stability and fitness of offspring. ● We employed hybrids F 1 and F 2 , six backcross generations BC 1 -BC 6 and BC 1 F 1 from transgenic Brassica napus with Bacillus thuringiensis (Bt) cry1Ac gene and its wild relative B. juncea through hand pollination. We detected exogenous gene copies, mRNA transcription, and protein expression by ddPCR, qRT-PCR and ELISA, and the fitness of hybrid and backcross generations. ● Exogenous genes followed Mendelian segregation expectations in hybrid and backcrossed generations, with stable gene copies, mRNA transcription and protein concentration of exogenous genes in offspring. Exogenous gene copies had no significant effects on plant fitness, but had positive effects on mRNA transcription and protein concentration that varied with growth stages in hybrid and backcross generations. ● Our study demonstrated inheritance and ecological effects of exogenous genes from transgenic plants to wild relatives, which will help ecological risk management of biotechnological plants released in the nature. • Performance of cry1Ac gene in offspring of transgenic Brassica napus and its wild relative Brassica juncea is studied. • Exogenous genes followed Mendelian segregation expectations in hybrid and backcrossed generations. • Exogenous gene copies had effects on mRNA transcription and protein concentration in hybrid and backcross generations. [ABSTRACT FROM AUTHOR]

Microplastic (MP) pollution is an emerging concern for soil health and crop productivity, yet its impacts remain poorly understood. This study addresses the critical issue by investigating the effects of MP contamination in agricultural soils through both field and pot experiments. Field analysis identified polyethylene terephthalate (PET) and high-density polyethylene in soil and plant roots, with MPs primarily concentrated in the upper soil layers. In controlled pot experiments, Brassica juncea (mustard) and Lycopersicum solanaceae (tomato) were exposed to varying concentrations of PET, polystyrene (PS) and nylon (NL). The results demonstrated dose-dependent effects on soil properties, including significant reductions in pH and available nitrogen levels at 5 % and 10 % MP concentrations (p < 0.05). PS at 5 % notably suppressed the leaf area index, while 10 % NL reduced root length and chlorophyll content. PET was particularly detrimental to root growth in both species. Among the polymers, NL (10 %) emerged as the most hazardous, with a high Potential Hazard Index score and a hazard category IV. The findings underscore the potential risks of MP pollution in the terrestrial environment, highlighting its adverse effects on soil health and plant growth. Future research should focus on enzyme activity and biochemical responses to provide deeper insights. Additionally, exploring phytoremediation techniques could offer innovative solutions to mitigate MP contamination in soils. [Display omitted] • Field study revealed polyethylene terephthalate and high-density polyethylene pollution in upper soil layers and roots. • Pot experiments showed that 5% and 10% microplastic concentrations significantly reduced soil pH and available nitrogen. • 5% polystyrene and 10% nylon suppressed leaf area index and root length, chlorophyll content in mustard and tomato plants. • Nylon at 10 % concentration posed the highest risk, with a high hazard category rating of IV. [ABSTRACT FROM AUTHOR]

Several MICRORNA genes belonging to same family or different families are often found in homologous or non-homologous clusters. Among the various classes, head-to-head arranged genes form one of the largest categories of non-canonically organized genes. Such head-to-head arranged, non-canonically organized genes possibly share cis-regulatory region with the intergenic sequence having the potential to function as bi-directional promoter (BDP). The transcriptional regulation of head-to-head arranged genes, especially with bidirectional promoters, remains an enigma. In the past, bidirectional promoters have been characterized for a small set of protein-coding gene pairs in plants; however, to the best of our knowledge, no such study has been carried so far for MICRORNA genes. The present study thus functionally characterizes bidirectional promoters associated with members of MIR395 family, which is evolutionary conserved and is most frequently occurring cluster across plant kingdom. In Arabidopsis thaliana , the MIR395 gene family contains six members with two head-to-head arranged gene pairs- MIR395A-B and MIR395E-F. This organization was found to be conserved at seven loci for MIR395A-B , and eleven loci for MIR395E-F in five Brassica sps. Sequence analysis of the putative bidirectional promoters revealed variation in length, GC content and distribution of strict TATA-box. Comparatively higher level of conservation at both the ends of the bidirectional promoters, corresponding to ca. 250 bp upstream of 5'end of the respective MIRNA precursor, was observed. These conserved regions harbour several abiotic stress (nutrient, salt, drought) and hormone (ABA, ethylene) responsive cis-motifs. Functional characterization of putative bidirectional promoters associated with MIR395A-B and MIR395E-F from Arabidopsis and their respective orthologs from Brassica juncea (Bj_A08 MIR395A-B, Bj_B03 MIR395A-B, Bj_A07.1 MIR395E-F and Bj_A07.2 MIR395E-F) was carried out using a dual-reporter vector with β-glucuronidase (GUS) and Green Fluorescent Protein (GFP). Analysis of transcriptional regulation of the two reporter genes - GUS and GFP during developmental stages confirmed their bidirectional nature. Orientation-dependent differential reporter activity indicated asymmetric nature of the promoters. Comparison of the reporter activity amongst orthologs, paralogs and homeologs revealed regulatory diversification, an outcome expected in polyploid genomes. Interestingly, reporter gene activities driven by selected bidirectional promoters were also observed in anther and siliques apart vegetative tissues indicating role of miR395 in anther and fruit development. Finally, we evaluated the activity of reporter genes driven under transcriptional regulation of bidirectional promoters under normal and sulfate-deprived conditions which revealed asymmetric inducibility under sulfate-starvation, in agreement with the known role of miR395 in sulfate homeostasis. [Display omitted] • Head-to-head arranged genes form one of the largest category of non-canonically organized gene. • Head-to-head arranged genes share cis-element and the intergenic sequence has the potential to act as bi-directional promoterr. • MIR395 associated bidirectional promoters exhibit orientation-dependent differential transcriptional activity. • Reporter activity analysis of orthologs, paralogs, homeologs from Arabidopsis, Brassica reveal regulatory diversification. [ABSTRACT FROM AUTHOR]

• Purple leaf mustard (ZC-Red) showed abundant anthocyanidins. • Green leaf mustard (ZC-2) exhibited higher phenolic acids and flavonoids content. • In ZC-Red, anthocyanins accumulation inhibited increased photosynthesis. • Seven key transcription factors were identified in color formation. Purple leaf mustard has garnered attention owing to its abundant anthocyanin content. This study investigated the phenolic metabolome and transcriptome of purple variety ZC-Red and green variety ZC-2. Each of the two varieties had three biological replicates, and each replicate consisted of ten leaves from five plants. Phenolic metabolome was analyzed using an ultra-high performance liquid chromatography system and tandem mass spectrometry, and the transcriptome sequencing platform was the Illumina NovaSeq 6000 platform. The green variety ZC-2 was used as the control for differential analysis. All differential metabolites and genes were significant. Findings revealed the accumulation of anthocyanins in ZC-Red, whereas ZC-2 exhibited higher levels of phenolic acids and flavonoids, excluding anthocyanins. This could be attributed to the directional preference of the phenylpropanoid metabolic flow. Transcriptome analysis highlighted the pivotal roles of phenylpropanoid metabolism and photosynthesis in color formation. In ZC-Red, nearly all anthocyanin biosynthesis genes exhibited up-regulation, with multiple genes in late biosynthesis showing increases thousands of times greater than ZC-2. Conversely, the expression of genes associated with photosystem subunits and light-harvesting complexes was down-regulated. Key candidate transcription factors, including MYB113, TT8, WRKY44, and MYB30, were identified as regulators of anthocyanin and photosynthesis. Findings contribute valuable insights into the molecular mechanisms underlying anthocyanin accumulation. Core differential metabolites such as anthocyanins and phenolic acids are of great significance to the utilization of diversified functional compounds and targeted breeding of leaf mustard. [Display omitted] [ABSTRACT FROM AUTHOR]

• The metabolome and transcriptome of B. juncea roots under Cd stress were analyzed. • A total of 104 DEGs related to lignin and pectin synthesis were screened out. • Eight DEMs related to lignin and pectin synthesis were observed. • Synthesis of lignin and pectin plays vital roles in response to Cd stress. Brassica juncea possesses the capacity to tolerate and accumulate various heavy metals. The cell wall is the first barrier to inhibit heavy metals into the plant cytoplasm. In this work, we investigated the effects of cadmium (Cd) exposure on lignin and pectin synthesis in B. juncea roots by integrated metabolomic and transcriptomic analyses. The findings showed that the contents of lignin and pectin, the activities of phenylalanine ammonia-lyase, laccase, superoxide dismutase, and peroxidase significantly increased following Cd stress, while the activity of pectinesterase decreased. Transcriptome analysis revealed 94 (20 upregulated and 74 downregulated) and 10 (7 upregulated and 3 downregulated) differentially expressed genes in the phenylpropane metabolic pathway and galactose metabolic pathway, respectively. Additionally, metabolomic analyses revealed differentially expressed metabolites related to lignin and pectin synthesis, including N-acetyl-D-phenylalanine, sinapyl alcohol, eugenol, galactosylglycerol, 2-keto-3-deoxy-D-gluconic acid, sucrose, and lactose 6-phosphate, implying lignin and pectin in cell wall play vital roles in Cd detoxification. Our results offer a molecular-scale perspective for further exploring the adaptability in Cd-exposed plants. [Display omitted] [ABSTRACT FROM AUTHOR]

Carrier-based biofertilizers, which involve the introduction of plant growth-promoting bacteria into agricultural industry waste materials, are gaining increasing attention due to their profoundly positive impacts on soil health. In current study, various carriers, including dry leaves, sugarcane husk (SCH), rice husk (RH), and a combination/Mixture (MIX) of all three, were employed to support the bacterial strain, Bacillus safensis (SCAL1). This strain was utilized as a bioinoculant in the production of carrier-based biofertilizers. This particular strain was sourced from the Environmental and Microbial Botany Lab at Quaid-i-Azam University in Islamabad. Bacillus safensis strain exhibited notable improvements, evident in a high rate of longevity and larger colony-forming units within the carriers, as confirmed by measurements taken after 21 days of incubation. The drought stress was induced using polyethylene glycol solutions with concentrations of 15% and 25% administrated 6 and 15 days after germination, respectively. Four distinct biofertilizer types were formulated in the study: DL + SCAL1, RH + SCAL1, SCH + SCAL1, and MIX + SCAL1. When applied to Brassica juncea L. plants, all of these biofertilizers demonstrated the ability to mitigate drought stress and enhance the quality of Brassica juncea L. The immobilization onto SCAL1 by using SCH showed the best result compared to the liquid inoculum as carrier materials improve stability and shelf life. Furthermore, our results affirmed that carrier-based biofertilizers improved biochemical and physiological attributes of plants during drought conditions, highlighting their importance as a valuable resource for mitigating drought stress in Brassica juncea L. The findings revealed that carrier-based biofertilizers are an excellent source of diverse phytohormones, which are pivotal in alleviating drought stress. [Display omitted] • Brassica juncea L growth was impacted by biofertilizers. • It depended upon the specific type of Carrier Material used. • Physiological traits of Brassica juncea were improved by different types of biofertilizers. • Biofertilizers had very influential impact on morphology and its parameters. [ABSTRACT FROM AUTHOR]

The involvement of auxin (IAA) in growth and development of plants is well known, but its role in the mitigation of metal stress, especially copper (Cu), is not fully understood; therefore, it is time to explore its involvement in minimizing the stress. A pot experiment was conducted to assess the protective function of IAA, applied to the foliage, on photosynthetic machinery, carbohydrate metabolism, and growth of Brassica juncea, grown with Cu (30 or 60 mg kg−1 of soil). Among the different concentrations (10−10, 10−8, or 10−6 M), 10−8 M of IAA alone enhanced the photosynthetic characteristics, sugar accumulation and vegetative growth with minimal cellular oxidative stress level. Moreover, the same concentration of auxin was most effective in decreasing the stress levels generated by Cu and maintained it nearly to that of the control in terms of photosynthetic attributes, gas exchange parameters, PSII activity, electron transport rate, and growth attributes. Auxin also maintained the membrane stability and ultrastructure of chloroplast, stomatal morphology with a reduction in malondialdehyde (MDA), electrolyte leakage (EL) and cell death in test plants even under Cu stress. IAA also improved the translocation of Cu from root to the aerial parts, thus enhanced the Cu-reclamation in metal contaminated soils. Our findings suggest that the application of 10−8 M of IAA maintains the overall growth of plants and may be used as an effective agent to improve growth, photosynthesis and phyto-remediation potential of B. juncea in Cu contaminated soil. • Excess Cu induces toxicity by inducing oxidative stress. • IAA spray improves the growth, photosynthesis and stress resilience. • IAA spray maintained chloroplast and stomata organization in Cu stressed plants. • IAA mitigated Cu generated toxicities by improving stress tolerance mechanisms. • IAA enhances Cu-remediation efficiency in mustard plants. [ABSTRACT FROM AUTHOR]

• All three agents significantly improved stress tolerability in mustard plants. • Heavy metal attenuating capacity of ethanol and acetic acid depicted. • Putresciene pretreatment showed best tolerance closely followed by acetic acid. Rapid increase in the accumulation of Cd in plants interrupt with metabolism thus inhibiting the growth and development. Hence, finding a remedy for Cd detoxification and improvement of plant's tolerability to metal toxicity is crucial. In this report, we investigate the differential responses to putrescine (PU), ethanol (ET) and acetic acid (AA) on Cd toxicity in conditions of metal aggregation and stress permissiveness in Indian mustard (Brassica juncea L.). Brassica juncea seedlings (12 day-old) were dealt with Cd (0.5 mM CdCl 2) alone and in combinations of PU (0.5 mM), 0.3% and 0.6% of ET and AA (250 µM) in hydroponic culture for three days. The enhancement in Cd concentration in the plant system induced reduction in growth, biomass and chlorophyll content due to heightened oxidative damage (raised malondialdehyde, MDA content; hydrogen peroxide, H 2 O 2 level) and gradual decrease in antioxidant defense. Cd treatment along with application of PU, ET and AA enhances growth of the plants as quantitated by biomass; represses oxidative damage; increases ascorbate-glutathione (AsA-GSH) and the activities of antioxidant enzymes (superoxide dismutase, SOD; glutathione reductase, GR; glutathione-S-transferase, GST; catalase, CAT; guaciol peroxidase, POX; glutathione peroxidase, GPx; polyphenol peroxidase, PPO; ascorbate peroxidase, APx; monodehydroascorbate reducatase, MDHAR and dehydroascorbate reductase, DHAR. Results indicated that PU, ET and AA plays a vital role in mustard seedlings by mitigating Cd toxicity and increasing the stress tolerance, through upregulating the antioxidant defense systems. Amelioration of heavy metal stress by ET and AA is being reported for the first time in this report. Although PU has been found to be the best among the three the ameliorating capacity of PU and AA are almost similar. [ABSTRACT FROM AUTHOR]

Interspecific hybridization has been a crucial technique in improving genetic diversity and developing novel traits in cultivated plants. The apetalous Brassica juncea (AABB, 2n = 36) represents a valuable germplasm resource with highly desirable characteristics for breeding purposes. Nevertheless, apetalous germplasm resources in B. juncea are currently scarce. In this study, we successfully established a stable apetalous B. juncea (ABJ) line by crossing B. juncea "YLC" with B. carinata "Bc216" (BBCC, 2n = 34). Cytological analysis indicates that ABJ exhibited normal chromosome behavior, with 18 bivalents in diakinesis and an equal number of chromosome segregations at anaphase I in the pollen mother cells during meiosis. Fluorescence in situ hybridization (FISH) confirmed that the chromosome configuration of ABJ is similar to that of its parental line B. juncea , containing 20 chromosomes from A-subgenome and 16 chromosomes from B-subgenome constituting the chromosome number of 36. However, a 15.8 Mb deletion in chromosome A03 of ABJ was uncovered by RNA-seq and molecular markers. Twenty-four differentially expressed genes which may be associated with the regulation of ABJ's apetalous traits were identified by gene expression profiling. To our knowledge, there have been no other documented instances of producing apetalous mustard with missing chromosome segments through interspecific hybridization. Consequently, this study not only verifies the feasibility of generating apetalous mustard with missing chromosome segments through interspecific hybridization but also provides a new insight into the mechanisms underlying the formation of apetalous trait. • A stable apetalous Brassica juncea (AABB, 2n = 36) line was established by crossing B. juncea 'YLC' with Brassica carinata 'Bc216' (BBCC, 2n = 34). • A 15.8 Mb deletion in chromosome A03 of ABJ was uncovered using RNA-seq and molecular markers. • Twenty-four differentially expressed genes associated with the regulation of ABJ's apetalous traits were identified. [ABSTRACT FROM AUTHOR]

Biochar amendment is a promising strategy for mitigating antibiotic resistance genes (ARGs) in soil and plants, but its effects on ARGs at field scale are not fully understood. Here, field trials were executed utilizing two plant varieties, Brassica juncea and Lolium multiflorum , with four types of biochar to investigate changes in ARGs and microbiome in soil, rhizosphere, root endophytes, and leaf endophytes. Results showed that biochar altered ARG distribution in soil and plant, and restrained their transmission from soil and rhizosphere to endophytes. A reduction of 1.2–2.2 orders of magnitude in the quantity of ARGs was observed in root and leaf endophytes following biochar addition, while no significant changes were observed in soil and rhizosphere samples. Procrustes and network analyses revealed significant correlations between microbial communities and mobile genetic elements with ARGs (P < 0.05). Besides, redundancy and variation partitioning analysis indicated that bacterial communities may play a dominant role in shaping the ARGs profile, contributing to 43 % of the variation observed in ARGs. These field results suggest that biochar amendment alone may not fully alleviate ARGs in soil, but it has a significant beneficial impact on food safety and human health by effectively reducing ARGs in plant endophytes. [Display omitted] • Biochar restrained ARGs transmission from the soil and rhizosphere to endophytes. • Biochar amendments reduced 1.2-2.2 orders of magnitude in ARGs in endophytes. • Bacterial communities could play a dominant role in shaping the ARGs profile. [ABSTRACT FROM AUTHOR]

[Display omitted] • Biosynthesis of ZnONPs using extract of Sida cordifolia. • Physicochemical characterization of ZnONPs by using UV–Visible, FTIR, TEM, Zeta-Potential, EDX techniques and an average nanoparticle size is 19 nm. • ZnONPs are examined for their effectiveness against different reference strains and they showed inhibition zones ranging from 3 to 16 nm. • ZnONPs boosted Seed germination percent, Shoot length, Root length, Fresh weight of Brassica juncea L. and the 80 % seed germination was achieved at a 20 ppm. • The highest antioxidant efficacy on DPPH was 36.05 % at 100 ppm and ZnONPs quench the DPPH radical more than Sida cordifolia extract. The present work involves the synthesis of zinc oxide nanoparticles(ZnONPs) using an extract obtained from the seeds of Sida cordifolia. UV–visible, FTIR, zeta potential, EDX and TEM analyses characterized the nanoparticles. The ZnONPs are stable with a zeta potential of −3.5 mV and an average nanoparticle size of 19 nm. These nanoparticles and Sida cordifolia extract are examined for their effectiveness against bacteria (E. coli and B. subtilis) and fungi (A. niger and P. chrysogenum) and they showed inhibition zones ranging from 3 to 16 nm. Furthermore, DPPH's radical quenching activities test the antioxidant potential of ZnONPs and plant extract. DPPH radical inhibition for Sida cordifolia plant extract ranges from a minimum of 17.66 % at 10 ppm to a maximum of 34.16 % at 100 ppm while for ZnONPs, it is from a minimum of 22.48 % at 10 ppm to a maximum of 36.05 % at 100 ppm. ZnONPs and plant extract both show concentration dependent antimicrobial activities and antioxidant activities. In addition, the seed germination of Brassica juncea L. was also tested at various concentrations using the produced ZnONPs. Observed results show maximum seed germination of 80 %, shoot length 1.86 cm, root length 1.35 cm at 20 ppm. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Heatwaves caused the most pronounced B. juncea biomass decrease at 250 ppm CO 2. • Elevated CO 2 levels enhanced Cd phytoremediation effect, especially at 550 ppm CO 2. • Heatwaves and CO 2 interaction increased antioxidant activity but decreased Cd removal effect. [ABSTRACT FROM AUTHOR]

The present research primarily focuses on Bras s ic a 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. • Fluctuation in temperature cause chromosome stickiness, irregularities in spindle formation, unoriented chromatins. • Spindle abnormalities produce sporads micronuclei, polyads, triads that irrupted the productiveness of pollen grains. • Presowing treatment of 28-homobrassinolide regulates the temperature induced clastogenic aberrations of Brassica juncea L. • 28-homoBL showed remarkable attenuation in meiotic irregularities and altered the concentration of soluble sugars. • Saccharides accumulation showed protective nature through maintenance of cell and acting as osmoprotectant. [ABSTRACT FROM AUTHOR]

Chromium (Cr) contamination in soils reduces crop yields and poses a remarkable risk to human and plant system. The main objective of this study was to observe the protective mechanisms of exogenously applied melatonin (Mel- 0.05, 0.1, and 0.15 μM) in seedlings of Brassica juncea L. under Cr (0.2 mM) stress. This was accomplished by analysing the plant's morpho-physiological, biochemical, nuclear, membrane, and cellular characteristics, as well as electrolyte leakage. Superoxide, malondialdehyde, and hydrogen peroxide increased with Cr toxicity. Cr also increased electrolyte leakage. Seedlings under Cr stress had 86.4% more superoxide anion and 27.4% more hydrogen peroxide. Electrolyte leakage increased 35.7% owing to Cr toxicity. B. juncea L. cells with high radical levels had membrane and nuclear damage and decreased viability. Besides this, the activities of the antioxidative enzymes, as POD, APOX, SOD, GST, DHAR, GPOX and GR also elevated in the samples subjected to Cr toxicity. Conversely, the activity of catalase was downregulated due to Cr toxicity. In contrast, Mel reduced oxidative damage and conserved membrane integrity in B. juncea seedlings under Cr stress by suppressing ROS generation. Moreover, the activity of antioxidative enzymes that scavenge reactive oxygen species was substantially upregulated by the exogenous application of Mel. The highest concentration of Mel (Mel c- 0.15 μM) applied showed maximum ameliorative effect on the toxicity caused by Cr. It causes alleviation in the activity of SOD, CAT, POD, GPOX, APOX, DHAR, GST and GR by 51.32%, 114%, 26.44%, 48.91%, 87.51%, 149%, 42.30% and 40.24% respectively. Histochemical investigations showed that Mel increased cell survival and reduced ROS-induced membrane and nuclear damage. The findings showed that Mel treatment upregulated several genes, promoting plant development. Its supplementation decreased RBOH1 gene expression in seedling sunder stress. The results supported the hypothesis that Mel concentrations reduce Cr-induced oxidative burst in B. juncea. [Display omitted] • Chromium (Cr) toxicity negatively affects ecosystem and crop production. • Cr toxicity affects physio-biochemical activities and causes oxidative burst. • Melatonin (Mel) supplementation decreases accumulation of reactive oxygen species. • Mel enhanced activity of antioxidant enzymes and preserved the membrane integrity. • Mel regulated gene expression which helped in minimizing oxidative stress biomarkers. [ABSTRACT FROM AUTHOR]

[Display omitted] • Synthesis of efficient nonionic gemini surfactants from Brassica Juncea. • Synthesized surfactants possess very low CMC with excellent interfacial properties. • Surfactants excel in emulsification, offering versatility for industrial applications. • Superior interfacial properties and enhanced self-aggregation ability observed. • Surfactants are biodegradable, making them environmentally friendly. This study investigates the synthesis and characterization of four non-ionic surfactants of different spacer lengths derived from Brassica Juncea (mustard oil) and explore their potential applications. The chemical structures of the surfactants were confirmed by FTIR, 1HNMR spectroscopy and GC. Surface tension values of the surfactants were measured at various temperatures, unveiling temperature-dependent behaviour and associated thermodynamics. The critical micelle concentrations (CMC) of different surfactants were found to be very low (0.064–0.096 mmol/l at 303 K) compared to conventional surfactants. The CMC and the surface tension further decrease significantly with an increase in temperature. Surface-active thermodynamic parameters of the surfactants demonstrate their lower interfacial characteristics and enhanced self-aggregation capability. At CMC, the synthesized surfactants effectively decrease the interfacial tension (IFT) between water and dodecane to a notably low range of 9.5 to 12.1 mN/m, which is further reduced to a range of 3.2 to 6.1 mN/m at optimal salinity. The surfactants also show good emulsification properties, the emulsions formed in the water-dodecane system using the gemini surfactants as emulsifiers were extensively characterized, encompassing phase behaviour and droplet size distribution analysis. These emulsions exhibit favourable viscous properties, with viscosities ranging from 5 to 20 mPa.s at a 5 s-1 shear rate. This study establishes that the synthesized Gemini surfactants are viable candidates for Enhanced Oil Recovery (EOR), showcasing robust stability, interfacial activity, and favourable rheological properties, alongside biodegradability. [ABSTRACT FROM AUTHOR]

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. • A total of 123 RAV genes, classified into three groups, were identified from six Brassica species. • BnaRAV genes showed diverse expression across tissues, except for two seed-specific genes in Group 3. • BnaA06. RAV3L was a negative regulator of seed development. [ABSTRACT FROM AUTHOR]

Brassica juncea var. tumida Tsen et Lee (Tumorous stem mustard) is an unique vegetable in China. Its enlarged tumorous stem was used as main raw material to produce pickle (Zhacai). In practice, early-bolting happens around 15% of planting area all year and inhibits its production. Here, about 209 PP2C proteins were identified through HMMER software and divided into 13 sub-families in B. juncea. BjuPP2C52 belongs to E sub-family, was up-regulated at reproductive growth stages and interacts with BjuFKF1, a key protein in regulating plant photoperiod flowering, in vitro and in vivo. To explore interactive proteins, BjuPP2C52 was used as bait, 12 potential interactive proteins were screened from yeast library, and they are BjuCOL3, BjuCOL5, BjuAP2, BjuAP2-1, BjuSVP-1, BjuFLC-2, BjuSKP1f, BjuA014572, BjuA008686, BjuO002119, BjuB036787 and BjuA019268. Further study verified that 10 out of the 12 screened proteins interacted with BjuPP2C52 in vivo. qRT-PCR was conducted to understand the expression pattern of those 10 interactive proteins in different tissues and development stages in B. juncea. The results showed that BjuCOL3 , BjuCOL5 , BjuB036787 and BjuA019268 were significantly up-regulated, while BjuA008686 and BjuO002119 were down-regulated in flowers compared with other four tissues. In developmental stages, BjuCOL5 , BjuAP2 , BjuAP2-1 , BjuA014572 , BjuB036787 and BjuA019268 were significantly up-regulated, while BjuSVP-1 , BjuA008686 and BjuO002119 were down-regulated at reproductive stages. Based on the results, BjuCOL5, BjuAP2 , BjuAP2-1 , BjuSVP-1 , BjuA014572 , BjuB036787 and BjuA019268 may function in regulating flowering time in B. juncea. • 209 PP2C proteins were identified and divided into 13 sub-families in B. juncea. • 12 potential interactive proteins of BjuPP2C52 were screened from yeast library. • 10 out of the 12 screened proteins interacted with BjuPP2C52 in vivo. • Four of those interactive proteins were significantly up-regulated, while two down-regulated in flowers. • Six of those interactive proteins were significantly up-regulated, while three down-regulated at reproductive stages. [ABSTRACT FROM AUTHOR]

• Differential response of 12 Indian mustard genotypes under rainfed and irrigated conditions were studied. • The results revealed a significant effect of drought on indicators of chlorophyll content, plant water status, sugars, proline concentration, antioxidative enzymes, malondialdehyde, oil yield and seed yield. • Relationship of proline, reducing sugars and oil yield with seed yield was relatively better under rainfed than irrigated condition. • JC 210-335, CSR 1163, MCN 09-40 and CJRD 1261 are the promising genotypes for developing drought tolerant mustard varieties. Global warming is elevating the risk of climate change which is increasing water scarcity, along with the population explosion and ever-expanding demands for food security resulting in the need for screening of drought tolerant genotypes which can be suitably grown both under rainfed and irrigated conditions. Brassica juncea (L.) Czern & Coss (Indian mustard) is one of the most important oilseed crops and is considerably affected by drought stress. In the present investigation, the impact of water stress on physiological and biochemical traits of advanced Indian mustard genotypes was studied. The experiment was conducted in factorial randomized block design accommodating 12 genotypes grown with two different irrigation modules, with only pre-sowing irrigation referred as rainfed /moisture stress and with two irrigations (35 and 65 days after sowing) referred as irrigated/normal. The results revealed a significant effect of drought on indicators of plant water status, sugars and proline concentration. The photosynthetic pigments (chlorophyll a and b, total chlorophyll and carotenoids) declined under rainfed significantly over irrigated condition. Relative water content decreased whereas water saturation deficit and relative saturation deficit increased under moisture stress as compared to normal condition. The proline, sugar levels and malondialdehyde content along with the antioxidative enzymes increased in all genotypes under stress. Seed yield, oil content and oil yield declined significantly due to moisture deficit. [ABSTRACT FROM AUTHOR]

Beginning of industrialization accelerates the heavy metal pollution in the biosphere. Plant being the immovable entity utilizes different mechanisms to flee from unfavourable conditions. To alleviate toxic impact of metals like cadmium (Cd) and lead (Pb), phytohormones such as indole acetic acid (IAA) has been applied exogenously. This manuscript aims to evaluate the significant change occurring in biochemical parameters of Indian mustard (Brassica napus) grown under individual and combined treatments of IAA with Cd and Pb. Herbicidal potential of treated Brassica extracts were evaluated on growth and development of Anagallis arvensis. Quantum yield parameters were more sensitive to Cd than Pb stress resulted in reduced photosynthetic pigments. However, exogenously applied IAA together with Cd and Pb considerably improved the level of photosynthetic attributes along with reduced accumulation of Cd and Pb in Brassica plant. Cd and Pb enhanced the activities of reactive oxygen species (ROS) and antioxidant machinery. However, addition of IAA with Cd and Pb mitigated the effect of heavy metals on antioxidant system. Moreover, activity of the phenylalanine ammonia lyase enzyme and the defensive metabolites (phenolic, flavonoid and anthocyanin compounds) were boosted under individual treatments of Cd and Pb responsible for increasing herbicidal potential of Brassica plant. Our results exhibited essentiality of IAA in mitigating Cd and Pb stress in Brassica through up-regulated mechanisms of the antioxidant system for balancing ROS related injuries. Increased metabolites enhancing herbicidal potential of Brassica napus against Anagallis weed were also observed. [Display omitted] • Herbicidal potential of Brassica napus on Anagallis arvensis has been evaluated under the influence of Cd and Pb along with IAA. • Adverse effects caused by Cd and Pb as well as their uptake were mitigated by IAA treatment. • Energy of Brassica napus plant exposed to heavy metals is diverted to synthesize antioxidants and other phytochemicals. • Cd and Pb stimulate the secondary metabolites which increases the efficiency of Brassica napus to control the growth of weed. • Soil contaminated with Pb and Cd may buttress the herbicidal potential of Brassica which can control the invasiveness of Anagallis. [ABSTRACT FROM AUTHOR]

• Ten Indian mustard genotypes were hydroponically screened for their ability to accumulate Hg. • All genotypes exhibited dose responses in DW and germination due to increasing Hg content. • Plants root accumulation of Hg in ranged from18.4 to 269.9 μg g−1 DW. • Pusa Jai Kisan was the most Hg-tolerant genotype, accumulating up to 269.9 μg Hg g−1 dry weight (DW) in its roots and up to 61.7 μg Hg g−1 DW in its aerial parts. • The Pusa Jai Kisan genotype had the greatest potential for phytoremediation of Hg-contaminated sites. Phytoremediation is an environment-friendly remediation technology, which has become widely accepted and is regarded as an easy and safe technique for removal of contaminants. Mercury (Hg) is one of the most toxic soil and water contaminants, which has not yet been widely treated via phytoremediation technology because Hg-accumulating plants were hitherto unknown. In the present study, ten genotypes (viz. Agrini, BTO, Kranti, Pusa Bahar, Pusa Basant, Pusa Bold, Pusa Jai Kisan, Vaibhav Vardhan and Varuna) of Indian mustard (Brassica juncea L.) were grown hydroponically under five different Hg concentrations (0–50 μM), and the changes in Hg uptake, plant growth and plant biomass were evaluated in order to determine the suitability of these genotypes, if any, for being used as Hg-phytoremediators. Genotype Pusa Jai Kisan was identified as the most Hg-tolerant genotype, accumulating up to 269.9 μg Hg g−1 dry weight (DW) in its subterranean parts and up to 61.7 μg Hg g−1 DW in its aerial parts. This genotype, therefore, seems to be quite suitable for use as a viable candidate for phytoremediation of Hg-contaminated sites. [ABSTRACT FROM AUTHOR]

Effect of triacontanol on drought-induced stress was studied in Brassica juncea L. Foliage of sixteen-days-old plants was sprayed with concentrations (0, 10, 20 and 30 μM) of triacontanol (TRIA) for 7 days. Subsequently, plants were subjected to drought stress (10% polyethylene glycol, PEG6000) for 7 days. Drought stress increased oxidative stress (TBARS, O 2 ●- and H 2 O 2), however, their contents were reduced by TRIA. Total soluble sugars, reduced glutathione, and proline content in stressed plants were increased by TRIA. Activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), catalase (CAT), and phenylalanine ammonia-lyase (PAL) activity were significantly increased in a dose-dependent manner with TRIA. Potassium (K+) level declined, while magnesium (Mg2+) and calcium (Ca2+) contents increased. The elevated level of lignin under drought with TRIA was significantly associated with MYB46 and PAL gene expression patterns. Altogether, our results suggest that foliar spray of 20 μM TRIA was more operative in reducing the negative impact of drought stress in B. juncea by regulating the antioxidant system, calcium, and lignification. [Display omitted] • Negative impact of drought attenuated by exogenous Triacontanol (TRIA) in Brassica juncea. • TRIA upholds the equilibrium between ROS generation and scavenging. • TRIA promote glutathione reductase activity, proline and calcium accumulation. • TRIA was able to increase phenylalanine ammonia lyase (PAL) activity and lignin content. • Under drought, TRIA increased MYB46 and PAL gene transcripts. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • The alginate-calcium loaded natural clinoptilolites (Alg-Ca-NCPs) were fabricated. • The SAXS patterns demonstrated the surface fractal evolution of the Alg-Ca-NCPs. • Micropore in Alg-Ca-NCP favored to improve higher loading and sustainable releasing. • Alg-Ca-NCPs presented a better release behavior on malondialdehyde drought stress. • Alg-Ca-NCPs had an anti-aging effect on peroxidase drought stress of mustard leaves. [ABSTRACT FROM AUTHOR]

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 Bj PRFs 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 Bj PRFs 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. • Cytoskeletal profilins are actin-polymerizing, structural proteins, having vital roles in maintaining cellular homeostasis. • For the first time, at least 23 individual Bj PRF proteins (131–176 aa) were identified in B. juncea genome. • Evolution of BjPRF genes in Brassica genomes is attributed to the formation of both paralogous and orthologous genes. • Spatiotemporal RNA-seq profiles emphasized the roles of BjPRF s in silique development and stress conditions in Brassica. [ABSTRACT FROM AUTHOR]

The adoption of bio-lubricants derived from vegetable oils is pivotal in advancing environmental sustainability and mitigating the ecological impact of industrial machinery. This study introduces a groundbreaking bio-lubricant formulated with mustard oil as the base, augmented by CaO nanoparticles synthesized from waste eggshells. The CaO nanoparticles were synthesized through a sol-gel method and meticulously characterized via scanning electron microscopy (SEM) and X-ray diffraction (XRD). This investigation delves into the tribological, rheological, and thermal properties of the developed bio-lubricant, highlighting its potential as a sustainable alternative. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) were employed to evaluate thermal stability and cold flow properties, revealing promising results. A comparative study with commercially available engine oil (SAE20W40) underscores the superior performance of the bio-lubricant. The key innovation lies in the incorporation of CaO nanoparticles, facilitated by a surfactant, which substantially enhances wear and friction reduction, lubrication qualities, shear stability, viscosity-temperature behavior, and thermal stability. These advancements position our bio-lubricant as a high-performance, environmentally friendly substitute for conventional lubricants. The study's findings underscore its potential for widespread industrial adoption, promising a transformative shift towards sustainable and eco-conscious lubrication practices. [Display omitted] [ABSTRACT FROM AUTHOR]

• Vanadium (V) toxicity impairs plant growth through severe oxidative damage. • Melatonin reduced v accumulation in both mustard genotypes against v stress. • Melatonin increased both chlorophyll and anthocyanin contents in mustard plants. • Melatonin upregulated the key enzymatic activities and genes expression of anthocyanin biosynthetic pathway. • Melatonin induced more stress tolerance in purple than green mustard genotype. Heavy metal vanadium (V) is poisonous to both plants and animals. Plant resistance to heavy metals stress can be conferred by the multifunctional signaling molecule melatonin, although the underlying mechanisms are still largely understood. We elucidated the significant contribution of the secondary metabolite anthocyanin to the enhancement of V stress tolerance by melatonin in both green and purple mustard genotypes. The effects of excessive V were leaf yellowing, slowed development, decreased photosynthetic activity, and an increase in the buildup of reactive oxygen species and malondialdehyde in both mustard genotypes. It interesting to note that during V stress, key anthocyanin biosynthetic pathway enzymes activities and leaf anthocyanin concentration increased while exogenous melatonin applied as foliar further boosted activities of key anthocyanin biosynthetic pathway enzymes and leaf anthocyanin concentration in both green and purple mustards. Furthermore, the application of exogenous melatonin increased the levels of endogenous melatonin and mitigated V toxicity. Conversely, the suppression of melatonin intensified V-induced phytotoxicity and delayed the accumulation of anthocyanin by downregulating the transcript levels of essential structural genes. Under V stress conditions, the genes expression related to anthocyanin production, antioxidant enzymes activities, and stress resistance was significantly higher in purple mustard than green mustard genotype. Additionally, melatonin decreased V deposition in roots and leaves. The results recommended that melatonin foliar application to mustard plants inhibited the availability of V to plant and promote V stress tolerance. [Display omitted] [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Cr stress reduced growth and physiological attributes of B. juncea. • Cr stress increased MDA, H 2 O 2 , EL and O 2.•‾ ions, hence imposed oxidative stress in B. juncea. • Co-supplementation of ZnONPs and Klebsiella sp. SBP-8 enhanced gene expression for SOD, CAT, POD and APX enzymes. • Cr stress alleviation in B. juncea is credited to enhanced activity of antioxidant enzymes. [ABSTRACT FROM AUTHOR]

The present study was designed to elucidate whether exogenously applied sulphur (S) (S1; 100 ppm, S2; 200 ppm S) alleviate lead (Pb)-induced (Pb1; 100 ppm, Pb2; 200 ppm and Pb3; 300 ppm Pb) stress in the leaves of Brassica juncea. To study this effect, oxidative stress biomarkers (hydrogen peroxide and malondialdehyde), enzymatic antioxidants (ascorbate peroxidase (APX), glutathione reductase (GR)), non-enzymatic antioxidants (chlorophyll and carotenoids) and S-metabolic enzymes like adenosine triphosphate sulfurylase (ATPS), O-acetylserine(thiol)lyase (OASTL) were studied. Pb treatment dramatically inhibited plant development by increasing lipid peroxidation and hydrogen peroxide (H 2 O 2) accumulation by 34.82 and 238.89 % respectively at 90 days plant. The overproduced oxidative biomarkers alter plant cell homeostasis. In contrast, APX (25.56 %) and GR (32.45 %) activities, chlorophyll (15.51 %) and carotenoids (30 %) contents increased under similar Pb treatments and regulated ROS-antioxidant balance. Exogenous S application, maintained the redox status of cell by further increasing the enzymatic activities. Pb treatment increased S assimilation in Brassica by elevating enzyme activities of ATPS and OASTL, which was further augmented by S supplementation to Pb-stressed plants. Besides gene expression of BjSULT , BjOASTL and BjGR1 exhibited slight increase under Pb stress, S application to the stressed plants doubled the expression and assisted the stress resistance. Overall, our findings demonstrate that S protect Pb-stressed Brassica seedlings, implying that S could be an ideal choice for reducing Pb toxicity in crops. [ABSTRACT FROM AUTHOR]

Mustard seeds is a traditional Chinese natural product of pharmaceutical and dietary origin, rich in polyphenols, flavonoids, and other active ingredients, exhibiting multiple bioactivities. In order to improve the yield, polyphenols from brown and white mustard seeds were extracted under better circumstances using Response Surface Methodology (RSM). Subsequently, components were assessed using LC-MS/MS. A total of 8 phenolic compounds and 3 flavonoid compounds were identified in brown mustard seeds extracts (BMSE), and 14 phenolic compounds and 4 flavonoid compounds were identified in white mustard seeds extracts (WMSE). Meanwhile, the biological activities of BMSE and WMSE were compared via in vitro free radical scavenging, hepatocyte oxidative injury protection, and macrophage immunoregulation models. The results showed that both BMSE and WMSE exhibited antioxidant activity and oxidative damage protection activity on HL-7702 cells, and BMSE was more effective. The viability of HL-7702 cells induced oxidative damage by hydrogen peroxide treated with BMSE and WMSE was increased by 36.7% and 32.8%, respectively. WMSE exhibited better in vitro immunomodulatory activities than BMSE, including proliferation promoting, phagocytosis, and NO release abilities on RAW264.7 cells. This study can contribute to the application of mustard seeds polyphenols in the field of functional foods, as well as provide a direction for the high-value utilization of mustard seeds. • The extraction conditions for two types of mustard seeds polyphenols were optimized. • Phenolic compounds in the extracts were identified by LC-MS/MS. • Brown mustard seeds polyphenols had higher antioxidant activity. • White mustard seeds polyphenols had superior immunomodulatory activity. [ABSTRACT FROM AUTHOR]

• Solanum sisymbriifolium is a resistant trap crop for both the species of potato cyst nematodes and able to reduce more than 97% of the nematode populations in next season of potato crop. • Comparing with the antagonistic crops, the trap crop, S. sisymbriifolium was found to minimize populations of both the species of potato cyst nematodes in two successive seasons. • Potato cyst nematodes can be managed effectively by growing of the trap crop, S. sisymbriifolium one season before the potato crop. The potato cyst nematodes are the most wilful and difficult to manage pests of potato crop. Due to the regulatory considerations and potential risk involved in environmental and health hazards, the options for controlling this nematode are becoming increasingly limited. Among the available management options, trap and antagonistic crops offer viable alternative for controlling PCN species. Therefore the present experiments were conducted to determine the effect of locally available nematode trap crop, Solanum sisymbriifolium and antagonistic crops against potato cyst nematodes. Initially, S. sisymbriifolium was evaluated for its trap crop potential against PCN and the results showed that S. sisymbriifolium acts as a resistant crop for both the species of PCN, as no females were observed on the roots of the trap crop up to the end of the crop season. In addition to this, in the subsequent season negligible populations of PCN females with improved plant growth parameters were observed in susceptible potato cv. Kufri Jyoti. In another experiment, its efficiency was compared with antagonistic crop species viz., Tagetes patula (marigold), Allium sativum (garlic) and Brassica juncea (mustard) on the population decline of PCN. From the study, it was observed that minimum populations of both the species of PCN (0.6 and 33.4 female's plant−1 respectively) were supported in two succeeding seasons of susceptible potato cultivar where S. sisymbriifolium was grown in the preceding season which was followed by marigold, garlic and mustard-based sequences. Therefore, S. sisymbriifolium can be used as a potential trap crop for the management of PCN; however, future studies on its suitability in field conditions are required for its effective utilization under field condition for the management of this nematode. [ABSTRACT FROM AUTHOR]

Plant cuticle, the site of perception of stress signals, is an extracellular hydrophobic barrier that covers the epidermis of the above-ground parts. This lipidic layer has been explored for its cutin and wax composition. However, reports on the cuticle proteins are scanty. Therefore, leaf cuticle proteins of Brassica juncea isolated using organic solvents (chloroform-methanol, 2:1(v/v)) were analyzed using gel based and quantitative shotgun proteomics. Out of 615 proteins identified, 27% (169) had signal peptides supporting extracellular localization. Bioinformatics tool, QuickGO predicted the involvement of these proteins in catabolism (21%), peptidase activity (13%), oxidoreductase (12%), defense response (9%), fatty acid binding (9%), nutrient reservoir activity (8%), chitin binding (7%) and lipid transport (2%). Myrosinase-catalyzed glucosinolate hydrolysis releases bioactive compounds, which contribute to plant defense. This system is termed as "mustard oil bomb". Myrosinase and its associating protein, GDSL esterase/lipase ESM1 (involved in cuticle structuring and defense) were detected in the cuticle. GDSL-esterase/lipase ESM1 and β-glucanase (an antifreeze protein) showed in vitro activity. Analysis of cuticle extract by nanoliter osmometer-phase contrast microscopy detected antifreeze activity due to non-protein component. Post-translational modification analysis using PTM viewer predicted N-glycosylation (66%), N-terminal proteolysis (40%), and phosphorylation (32%) to be the dominant modification in the classical secretory proteins. N-glycosylation of myrosinase and GDSL esterase/lipase, ESM1 was confirmed by Con A affinoblotting. This study not only identified leaf cuticle proteins, but also laid the foundation for exploring the extracellular glucosinolate-myrosinase system, PTM crosstalk, and antifreeze activity as stress adaptive strategies in B. juncea. Image 1 • Classical secretory proteins, Myrosinase, GDSL lipase/esterase ESM1, and β- glucanase were identified as abundant targets. • GDSL lipase/esterase ESM1, and β- glucanase showed in vitro activity. • Nanolitre osmometry detected antifreeze activity in the cuticle protein enriched fraction supernatant. • GPS-lipid server predicted lipid modifications in 59% classical secretory proteins. • Many (66%) classical secretory proteins were predicted N-glycosylated by PTM viewer. [ABSTRACT FROM AUTHOR]

• Glucose (Glc) promotes growth of Brassica juncea by maintaining dynamics between formation and utilization of photosynthates. • Glc enhance primary photochemistry, net photosynthetic rate and activity of photosynthetic as well as respiratory enzymes. • Glc reduces hydrogen peroxide and superoxide content by enhancing the antioxidant enzyme activity. • Glc improve the elemental status and also enhance root cell viability. • Glc-mediated effect was of the following order: Glc 30 > Glc 20 > Glc 40 > Glc 10 > Glc 50 > Glc 0 mM. Glucose (Glc), a monosaccharide, accumulates in plants as a minor product of photosynthesis as well as a breakdown product of sucrose. Glucose regulates various physiological processes in plants; therefore, the present experiment was carried out to assess its role in mustard plants. Seeds of Brassica juncea were sown into pots filled with soil and farmyard manure. Glc treatments were provided as a foliar spray at the 25-day stage of growth for 5 days consecutively. Plants were sampled at the 30, 45 and 60-day stage of growth to assess various biological and biochemical parameters. Glc promoted the growth of plants in the following order of concentration: 30 mM > 20 mM > 40 mM > 10 mM > 50 mM > 0 mM. Glc treatment augmented photosynthetic efficiency by elevating primary photochemistry, stomatal movement and conductance, internal CO 2 concentration, transpiration rate, and net photosynthetic rate leading to enhanced synthesis of carbohydrate. A decline in photosynthesis at higher sugar accumulation was prevented by its utilization during respiration. Moreover, the activity of photosynthetic (rubisco and carbonic anhydrase) and respiratory enzymes (hexokinase, succinate dehydrogenase, and fumarase) also increased, which suggests that Glc-mediated growth promotion depends on the dynamics between anabolic and catabolic pathways. Generation of reactive oxygen species, antioxidant enzyme activity, and elemental status, which affect the rate of photosynthesis, were also regulated in Glc-treated plants. [ABSTRACT FROM AUTHOR]

Soil salinity and drought stress (DS) are the massive problem for worldwide agriculture. Both stresses together become more toxic to the plant growth and development. Silicon (Si) being the second most abundant element in the earth's crust, exerts beneficial effects on plants under both stress and non-stress conditions. However, limited information is available to substantiate the beneficial role of Si in delaying the premature leaf senescence and imparting tolerance of mustard (Brassica juncea L.) plants to salinity and DS. Therefore, the present study aimed to explore the role of Si (source K 2 SiO 3) in chlorophyll (Chl) biosynthesis, nutrients uptake, relative water content (RWC), proline (Pro) metabolism, antioxidant system and delaying of premature leaf senescence in mustard plants under sodium chloride (NaCl) and DS conditions. Results of this study show that exogenous Si (1.7 mM) significantly delayed the salt plus DS-induced premature leaf senescence. This was further accompanied by the enhanced nutrients accumulation and activity of chlorophyll metabolizing enzymes [ δ -aminolevulinic acid (δ -ALA) dehydratase and porphobilinogen deaminase] and levels of δ -ALA, and Chls a and b and also by decreased the Chl degradation and Chl degrading enzymes (Chlorophyllase, Chl-degrading peroxidase, pheophytinase) activity. Exogenous Si treatment induced redox homoeostasis in B. juncea L. plants, which is evident by a reduced generation of reactive oxygen species (ROS) resulting due to suppressed activity of their generating enzymes (glycolate oxidase and NADPH oxidase) and enhanced defence system. Furthermore, application of Si inhibited the activity of protease and triggered the activity of antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) and plasma membrane H+-ATPase activity. In conclusion, all these results reveal that Si could help in the modulation of Chl metabolism, redox hemostasis, and the regulation of nutrients (nitrogen, phosphorus, Si and potassium) uptake in the mustard plants that lead to the postponement of premature leaf senescence under salinity plus DS. Image 1 • Salinity and drought stress induce premature leaf senescence. • Si induces delay in premature leaf senescence. • Si improves nutrient acquisition in mustard plants. • Si modulates chlorophyll and proline metabolism in plant. • Si regulates antioxidant enzymes activity in salinity and drought stressed-plant. [ABSTRACT FROM AUTHOR]

• 5-aminolevulinic acid (ALA) regulates Krebs cycle under lead (Pb) stress. • ALA reduces oxidative stress by regulating gene expression on antioxidant system. • ALA regulates phenol biosynthesis by modulating expression of key enzymes. • ALA confers Pb tolerance in Brassica juncea accompanied by less Pb accumulation. Heavy metal pollution seriously impairs crop production and poses serious concerns for human health. Exogenous application of biomolecules has been efficiently tested for enhancing plant resistance to metal toxicity. Current study evaluates the possible effect of 5-aminolevulinic acid (ALA) in Brassica juncea L. seedlings subjected to lead (Pb) stress. Our results showed that shoot length, root length and chlorophyll contents were significantly recovered in Pb stressed seedlings after ALA application, accompanied by reduction in the Pb accumulation. Significant reduction in the contents of reactive oxygen species (ROS) like superoxide anion, hydrogen peroxide and malondialdehyde were also observed in ALA treated seedlings under Pb stress. Furthermore, we also noticed enhancement in the activities of antioxidative enzymes like superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), glutathione reductase (GR), glutathione-S-transferase (GST) and dehydroascorbate reductase (DHAR). We further noticed that ALA upregulated the expression of SOD (7.30 folds), POD (6.11 folds), CAT (3.52 folds), DHAR (6.42 folds), GR (6.04 folds), and GST (5.58 folds) under the Pb stress. However, RBOH1 (gene involved in ROS generation) and CHLASE (chlorophyllase) expressions were reduced in ALA treated seedlings grown under Pb stress (RBOH1 expression decreased to 3.44 from 6.50 fold and CHLASE expression decreased to 2.97 from 5.58 fold). Phenolic contents were increased in the presence of ALA and expression of genes like CHS (chalcone synthase; 7.50 fold) and PAL (phenylalanine ammonia lyase; 4.77 fold) was also stimulated by ALA under Pb stress. Furthermore, contents of the Krebs cycle metabolites (fumarate, succinate, malate and citrate) were also enhanced accompanied by upregulated expression of genes like CS (citrate Synthase; 8.13 fold), SUCLG1 (succinyl CoA ligase 1; 7.40 fold), SDH (succinate dehydrogenase; 5.10 fold) and FH (fumarate hydratase; 5.65 fold). In conclusion, current investigation revealed that ALA attenuated Pb toxicity by modulating the transcription patterns of key enzymes involved in plant defense system. [ABSTRACT FROM AUTHOR]

Brassica genus comprises many prominent species valuable for human nutrition including vegetable crops and oilseed. Production of B. juncea is challenged by many abiotic and biotic stresses, Alternaria blight caused by a necrotrophic fungal pathogen Alternaria brassicae is one of the most prominent diseases of cruciferous crops including B. juncea. However, some closely related wild species like Sinapis alba and Camelina sativa exhibit a variable level of resistance towards the pathogen. Apart from the host resistance, intra-specific pathogen variability also influences disease severity to a larger extent. In this study, we identified and isolated two strains of A. brassicae viz ABS1 and ABS2 exhibiting morphological and pathological variability. These isolates were further used to artificially inoculate B. juncea and two of its wild relatives under in-vitro as well as in-vivo conditions to inspect their pathogenicity in a susceptible, a moderately resistant and a highly resistant host. virulent isolate (ABS2) was able to readily establish infection in all the three species whereas the less virulent isolate (ABS1) readily infected susceptible species B. juncea but delayed and mild infection was noticed in tolerant hosts. Variable physiological and molecular host response towards the differential level of virulence of pathogen were established with many confirmatory experiments like DAB staining study, Disease severity index and microscopic analysis. Real-time PCR results confirm that these two isolates induce a variable level of induction in genes PR1 and PDF1.2 within 48 h of the artificial inoculation in B. juncea and its wild relatives. Image 1 • Identification and characterization of isolates of A. brassicae in terms of morphological and pathogenicity parameters. • Diverse responses of three host species- B. juncea , S. alba and C. sativa towards variable pathogen virulence. • Assessment of disease severity in all three host species in response to virulent isolate of A. brassicae. • Differential gene expressionof defence related genes in three host species in response to variable virulence of the pathogen. [ABSTRACT FROM AUTHOR]

In the present study an effort has been made to optimize the in vitro regeneration protocol for Agrobacterium -mediated transformation of Brassica juncea, because of its importance as oilseed crops. The highest callus induction frequency of 87% was observed on MS (Murashige and Skoog, 1962) medium supplemented with 4 µM 6-benzyladenine (BA) after four weeks of culture period. Subculturing of organogenic calli in MS media with a similar hormonal composition resulted in shoot organogenesis after six weeks of culture cultivation. The highest shoot induction frequency (92%) was recorded on MS medium containing 4 µM BA in combination with 1 µM of α-naphthalene acetic acid (NAA). Further, well-developed roots were formed in MS media augmented with 6 µM of Indole acetic acid (IAA) in combination with 1 µM Kinetin (Kn). Cotyledon explants were exploited in vitro for the successful transformation of B. juncea. A binary vector comprised of the Euonymus alatus diacylglycerol acetyltransferase (Ea DAcT) gene under the transcriptional control of a glycinin promoter and with a basta selection marker was introduced into A. tumefaciens strain GV3101 via electroporation. Ea DAcT gene is responsible for unusual triacylglycerol's production where the sn-3 position is esterified with acetate instead of the long-chain fatty acid found in the triacylglycerol's. The highest regeneration frequency (100%) of transgenic shoots was observed on MS medium supplemented with 4 µM BA plus 1 µM NAA in the presence of 25 mg l−1 basta and 160 mg l−1 timintin. The efficiency of stable transformation was found to be approximately 7% in the transgenic plants. Moreover, the transformed regenerated shoots were confirmed by PCR analysis using Ea DAcT gene-specific primers. [ABSTRACT FROM AUTHOR]