Search

Low temperature is one of the environmental factors that restrict the growth and geographical distribution of Brassica. To investigate the effects of exogenous calcium and calcium inhibitor on the ability of winter turnip rapeseed (Brassica rapa L.) to withstand low temperatures (4℃), we used a strong cold-resistant variety Longyou 7 (L7) and a weak cold-resistant variety Longyou 99 (L99) as the materials. The seedlings were treated with CaCl 2 (20 mmol·L -1 ) and calcium inhibitor LaCl 3 (10 mmol·L -1 ) at 0 h (CK), 6 h, 12 h, 24 h and 48 h after 4℃ treatments. Physiological characteristics, Ca 2+ flux and Ca 2+ concentration in roots after treatments were analyzed. Results illustrated that under 4℃ treatment, activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) increased by both CK and exogenous CaCl 2 treatments. Contents of soluble protein (SP) and proline (Pro) increased, while contents of malondialdehyde (MDA) decreased, resulting in reduced membrane lipid peroxidation. But enzyme activity decreased and MDA content increased following treatment with exogenous LaCl 3 . The rate of Ca 2+ flow showed a higher uptake in L7 roots compared with L99. L99 showed Ca 2+ efflux with a rate of 30.21 pmol‧cm -2 ‧s -1 , whereas L7 showed short efflux then returned to influx. Calcium ion content in roots decreased in both cultivars after CaCl 2 treatment. Results of RNA-seq revealed that genes were differentially expressed in response to low temperatures, hormones, photosystem II, chloroplasts, DNA replication, ribosomal RNA processing, and translation. This study found significant expression genes related to cellular signal transduction (MAPK signaling pathway) and material metabolism (nitrogen metabolism, glycerol ester metabolism).It was also analyzed by WGCNA that two modules had the strongest correlation with physiological indicators. Eight candidate genes were identified among MAPK signaling pathway and the two modules., (© 2024. The Author(s).)

Drought is a major abiotic stress factor that reduces agricultural productivity. Understanding the molecular regulatory network of drought response in winter rape is of great significance for molecular Brassica rapa . In order to comprehensively analyze the network expression of DEGs and DEMIs in winter rape under drought stress, in this study we used Longyou 7 as the experimental material to identify DEGs and DEMIs related to drought stress by transcriptome and miRNA sequencing. A total of 14-15 key differential mRNA genes related to drought stress and biological stress were screened out under different treatments in the three groups. and 32 differential miRNAs were identified through targeted regulatory relationships, and the mRNA expression of 20 target genes was negatively regulated by the targeting regulatory relationship. It is mainly enriched in starch and sucrose metabolism, carbon metabolism and other pathways. Among them, gra-MIR8731-p3_2ss13GA18GA regulated the expression of multiple mRNAs in the three treatments. miRNA is mainly involved in the drought resistance of Chinese cabbage winter rape by regulating the expression of target genes, such as starch and sucrose metabolism, amino acid biosynthesis, and carbon metabolism. These miRNAs and their target genes play an indispensable role in winter rapeseed drought stress tolerance regulation.

The global expansion of domesticated plant and animal species has profoundly impacted biodiversity and ecosystem functions. However, the spillover effect of non-native honey bees from mass-flowering crops into adjacent natural vegetation on pollination function within plant communities remains unclear. To address this, we conduct field experiments to investigate the ecological impacts of honey bees (Apis mellifera) and a mass-flowering crop (Brassica rapa var. oleifera) on pollinator communities, plant-pollinator interactions, and reproductive performance of wild plants in 48 pollinator-limited alpine grasslands. Our findings indicate that the transition of dominant pollinators from flies to honey bees enhances visitation fidelity of pollinator species and reconfigures pollination interactions due to an increase in competition between honey bees and native pollinator species. Additionally, honey bees increase, decrease or do not alter plant reproductive success, depending on the plant species. Here, we report the mixed effects of honey bees on pollination function in pollinator-limited alpine grasslands., (© 2024. The Author(s).)

Gibberellic acid-stimulated Arabidopsis sequences ( GASAs ) are a subset of the gibberellin (GA)-regulated gene family and play crucial roles in various physiological processes. However, the GASA genes in Brassica rapa have not yet been documented. In this study, we identified and characterized 16 GASA genes in Chinese cabbage ( Brassica rapa L. ssp. pekinensis ). Analysis of the conserved motifs revealed significant conservation within the activation segment of BraGASA genes. This gene family contains numerous promoter elements associated with abiotic stress tolerance, including those for abscisic acid (ABA) and methyl jasmonate (MeJA). Expression profiling revealed the presence of these genes in various tissues, including roots, stems, leaves, flowers, siliques, and callus tissues. When plants were exposed to drought stress, the expression of BraGASA3 decreased notably in drought-sensitive genotypes compared to their wild-type counterparts, highlighting the potentially crucial role of BraGASA3 in drought stress. Additionally, BraGASAs exhibited various functions in sexual reproduction dynamics. The findings contribute to the understanding of the function of BraGASAs and provide valuable insights for further exploration of the GASA gene function of the BraGASA gene in Chinese cabbage.

Waterlogging stress, particularly during seed germination, significantly affects plant growth and development. However, the physiological and molecular mechanisms underlying waterlogging stress responses during rapeseed germination remain unclear. In this study, two rapeseed cultivars, Xiangzayou518 (waterlogging-sensitive) and Dadi199 (waterlogging-tolerant), were used to explore the physiological mechanisms underlying rapeseed response to waterlogging stress during germination. Our results showed that waterlogging significantly decreased the emergence percentage and seedling growth rate. During the radicle elongation period (from 48 to 96 h post-germination), the most sensitive period to waterlogging during germination, sugar content, and glycolysis efficiency were significantly decreased, but anaerobic fermentation was enhanced. In tolerant cultivars, when the energy supply was insufficient, the conversion efficiency of lipids into sugar increased, and the activities of isocitrate lyase, malate synthase, and fructose-1, 6-diphosphatase were enhanced by 11.63, 19.06, and 20.37%, respectively, at 72 h post-germination under waterlogging stress. Transcriptome data showed that the differentially expressed genes were significantly enriched in glucose and lipid metabolism pathways when comparing waterlogged stress and normal conditions. These results indicate that waterlogging affects seed germination in rapeseed by inhibiting carbohydrate metabolism, and the conversion capacity of lipids into sugar under waterlogging stress was stronger in tolerant cultivars than in sensitive cultivars, thus rescuing the insufficient energy supply in seed germination and seedling growth. This study reveals the physiological mechanism of rapeseed response to waterlogging stress during seed germination and provides a valuable reference for improving waterlogging tolerance., (© 2024 Scandinavian Plant Physiology Society. Published by John Wiley & Sons Ltd.)

The current agricultural system is in search of new strategies to achieve a more sustainable production while keeping or even increasing crop yield and quality. In this scenario, the application of biostimulants constitutes a potent solution. In the current study, the impact of a blue-green microalgal extract (MB) and a pig tissue hydrolysate (PTH) on rapeseed plants' development was characterized. Obtained results revealed a positive effect on yield parameters of plants treated with MB and, especially, PTH; this was associated to an improvement on the photosynthetic performance. Moreover, this study remarked the effects of biostimulants on plant phenology through their pivotal role in modulating developmental processes. More specifically, proteomic, metabolomic, and hormone content analyses revealed distinct alterations associated with the acceleration of phenology induced by biostimulant application. Additionally, some antioxidant enzymes and stress-related compounds were up-regulated upon MB and PTH treatments, indicating enhanced plant defense mechanisms in response to accelerated phenological transitions. Such findings highlight the intricate interplay between biostimulants and plant physiology, wherein biostimulants orchestrate rapid developmental changes, ultimately influencing growth dynamics. Altogether, the current study reveals that the application of both MB and PTH biostimulants promoted rapeseed plant phenology and productivity associated with an improvement in the photosynthetic machinery while boosting other physiological and molecular mechanisms., (© 2024 The Author(s). Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

Shifts in pollinator occurrence and their pollen transport effectiveness drive the evolution of mating systems in flowering plants. Understanding the genomic basis of these changes is essential for predicting the persistence of a species under environmental changes. We investigated the genomic changes in Brassica rapa over nine generations of pollination by hoverflies associated with rapid morphological evolution toward the selfing syndrome. We combined a genotyping-by-sequencing (GBS) approach with a genome-wide association study (GWAS) to identify candidate genes, and assessed their functional role in the observed morphological changes by studying mutations of orthologous genes in the model plant Arabidopsis thaliana. We found 31 candidate genes involved in a wide range of functions from DNA/RNA binding to transport. Our functional assessment of orthologous genes in A. thaliana revealed that two of the identified genes in B. rapa are involved in regulating the size of floral organs. We found a protein kinase superfamily protein involved in petal width, an important trait in plant attractiveness to pollinators. Moreover, we found a histone lysine methyltransferase (HKMT) associated with stamen length. Altogether, our study shows that hoverfly pollination leads to rapid evolution toward the selfing syndrome mediated by polygenic changes., (© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.)

Increased temperature can induce plastic changes in many plant traits. However, little is known about how these changes affect plant interactions with insect pollinators and herbivores, and what the consequences for plant fitness and selection are. We grew fast-cycling Brassica rapa plants at two temperatures (ambient and increased temperature) and phenotyped them (floral traits, scent, colour and glucosinolates). We then exposed plants to both pollinators (Bombus terrestris) and pollinating herbivores (Pieris rapae). We measured flower visitation, oviposition of P. rapae, herbivore development and seed output. Plants in the hot environment produced more but smaller flowers, with lower UV reflectance and emitted a different volatile blend with overall lower volatile emission. Moreover, these plants received fewer first-choice visits by bumblebees and butterflies, and fewer flower visits by butterflies. Seed production was lower in hot environment plants, both because of a reduction in flower fertility due to temperature and because of the reduced visitation of pollinators. The selection on plant traits changed in strength and direction between temperatures. Our study highlights an important mechanism by which global warming can change plant-pollinator interactions and negatively impact plant fitness, as well as potentially alter plant evolution through changes in phenotypic selection., (© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.)

Pollination is crucial for biodiversity and food security. Heterogeneous agricultural landscapes have a positive effect on pollinator abundance and enhance crop production and quality. In this study, we explored the effects of three landscape features (past crop diversity measured as the Equivalent Richness of crop functional Groups in the previous year [ERGp], semi-natural habitat percentage [SNH], and mean field size [MFS]) and pollinator densities (wild bees [WB] and honey bees [HB]) on pollination and seed quantity and quality in rapeseed crops. Surveying the pollinator density in 20 rapeseed fields revealed a positive relationship with ERGp in the landscape. A pollinator exclusion experiment compared bagged and open-pollinated self-compatible rapeseed plants and revealed insect pollination effectiveness (fruits per flower and number of seeds per pod) and seed quality (oil content). Seed parameters were evaluated in relation to pollinator density (WB-HB) and landscape characteristics. The ERGp emerged as a crucial landscape feature that positively impacted WB density. When insect pollinators were excluded, plants exhibited reduced pollination effectiveness and seed quality. Analysis of open-pollinated plants highlighted ERGp as the most influential variable, positively affecting both sets of parameters. The MFS and SNH showed different but important relationships. Total tocopherol and α-tocopherol were positively correlated with pollinator density in HB, whereas WB showed a positive correlation with γ-tocopherol levels. Increased ERGp positively affected pollinator density and pollination effectiveness, thereby improving oilseed rape production quantity and quality. This study provides new insights into agroecosystem management and pollinator-friendly practices., Competing Interests: Declaration of competing interest The authors declare no competing financial interests or personal relationships that may have influenced the work reported in this study., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

The increasing use of antibiotics has attracted widespread attention to their environmental risks. However, the phototoxicity of sulfonamide antibiotics to plants remain unclear. In this study, the mechanism of the effect of sulfamethoxazole on photosynthesis of pakchoi cabbage (Brassica rapa var. chinensis) was investigated. The results showed that sulfamethoxazole inhibited the growth of pakchoi cabbage and produced photosynthetic toxicity. The growth inhibition rates increased with concentration, the root and shoot weight were 76.02 % and 47.04 % of the control, respectively, with stay-greens phenomenon in 4 mg·L -1 sulfamethoxazole treatment. Chlorophyll precursors (protoporphyrin IX (Proto IX), Mg-proto IX, and protochlorophyllide (Pchlide), 5-aminolevulinic acid (ALA), and porphobilinogen (PBG)) were 1.38-, 1.26-, 1.12-, 1.71-, and 0.96-fold of the control, respectively; photosynthetic pigments (chlorophyll a, chlorophyll b, and carotenoids) were 1.26-, 1.39-, and 1.03-fold of the control, respectively. Respiration rate was 271.42 % of the control, whereas the net photosynthetic rate was 50.50 % of the control. The maximum photochemical quantum yield of PSII (F v /F m ), the actual photosynthetic efficiency (Y(II)), the quantum yield of non-regulated energy dissipation (Y(NO)), the apparent electron transfer efficiency of PSII (ETR) under actual light intensity were affected, and chloroplast swelling was observed. Proteomic analysis showed that photosynthesis-related pathways were significantly up-regulated, biological processes such as light response, carbohydrates, and reactive oxygen species were activated. Metabolomic analysis revealed that the tricarboxylic acid cycle (TCA cycle) and carbohydrate catabolism were stimulated significantly (p < 0.05), sugars and amino acids were increased to regulate and enhance the resilience of photosynthesis. While folate biosynthesis and ribosomal pathways were significantly down-regulated, the synthesis and translation processes of amino acids and nucleotides were inhibited., 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.)

Global warming has a severe impact on the flowering time and yield of crops. Histone modifications have been well-documented for their roles in enabling plant plasticity in ambient temperature. However, the factor modulating histone modifications and their involvement in habitat adaptation have remained elusive. In this study, through genome-wide pattern analysis and quantitative-trait-locus (QTL) mapping, we reveal that BrJMJ18 is a candidate gene for a QTL regulating thermotolerance in thermotolerant B. rapa subsp. chinensis var. parachinensis (or Caixin, abbreviated to Par). BrJMJ18 encodes an H3K36me2/3 Jumonji demethylase that remodels H3K36 methylation across the genome. We demonstrate that the BrJMJ18 allele from Par (BrJMJ18 Par ) influences flowering time and plant growth in a temperature-dependent manner via characterizing overexpression and CRISPR/Cas9 mutant plants. We further show that overexpression of BrJMJ18 Par can modulate the expression of BrFLC3, one of the five BrFLC orthologs. Furthermore, ChIP-seq and transcriptome data reveal that BrJMJ18 Par can regulate chlorophyll biosynthesis under high temperatures. We also demonstrate that three amino acid mutations may account for function differences in BrJMJ18 between subspecies. Based on these findings, we propose a working model in which an H3K36me2/3 demethylase, while not affecting agronomic traits under normal conditions, can enhance resilience under heat stress in Brassica rapa., (© 2024. The Author(s).)

Although different ecological factors shape adaptative evolution in natural habitats, we know little about how their interactions impact local adaptation. Here we used eight generations of experimental evolution with outcrossing Brassica rapa plants as a model system, in eight treatment groups that varied in soil type, herbivory (with/without aphids), and pollination mode (hand- or bumblebee-pollination), to study how biotic interactions affect local adaptation to soil. First, we show that several plant traits evolved in response to biotic interactions in a soil-specific way. Second, using a reciprocal transplant experiment, we demonstrate that significant local adaptation to soil-type evolved in the "number of open flowers", a trait used as a fitness proxy, but only in plants that evolved with herbivory and bee pollination. Whole genome re-sequencing of experimental lines revealed that biotic interactions caused a 10-fold increase in the number of SNPs across the genome with significant allele frequency change, and that alleles with opposite allele frequency change in different soil types (antagonistic pleiotropy) were most common in plants with an evolutionary history of herbivory and bee pollination. Our results demonstrate that the interaction with mutualists and antagonists can facilitate local adaptation to soil type through antagonistic pleiotropy., (© 2024. The Author(s).)

Main Conclusion: In Brassica rapa, the epigenetic modifier BraA.CLF orchestrates flowering by modulating H3K27me3 levels at the floral integrator genes FT, SOC1, and SEP3, thereby influencing their expression. CURLY LEAF (CLF) is the catalytic subunit of the plant Polycomb Repressive Complex 2 that mediates the trimethylation of histone H3 lysine 27 (H3K27me3), an epigenetic modification that leads to gene silencing. While the function of CURLY LEAF (CLF) has been extensively studied in Arabidopsis thaliana, its role in Brassica crops is barely known. In this study, we focused on the Brassica rapa homolog of CLF and found that the loss-of-function mutant braA.clf-1 exhibits an accelerated flowering together with pleiotropic phenotypic alterations compared to wild-type plants. In addition, we carried out transcriptomic and H3K27me3 genome-wide analyses to identify the genes regulated by BraA.CLF. Interestingly, we observed that several floral regulatory genes, including the B. rapa homologs of FT, SOC1 and SEP3, show reduced H3K27me3 levels and increased transcript levels compared to wild-type plants, suggesting that they are direct targets of BraA.CLF and key players in regulating flowering time in this crop. In addition, the results obtained will enhance our understanding of the epigenetic mechanisms regulating key developmental traits and will aid to increase crop yield by engineering new Brassica varieties with different flowering time requirements., (© 2024. The Author(s).)

Increased exposure to environmental stresses due to climate change have adversely affected plant growth and productivity. Upon stress, plants activate a signaling cascade, involving multiple molecules like H 2 O 2, and plant hormones such as salicylic acid (SA) leading to resistance or stress adaptation. However, the temporal ordering and composition of the resulting cascade remains largely unknown. In this study we developed a nanosensor for SA and multiplexed it with H 2 O 2 nanosensor for simultaneous monitoring of stress-induced H 2 O 2 and SA signals when Brassica rapa subsp. Chinensis (Pak choi) plants were subjected to distinct stress treatments, namely light, heat, pathogen stress and mechanical wounding. Nanosensors reported distinct dynamics and temporal wave characteristics of H 2 O 2 and SA generation for each stress. Based on these temporal insights, we have formulated a biochemical kinetic model that suggests the early H 2 O 2 waveform encodes information specific to each stress type. These results demonstrate that sensor multiplexing can reveal stress signaling mechanisms in plants, aiding in developing climate-resilient crops and pre-symptomatic stress diagnoses., (© 2024. The Author(s).)

Melatonin is a master regulator of diverse biological processes, including plant's abiotic stress responses and tolerance. Despite the extensive information on the role of melatonin in response to abiotic stress, how plants regulate endogenous melatonin content under stressful conditions remains largely unknown. In this study, we computationally mined Expressed Sequence Tag (EST) libraries of salinity-exposed Chinese cabbage (Brassica rapa) to identify the most reliable differentially expressed miRNA and its target gene(s). In light of these analyses, we found that miR168a potentially targets a key melatonin biosynthesis gene, namely O-METHYLTRANSFERASE 1 (OMT1). Accordingly, molecular and physiochemical evaluations were performed in a separate salinity experiment using contrasting B. rapa genotypes. Then, the association between B. rapa salinity tolerance and changes in measured molecular and physiochemical characteristics was determined. Results indicated that the expression profiles of miR168a and OMT1 significantly differed between B. rapa genotypes. Moreover, the expression profiles of miR168a and OMT1 significantly correlated with more melatonin content, robust antioxidant activities, and better ion homeostasis during salinity stress. Our results suggest that miR168a plausibly mediates melatonin biosynthesis, mainly through the OMT1 gene, under salinity conditions and thereby contributes to the salinity tolerance of B. rapa. To our knowledge, this is the first report on the role of miR168a and OMT1 in B. rapa salinity response., (© 2022 Scandinavian Plant Physiology Society.)

Key Message: Nonsense-mediated mRNA decay (NMD)-mediated degradation of BrFLC2 transcripts is the main cause of rapid flowering of oilseed-type B. rapa 'LP08' plants. Many Brassica species require vernalization (long-term winter-like cooling) for transition to the reproductive stage. In the past several decades, scientific efforts have been made to discern the molecular mechanisms underlying vernalization in many species. Thus, to identify the key regulators required for vernalization in Brassica rapa L., we constructed a linkage map composed of 7833 single nucleotide polymorphism markers using the late-flowering Chinese cabbage (B. rapa L. ssp. pekinensis) inbred line 'Chiifu' and the early-flowering yellow sarson (B. rapa L. ssp. trilocularis) line 'LP08' and identified a single major QTL on the upper-arm of the chromosome A02. In addition, we compared the transcriptomes of the lines 'Chiifu' and 'LP08' at five vernalization time points, including both non-vernalized and post-vernalization conditions. We observed that BrFLC2 was significantly downregulated in the early flowering 'LP08' and had two deletion sites (one at 4th exon and the other at 3' downstream region) around the BrFLC2 genomic region compared with the BrFLC2 genomic region in 'Chiifu'. Large deletion at 3' downstream region did not significantly affect transcription of both sense BrFLC2 transcript and antisense transcript, BrFLC2as along vernalization time course. However, the other deletion at 4th exon of BrFLC2 resulted in the generation of premature stop codon in BrFLC2 transcript in LP08 line. Cycloheximide treatment of LP08 line showed the de-repressed level of BrFLC2 in LP08, suggesting that low transcript level of BrFLC2 in LP08 might be caused by nonsense-mediated mRNA decay removing the nonsense transcript of BrFLC2. Collectively, this study provides a better understanding of the molecular mechanisms underlying floral transition in B. rapa., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Arabinogalactan proteins (AGPs) are a superfamily of hydroxyproline-rich glycoproteins that are massively glycosylated, widely implicated in plant growth and development. No comprehensive analysis of the AGP gene family has been performed in Chinese cabbage ( Brassica rapa ssp. chinensis ). Here, we identified a total of 293 putative AGP-encoding genes in B. rapa , including 25 classical AGPs, three lysine-rich AGPs, 30 AG-peptides, 36 fasciclin-like AGPs (FLAs), 59 phytocyanin-like AGPs, 33 xylogen-like AGPs, 102 other chimeric AGPs, two non-classical AGPs and three AGP/extensin hybrids. Their protein structures, phylogenetic relationships, chromosomal location and gene duplication status were comprehensively analyzed. Based on RNA sequencing data, we found that 73 AGP genes were differentially expressed in the floral buds of the sterile and fertile plants at least at one developmental stage in B. rapa , suggesting a potential role of AGPs in male reproductive development. We further characterized BrFLA2 , BrFLA28 and BrFLA32 , three FLA members especially expressed in anthers, pollen grains and pollen tubes. BrFLA2 , BrFLA28 and BrFLA32 are indispensable for the proper timing of pollen germination under high relative humidity. Our study greatly extends the repertoire of AGPs in B. rapa and reveals a role for three members of the FLA subfamily in pollen germination.

Salinity is an important environmental factor that reduces plant productivity in many world regions. It affects negatively photosynthesis causing a growth reduction. Likewise, calcium (Ca 2+ ) is crucial in plant stress response. Therefore, the modification of Ca 2+ cation exchangers (CAX) transporters could be a potential strategy to increase plant tolerance to salinity. Using Targeting Induced Local Lesions in Genomes (TILLING), researchers generated three mutants of Brassica rapa CAX1a transporter: BraA.cax1a-7, BraA.cax1a-4, and BraA.cax1a-12. The aim of this study was to test the effect of those mutations on salt tolerance focusing on the response to the photosynthesis process. Thus, the three BraA.cax1a mutants and the parental line (R-o-18) were grown under salinity conditions, and parameters related to biomass, photosynthesis performance, glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49), and soluble carbohydrates were measured. BraA.cax1a-4 provided higher biomass and a better photosynthetic performance manifested by higher water use efficiency (WUE), Fv/Fm, electron fluxes, and Rubisco (EC 4.1.1.39) values. In addition, BraA.cax1a-4 presented increased osmotic protection through myo-inositol accumulation. On the other hand, BraA.cax1a-7 produced some negative effects on photosynthesis performance and lower G6PDH and Rubisco accumulations. Therefore, this study points out BraA.cax1a-4 as a useful mutation to improve photosynthetic performance in plants grown under saline conditions., (Copyright © 2021 Elsevier B.V. All rights reserved.)

With the discovery of essential genes regulating tillering, such as MONOCULM 1 (MOC1) in rice and LATERAL SUPPRESSOR (LAS in Arabidopsis, LS in tomato), research on tillering mechanisms has made great progress; however, the study of tillering in non-heading Chinese cabbage (NHCC) is rare. Here, we report that BcLAS, as a member of the GRAS family, plays an important role in the tillering of NHCC during its vegetative growth. BcLAS was almost not expressed in other examed parts except leaf axils throughout life. When the expression of BcLAS was silenced utilizing virus-induced gene silencing (VIGS) technology, we found that the tiller number of 'Maertou' decreased sharply. In 'Suzhouqing', overexpression of BcLAS significantly promoted tillering. BcCCS52, the orthologue to CELL CYCLE SEITCH 52 (CCS52), interacts with BcLAS. Downregulation of the expression of BcCCS52 promoted tillering of 'Suzhouqing'; therefore, we conclude that BcCCS52 plays a negative role in tillering regulation. Our findings reveal the tillering regulation mechanism of NHCCs at the vegetative stage and report an orthologue of CCS52 regulating tillering in NHCC., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Plant phosphoprotein phosphatases are ubiquitous and multifarious enzymes that respond to developmental requirements and stress signals through reversible dephosphorylation of target proteins. In this study, we investigated the hitherto unknown functions of Brassica rapa protein phosphatase 5.2 ( BrPP5.2 ) by transgenic overexpression of B. rapa lines. The overexpression of BrPP5.2 in transgenic lines conferred heat shock tolerance in 65-89% of the young transgenic seedlings exposed to 46 °C for 25 min. The examination of purified recombinant BrPP5.2 at different molar ratios efficiently prevented the thermal aggregation of malate dehydrogenase at 42 °C, thus suggesting that BrPP5.2 has inherent chaperone activities. The transcriptomic dynamics of transgenic lines, as determined using RNA-seq, revealed that 997 and 1206 (FDR < 0.05, logFC ≥ 2) genes were up- and down-regulated, as compared to non-transgenic controls. Statistical enrichment analyses revealed abiotic stress response genes, including heat stress response (HSR), showed reduced expression in transgenic lines under optimal growth conditions. However, most of the HSR DEGs were upregulated under high temperature stress (37 °C/1 h) conditions. In addition, the glucosinolate biosynthesis gene expression and total glucosinolate content increased in the transgenic lines. These findings provide a new avenue related to BrPP5.2 downstream genes and their crucial metabolic and heat stress responses in plants.

Precise flowering timing is critical for the plant life cycle. Here, we examined the molecular mechanisms and regulatory network associated with flowering in Chinese cabbage ( Brassica rapa L.) by comparative transcriptome profiling of two Chinese cabbage inbred lines, "4004" (early bolting) and "50" (late bolting). RNA-Seq and quantitative reverse transcription PCR (qPCR) analyses showed that two positive nitric oxide (NO) signaling regulator genes, nitrite reductase ( BrNIR ) and nitrate reductase ( BrNIA ), were up-regulated in line "50" with or without vernalization. In agreement with the transcription analysis, the shoots in line "50" had substantially higher nitrogen levels than those in "4004". Upon vernalization, the flowering repressor gene Circadian 1 ( BrCIR1 ) was significantly up-regulated in line "50", whereas the flowering enhancer genes named SUPPRESSOR OF OVEREXPRESSION OF CONSTANCE 1 homologs ( BrSOC1s ) were substantially up-regulated in line "4004". CRISPR/Cas9-mediated mutagenesis in Chinese cabbage demonstrated that the BrSOC1-1/1-2/1-3 genes were involved in late flowering, and their expression was mutually exclusive with that of the nitrogen signaling genes. Thus, we identified two flowering mechanisms in Chinese cabbage: a reciprocal negative feedback loop between nitrogen signaling genes ( BrNIA1 and BrNIR1 ) and BrSOC1s to control flowering time and positive feedback control of the expression of BrSOC1s .

Background: Leaf color is an important trait in breeding of leafy vegetables. Y-05, a pakchoi (Brassica rapa ssp. chinensis) cultivar, displays yellow inner (YIN) and green outer leaves (GOU) after cold acclimation. However, the mechanism of this special phenotype remains elusive., Results: We assumed that the yellow leaf phenotype of Y-05 maybe caused by low chlorophyll content. Pigments measurements and transmission electron microscopy (TEM) analysis showed that the yellow phenotype is closely related with decreased chlorophyll content and undeveloped thylakoids in chloroplast. Transcriptomes and metabolomes sequencing were next performed on YIN and GOU. The transcriptomes data showed that 4887 differentially expressed genes (DEGs) between the YIN and GOU leaves were mostly enriched in the chloroplast- and chlorophyll-related categories, indicating that the chlorophyll biosynthesis is mainly affected during cold acclimation. Together with metabolomes data, the inhibition of chlorophyll biosynthesis is contributed by blocked 5-aminolevulinic acid (ALA) synthesis in yellow inner leaves, which is further verified by complementary and inhibitory experiments of ALA. Furthermore, we found that the blocked ALA is closely associated with increased BrFLU expression, which is indirectly altered by cold acclimation. In BrFLU-silenced pakchoi Y-05, cold-acclimated leaves still showed green phenotype and higher chlorophyll content compared with control, meaning silencing of BrFLU can rescue the leaf yellowing induced by cold acclimation., Conclusions: Our findings suggested that cold acclimation can indirectly promote the expression of BrFLU in inner leaves of Y-05 to block ALA synthesis, resulting in decreased chlorophyll content and leaf yellowing. This study revealed the underlying mechanisms of leaves color change in cold-acclimated Y-05.

Glucosinolates (GLs), found in Brassicaceae family, are precursor metabolites with anti-cancer properties. Increased GLs have been studied under various environmental growth conditions. Pak choi (Brassica rapa subsp. chinensis) is a GL-rich vegetable. We hypothesize that long exposure to light and drought will increase the biomass of, and GL production in, pak choi. The experiment was conducted for 6 weeks. Long light exposure (20 h/day) increased, whilst drought exposure (12 h/week) decreased the plant growth. The plants exposed to a combination of drought and long light conditions showed similar growth pattern as control plants. GL production increased at week 6 in plants exposed to long light, while drought exposure had no impact on GL production, with the exception of glucoraphanin. Significant positive correlations were observed between plant growth and GL yield with accumulated light exposure time. Our findings suggest that long exposure to light can be used to increase both the biomass and GL production in pak choi., (Copyright © 2020. Published by Elsevier Ltd.)

Macrophages can initiate innate immune responses against microbes and cancer. The aim of this study was to elucidate the effects of Brassica rapa L. on macrophages. The production of interleukin (IL)-6, tumor necrosis factor (TNF)-α, and interferon-γ induced by the insoluble fraction of B. rapa L. was decreased in macrophage-depleted spleen cells compared with controls. The insoluble fraction of B. rapa L. induced expression of H-2Kb, I-Ab, CD40, and CD86, production of cytokines and nitric oxide, and phagocytic activity in RAW264 cells. After treatment with the insoluble fraction, IL-6 and TNF-α production was significantly decreased by anti-Toll-like receptor (TLR)2 mAb or polymyxin B compared with the control. Furthermore, insoluble fraction-mediated cytokine production was significantly lower in peritoneal macrophages from TLR2-/- and TLR4-/- mice compared with wild-type mice. These results suggest that B. rapa L. is a potentially effective immunomodulator for activating macrophages to prevent infections., (© The Author(s) 2021. Published by Oxford University Press on behalf of Japan Society for Bioscience, Biotechnology, and Agrochemistry.)

The production of haploid and doubled haploid plants is a biotechnological tool that shortens the breeding process of new cultivars in many species. Doubled haploid plants are homozygous at every locus and they can be utilized as parents to produce F1 hybrids. In this chapter, we describe a protocol for the production of doubled haploid plants in Brassica rapa L. subsp. pekinensis using androgenesis induced by isolated microspore cultures., (© 2021. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Strigolactone (SL) is a plant hormone that can improve plant stress resistance by regulating physiological processes and gene expression. GR24 is a synthetic strigolactone, which can also be used as a plant growth regulator. In this paper, the effects of exogenous GR24 on the growth and development of rape (Brassica rapa L.) under low temperature (4 °C) were studied. The results showed that low temperature (4 °C) inhibited the growth of rape seedlings, and exogenous GR24 significantly alleviated the effect of low temperature stress on rape seedlings. Compared with 4 °C treatment, GR24 + 4 °C treatment can increase the cell viability, soluble protein and proline content, enhance antioxidant enzyme activity, inhibit the production of reactive oxygen species (ROS), improve photosynthesis, and reduce the relative conductivity of rape seedlings. Further research shows that H 2 O 2 plays a central role in improving the cold resistance of rape seedlings by GR24. qRT-PCR results indicated that GR24 significantly increased the expression of genes. Mainly includes antioxidant enzyme genes, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase genes, mitogen-activated protein kinase (MAPK) genes and cold-regulated genes. These results indicate that GR24 improves the cold tolerance of plants by regulating the expression of related genes. RNA-seq analysis revealed that there were 152 differentially expressed genes (DGEs) in T (4 °C)&#95;vs&#95;ST (GR24 + 4 °C), including 100 up-regulated genes and 52 down-regulated genes. These DEGs play an important role in carbon metabolism pathway, oxidative phosphorylation pathway, antioxidant activity and photosynthesis pathways. We selected 11 differentially expressed genes for qRT-PCR verification, and the verification results were consistent with RNA-seq results., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Key Message: The DEAD-Box RNA helicase OsTOGR1 positively regulates heat stress tolerance in Chinese cabbage. Non-heading Chinese cabbage (Brassica rapa L. ssp. chinensis) is primarily cultivated vegetable crop in Asian countries. Heat stress is one of the major threats for its growth and yield. Numerous regulatory genes in various crops have shown to contribute thermotolerance. Among them, Thermotolerant growth required 1 (TOGR1) is an important DEAD-box RNA helicase. To examine whether its role is conserved in other crops, we constructed pCAMBIA1300-pHSP:OsTOGR1 expression vector driven by the rice small heat shock protein promoter (pHSP17.9) and successfully produced transgenic non-heading Chinese cabbage plants expressing OsTOGR1 gene via Agrobacterium-mediated vacuum infiltration transformation. In total, we generated three independent transgenic cabbage lines expressing TOGR1 gene. Expression and integration of TOGR1 was confirmed by PCR, RT-PCR and qPCR in T 1 and T 2 generations. The relative leaf electrical conductivity of transgenic seedlings was reduced subjected to high temperature (38 °C) compared to heat shock treatment (46 °C). In addition, hypocotyl length of transgenic seedlings increased compared to wild-type plants under high temperature and heat shock treatment. Furthermore, the transgenic plants exhibited higher chlorophyll content than wild-type plants under high temperature and heat shock treatment. The transgenic seeds displayed better germination under heat shock treatment. Tested heat stress-responsive genes were also up-regulated in the transgenic plants subjected to high temperature or heat shock treatment. To the best of our knowledge, this is the first report on describing the role of DAED-Box RNA helicases in improving heat stress tolerance of transgenic plants.

MicroRNAs (miRNAs) and natural antisense transcripts (NATs) control many biological processes and have been broadly applied for genetic manipulation of eukaryotic gene expression. Still unclear, however, are whether and how NATs regulate miRNA production. Here, we report that the cis-NATs of MIR398 genes repress the processing of their pri-miRNAs. Through genome-wide analysis of RNA sequencing data, we identify cis-NATs of MIRNA genes in Arabidopsis and Brassica. In Arabidopsis, MIR398b and MIR398c are coexpressed in vascular tissues with their antisense genes NAT398b and NAT398c, respectively. Knock down of NAT398b and NAT398c promotes miR398 processing, resulting in stronger plant thermotolerance owing to silencing of miR398-targeted genes; in contrast, their overexpression activates NAT398b and NAT398c, causing poorer thermotolerance due to the upregulation of miR398-targeted genes. Unexpectedly, overexpression of MIR398b and MIR398c activates NAT398b and NAT398c. Taken together, these results suggest that NAT398b/c repress miR398 biogenesis and attenuate plant thermotolerance via a regulatory loop.

Self-incompatibility (SI) is a breeding system that promotes cross-fertilization. In Brassica, pollen rejection is induced by a haplotype-specific interaction between pistil determinant SRK (S receptor kinase) and pollen determinant SP11 (S-locus Protein 11, also named SCR) from the S-locus. Although the structure of the B. rapa S 9 -SRK ectodomain (eSRK) and S 9 -SP11 complex has been determined, it remains unclear how SRK discriminates self- and nonself-SP11. Here, we uncover the detailed mechanism of self/nonself-discrimination in Brassica SI by determining the S 8 -eSRK-S 8 -SP11 crystal structure and performing molecular dynamics (MD) simulations. Comprehensive binding analysis of eSRK and SP11 structures reveals that the binding free energies are most stable for cognate eSRK-SP11 combinations. Residue-based contribution analysis suggests that the modes of eSRK-SP11 interactions differ between intra- and inter-subgroup (a group of phylogenetically neighboring haplotypes) combinations. Our data establish a model of self/nonself-discrimination in Brassica SI.

Plant cell wall-associated polygalacturonase-inhibiting proteins (PGIPs) are widely distributed in the plant kingdom. They play a crucial role in plant defense against phytopathogens by inhibiting microbial polygalacturonases (PGs). PGs hydrolyze the cell wall polysaccharide pectin and are among the first enzymes to be secreted during plant infection. Recent studies demonstrated that herbivorous insects express their own PG multi-gene families, raising the question whether PGIPs also inhibit insect PGs and protect plants from herbivores. Preliminary evidence suggested that PGIPs may negatively influence larval growth of the leaf beetle Phaedon cochleariae (Coleoptera: Chrysomelidae) and identified BrPGIP3 from Chinese cabbage ( Brassica rapa ssp. pekinensis ) as a candidate. PGIPs are predominantly studied in planta because their heterologous expression in microbial systems is problematic and instability and aggregation of recombinant PGIPs has complicated in vitro inhibition assays. To minimize aggregate formation, we heterologously expressed BrPGIP3 fused to a glycosylphosphatidylinositol (GPI) membrane anchor, immobilizing it on the extracellular surface of insect cells. We demonstrated that BrPGIP3&#95;GPI inhibited several P. cochleariae PGs in vitro , providing the first direct evidence of an interaction between a plant PGIP and an animal PG. Thus, plant PGIPs not only confer resistance against phytopathogens, but may also aid in defense against herbivorous beetles., Competing Interests: Conflict of interest—The authors declare that they have no conflicts of interest with the contents of this article., (© 2020 Haeger et al.)

Brassica rapa L. is a valuable and widespread species, but its cultivation in risk farming areas requires high-quality cold-hardy varieties to be developed. Mechanisms of the cold stress response in plants involve expression of numerous genes, including ribosomal ones, and are related to plant chromosome variability. FISH- and PCR-based methods were used to study intraspecific chromosome variability in the number and localization of 45S and 5S rDNA clusters and also to examine a set of molecular markers associated with cold-hardiness in winter B. rapa cultivars from high-risk farming areas. Several SSR (Na10-CO3 and BrgMS5339-1) and SCAR (BoCCA1-F/BoCCA1-1R1 and BoCCA1-F/BoCCA1-2R1) markers were identified as suitable for diagnosing cold-resistant and cold-susceptible genotypes in B. rapa. Compared with fodder cultivars, oilseed and leaf cultivars were shown to have more molecular markers associated with cold-hardiness and a higher level of polymorphism for the chromosomal distribution of 45S and 5S rDNAs, including chromosome heteromorphism. Thus, the least cold-resistant genotypes were found to display the lowest level of chromosome variability in the distribution of the 45S and 5S rDNA clusters and vice versa. The findings could be useful for the development of new cold-tolerant B. rapa varieties.

Self-incompatibility (SI) is a genetic mechanism most flowering plants adopted to reject self-pollen thus avoid inbreeding. In the Brassicaceae, self-pollen recognition triggers downstream signaling pathways to reject self-pollen. However, the downstream signaling pathways are not very clear. Here we show that ethylene negatively mediates self-incompatibility response of Chinese cabbage (Brassica rapa L. ssp. Pekinensis) via PCD in papilla cells. We found that ethylene signaling genes were upregulated after cross-pollination. Treating stigmas with ethylene, or suppressing the expression of a negative regulator of ethylene signaling, CONSTITUTIVE TRIPLE RESPONSE 1 (CTR1), caused PCD in papilla cells and broke down the self-incompatibility. On the other hand, treating stigmas with ethylene inhibitors, or suppressing the expression of ethylene-responsive factors (ERFs), inhibited PCD in papilla cells and the compatible pollination. Our study identified an additional signaling pathway mediating self-incompatibility responses in the Brassicaceae and also developed a new method in overcoming self-incompatibility to improve the efficiency of inbred line propagation in agriculture practice., Competing Interests: Declaration of competing interest All authors declare no financial interests relevant to this study., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Several members of the Brassicaeae family are known to possess beneficial properties which positively impact human diet, thanks to the presence of antioxidants, bioactive polyphenols and amino acids. B. rapa , one of the most widespread and economically relevant species, represents an outstanding example. The aim of this study is to investigate, at the molecular level, the effect of plant aging on the concentration of some biologically relevant compounds in different parts of the plant. Using HPLC and NMR techniques, the quantification of polyphenolic species (caffeic acid, quercetin and rutin), succinic acid and alanine was performed in flowers and leaves of young and mature B. rapa plants.

Background: A leaf of Chinese cabbage (Brassica rapa ssp. pekinensis) is composed of a photosynthetic blade and a non-photosynthetic large midrib; thus each leaf contains both source and sink tissues. This structure suggests that, unlike in other plants, source-sink metabolism is present in a single leaf of Chinese cabbage., Objective: This study was designed to identify the transport route of photosynthetic carbon and to determine whether both source and sink tissues were present in a leaf., Methods: Plant samples were collected diurnally. Their carbohydrate contents were measured, and a genome-wide transcriptome analysis was performed using the Br300K microarray. Expression profiles of selected genes were validated using qRT-PCR analysis., Results: The presence of two contrasting tissues (blade as source and midrib as sink) in a leaf was demonstrated by (1) diurnal distribution patterns of starch and sucrose content; (2) Gene Ontology (GO) enrichment analysis of microarray data; (3) expression profiles of photosynthetic and sucrose biosynthetic genes; and (4) expression patterns of a variety of sugar transporter genes., Conclusion: Source and sink tissues were both present in Chinese cabbage leaves, but the midrib functioned as a sink tissue as well as a site exporting to roots and other sink tissues. Function of most genes discriminating between source and sink tissue appeared to be regulated largely at the post-transcriptional level, not at the transcriptional level.

Background: Plant homeodomain (PHD) finger proteins are widely present in all eukaryotes and play important roles in chromatin remodeling and transcriptional regulation. The PHD finger can specifically bind a number of histone modifications as an "epigenome reader", and mediate the activation or repression of underlying genes. Many PHD finger genes have been characterized in animals, but only few studies were conducted on plant PHD finger genes to this day. Brassica rapa (AA, 2n = 20) is an economically important vegetal, oilseed and fodder crop, and also a good model crop for functional and evolutionary studies of important gene families among Brassica species due to its close relationship to Arabidopsis thaliana., Results: We identified a total of 145 putative PHD finger proteins containing 233 PHD domains from the current version of B. rapa genome database. Gene ontology analysis showed that 67.7% of them were predicted to be located in nucleus, and 91.3% were predicted to be involved in protein binding activity. Phylogenetic, gene structure, and additional domain analyses clustered them into different groups and subgroups, reflecting their diverse functional roles during plant growth and development. Chromosomal location analysis showed that they were unevenly distributed on the 10 B. rapa chromosomes. Expression analysis from RNA-Seq data showed that 55.7% of them were constitutively expressed in all the tested tissues or organs with relatively higher expression levels reflecting their important housekeeping roles in plant growth and development, while several other members were identified as preferentially expressed in specific tissues or organs. Expression analysis of a subset of 18 B. rapa PHD finger genes under drought and salt stresses showed that all these tested members were responsive to the two abiotic stress treatments., Conclusions: Our results reveal that the PHD finger genes play diverse roles in plant growth and development, and can serve as a source of candidate genes for genetic engineering and improvement of Brassica crops against abiotic stresses. This study provides valuable information and lays the foundation for further functional determination of PHD finger genes across the Brassica species.

Cuticular wax accumulation in plants contributes to drought tolerance. Here, we compared the drought levels on two varieties with different genotypes in turnip ( Brassica rapa var. rapa) and found that the drought tolerance was higher in the waxy KTRG-B48a than in the wax-free KTRG-B48b. A combination of PacBio and Illumina sequencing analyses revealed that differential transcripts were mainly enriched in the wax synthesis pathway, and a splice variant ( BrrWSD1-X2 ) was identified in the waxy KTRG-B48a. BrrWSD1-X2 had a stronger ability to synthesize wax esters than BrrWSD1-X1 using heterologous expression in yeast ( Saccharomyces cerevisiae ) mutant H1246a. Then, we speculated that the T to C transversion of the third intron and the higher number of TA repeats in the third intron of BrrWSD1 DNA in the waxy KTRG-B48a may result in a lower efficiency of splicing recognition of the third intron, resulting in the emergence of BrrWSD1-X2 in waxy varieties.

Nowadays, Zinc (Zn) deficiency is the most widespread micronutrient deficiency but simultaneously Zn toxicity is produced due to environmental pollution. A potential method to alleviate Zn deficiency and to reduce Zn concentration in soils is through the generation of plants with enhanced capacity for Zn accumulation and higher tolerance. This could be achieved through the modification of HMA4 transporter. BraA.hma4a-3 is a TILLING mutant plant that presents one modification in HMA4 transporter. Thus, in this study we analyzed the potential of BraA.hma4a-3 for Zn accumulation and Zn deficiency and toxicity tolerance. BraA.hma4a-3 and parental R-o-18 plants were grown with different Zn doses: 1 μM ZnSO 4 (Control), 0.01 μM ZnSO 4 (Zn deficiency) and 100 μM ZnSO 4 (Zn toxicity). Parameters of biomass, Zn concentration, photosynthesis, oxidative stress, N metabolism and amino acids (AAs) were measured. BraA.hma4a-3 did not affect plant biomass but did increase Zn accumulation in leaves under an adequate Zn supply and Fe under control and Zn deficiency doses. Regarding stress tolerance parameters and N metabolism, BraA.hma4a did not produce alterations under control conditions. In addition, under Zn toxicity, parameters suggest a greater tolerance. Briefly, the obtained results point to BraA.hma4a-3 as a useful mutant to increase Zn accumulation., (Copyright © 2019 Elsevier B.V. All rights reserved.)

There is a wide variation of flowering time among lines of Brassica rapa L. Most B. rapa leafy (Chinese cabbage etc.) or root (turnip) vegetables require prolonged cold exposure for flowering, known as vernalization. Premature bolting caused by low temperature leads to a reduction in the yield/quality of these B. rapa vegetables. Therefore, high bolting resistance is an important breeding trait, and understanding the molecular mechanism of vernalization is necessary to achieve this goal. In this study, we demonstrated that BrFRIb functions as an activator of BrFLC in B. rapa. We showed a positive correlation between the steady state expression levels of the sum of the BrFLC paralogs and the days to flowering after four weeks of cold treatment, suggesting that this is an indicator of the vernalization requirement. We indicate that BrFLCs are repressed by the accumulation of H3K27me3 and that the spreading of H3K27me3 promotes stable FLC repression. However, there was no clear relationship between the level of H3K27me3 in the BrFLC and the vernalization requirement. We also showed that if there was a high vernalization requirement, the rate of repression of BrFLC1 expression following prolonged cold treatments was lower.

Mutualistic (e.g. pollination) and antagonistic (e.g. herbivory) plant-insect interactions shape levels of plant fitness and can have interactive effects. By using experimental plots of Brassica rapa plants infested with generalist (Mamestra brassicae) and specialised (Pieris brassicae) native herbivores and with a generalist invasive (Spodoptera littoralis) herbivore, we estimated both pollen movement among treatments and the visiting behaviour of honeybees versus other wild pollinators. Overall, we found that herbivory has weak effects on plant pollen export, either in terms of inter-treatment movements or of dispersion distance. Plants infested with the native specialised herbivore tend to export less pollen to other plants with the same treatment. Other wild pollinators preferentially visit non-infested plants that differ from those of honeybees, which showed no preferences. Honeybees and other wild pollinators also showed different behaviours on plants infested with different herbivores, with the former tending to avoid revisiting the same treatment and the latter showing no avoidance behaviour. When taking into account the whole pollinator community, i.e. the interactive effects of honeybees and other wild pollinators, we found an increased avoidance of plants infested by the native specialised herbivore and a decreased avoidance of plants infested by the invasive herbivore. Taken together, our results suggest that herbivory may have an effect on B. rapa pollination, but this effect depends on the relative abundance of honeybees and other wild pollinators., (© 2019 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

Main Conclusion Root gravitropism of primary roots is assisted by curvature of the hypocotyl base. Root gravitropism is typically described as the sequence of signal perception, signal processing, and response that causes differential elongation and the establishment of a new gravitropic set-point angle. We describe two components of the graviresponse of Brassica seedlings that comprise a primary curvature of the root tip and a later onset but stronger curvature that occurs at the base of the hypocotyl. This second curvature is preceded by straightening of the tip region but leads to the completion of the alignment of the root axis. Curvature in both regions require a minimum displacement of 20 deg. The rate of tip curvature is a function of root length. After horizontal reorientation, tip curvature of five mm long roots curved twice as fast as 10 mm long roots (33.6 ± 3.3 vs. 14.3 ± 1.5 deg hr-1). The onset of curvature at the hypocotyl base is correlated with root length, but the rate of this curvature is independent of seedling length. Decapping of roots prevented tip curvature but the curvature at base of hypocotyl was unaffected. Endodermal cells at the root shoot junction show numerous, large and sedimenting amyloplasts, which likely serve as gravity sensors (statoliths). The amyloplasts at the hypocotyl were 3-4 μm in diameter, similar in size to those in the root cap, and twice the size of starch grains in the cortical layers of hypocotyl or elsewhere in the root. These data indicate that the root shoot reorientation of young seedlings is not limited to the root tip but includes more than one gravitropically responsive region., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Main Conclusion: Compared with its parents, Brassica hexaploid underwent significant AS changes, which may provide diversified gene expression regulation patterns and could enhance its adaptability during evolution Polyploidization is considered a significant evolution force that promotes species formation. Alternative splicing (AS) plays a crucial role in multiple biological processes during plant growth and development. To explore the effects of allopolyploidization on the AS patterns of genes, a genome-wide AS analysis was performed by RNA-seq in Brassica hexaploid and its parents. In total, we found 7913 (27540 AS events), 14447 (70179 AS events), and 13205 (60804 AS events) AS genes in Brassica rapa, Brassica carinata, and Brassica hexaploid, respectively. A total of 920 new AS genes were discovered in Brassica hexaploid. There were 56 differently spliced genes between Brassica hexaploid and its parents. In addition, most of the alternative 5' splice sites were located 4 bp upstream of the dominant 5' splice sites, and most of the alternative 3' splice sites were located 3 bp downstream of the dominant 3' splice sites in Brassica hexapliod, which was similar to B. carinata. Furthermore, we cloned and sequenced all amplicons from the RT-PCR products of GRP7/8, namely, Bol045859, Bol016025 and Bol02880. The three genes were found to produce AS transcripts in a new way. The AS patterns of genes were diverse between Brassica hexaploid and its parents, including the loss and gain of AS events. Allopolyploidization changed alternative splicing sites of pre-mRNAs in Brassica hexaploid, which brought about alterations in the sequences of transcripts. Our study provided novel insights into the AS patterns of genes in allopolyploid plants, which may provide a reference for the study of polyploidy adaptability.

Nickel oxide nanoparticles (NiO NPs) are utilized in various industries and their release into the environment may lead to the pollution of agricultural areas. However, assessing the toxicity of NiO NPs in major food crops is difficult due to the limited information available on their toxicity. The present investigation was carried out to evaluate how NiO NPs affect plant growth, photosynthetic efficiency, and phytochemical content, as well as changes at the transcriptional level of these phytochemicals in Chinese cabbage seedlings. Chlorophyll, carotenoid, and sugar contents were reduced, while proline and the anthocyanins were significantly upregulated in NiO NPs-treated seedlings. The levels of malondialdehyde, hydrogen peroxide, and reactive oxygen species, as well as peroxidase (POD) enzyme activity, were all enhanced in seedlings exposed to NiO NPs. The levels of glucosinolates and phenolic compounds were also significantly increased in NiO NPs-treated seedlings compared to control seedlings. The expression of genes related to oxidative stress (CAT, POD, and GST), MYB transcription factors (BrMYB28, BrMYB29, BrMYB34, and BrMYB51), and phenolic compounds (ANS, PAP1, and PAL) were significantly upregulated. We suggest that NiO NPs application stimulates toxic effects and enhances the levels of phytochemicals (glucosinolates and phenolic compounds) in Chinese cabbage seedlings., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Biodiversity influences ecosystem function, but there is limited understanding of the mechanisms that support this relationship across different land use types in mosaic agroecosystems. Network approaches can help to understand how community structure influences ecosystem function across landscapes; however, in ecology, network analyses have largely focused on species-species interactions. Here, we use bipartite network analysis in a novel way: to link pollinator communities to sites in a tropical agricultural landscape. We used sentinel plants of Brassica rapa to examine how the structure of the community network influences plant reproduction. Diptera was the most common order of flower visitors at every site. Syrphidae visits were the strongest contributor to the number of fertilized pods, while visits by Syrphidae, Hymenoptera and Lepidoptera had the strongest effect on the number of seeds per pod. Sentinel pots at forest sites were visited by more unique species (i.e. species with higher d') than sites in other land uses, and dairy sites had more visitors that were common across the network. Participation coefficients, which indicate how connected a single node is across network modules, were strong predictors of ecosystem function: plant reproduction increased at sites with higher participation coefficients. Flower visitor taxa with higher participation coefficients also had the strongest effect on plant reproduction. Hymenoptera visits were the best predictor for participation coefficients but an Allograpta sp. (Diptera: Syrphidae) was the most influential flower visitor species in the landscape network. A diverse insect community contributed to plant reproduction and connection among nodes in this system. Identifying the 'keystone' flower visitor species and sites that have a strong influence on network structure is a significant step forward to inform conservation priorities and decision-making in diverse agroecosystems.

Background: Abiotic stresses like drought, heat, cold and salinity cause major productivity loss in the rapeseed-mustard crops (Brassica). Major efforts have been made in the past to identify genes that provide resistance against such stresses. Superoxide dismutase (SOD) proteins, member of the metallo-enzyme family play vital role in protecting plants against abiotic stresses. In the present study, genome-wide analysis of abiotic stress responsive SOD gene family has been done in B. juncea and B. rapa., Results: A total of 29 and 18 SOD genes were identified in B. juncea and B. rapa respectively and chromosome location mapping indicated their wide distribution across genome. On the basis of domain composition, the SODs were phylogenetically classified into sub-groups which was also substantiated by the gene structure and sub-cellular locations of SOD proteins. Functional annotation of SODs was also done by Gene Ontology (GO) mapping and the result was corroborated by the identified cis-regulatory elements in the promoter region of SOD genes. Based on FPKM analysis of SRA data available for drought, heat and salt stress, we identified 14 and 10 abiotic stress responsive SOD genes in B. rapa and B. juncea respectively. The differential expression analysis under drought and heat stress of identified abiotic-stress responsive SOD genes was done through quantitative Real Time PCR., Conclusion: We identified abiotic-stress responsive genes that could help in improving the plant tolerance against abiotic stresses. This was the first study to describe the genome-wide analysis of SOD gene family in B. rapa and B. juncea, and the results will help in laying basic ground for future work of cloning and functional validation of SOD genes during abiotic stresses leading to Brassica crop improvement.

Epigenetic regulations in the form of changes in differential expression of noncoding RNAs (ncRNAs) are an essential mechanism of stress response in plants. Previously we showed that heat treatment in L. results in the differential processing and accumulation of ncRNA fragments (ncRFs) stemming from transfer RNAs (tRNAs), ribosomal RNAs (rRNAs), small nuclear RNAs (snRNAs), and small nucleolar RNAs (snoRNAs). In this work, we analyzed whether ncRFs are differentially expressed in the progeny of heat-stressed plants. We found significant changes in the size of tRF reads and a significant decrease in the percentage of tRFs mapping to tRNA-Ala, tRNA-Arg, and tRNA-Tyr and an increase in tRFs mapping to tRNA-Asp. The enrichment analysis showed significant differences in processing of tRFs from tRNA, tRNA, tRNA, tRNA, tRNA, and tRNA isoacceptors. Analysis of potential targets of tRFs showed that they regulate brassinosteroid metabolism, the proton pump ATPase activity, the antiporter activity, the mRNA decay activity as well as nucleosome positioning and the epigenetic regulation of transgenerational response. Gene ontology term analysis of potential targets demonstrated a significant enrichment in tRFs that potentially targeted a cellular component endoplasmic reticulum (ER) and in small nucleolar RNA fragments (snoRFs), the molecular function protein binding. To summarize, our work demonstrated that the progeny of heat-stressed plants exhibit changes in the expression of tRFs and snoRFs but not of small nuclear RNA fragments (snRFs) or ribosomal RNA fragments (rRFs) and these changes likely better prepare the progeny of stressed plants to future stress encounters., (Copyright © 2019 Crop Science Society of America.)

Salinity stress is a major abiotic stress that affects plant growth and limits crop production. Roots are the primary site of salinity perception, and salt sensitivity in roots limits the productivity of the entire plant. To better understand salt stress responses in canola, we performed a comparative proteomic analysis of roots from the salt-tolerant genotype Safi-7 and the salt-sensitive genotype Zafar. Plants were exposed to 0, 150, and 300 mM NaCl. Our physiological and morphological observations confirmed that Safi-7 was more salt-tolerant than Zafar. The root proteins were separated by two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry was applied to identify proteins regulated in response to salt stress. We identified 36 and 25 protein spots whose abundance was significantly affected by salt stress in roots of plants from the tolerant and susceptible genotype, respectively. Functional classification analysis revealed that the differentially expressed proteins from the tolerant genotype could be assigned to 14 functional categories, while those from the susceptible genotype could be classified into 9 functional categories. The most significant differences concerned proteins involved in glycolysis (Glyceraldehyde-3-phosphate dehydrogenase, Fructose-bisphosphate aldolase, Phosphoglycerate kinase 3), stress (heat shock proteins), Redox regulation (Glutathione S-transferase DHAR1, L-ascorbate peroxidase), energy metabolism (ATP synthase subunit B), and transport (V-type proton ATPase subunit B1) which were increased only in the tolerant line under salt stress. Our results provide the basis for further elucidating the molecular mechanisms of salt-tolerance and will be helpful for breeding salt-tolerant canola cultivars., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The Brassica rapa (B. rapa) species displays enormous phenotypic diversity, with leafy vegetables, storage root vegetables and oil crops. These different crops all have different flowering time, which determine their growing season and cultivation area. Little is known about the effects of diverse temperature and day-lengths on flowering time QTL associated with FLC paralogues. We phenotyped the flowering time of a doubled haploid population, established from a cross between Yellow sarson and Pak choi under diverse environmental conditions. We identified flowering-time QTL (fQTL) in different photoperiod and temperature regimes in the greenhouse, and studied their colocation with known flowering time genes. As several fQTL colocalized with FLC paralogues, we studied the expression patterns of four FLC paralogues during the course of vernalization in parental lines. Under all environmental conditions tested the major fQTL that mapped to the BrFLC2&#95;A02 locus was detected, however its effect decreased when plants were grown at low temperatures. Another fQTL that mapped to the FLC paralogue, BrFLC5&#95;A03 was also identified under all tested environments, while no fQTL colocated with BrFLC1&#95;A10 or BrFLC3&#95;A03. Furthermore, the vernalization treatment decreased expression of all BrFLC paralogues in the parental lines, and showed the lowest transcript level after 28 days of vernalization. Transcript abundance stayed low after returning the plants for seven days to normal growth temperature. Interestingly, transcript abundance of BrFLC3&#95;A03 and BrFLC5&#95;A03 was repressed much stronger and already reached lowest levels after 14d in the early-flowering type YS-143. This study improves understanding of the effects of daylength and vernalization on flowering time in B. rapa and the role of the different BrFLC paralogues therein., (Copyright © 2018. Published by Elsevier B.V.)

The objective of this study was to investigate the effect of addition of sawdust co-composted with sewage sludge and wheat straw on seeds germination. Two mixtures were piled and composted over 90 days. The first mixture (C1) was composed of sewage sludge and wheat straw, while the second mixture (C2) was composed of sewage sludge, wheat straw and wood sawdust. The results showed that the physicochemical parameters of both composts, i.e., temperature (> 55 °C in the thermophilic phase), moisture content (~ 30%). pH (6.73 for C1 and 7.19 for C2) and EC values (1.81 mS cm -1 for C1 and 1.32 mS cm -1 for C2) reached the required maturity standard. The values of C/N were below 12 indicating a high degree of maturity. Also, no bacterial pathogens were detected in the final composts. The concentration of total heavy metals has been reduced allowing the elimination of sewage sludge toxicity, confirmed by the germination index, which reached over 80%. Strong positive correlations were noticed between total Kjeldahl nitrogen of C2 and germination indexes of all the studied species. The obtained results indicate that the addition of wood sawdust increases the nitrogen content leading to slightly alkaline compost which influences seeds germination by reducing the phytotoxicity of sewage sludge., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Microspore embryogenesis is a powerful biotechnological tool that is very useful in crop breeding for the rapid production of haploid and double-haploid embryos and plants. In this in vitro system, the haploid microspore is reprogrammed by the application of specific stress treatments. A high level of cell death after the stress is a major factor that greatly reduces embryogenesis yield at its initial stages. Autophagy is a degradation pathway that is present in all eukaryotes and plays key roles in a range of processes, including stress responses. Many proteases participate in autophagy and cell death; among them, cathepsins are the most abundant enzymes with a role in plant senescence and programmed cell death (PCD). Moreover, although plant genomes do not contain homologues of caspases, caspase 3-like activity (main executioner protease of animal cell death) has been detected in many plant PCD processes. Recent studies by our group in barley microspore cultures reported that the stress treatment required for inducing microspore embryogenesis (cold treatment), also produced reactive oxygen species (ROS) and cell death, concomitantly with the induction of autophagy, as well as cathepsin-like and caspase 3-like proteolytic activities. In the present study, we report new data on microspore embryogenesis of rapeseed that indicate, as in barley, activation of cell death and autophagy processes after the inductive stress. The results revealed that treatments modulating autophagy and proteases produced the same effect in the two plant systems, regardless of the stress applied, cold in barley or heat in rapeseed. Pharmacological treatments with small bioactive compounds that inhibit ROS, autophagy and specific cell death-proteases led to reduced cell death and an increased embryogenesis initiation rate in both, barley and rapeseed. Taken together, these findings open up new intervention pathways by modulating autophagy and proteases, which are very promising in terms of increasing the efficiency of in vitro microspore embryogenesis systems for biotechnological applications in crop breeding.

Background: Ubiquitous CCCH nucleic acid-binding motif is found in a wide-variety of organisms. CCCH genes are involved in plant developmental processes and biotic and abiotic stress responses. Brassica rapa is a vital economic crop and classical model plant of polyploidy evolution, but the functions of CCCH genes in B. rapa are unclear., Results: In this study, 103 CCCH genes in B. rapa were identified. A comparative analysis of the chromosomal position, gene structure, domain organization and duplication event between B. rapa and Arabidopsis thaliana were performed. Results showed that CCCH genes could be divided into 18 subfamilies, and segmental duplication might mainly contribute to this family expansion. C-X 7/8 -C-X 5 -C 3 -H was the most commonly found motif, but some novel CCCH motifs were also found, along with some loses of typical CCCH motifs widespread in other plant species. The multifarious gene structures and domain organizations implicated functional diversity of CCCH genes in B. rapa. Evidence also suggested functional redundancy in at least one subfamily due to high conservation between members. Finally, the expression profiles of subfamily-IX genes indicated that they are likely involved in various stress responses., Conclusion: This study provides the first genome-wide characterization of the CCCH genes in B. rapa. The results suggest that B. rapa CCCH genes are likely functionally divergent, but mostly involved in plant development and stress response. These results are expected to facilitate future functional characterization of this potential RNA-binding protein family in Brassica crops.