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Lunar farming, often regarded as a myth, is regularly practiced in many places around the world (e.g., India) where framers organized their agricultural activities according to moon phases. Early and recent work showed that exposure to moonlight affects the life cycle of plants, from seed germination and vegetative growth to fruit maturation and dispersal. Here we addressed the long-term effect of short exposure to full moonlight (FML) on cellular activities in Brassica juncea by analyzing protein and metabolite profiles immediately after 3-night-exposure (3NE) or 7 and 15 days after exposure (DAE) to FML. This study shows an increase in nuclear size following 3NE to FML, which was accompanied by changes in protein and metabolite profiles. We identified significant alterations in protein and metabolite profiles between FML and dark-treated plants in conjunction with developmental stages, which persisted long after exposure to FML. Most notable are the changes in composition of metabolite interconversion enzymes (MIEs) at various developmental stages which were intensified in FML-treated plants. Changes in MIEs were accompanied by significant alterations in metabolite composition and level, particularly at 15DAE, including branched-chain amino acids (e.g., valine, leucine), multiple sugars (raffinose, glucose, sucrose) as well as the tricarboxylic acid (TCA) cycle intermediates malic acid and citric acid. Thus, our results show that short-term exposure to FML triggers a developmental switch resulting in a long-term impact on plant performance that brings about an increase in cell activities and consequently enhanced growth. Our results call for meticulous research on this lunar phenomenon and its potential to enhance crop plant growth and development., (© 2024 The Author(s). Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Introduction: Brassica juncea is a major oilseed crop of Brassica . The seed weight is one of yield components in oilseed Brassica crops. Research on the genetic mechanism of seed weight is not only directly related to the yield and economic value of Brassica juncea but also can provide a theory foundation for studying other Brassica crops., Methods: To map the genes for seed weight, the parental and F 2 extreme bulks derived were constructed from the cross between the heavy-seeded accession 7981 and the light-seeded one Sichuan yellow (SY) of B. juncea , and used in bulk segregant sequencing (BSA-seq). Meanwhile, RNA-sequencing (RNA-seq) was performed for both parents at six seed development stages., Results: Our results showed that a total of thirty five SNPs were identified in thirty two genes located on chromosomes A02 and A10, while fifty eight InDels in fifty one genes located on A01, A03, A05, A07, A09, A10, B01, B02 and B04. The 7,679 differentially expressed genes were identified in developing seeds between the parents. Furthermore, integrated analysis of BSA-seq and RNA-seq data revealed a cluster of nine genes on chromosome A10 and one gene on chromosome A05 that are putative candidate genes controlling seed weight in B. juncea ., Discussion: This study provides a new reference for research on Brassica seed weight and lays a solid foundation for the examination of seed in other Brassica crops., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Yang, Yang, Kang, You, Chen, Xiao, Qian, Rao and Liu.)

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

Most of the released high-yielding hybrids of Brassica juncea have a low 1000-seed weight (TSW) with no increment in the percent oil content (OC), and, therefore, these hybrids have poor acceptance among the farmers in India. It is, thus, imperative to understand the genetic basis of these traits and deploy them in commercial hybrid breeding programs. The present study utilized a set of 15 diverse B. juncea genotypes with TSW and OC ranging from 1.32 to 8.26 g and 31.93-43.39 %, respectively, to generate 210 hybrids following a full diallel mating scheme. These hybrids along with their parents, were evaluated in three different environments. Inheritance of TSW suggests the predominance of additive gene action, whereas non-additive gene action was observed to regulate OC. Further, TSW and OC were reported to be influenced by maternal and non-maternal effects, respectively. Parents with bold seeds viz., NPJ 253, RH 761 and EC 223389 were identified as good general combiners for both the traits. Hybrid generated from the cross NPJ 253 x NPJ 161, with both parents having high seed weights, exhibited the highest mean values (8.43 g) and heterobeltiosis (17.2 %) for TSW. Whereas, hybrid between parents NPJ 253 and IC 426372, possessing high and low seed weights, respectively, observed the highest mean value (44.95 %) and heterobeltiosis (14.89 %) for OC. Keeping both TSW and OC together, hybrids viz., NPJ 253 x EC 223389 (H1), NPJ 253 x NPJ 161 (H2) and NPJ 253 x Pusa Tarak (H4) were identified as promising using the Multi-trait Genotype-Ideotype Distance Index. Lines with higher TSW and better combining ability for OC shall be converted to male and/or female lines for generating commercial hybrids. The scheme for deploying higher seed weight and improved oil content in B. juncea hybrids is discussed., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Naveen Singh reports financial support and administrative support were provided by 10.13039/501100001503ICAR - 10.13039/501100005384Indian Agricultural Research Institute. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

Indian Mustard (Brassica juncea L.) is a globally cultivated winter oilseed crop of the rapeseed-mustard group. It is predominantly grown in the semi-arid northwest agroclimatic zone of India, characterized by high soil salinity. Enhancing tolerance to salt stress in B. juncea is therefore crucial for sustaining its production in this region. Long non-coding RNAs (lncRNAs) play critical roles in coordinating gene expression under various abiotic stresses, including salt stress, but their involvement in the salt stress response in B. juncea remains largely unknown. In this study, we conducted RNA-seq analysis on control, salt-stressed, and salt-shocked young leaves of the salt-tolerant B. juncea cv CS-52. We identified a total of 3,602 differentially expressed transcripts between stress versus control and shock versus control samples. Among these, 61 were identified as potential lncRNAs, with 21 specific to salt stress and 40 specific to salt shock. Of the 21 lncRNAs specific to salt stress, 15 were upregulated and six were downregulated, while all 40 lncRNAs unique to salt shock were downregulated. Chromosomal distribution analysis of the lncRNAs revealed their uneven placement across 18 chromosomes in B. juncea. RNA-RNA interaction analysis between salt stress-upregulated lncRNAs and salt stress-related miRNAs identified 26 interactions between 10 lncRNAs and 23 miRNAs and predicted 13 interactions between six miRNAs and 13 mRNAs. Finally, six lncRNA-miRNA-mRNA interaction networks were established, involving five lncRNAs, 13 miRNAs, and 23 mRNAs. RT-qPCR analysis revealed the upregulation of four out of five lncRNAs, along with their target mRNAs, supporting their involvement in the salt stress response in B. juncea., Competing Interests: Declarations. Ethics approval and consent to participate: This study was conducted following all ethical guidelines and principles and was approved by the competent authority of the Institute. All participants provided informed consent before participating in the study. Consent for publication: All authors have agreed to the publication of this manuscript. The authors declare that they have no competing interests. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Chromium (Cr) toxicity impairs essential morphological and metabolic activities in plants. The present investigation was carried out to evaluate the beneficial role of plant growth promoting rhizobacterial strains namely Pseudomonas aeruginosa (M1), Burkholderia gladioli (M2) and earthworms (Eisenia fetida) in alleviating Cr toxicity in 10 days old Brassica juncea L. The findings delineated that addition of earthworms and PGPR restored growth, boosted Cr uptake and showed upregulation of metal transporter genes (SULTR 1-4). Supplementation of rhizospheric amendments reinstated Cr induced impairment in photosynthetic attributes. Gaseous exchange attributes, the efficiency of PS II, the content of total phenols, anthocyanin and flavonoids was enhanced with application of earthworms along with PGPR. Confocal imaging of primary photosynthetic pigment (chlorophyll), accessory photosynthetic pigment (carotenoids) and total phenols showed maximum fluorescence with combined inoculation of earthworms and both microbial strains (M1M2). The gene expression analysis revealed that Phyotene synthase (PSY), Photosystem II core protein psb A, psb B were down regulated in Cr stressed seedlings which upon supplementation with earthworms and PGPR were upregulated. Further, Phenylalanine ammonialyase (PAL), chalcone synthase (CHS) were upregulated with addition of earthworms and PGPR. Increased nitric oxide content, enhanced activity and upregulation of nitrate reductase (NR) gene was observed with addition of PGPR and earthworms., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Brassica juncea L. (family Brassicaceae) or Indian mustard is a fast-growing oilseed crop. Climate changes mean that it is very important to evaluate the effects of salinity stress on B. juncea. The aim of this study was therefore to show the metabolic effect of salinity stress on shoots and roots using two cultivation models - hydroponic and microplot - in different cultivars, including RH-725 and RH-761. Salinity levels of 5, 7.5, and 10 dS m⁻¹ were investigated, and compared with a control of 0 dS m⁻¹, using untargeted metabolomics with gas chromatography-mass spectrometry (GC-MS) post-silylation, focusing on metabolic markers such as proline and glycine-betaine., Results: A total of 56 metabolites were identified, with the most prevalent classes belonging to sugars (8), followed by organic acids (13), amino acids (11), and fatty acids/esters (11). Shoots were found to have a higher sugar content than roots. Increases in unsaturated fatty acids were also associated with salinity stress, compared with a decrease in saturated fatty acids. Absolute levels of proline and glycine-betaine correlated with salinity stress, with the largest increases detected in shoots grown under hydroponic conditions, particularly for the RH-761 cultivar. Multivariate data analyses revealed that roots were more affected than shoots, regardless of cultivation model., Conclusion: These findings might explain the different metabolic behavior of B. juncea's roots and shoots under various levels of salinity, associated with higher levels of free sugars in shoots and lipids in roots. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

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., Competing Interests: Declaration of Competing Interest All the authors declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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

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

The WRKY transcription factors comprise one of the most extensive gene families and serve as pivotal regulators of plant responses to heavy metal stress. They contribute significantly to maintaining plant growth and development by enhancing plant tolerance. However, research on the role of WRKY genes in response to cadmium (Cd) stress in mustard is minimal. In this study, we conducted a genome-wide analysis of the mustard WRKY gene family using bioinformatics. The results revealed that 291 WRKY putative genes ( BjuWRKY s) were identified in the mustard genome. These genes were categorized into seven subgroups (I, IIa-e and III) through phylogenetic analysis, with differences in motif composition between each subgroup. Homology analysis indicated that 31.62% of the genes originated from tandem duplication events. Promoter analysis revealed an abundance of abiotic stress-related elements and hormone-related elements within the BjuWRKY genes. Transcriptome analysis demonstrated that most BjuWRKY genes exhibited differential expression patterns at different Cd treatment stages in mustard. Furthermore, 10 BjuWRKY genes were confirmed to respond to Cd stress through the construction of a BjuWRKY protein interaction network, prediction of hub genes, and real-time fluorescence quantitative PCR analysis, indicating their potential involvement in Cd stress. Our findings provide a comprehensive insight into the WRKY gene family in mustard and establish a foundation for further studies of the functional roles of BjuWRKY genes in Cd stress response., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Li, Ji, An, Chen, Luo, Liu and Zou.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Masson SAS.)

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

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

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

Main Conclusion: Using octoploid somatic hybrids with excessive C genome sets, AABBCCCC, a diverse allohexaploid, AABBCC, was produced by C genome reduction through subsequent crossing with various AABB cultivars. Even when somatic hybrids are produced, the plants that are produced are rarely in themselves an innovative crop. In this study, we used somatic hybrids of Brassica juncea (AABB) and B. oleracea (CC) as model cases for the genetic diversification of the somatic hybrids. One cell of 'Akaoba Takana' (B. juncea) and two cells of 'Snow Crown' (B. oleracea) were fused to create several somatic hybrids with excessive C genomes, AABBCCCC. Using AABBCCCC somatic hybrids as mother plants and crossing with 'Akaoba Takana', the AABBCC progenies were generated. When these AABBCC plants were self-fertilized, and flow cytometric (FCM) analysis was performed on the next generations, differences in the relative amount of genome size variation were observed, depending on the different AABBCCCC parents used for AABBCC creation. Further self-progeny was obtained for AABBCC plants with a theoretical allohexaploid DNA index by FCM. However, as the DNA indices of the progeny populations varied between plants used and aneuploid individuals still occurred in the progeny populations, it was difficult to say that the allohexaploid genome was fully stabilized. Next, to obtain genetic diversification of the allohexaploid, different cultivars of B. juncea were crossed with AABBCCCC, resulting in diverse AABBCC plants. Genetic diversity can be further expanded by crossbreeding plants with different AABBCC genome sets. Although genetic stability is necessary to ensure in the later generations, the results obtained in this study show that the use of somatic hybrids with excess genomes is an effective strategy for creating innovative crops., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

White rust disease caused by a biotrophic oomycete Albugo candida is one of the most serious impediments in realizing the production potential of Brassica juncea . Due to the obligate nature of the pathogen, R-gene-based resistance is unstable as the newer virulent races emerge quickly. For this, a deep understanding of the molecular basis of resistance is essential for developing durable resistant varieties. In this study, we selected one susceptible cultivar, 'Pusa Jaikisan' and its single R gene based resistant NIL, 'Pusa Jaikisan WRR as the source of understanding the defense mechanism in B. juncea against A. candida . Comparative histochemical analysis at 12 dpi showed higher callose deposition in the resistant cultivar than in the susceptible which hints towards its possible role in defense mechanism. Based on the biochemical markers observation, total protein was found to have a negative correlation with the resistance. The antioxidant enzymes (POX, CAT, and SOD) and non-enzymatic ROS scavenging compounds such as polyphenols and proline showed a positive correlation with the white rust resistance. Polyphenol Oxidase (PPO) total chlorophyll and total carotenoids were also found to be more abundant in the 'Pusa Jaikisan WRR'. Based on the heat map analysis, PAL was identified to be the comparatively most induced enzyme involved in the defense mechanism. The polyphenol oxidase, total chlorophyll and total carotenoids were also found to show higher activity in the 'Pusa Jaikisan WRR '. Furthermore, to study the defense response of 'Pusa Jaikisan WRR ' compared to 'Pusa Jaikisan' against A. candida infection, the gene expression analyses of salicylic acid (SA)-marker PR protein genes ( PR1 and PR2 ) and jasmonic acid (JA)-marker PR protein genes ( PR3 and PR12 ) were done by qRT-PCR. Based on the results, PR2 emerged as the best possible gene for defense against A. candida followed by PR1 . PR3 and PR12 also showed positive correlation with the disease resistance which may be due to the JA pathway acting complementary to the SA pathway in case of B. juncea - A. candida interaction. This provides evidence for the JA-SA hormonal crosstalk to be synergistic in case of the white rust resistance., Competing Interests: PR is Ph.D. student in Division of Plant Pathology, ICAR-IARI, New Delhi. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Rai, Prasad, Mehta, Yadav, Sharma and Mishra.)

Pre-experimental one-group pretest posttest research design was adopted to investigate the effectiveness of Brassica juncea (mustard) plaster on reduction of knee pain and inability among 60 geriatrics with osteoarthritis. The inability was assessed using the Western Ontario McMaster University (WOMAC) Index, which includes 24 parameters under three major headings: joint pain, joint stiffness, and difficulty in performing daily activities, and the severity of pain level was assessed using a numerical pain rating scale. Mustard plaster is applied against the knee for 15 minutes once a day for seven days. On day 8, study participants were re-assessed using the WOMAC Osteoarthritis Index in the interventional group. The pretest mean score of pain was 77.80 ± 5.65, and the posttest mean score was 38.88 ± 11.76. The mean difference score was 38.92. The calculated paired 't' test value of t = 24.428 was statistically significant at P < 0.001 level., Competing Interests: There are no conflicts of interest., (Copyright: © 2024 Journal of Pharmacy and Bioallied Sciences.)

Brassica carinata (BBCC) commonly referred to as Ethiopian mustard is a natural allotetraploid containing the genomes of Brassica nigra (BB) and Brassica oleracea (CC). It is an oilseed crop endemic to the northeastern regions of Africa. Although it is under limited cultivation, B. carinata is valuable as it is resistant/highly tolerant to most of the pathogens affecting widely cultivated Brassica species of the U's triangle. We report a chromosome-scale genome assembly of B. carinata accession HC20 using long-read Oxford Nanopore sequencing and Bionano optical maps. The assembly has a scaffold N50 of ~39.8 Mb and covers ~1.11 Gb of the genome. We compared the long-read genome assemblies of the U's triangle species and found extensive gene collinearity between the diploids and allopolyploids with no evidence of major gene losses. Therefore, B. juncea (AABB), B. napus (AACC), and B. carinata can be regarded as strict allopolyploids. We cataloged the nucleotide-binding and leucine-rich repeat immune receptor (NLR) repertoire of B. carinata and, identified 465 NLRs, and compared these with the NLRs in the other Brassica species. We investigated the extent and nature of early-generation genomic interactions between the constituent genomes of B. carinata and B. juncea in interspecific crosses between the two species. Besides the expected recombination between the constituent B genomes, extensive homoeologous exchanges were observed between the A and C genomes. Interspecific crosses, therefore, can be used for transferring disease resistance from B. carinata to B. juncea and broadening the genetic base of the two allotetraploid species., (© 2024 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Most Fusarium species are known as endophytes and/or phytopathogens of higher plants and have a worldwide distribution. Recently, information discovered with molecular tools has been also published about the presence of these fungi in the microbiome of truffle fruiting bodies. In the present work, we isolated and identified three Fusarium strains from truffle fruiting bodies. All isolates were assigned to the same species, F. commune , and the strains were deposited in the All-Russian Collection of Microorganisms under accession numbers VKM F-5020, VKM F-5021, and VKM F-5022. To check the possible effects of the isolated strains on the plants, the isolates were used to infect sterile seedlings of Sarepta mustard ( Brassica juncea L.). This model infection led to a moderate suppression of the photosynthetic apparatus activity and plant growth. Here, we present characteristics of the F. commune isolates: description of the conidial morphology, pigmentation, and composition of the mycelium fatty acids. Overall, this is the first description of the Fusarium cultures isolated from truffle fruiting bodies. Possible symbiosis of the F. commune strains with truffles and their involvement in the cooperative fatty acid production are proposed.

Plant architecture is a crucial determinant of crop yield. The number of primary (PB) and secondary branches (SB) is particularly significant in shaping the architecture of Indian mustard. In this study, we analyzed a panel of 86 backcross introgression lines (BCILs) derived from the first stable allohexaploid Brassicas with 170 Sinapis alba genome-specific SSR markers to identify associated markers with higher PB and SB through association mapping. The structure analysis revealed three subpopulations, i.e., P1, P2, and P3, in the association panel containing a total of 11, 33, and 42 BCILs, respectively. We identified five novel SSR markers linked to higher PB and SB. Subsequently, we explored the 20 kb up- and downstream regions of these SSR markers to predict candidate genes for improved branching and annotated them through BLASTN. As a result, we predicted 47 complete genes within the 40 kb regions of all trait-linked markers, among which 35 were identified as candidate genes for higher PB and SB numbers in BCILs. These candidate genes were orthologous to ANT , RAMOSUS , RAX , MAX , MP , SEU , REV , etc., branching genes. The remaining 12 genes were annotated for additional roles using BLASTP with protein databases. This study identified five novel S. alba genome-specific SSR markers associated with increased PB and SB, as well as 35 candidate genes contributing to plant architecture through improved branching numbers. To the best of our knowledge, this is the first report of introgressive genes for higher branching numbers in B. juncea from S. alba ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Singh, Kumari and Rai.)

The addition of plant-growth-promoting bacteria (PGPB) to heavy-metal-contaminated soils can significantly improve plant growth and productivity. This study isolated heavy-metal-tolerant bacteria with growth-promoting traits and investigated their inoculation effects on the germination rates and growth of millet (Panicum miliaceum) and mustard (Brassica juncea) in Cd- and Zn-contaminated soil. Leifsonia sp. ZP3, which is resistant to Cd (0.5 mM) and Zn (1 mM), was isolated from forest soil. The ZP3 strain exhibited plant-growth-promoting activity, including indole-3-acetic acid production, phosphate solubilization, catalase activity, and 2,2-diphenyl-1-picrylhydrazyl radical scavenging. In soil contaminated with low concentrations of Cd (0.232 ± 0.006 mM) and Zn (6.376 ± 0.256 mM), ZP3 inoculation significantly increased the germination rates of millet and mustard 8.35- and 31.60-fold, respectively, compared to the non-inoculated control group, while the shoot and root lengths of millet increased 1.77- and 4.44-fold (p < 0.05). The chlorophyll content and seedling vigor index were also 4.40 and 18.78 times higher in the ZP3-treated group than in the control group (p < 0.05). The shoot length of mustard increased 1.89-fold, and the seedling vigor index improved 53.11-fold with the addition of ZP3 to the contaminated soil (p < 0.05). In soil contaminated with high concentrations of Cd and Zn (0.327 ± 0.016 and 8.448 ± 0.250 mM, respectively), ZP3 inoculation led to a 1.98-fold increase in the shoot length and a 2.07-fold improvement in the seedling vigor index compared to the control (p < 0.05). The heavy-metal-tolerant bacterium ZP3 isolated in this study thus represents a promising microbial resource for improving the efficiency of phytoremediation in Cd- and Zn-contaminated soil., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Main Conclusion: The study evaluates the potential of Spray-Induced Gene Silencing and Host-Induced Gene Silencing for sustainable crop protection against the broad-spectrum necrotrophic fungus Sclerotinia sclerotiorum. Sclerotinia sclerotiorum (Lib.) de Bary, an aggressive ascomycete fungus causes white rot or cottony rot on a broad range of crops including Brassica juncea. The lack of sustainable control measures has necessitated biotechnological interventions such as RNA interference (RNAi) for effective pathogen control. Here we adopted two RNAi-based strategies-Spray-Induced Gene Silencing (SIGS) and Host-Induced Gene Silencing (HIGS) to control S. sclerotiorum. SIGS was successful in controlling white rot on Nicotiana benthamiana and B. juncea by targeting SsPac1, a pH-responsive transcription factor and SsSmk1, a MAP kinase involved in fungal development and pathogenesis. Topical application of dsRNA targeting SsPac1 and SsSmk1 delayed infection initiation and progression on B. juncea. Further, altered hyphal morphology and reduced radial growth were also observed following dsRNA application. We also explored the impact of stable dsRNA expression in A. thaliana against S. sclerotiorum. In this report, we highlight the utility of RNAi as a biofungicide and a tool for preliminary functional genomics., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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

This research paper investigates the variability in seed oil content (SOC) in Indian mustard ( Brassica juncea  L.) under terminal heat stress (THS) conditions. A genetic stock of 488 genotypes of B . juncea was evaluated over two years and grouped into five classes based on the reduction in oil content under THS compared to normal sown crop. Based on heat susceptibility index (HSI), a diverse panel of 96 genotypes was selected and evaluated under THS. Twenty-two heat-tolerant donor genotypes were identified, including introgression lines derived from B . tournefortii , B . carinata and Erucastrum cardaminoides . This study is the first to report on marker-trait associations for SOC in B. juncea under THS using a GWAS approach. Furthermore, candidate genes associated with abiotic stress tolerance and lipid metabolism were identified near the significant SNPs, emphasizing their role in SOC regulation under stress. Notable candidate genes include BjuA003240 (encoding for alcohol-forming fatty acyl-CoA reductase), BjuA003242 (involving in lipid biosynthesis), BjuA003244 (associated with mitochondrial functions and stress tolerance), and BjuA003245 (related to MYB transcription factors regulating lipid biosynthesis). This study provides valuable insights into the genetic basis of SOC variation under THS in B. juncea , highlighting potential breeding targets for improved heat stress resilience in Indian mustard cultivation., Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-03985-w., Competing Interests: Conflict of interestThe authors declare that no conflict of interest exists., (© King Abdulaziz City for Science and Technology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

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

This paper explores the potential of Technosols made from non-hazardous industrial wastes as a sustainable solution for highly acidic iron-rich soils at the Rio Tinto mining site (Spain), a terrestrial Mars analog. These mine soils exhibit extreme acidity (pH H2O  = 2.1-3.0), low nutrient availability (non-acid cation saturation < 20 %), and high levels of Pb (3420 mg kg -1 ), Cu (504 mg kg -1 ), Zn (415 mg kg -1 ), and As (319 mg kg -1 ), hindering plant growth and ecosystem restoration. To address these challenges, the study systematically analyzed selected waste materials, formulated them into Technosols, and conducted a four-month pot trial to evaluate the growth of Brassica juncea under greenhouse conditions. Technosols were tailored by adding varying weight percentages of waste amendments into the mine Technosol, specifically 10 %, 25 %, and 50 %. The waste amendments comprised a blend of organic waste (water clarification sludge, WCS) and inorganic wastes (white steel slag, WSS; and furnace iron slag, FIS). The formulations included: (T0) exclusively mine Technosol (control); (T1) 60 % WCS + 40 % WSS; (T2) 60 % WCS + 40 % FIS; and (T3) 50 % WCS + 16.66 % WSS + 33.33 % FIS. The analyses covered leachate quality, soil pore water chemistry, and plant response (germination and survival rates, plant height, and leaf number). Results revealed a significant reduction in leachable contaminant concentrations, with Pb (26.16 mg kg -1 ), Zn (4.94 mg kg -1 ), and Cu (2.29 mg kg -1 ) dropping to negligible levels and shifting towards less toxic species. These changes improved soil conditions, promoting seed germination and seedling growth. Among the formulations tested, Technosol T1 showed promise in overcoming mine soil limitations, enhancing plant adaptation, buffering against acidification, and stabilizing contaminants through precipitation and adsorption mechanisms. The paper stresses the importance of tailoring waste amendments to specific soil conditions, and highlights the broader implications of the Technosol approach, such as waste valorization, soil stabilization, and insights for Brassica juncea growth in extreme environments, including Martian soil simulants., Competing Interests: Declaration of competing interest Juan Carlos Fernandez-Caliani reports financial support, equipment, drugs, or supplies, and travel were provided by Regional Government of Andalusia (Spain). Juan Carlos Fernandez-Caliani reports equipment, drugs, or supplies and travel were provided by DSM Soluciones Medioambientales. Pablo J. Hidalgo reports financial support provided by EU project 101071300 Sustainable Horizons. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

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

Background: Brassica species is the second most important edible oilseed crop in India. Albugo candida (Pers.) Kuntze, a major oomycete disease of oilseed brassica causing white rust, leads to 60% yield loss globally. The prevalence of A. candida race 2 (Ac2V) that specifically infects B. juncea, coupled with limitations of conventional methods has resulted in a dearth of white rust resistance resources in cultivated varieties., Methods and Results: In an effort to develop resistant plants, Agrobacterium mediated genetic transformation of three B. juncea genotypes viz., susceptible host var. Varuna, along with its doubled haploid mutant lines C66 and C69 (showing moderate tolerance to field isolates of A. candida) was initiated to transfer resistance genes (WRR8 Sf-2 and WRR9 Hi-0 ) identified in Arabidopsis thaliana against race Ac2V, that encode for Toll-like/interleukin-1 receptor-nucleotide binding-leucine-rich repeat proteins that recognize effectors of the pathogen races., Conclusions: Our results demonstrate that introduction of resistance genes from a tertiary gene pool by genetic transformation enhances disease resistance in B. juncea genotypes to a highly virulent Ac2V isolate., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

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

Key Message: Imbalanced chromosomes and cell cycle arrest, along with down-regulated genes in DNA damage repair and sperm cell differentiation, caused pollen abortion in synthetic allodiploid Brassica juncea hybrids. Interspecific hybridization is considered to be a major pathway for species formation and evolution in angiosperms, but the occurrence of pollen abortion in the hybrids is common, prompting us to recheck male gamete development in allodiploid hybrids after the initial combination of different genomes. Here, we investigated the several key meiotic and mitotic events during pollen development using the newly synthesised allodiploid B. juncea hybrids (AB, 2n = 2× = 18) as a model system. Our results demonstrated the partial synapsis and pairing of non-homologous chromosomes concurrent with chaotic spindle assembly, affected chromosome assortment and distribution during meiosis, which finally caused difference in genetic constitution amongst the final tetrads. The mitotic cell cycle arrest during microspore development resulted in the production of anucleate pollen cells. Transcription analysis showed that sets of key genes regulating cyclin (CYCA1;2 and CYCA2;3), DNA damage repair (DMC1, NBS1 and MMD1), and ubiquitin-proteasome pathway (SINAT4 and UBC) were largely downregulated at the early pollen meiosis stages, and those genes involved in sperm cell differentiation (DUO1, PIRL1, PIRL9 and LBD27) and pollen wall synthesis (PME48, VGDH11 and COBL10) were mostly repressed at the late pollen mitosis stages in the synthetic allodiploid B. juncea hybrids (AB). In conclusion, this study elucidated the related mechanisms affecting pollen fertility during male gametophyte development at the cytological and transcriptomic levels in the synthetic allodiploid B. juncea hybrids., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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

Soil heavy metal contamination and salinity constitute a major environmental problem worldwide. The affected area and impact of these problems are increasing day by day; therefore, it is imperative to restore their potential using environmentally friendly technology. Plant growth-promoting rhizobacteria (PGPR) provides a better option in this context. Thirty-seven bacteria were isolated from the rhizosphere of maize cultivated in metal- and salt-affected soils. Some selected bacterial strains grew well under a wide range of pH (4-10), salt (5-50 g/L), and Cd (50-1000 mg/L) stress. Three bacterial strains, Exiguobacterium aestuarii (UM1), Bacillus cereus (UM8), and Bacillus megaterium (UM35), were selected because of their robust growth and high tolerance to both stress conditions. The bacterial strains UM1, UM8, and UM35 showed P-solubilization, whereas UM8 and UM35 exhibited 1-aminocyclopropane-1-carboxylate deaminase activity and indole acetic acid (IAA) production, respectively. The bacterial strains were inoculated on Brassica juncea plants cultivated in Cd and salt-affected soils due to the above PGP activities and stress tolerance. Plants inoculated with the bacterial strains B. cereus and B. megaterium significantly (p < 0.05) increased shoot fresh weight (17 ± 1.17-29 ± 0.88 g/plant), shoot dry weight (2.50 ± 0.03-4.40 ± 0.32 g/plant), root fresh weight (7.30 ± 0.58-13.30 ± 0.58 g/plant), root dry weight (0.80 ± 0.04-2.00 ± 0.01 g/plant), and shoot K contents (62.76 ± 1.80-105.40 ± 1.15 mg/kg dwt) in normal and stressful conditions. The bacterial strain B. megaterium significantly (p < 0.05) decreased shoot Na + and Cd ++ uptake in single and dual stress conditions. Both bacterial strains, E. aestuarii and B. cereus, efficiently reduced Cd ++ translocation and bioaccumulation in the shoot. Bacterial inoculation improved the uptake of K + and Ca ++ , while restricted Na + and Cd ++ in B. juncea shoots indicated their potential to mitigate the dual stresses of salt and Cd in B. juncea through ion homeostasis., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Iftikhar Ahmad reports was provided by COMSATS University, Islamabad, Vehari Campus., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Phytophthora blight, caused by Phytophthora capsici , is a serious disease of many vegetable crops worldwide. In New Mexico, U.S.A., the disease affects chile pepper ( Capsicum annuum L.), a major crop in the state. There is no single tool that effectively controls the disease. Continuous research is needed in identifying combination of tools that can reduce the impact of Phytophthora blight. We explored the potential of combining cover crops and biocontrol agents to reduce soilborne diseases. This study aimed to evaluate the effects of Indian mustard ( Brassica juncea L.) cover crop on the antagonistic ability of Trichoderma harzianum against P. capsici in vitro and to quantify the impacts of combining soil amendment with residues from B. juncea and barley ( Hordeum vulgare L.) cover crops and plastic covering on infection of chile pepper seedlings by P. capsici under greenhouse conditions. Volatiles from macerated tissue of B. juncea significantly reduced P. capsici and T. harzianum growth in the absence of soil by 89.0 and 79.0%, respectively. When incorporated in soils, volatiles from macerated tissue of B. juncea significantly reduced P. capsici and T. harzianum by 33.4 and 7.8%, respectively. T. harzianum was more resilient to B. juncea biofumigation than P. capsici. Significant reduction in disease incidence was observed with B. juncea -fumigated soil, while no disease suppression was observed with soil incorporation of H. vulgare residues. Covering soil with plastic was necessary for increasing the efficacy of B. juncea biofumigation., Competing Interests: The author(s) declare no conflict of interest.

Brassica juncea (L.) Czern. & Coss. (Indian mustard) is an economically important edible oil crop. Over the years, plant breeders have developed many elite varieties of B. juncea with better yield traits, but research work on the introgression of stress resilience traits has largely been lagging due to scarcity of resistant donors. Crop wild relatives (CWRs) are the weedy relatives of domesticated plant species which are left unutilized in their natural habitat due to the presence of certain undesirable alleles which hamper their yield potential, and thus, their further domestication. CWRs of B. juncea namely include Sinapis alba L. (White mustard), B. tournefortii Gouan. (African mustard), B. fruticulosa Cirillo (Twiggy turnip), Camelina sativa L. (Gold-of-pleasure), Diplotaxis tenuisiliqua Delile (Wall rocket), D. erucoides L. (White wall rocket), D. muralis L. (Annual wall rocket), Crambe abyssinica R.E.Fr. (Abyssinian mustard), Erucastrum gallicum Willd. (Common dogmustard), E. cardaminoides Webb ex Christ (Dogmustard), Capsella bursa - pastoris L. (Shepherds purse), Lepidium sativum L. (Garden Cress) etc. These CWRs have withstood several regimes of biotic and abiotic stresses over the past thousands of years which led them to accumulate many useful alleles contributing in resistance against various environmental stresses. Thus, CWRs could serve as resourceful gene pools for introgression of stress resilience traits into Indian mustard. This review summarizes research work on the introgression of resistance against Sclerotinia stem rot (caused by Sclerotinia sclerotiorum ), Alternaria blight (caused by Alternaria brassicae ), white rust (caused by Albugo candida ), aphid attack, drought and high temperature from CWRs into B. juncea . However, various pre- and post-fertilization barriers due to different ploidy levels are major stumbling blocks in the success of such programmes, therefore, we also insightfully discuss how the advances made in -omics technology could be helpful in assisting various breeding programmes aiming at improvisation of stress resilience traits in B. juncea ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Verma, Dubey, Singh, Parmar, Singh, Sharma, Rana, Thakur, Vaidya and Thakur.)

Although Brassica juncea has demonstrated potential as a hyperaccumulator crop, it was not entirely clear how cadmium (Cd) accumulates in plants. Here, we found that BjNRAMP1 (Natural Resistance-Associated Macrophage Protein 1) plays a crucial role in the accumulation of Cd and manganese (Mn) through its expression in yeast and Arabidopsis thaliana . The high concentration of Cd exposure could induce the expression of BjNRAMP1 . The ectopic expression of BjNRAMP1 in yeast led to higher accumulation of Cd and Mn compared to the vector control. BjNARAMP1 was localized to the plasma membrane and expressed in the vascular system of roots, leaves, and flowers. The overexpression of BjNRAMP1 in A. thaliana resulted in an increased accumulation of Cd in both roots and shoots, which inhibited the normal growth of transgenic lines. Moreover, Mn uptake in roots was activated by the increase in Cd stress. Together, our results indicated that BjNRAMP1 significantly contributes to the uptake of Mn and Cd in B. juncea., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Li, Li, Wang, Peng, Liu, Deng, Zhang and Yan.)

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

Heavy metal (HM) contamination of soil due to anthropogenic activities has led to bioaccumulation and biomagnification, posing toxic effects on plants by interacting with vital cellular biomolecules such as DNA and proteins. Brassica species have developed complex physiological, biochemical, and molecular mechanisms for adaptability, tolerance, and survival under these conditions. This review summarizes the HM tolerance strategies of Brassica species, covering the role of root exudates, microorganisms, cell walls, cell membranes, and organelle-specific proteins. The first line of defence against HM stress in Brassica species is the avoidance strategy, which involves metal ion precipitation, root sorption, and metal exclusion. The use of plant growth-promoting microbes, Pseudomonas, Psychrobacter, and Rhizobium species effectively immobilizes HMs and reduces their uptake by Brassica roots. The roots of Brassica species efficiently detoxify metals, particularly by flavonoid glycoside exudation. The composition of the cell wall and callose deposition also plays a crucial role in enhancing HMs resistance in Brassica species. Furthermore, plasma membrane-associated transporters, BjCET, BjPCR, BjYSL, and BnMTP, reduce HM concentration by stimulating the efflux mechanism. Brassica species also respond to stress by up-regulating existing protein pools or synthesizing novel proteins associated with HM stress tolerance. This review provides new insights into the HM tolerance mechanisms of Brassica species, which are necessary for future development of HM-resistant crops., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Non-alcoholic fatty liver disease (NAFLD) is mainly characterized by excessive fat accumulation in the liver. It spans a spectrum of diseases from hepatic steatosis to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). Brassica juncea is rich in glucosinolates and has been proven to possess many potential pharmacological properties, including hypoglycemic, anti-oxidation, anti-inflammatory, and anti-carcinogenic activities. This study aims to investigate whether whole-plant Brassica juncea (WBJ) and its glucosinolates extracts (BGE) have hepatoprotective effects against a high-fat diet (HFD)-induced NAFLD and further explore the mechanism underlying this process in vivo and in vitro. WBJ treatment significantly reduced body fat, dyslipidemia, hepatic steatosis, liver injury, and inflammation; WBJ treatment also reversed the antioxidant enzyme activity to attenuate oxidative stress in HFD-fed rat liver. Moreover, WBJ and BGE enhanced the activation of AMPK to reduce SREBPs, fatty acid synthase, and HMG-CoA reductase but increased the expression of CPT-I and PPARα to improve hepatic steatosis. In addition, WBJ and BGE could ameliorate NAFLD by inhibiting TNF-α and NF-κB. Based on the above results, this study demonstrates that WBJ and BGE ameliorate HFD-induced hepatic steatosis and liver injury. Therefore, these treatments could represent an unprecedented hope toward improved strategies for NAFLD.

Heavy metal pollution of soil is a major concern due to its non-biodegradable nature, bioaccumulation, and persistence in the environment. To explore the probable function of EDTA in ameliorating heavy metal toxicity and achieve the sustainable development goal (SDG), Brassica juncea L. seedlings were treated with different concentrations of EDTA (0, 1.0, 2.0, 3.0, and 4.0 mM Kg -1 ) in heavy metal-polluted soil. Plant samples were collected 60 days after sowing; photosynthetic pigments, H 2 O 2 , monoaldehyde (MDA), antioxidant enzymes, and ascorbic acid content, as well as plant biomass, were estimated in plants. Soil and plant samples were also examined for the concentrations of Cd, Cr, Pb, and Hg. Moreover, values of the phytoremediation factor were utilized to assess the accumulation capacity of heavy metals by B. juncea under EDTA treatments. In the absence of EDTA, B. juncea seedlings accrued heavy metals in their roots and shoots in a concentration-dependent manner. However, the highest biomass of plants (roots and shoots) was recorded with the application of 2 mM kg -1 EDTA. Moreover, high levels (above 3 mM kg -1 ) of EDTA concentration have reduced the biomass of plants (roots and shoots), photosynthetic area, and chlorophyll content. The effect of EDTA levels on photosynthetic pigments (chlorophyll a and b) revealed that with an increment in EDTA concentration, accumulation of heavy metals was also increased in the plant, subsequently decreasing the chlorophyll a and b concentration in the plant. TLF was found to be in the order Pb> Hg> Zn> and >Ni, while TF was found to be in the order Hg>Zn>Ni>Pb, and the best dose was 3 mM kg -1 EDTA for Hg and 4 mM kg -1 for Pb, Ni, and Zn. Furthermore, hyperaccumulation of heavy metals enhanced the generation of hydrogen peroxide (H 2 O 2 ), superoxide anions (O 2 •- ), and lipid peroxidation. It also interrupts mechanisms of the antioxidant defense system. Furthermore, heavy metal stress reduced plant growth, biomass, and chlorophyll (chl) content. These findings suggest that the exogenous addition of EDTA to the heavy metal-treated seedlings increases the bioavailability of heavy metals for phytoextraction and decreases heavy metal-induced oxidative injuries by restricting heavy metal uptake and components of their antioxidant defense systems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Kamal, Perveen, Khan, Sayyed, Hock, Bhatt, Singh and Qamar.)

The health-beneficial effects of Brassica vegetables are mainly attributed to their high contents of glucosinolates and the products of their hydrolysis, especially isothiocyanates. Distribution of glucosinolates across plant organs can strongly vary. Here, we investigated the effect of leaf age on glucosinolate accumulation and hydrolysis in two leafy Brassica vegetables, pak choi and giant red mustard. We also evaluated the activity of the hydrolyzing enzyme myrosinase across the leaves. Finally, we assessed whether glucosinolates are transported from older leaves to younger leaves. Young leaves of both species contained more than 3-fold more glucosinolates than older ones. Accordingly, more isothiocyanates were released in the young leaves. Myrosinases fully hydrolyzed all of the amounts of glucosinolates regardless of the leaf age. Moreover, older leaves were observed to supply younger leaves with glucosinolates. Thus, this study suggests that consumers can improve the nutritional value of food by incorporating young leaves of leafy Brassicas in their diet.

Brassica species show varying levels of resistance to salt stress. To understand the genetics underlying these differential stress tolerance patterns in Brassicas, we exposed two widely cultivated amphidiploid Brassica species having different genomes, Brassica juncea (AABB, n = 18) and Brassica napus (AACC, n = 19), to elevated levels of NaCl concentration (300 mM, half the salinity of seawater). B. juncea produced more biomass, an increased chlorophyll content, and fewer accumulated sodium (Na + ) and chloride (Cl - ) ions in its photosynthesizing tissues. Chlorophyll fluorescence assays revealed that the reaction centers of PSII of B. juncea were more photoprotected and hence more active than those of B. napus under NaCl stress, which, in turn, resulted in a better PSII quantum efficiency, better utilization of photochemical energy with significantly reduced energy loss, and higher electron transport rates, even under stressful conditions. The expression of key genes responsible for salt tolerance ( NHX1 and AVP1 , which are nuclear-encoded) and photosynthesis ( psbA , psaA , petB , and rbcL , which are chloroplast-encoded) were monitored for their genetic differences underlying stress tolerance. Under NaCl stress, the expression of NHX1 , D1 , and Rubisco increased several folds in B. juncea plants compared to B. napus , highlighting differences in genetics between these two Brassicas. The higher photosynthetic potential under stress suggests that B. juncea is a promising candidate for genetic modifications and its cultivation on marginal lands.

The lack of resistance to Lipaphis erysimi in cultivated Brassicas makes caused this pest highly devastating resulting in significant loss of rapeseed-mustard productivity in India. B. fruticulosa , a wild crucifer is known as an excellent source of resistance to L. erysimi . Therefore, we planned to assess defense associated biochemical alterations and molecular components of B. juncea - B. fruticulosa ILs to mustard aphid. Phenotypic assessment of ILs on the basis of aphid population per plant (APP) categorized genotypes into resistant (7.15-18.50 APP), moderately susceptible (42.29-53.33 APP) and susceptible (70.00-77.07 APP) genotypes. Mustard aphid infested minimally B. fruticulosa (0.80 APP) among tested genotypes. The maximum increase in catalase (CAT) activity was determined in B. fruticulosa and resistant ILs after 48 h (2.03 and 1.76-fold, respectively) and one week (2.98 and 1.79-fold, respectively) of mustard aphid infestation. The strong induction of CAT2 transcripts (19.25-fold) and CAT activity (5.88-fold) along with low aphid count in resistant IL, Ad 4 -64 (13.85 APP) suggested the pivotal role of CAT in resistance to mustard aphid. Guaiacol peroxidase (GPX) was significantly decreased following pest infestation at both infestation stages. The ascorbate content was highest in resistant IL, ADV-6RD (2.14-fold) after one week of aphid infestation. H 2 O 2 content rapidly increased in B. juncea - B. fruticulosa derived lines after 48 h of aphid infestation. The negative and significant association between APP and CAT (- 0.56** and - 0.48*, respectively), glutathione (- 0.43* and - 0.40*, respectively), H 2 O 2 (- 0.57** and - 0.43*, respectively) at both 48 h and one week infestation stages signified their role in deterring mustard aphid infestation. The positive and significant association between total sugars (0.33* at 7 DPI), reducing sugars (0.33* at 7 DPI), sucrose (0.36** at 48 h) and APP indicated that higher the sugars content, higher will be mustard aphid infestation in B. juncea derived ILs. The information being generated and key candidates ( CAT2 , ascorbate and H 2 O 2 ) being identified may help in effective deployment of B. fruticulosa resistance in mustard breeding., Competing Interests: Conflict of interestThe research leading to these results received funding from Department of Science and Technology, New Delhi under INSPIRE Faculty Research Grant number no. DST/INSPIRE Faculty Award/2017/DST/INSPIRE/04/2016/001086 and DST/INSPIREFaculty/2018/P19053., (© Prof. H.S. Srivastava Foundation for Science and Society 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

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

Key Message: Overexpression of BjFLD in Brassica juncea imparts resistance against fungal pathogens and increases the yield. These transgenics could lower the use of fungicides, which have detrimental effects on the environment. Productivity of Indian mustard (Brassica juncea) is adversely affected by fungal phytopathogens, Alternaria brassicae and Sclerotinia sclerotiorum. Arabidopsis flowering locus D (FLD) positively regulates jasmonic acid signaling and defense against necrotrophic pathogens. In this study, the endogenous FLD (B. juncea FLD; BjFLD) in Indian mustard was overexpressed in B. juncea to determine its role in biotic stress tolerance. We report the isolation, characterization, and functional validation of BjFLD. The transgene expression was confirmed by qRT-PCR. The constitutive overexpression of BjFLD enhanced the tolerance of B. juncea to A. brassicae and S. sclerotiorum, which was manifested as delayed appearance of symptom, impeded disease progression, and enhanced percentage of disease protection. The transgenic lines maintained a higher photosynthetic capacity and redox potential under biotic stress and could detoxify reactive oxygen species (ROS) by modulating the antioxidant machinery and physiochemical attributes. The BjFLD-overexpressing lines showed enhanced SA level as well higher NPR1 expression. The overexpression of BjFLD induced early flowering and higher seed yield in the transgenic lines. These findings indicate that overexpression of BjFLD enhances the tolerance of B. juncea to A. brassicae and S. sclerotiorum by induction of systemic acquired resistance and mitigating the damage caused by stress-induced ROS., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The pale cyst nematode Globodera pallida is a highly specialized, economically important pest for potato production. The specialized hatching requirements, ability to adapt, and the loss of effective control strategies such as methyl bromide fumigation increase the challenge to eradicate G. pallida in Idaho. Without a suitable host, this nematode can remain dormant as encysted eggs in soil for up to 20 years. In this study, we first demonstrated that Sinapis alba seed meal extract (SME) or 4-hydroxybenzyl alcohol (HBA), under laboratory and greenhouse conditions, enhances G. pallida egg hatch rate when exposed to potato root diffusate (PRD). This hatch rate enhancement in the presence of PRD is speculated to be due to an increase in egg-shell permeability. We then tested the efficacy of (i) Solanum sisymbriifolium following prior treatment with S. alba SME (0 and 4.48 t/ha) or HBA (0 and 0.12 t/ha) and (ii) Brassica juncea SME (0, 0.14, 0.56, and 1.12 t/ha) following HBA treatment (0 and 4.48 t/ha) on egg viability, hatch rate, and reproduction of G. pallida encysted eggs. S. sisymbriifolium alone reduced the number of encysted eggs compared to the nontreated control by up to 67%, indicating that this trap crop triggered G. pallida eggs to hatch. When combined with S. alba SME or HBA, S. sisymbriifolium significantly reduced egg count, hatch rate, and viability more than S. sisymbriifolium alone. The combination of S. sisymbriifolium with HBA or S. alba SME eliminated G. pallida reproduction on the susceptible potato. All the tested rates of B. juncea SME alone or with HBA reduced egg hatch rate, viability, and reproduction compared to the nontreated control. Combining HBA and B. juncea SME further significantly reduced egg hatch rate, viability, and reproduction than those rates of B. juncea SME alone., Competing Interests: The author(s) declare no conflict of interest.

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

Rhythmic exposure to moonlight has been shown to affect animal behavior, but its effects on plants, often observed in lunar agriculture, have been doubted and often regarded as myth. Consequently, lunar farming practices are not well scientifically supported, and the influence of this conspicuous environmental factor, the moon, on plant cell biology has hardly been investigated. We studied the effect of full moonlight (FML) on plant cell biology and examined changes in genome organization, protein and primary metabolite profiles in tobacco and mustard plants and the effect of FML on the post-germination growth of mustard seedlings. Exposure to FML was accompanied by a significant increase in nuclear size, changes in DNA methylation and cleavage of the histone H3 C-terminal region. Primary metabolites associated with stress were significantly increased along with the expression of stress-associated proteins and the photoreceptors phytochrome B and phototropin 2; new moon experiments disproved the light pollution effect. Exposure of mustard seedlings to FML enhanced growth. Thus, our data show that despite the low-intensity light emitted by the moon, it is an important environmental factor perceived by plants as a signal, leading to alteration in cellular activities and enhancement of plant growth.

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