RESUMO
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.
Assuntos
Brassica rapa , Secas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , MicroRNAs , RNA Mensageiro , Estresse Fisiológico , Transcriptoma , MicroRNAs/genética , Brassica rapa/genética , Brassica rapa/fisiologia , Brassica rapa/metabolismo , Perfilação da Expressão Gênica/métodos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Estresse Fisiológico/genética , Redes Reguladoras de Genes , RNA de Plantas/genéticaRESUMO
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.
Assuntos
Brassica rapa , Secas , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Estresse Fisiológico , Brassica rapa/genética , Brassica rapa/fisiologia , Estresse Fisiológico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Filogenia , Reprodução/genética , Giberelinas/metabolismo , Oxilipinas/metabolismo , Perfilação da Expressão Gênica , Acetatos , CiclopentanosRESUMO
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.
Assuntos
Pradaria , Polinização , Animais , Polinização/fisiologia , Abelhas/fisiologia , Tibet , Brassica rapa/fisiologia , Biodiversidade , Ecossistema , Reprodução/fisiologia , Flores/fisiologia , Dípteros/fisiologiaRESUMO
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.
Assuntos
Agricultura , Produtos Agrícolas , Polinização , Animais , Agricultura/métodos , Abelhas/fisiologia , Biodiversidade , Brassica rapa/fisiologia , Brassica napus/fisiologia , Sementes/fisiologiaRESUMO
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 (Fv/Fm), 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.
Assuntos
Brassica rapa , Fotossíntese , Plântula , Sulfametoxazol , Sulfametoxazol/toxicidade , Brassica rapa/efeitos dos fármacos , Brassica rapa/fisiologia , Fotossíntese/efeitos dos fármacos , Plântula/efeitos dos fármacos , Clorofila/metabolismoRESUMO
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.
Assuntos
Brassica rapa , Borboletas , Flores , Aptidão Genética , Temperatura Alta , Polinização , Polinização/fisiologia , Animais , Flores/fisiologia , Abelhas/fisiologia , Brassica rapa/fisiologia , Borboletas/fisiologia , Herbivoria/fisiologia , Sementes/fisiologia , Compostos Orgânicos Voláteis/metabolismo , Compostos Orgânicos Voláteis/análise , Fenótipo , Oviposição/fisiologia , Temperatura , Característica Quantitativa HerdávelRESUMO
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.
Assuntos
Evolução Biológica , Brassica rapa , Genes de Plantas , Polinização , Polinização/genética , Brassica rapa/genética , Brassica rapa/fisiologia , Animais , Estudo de Associação Genômica Ampla , Autofertilização/genética , Flores/genética , Flores/fisiologia , Flores/anatomia & histologia , Reprodução/genética , Arabidopsis/genética , Arabidopsis/fisiologia , Mutação/genética , Dípteros/genética , Dípteros/fisiologia , Fenótipo , Pólen/genética , Pólen/fisiologiaRESUMO
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.
Assuntos
Brassica rapa , Flores , Regulação da Expressão Gênica de Plantas , Histonas , Brassica rapa/genética , Brassica rapa/fisiologia , Brassica rapa/crescimento & desenvolvimento , Flores/genética , Flores/crescimento & desenvolvimento , Flores/fisiologia , Histonas/metabolismo , Histonas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Epigênese Genética , Arabidopsis/genética , Arabidopsis/fisiologia , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Histona-Lisina N-Metiltransferase/genética , Histona-Lisina N-Metiltransferase/metabolismoRESUMO
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.
Assuntos
Adaptação Fisiológica , Brassica rapa , Herbivoria , Polinização , Solo , Solo/química , Animais , Herbivoria/fisiologia , Brassica rapa/genética , Brassica rapa/fisiologia , Abelhas/fisiologia , Abelhas/genética , Adaptação Fisiológica/genética , Polimorfismo de Nucleotídeo Único , Evolução Biológica , Flores/genética , Flores/fisiologia , Frequência do Gene , Afídeos/fisiologia , EcossistemaRESUMO
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 (BrJMJ18Par) 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 BrJMJ18Par can modulate the expression of BrFLC3, one of the five BrFLC orthologs. Furthermore, ChIP-seq and transcriptome data reveal that BrJMJ18Par 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.
Assuntos
Brassica rapa , Flores , Regulação da Expressão Gênica de Plantas , Histonas , Histona Desmetilases com o Domínio Jumonji , Proteínas de Plantas , Locos de Características Quantitativas , Brassica rapa/genética , Brassica rapa/metabolismo , Brassica rapa/crescimento & desenvolvimento , Brassica rapa/fisiologia , Flores/genética , Flores/crescimento & desenvolvimento , Histonas/metabolismo , Histona Desmetilases com o Domínio Jumonji/metabolismo , Histona Desmetilases com o Domínio Jumonji/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Temperatura , Termotolerância/genética , Metilação , Plantas Geneticamente Modificadas , Clorofila/metabolismoRESUMO
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 H2O2, 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 H2O2 nanosensor for simultaneous monitoring of stress-induced H2O2 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 H2O2 and SA generation for each stress. Based on these temporal insights, we have formulated a biochemical kinetic model that suggests the early H2O2 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.
Assuntos
Brassica rapa , Peróxido de Hidrogênio , Peróxido de Hidrogênio/farmacologia , Estresse Fisiológico , Brassica rapa/fisiologia , Reguladores de Crescimento de Plantas/farmacologia , Ácido SalicílicoRESUMO
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.
Assuntos
Brassica rapa , Melatonina , MicroRNAs , Brassica rapa/fisiologia , Tolerância ao Sal/genética , Regulação da Expressão Gênica de Plantas , Antioxidantes/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Metiltransferases/genéticaRESUMO
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.
Assuntos
Brassica rapa/genética , Brassica rapa/fisiologia , Códon de Terminação/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/metabolismo , Sequência de Bases , DNA de Plantas , Genoma de Planta , Mutação , Proteínas de Plantas/genética , Locos de Características QuantitativasRESUMO
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.
Assuntos
Brassica rapa/fisiologia , Perfilação da Expressão Gênica/métodos , Mucoproteínas/genética , Brassica rapa/genética , Clonagem Molecular , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Germinação , Filogenia , Infertilidade das Plantas , Proteínas de Plantas/genética , Análise de Sequência de RNARESUMO
Salinity is an important environmental factor that reduces plant productivity in many world regions. It affects negatively photosynthesis causing a growth reduction. Likewise, calcium (Ca2+) is crucial in plant stress response. Therefore, the modification of Ca2+ 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.
Assuntos
Brassica rapa/genética , Brassica rapa/fisiologia , Fotossíntese/genética , Fotossíntese/fisiologia , Tolerância ao Sal/efeitos dos fármacos , Tolerância ao Sal/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética , Genótipo , Mutação , Folhas de Planta/genética , Folhas de Planta/fisiologiaRESUMO
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.
Assuntos
Brassica rapa/fisiologia , Regulação da Expressão Gênica de Plantas , Glucosinolatos/biossíntese , Resposta ao Choque Térmico/genética , Proteínas Nucleares/genética , Fosfoproteínas Fosfatases/genética , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Biomarcadores , Biologia Computacional/métodos , Perfilação da Expressão Gênica , Proteínas Nucleares/metabolismo , Fosfoproteínas Fosfatases/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente ModificadasRESUMO
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.
Assuntos
Brassica rapa/crescimento & desenvolvimento , Brassica rapa/genética , Melhoramento Vegetal/métodos , Aclimatação/genética , Brassica rapa/fisiologia , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/fisiologia , Meios de Cultura/química , DNA de Plantas/genética , Diploide , Glucose-6-Fosfato Isomerase/genética , Haploidia , Homozigoto , Biologia Molecular/métodos , Pólen/genética , Pólen/crescimento & desenvolvimento , Reação em Cadeia da Polimerase , Regeneração/genética , Técnicas de Cultura de TecidosRESUMO
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.
Assuntos
Brassica rapa/genética , Proteínas de Ciclo Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Motivos de Aminoácidos , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Brassica rapa/crescimento & desenvolvimento , Brassica rapa/fisiologia , Ciclo Celular , Proteínas de Ciclo Celular/genética , Expressão Gênica , Inativação Gênica , Filogenia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/fisiologia , Proteínas de Plantas/genética , Domínios ProteicosRESUMO
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.
Assuntos
Brassica rapa/fisiologia , Flores/fisiologia , Proteínas de Domínio MADS/fisiologia , Nitrogênio/metabolismo , Proteínas de Plantas/fisiologia , Sistemas CRISPR-Cas , Retroalimentação Fisiológica , Redes Reguladoras de Genes , Nitrato Redutase/genética , Nitrato Redutase/metabolismo , Análise de Sequência de RNA , TranscriptomaRESUMO
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.