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1.
BMC Plant Biol ; 21(1): 372, 2021 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-34388971

RESUMO

BACKGROUND: Oilseed rape (B. napus L.) has great potential for phytoremediation of cadmium (Cd)-polluted soils due to its large plant biomass production and strong metal accumulation. Soil properties and the presence of other soluble compounds or ions, cause a heterogeneous distribution of Cd. RESULTS: The aim of our study was to reveal the differential responses of B. napus to different Cd abundances. Herein, we found that high Cd (50 µM) severely inhibited the growth of B. napus, which was not repressed by low Cd (0.50 µM) under hydroponic culture system. ICP-MS assays showed that the Cd2+ concentrations in both shoots and roots under 50 µM Cd were over 10 times higher than those under 0.50 µM Cd. Under low Cd, the concentrations of only shoot Ca2+/Mn2+ and root Mn2+ were obviously changed (both reduced); under high Cd, the concentrations of most cations assayed were significantly altered in both shoots and roots except root Ca2+ and Mg2+. High-throughput transcriptomic profiling revealed a total of 18,021 and 1408 differentially expressed genes under high Cd and low Cd conditions, respectively. The biological categories related to the biosynthesis of plant cell wall components and response to external stimulus were over-accumulated under low Cd, whereas the terms involving photosynthesis, nitrogen transport and response, and cellular metal ion homeostasis were highly enriched under high Cd. Differential expression of the transporters responsible for Cd uptake (NRAMPs), transport (IRTs and ZIPs), sequestration (HMAs, ABCs, and CAXs), and detoxification (MTPs, PCR, MTs, and PCSs), and some other essential nutrient transporters were investigated, and gene co-expression network analysis revealed the core members of these Cd transporters. Some Cd transporter genes, especially NRAMPs and IRTs, showed opposite responsive patterns between high Cd and low Cd conditions. CONCLUSIONS: Our findings would enrich our understanding of the interaction between essential nutrients and Cd, and might also provide suitable gene resources and important implications for the genetic improvement of plant Cd accumulation and resistance through molecular engineering of these core genes under varying Cd abundances in soils.


Assuntos
Brassica napus/genética , Brassica napus/metabolismo , Cádmio/metabolismo , Transporte Biológico , Brassica napus/crescimento & desenvolvimento , Quelantes/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Íons/metabolismo , Solo/química , Tetraploidia , Transcriptoma
2.
Mol Biol Rep ; 48(8): 5977-5992, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34327662

RESUMO

BACKGROUND: Nitrogen (N) is an essential macronutrient to maintain plant growth and development. Plants absorb nitrate-N or ammonium-N in the environment and undergo reduction reactions catalyzed by nitrate reductase (NR), nitrite reductase (NIR), glutamine synthetase (GS), and glutamine oxoglutarate aminotransferase (GOGAT) within plants. METHODS AND RESULTS: A total of 42 N assimilation-related genes (NAG) members were identified in rapeseed. Darwin's evolutionary pressure analysis showed that rapeseed NAGs underwent purification selection. Cis-element analysis revealed differences in the transcriptional regulation of NAGs between Arabidopsis and rapeseed. Expression analyses revealed that NRs were expressed mainly in old leaves, NIRs were expressed mainly in old leaves and lower stem peels, while the expression situation between different subfamilies of GSs and GOGATs was more complicated. CONCLUSIONS: Differential expression of NAGs suggested that they might be involved in abiotic stresses. The above results greatly enriched our understanding of NAGs' molecular characteristics and provided central gene resources for NAGs-mediated NUE improvement in rapeseed.

3.
BMC Plant Biol ; 21(1): 288, 2021 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-34167468

RESUMO

BACKGROUND: B-box (BBX) genes play important roles in plant growth regulation and responses to abiotic stresses. The plant growth and yield production of allotetraploid rapeseed is usually hindered by diverse nutrient stresses. However, no systematic analysis of Brassicaceae BBXs and the roles of BBXs in the regulation of nutrient stress responses have not been identified and characterized previously. RESULTS: In this study, a total of 536 BBXs were identified from nine brassicaceae species, including 32 AtBBXs, 66 BnaBBXs, 41 BoBBXs, 43 BrBBXs, 26 CrBBXs, 81 CsBBXs, 52 BnBBXs, 93 BjBBXs, and 102 BcBBXs. Syntenic analysis showed that great differences in the gene number of Brassicaceae BBXs might be caused by genome duplication. The BBXs were respectively divided into five subclasses according to their phylogenetic relationships and conserved domains, indicating their diversified functions. Promoter cis-element analysis showed that BBXs probably participated in diverse stress responses. Protein-protein interactions between BnaBBXs indicated their functions in flower induction. The expression profiles of BnaBBXs were investigated in rapeseed plants under boron deficiency, boron toxicity, nitrate limitation, phosphate shortage, potassium starvation, ammonium excess, cadmium toxicity, and salt stress conditions using RNA-seq data. The results showed that different BnaBBXs showed differential transcriptional responses to nutrient stresses, and some of them were simultaneously responsive to diverse nutrient stresses. CONCLUSIONS: Taken together, the findings investigated in this study provided rich resources for studying Brassicaceae BBX gene family and enriched potential clues in the genetic improvement of crop stress resistance.


Assuntos
Brassica napus/genética , Brassicaceae/genética , Genes de Plantas/genética , Fatores de Transcrição/genética , Brassica napus/fisiologia , Sequência Conservada , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/fisiologia , Estudo de Associação Genômica Ampla , Filogenia , Mapas de Interação de Proteínas , Estresse Fisiológico , Sintenia , Tetraploidia , Fatores de Transcrição/fisiologia
4.
J Exp Bot ; 72(15): 5687-5708, 2021 07 28.
Artigo em Inglês | MEDLINE | ID: mdl-33989425

RESUMO

The large size and complexity of the allotetraploid rapeseed (Brassica napus) genome present huge challenges for understanding salinity resistance in this important crop. In this study, we identified two rapeseed genotypes with significantly different degrees of salinity resistance and examined the underlying mechanisms using an integrated analysis of phenomics, ionomics, genomics, and transcriptomics. Under salinity, a higher accumulation of osmoregulation substances and better root-system architecture was observed in the resistant genotype, H159, than in the sensitive one, L339. A lower shoot Na+ concentration and a higher root vacuolar Na+ concentration indicated lower root-to-shoot translocation and higher compartmentation in H159 than in L339. Whole-genome re-sequencing (WGRS) and transcriptome sequencing identified numerous DNA variants and differentially expressed genes involved in abiotic stress responses and ion transport. Combining ionomics with transcriptomics identified plasma membrane-localized BnaC2.HKT1;1 and tonoplast-localized BnaC5.NHX2 as the central factors regulating differential root xylem unloading and vacuolar sequestration of Na+ between the two genotypes. Identification of polymorphisms by WGRS and PCR revealed two polymorphic MYB-binding sites in the promoter regions that might determine the differential gene expression of BnaC2.HKT1;1 and BnaC5.NHX2. Our multiomics approach thus identified core transporters involved in Na+ translocation and compartmentation that regulate salinity resistance in rapeseed. Our results may provide elite gene resources for the improvement of salinity resistance in this crop, and our multiomics approach can be applied to other similar studies.


Assuntos
Brassica napus , Brassica napus/genética , Brassica napus/metabolismo , Regulação da Expressão Gênica de Plantas , Salinidade , Sódio/metabolismo , Estresse Fisiológico
5.
BMC Plant Biol ; 20(1): 534, 2020 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-33228523

RESUMO

BACKGROUND: Salinity severely inhibit crop growth, yield, and quality worldwide. Allotetraploid rapeseed (Brassica napus L.), a major glycophyte oil crop, is susceptible to salinity. Understanding the physiological and molecular strategies of rapeseed salinity resistance is a promising and cost-effective strategy for developing highly resistant cultivars. RESULTS: First, early leaf senescence was identified and root system growth was inhibited in rapeseed plants under severe salinity conditions. Electron microscopic analysis revealed that 200 mM NaCl induced fewer leaf trichomes and stoma, cell plasmolysis, and chloroplast degradation. Primary and secondary metabolite assays showed that salinity led to an obviously increased anthocyanin, osmoregulatory substances, abscisic acid, jasmonic acid, pectin, cellulose, reactive oxygen species, and antioxidant activity, and resulted in markedly decreased photosynthetic pigments, indoleacetic acid, cytokinin, gibberellin, and lignin. ICP-MS assisted ionomics showed that salinity significantly constrained the absorption of essential elements, including the nitrogen, phosphorus, potassium, calcium, magnesium, iron, mangnese, copper, zinc, and boron nutrients, and induced the increase in the sodium/potassium ratio. Genome-wide transcriptomics revealed that the differentially expressed genes were involved mainly in photosynthesis, stimulus response, hormone signal biosynthesis/transduction, and nutrient transport under salinity. CONCLUSIONS: The high-resolution salt-responsive gene expression profiling helped the efficient characterization of central members regulating plant salinity resistance. These findings might enhance integrated comprehensive understanding of the morpho-physiologic and molecular responses to salinity and provide elite genetic resources for the genetic modification of salinity-resistant crop species.


Assuntos
Brassica napus/genética , Fotossíntese/efeitos dos fármacos , Reguladores de Crescimento de Plantas/metabolismo , Transcriptoma/efeitos dos fármacos , Brassica napus/efeitos dos fármacos , Brassica napus/fisiologia , Perfilação da Expressão Gênica , Homeostase/efeitos dos fármacos , Íons/metabolismo , Nitrogênio/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/fisiologia , Estômatos de Plantas/efeitos dos fármacos , Estômatos de Plantas/genética , Estômatos de Plantas/fisiologia , Salinidade , Cloreto de Sódio/farmacologia , Tricomas/efeitos dos fármacos , Tricomas/genética , Tricomas/fisiologia
6.
Int J Mol Sci ; 21(22)2020 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-33182819

RESUMO

Improving crop nitrogen (N) limitation adaptation (NLA) is a core approach to enhance N use efficiency (NUE) and reduce N fertilizer application. Rapeseed has a high demand for N nutrients for optimal plant growth and seed production, but it exhibits low NUE. Epigenetic modification, such as DNA methylation and modification from small RNAs, is key to plant adaptive responses to various stresses. However, epigenetic regulatory mechanisms underlying NLA and NUE remain elusive in allotetraploid B. napus. In this study, we identified overaccumulated carbohydrate, and improved primary and lateral roots in rapeseed plants under N limitation, which resulted in decreased plant nitrate concentrations, enhanced root-to-shoot N translocation, and increased NUE. Transcriptomics and RT-qPCR assays revealed that N limitation induced the expression of NRT1.1, NRT1.5, NRT1.7, NRT2.1/NAR2.1, and Gln1;1, and repressed the transcriptional levels of CLCa, NRT1.8, and NIA1. High-resolution whole genome bisulfite sequencing characterized 5094 differentially methylated genes involving ubiquitin-mediated proteolysis, N recycling, and phytohormone metabolism under N limitation. Hypermethylation/hypomethylation in promoter regions or gene bodies of some key N-metabolism genes might be involved in their transcriptional regulation by N limitation. Genome-wide miRNA sequencing identified 224 N limitation-responsive differentially expressed miRNAs regulating leaf development, amino acid metabolism, and plant hormone signal transduction. Furthermore, degradome sequencing and RT-qPCR assays revealed the miR827-NLA pathway regulating limited N-induced leaf senescence as well as the miR171-SCL6 and miR160-ARF17 pathways regulating root growth under N deficiency. Our study provides a comprehensive insight into the epigenetic regulatory mechanisms underlying rapeseed NLA, and it will be helpful for genetic engineering of NUE in crop species through epigenetic modification of some N metabolism-associated genes.


Assuntos
Brassica napus/genética , Brassica napus/metabolismo , Epigênese Genética , Nitrogênio/metabolismo , Adaptação Fisiológica , Brassica napus/crescimento & desenvolvimento , Metilação de DNA , Fertilizantes , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genoma de Planta , MicroRNAs/genética , MicroRNAs/metabolismo , Modelos Biológicos , RNA de Plantas/genética , RNA de Plantas/metabolismo , Tetraploidia
7.
Int J Mol Sci ; 21(10)2020 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-32408717

RESUMO

Soil salinity is a main abiotic stress in agriculture worldwide. The Na+/H+ antiporters (NHXs) play pivotal roles in intracellular Na+ excretion and vacuolar Na+ compartmentalization, which are important for plant salt stress resistance (SSR). However, few systematic analyses of NHXs has been reported in allotetraploid rapeseed so far. Here, a total of 18 full-length NHX homologs, representing seven subgroups (NHX1-NHX8 without NHX5), were identified in the rapeseed genome (AnAnCnCn). Number variations of BnaNHXs might indicate their significantly differential roles in the regulation of rapeseed SSR. BnaNHXs were phylogenetically divided into three evolutionary clades, and the members in the same subgroups had similar physiochemical characteristics, gene/protein structures, and conserved Na+ transport motifs. Darwin´s evolutionary pressure analysis suggested that BnaNHXs suffered from strong purifying selection. The cis-element analysis revealed the differential transcriptional regulation of NHXs between the model Arabidopsis and B. napus. Differential expression of BnaNHXs under salt stress, different nitrogen forms (ammonium and nitrate), and low phosphate indicated their potential involvement in the regulation of rapeseed SSR. Global landscapes of BnaNHXs will give an integrated understanding of their family evolution and molecular features, which will provide elite gene resources for the genetic improvement of plant SSR through regulating the NHX-mediated Na+ transport.


Assuntos
Brassica napus/genética , Resistência a Medicamentos/genética , Família Multigênica , Proteínas de Plantas/genética , Estresse Salino/genética , Trocadores de Sódio-Hidrogênio/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Brassica napus/classificação , Brassica napus/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Genoma de Planta/genética , Filogenia , Proteínas de Plantas/metabolismo , Salinidade , Seleção Genética , Sódio/metabolismo , Trocadores de Sódio-Hidrogênio/metabolismo , Estresse Fisiológico/genética
8.
BMC Plant Biol ; 20(1): 151, 2020 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-32268885

RESUMO

BACKGROUND: Nitrogen (N), referred to as a "life element", is a macronutrient essential for optimal plant growth and yield production. Amino acid (AA) permease (AAP) genes play pivotal roles in root import, long-distance translocation, remobilization of organic amide-N from source organs to sinks, and other environmental stress responses. However, few systematic analyses of AAPs have been reported in Brassica napus so far. RESULTS: In this study, we identified a total of 34 full-length AAP genes representing eight subgroups (AAP1-8) from the allotetraploid rapeseed genome (AnAnCnCn, 2n = 4x = 38). Great differences in the homolog number among the BnaAAP subgroups might indicate their significant differential roles in the growth and development of rapeseed plants. The BnaAAPs were phylogenetically divided into three evolutionary clades, and the members in the same subgroups had similar physiochemical characteristics, gene/protein structures, and conserved AA transport motifs. Darwin's evolutionary analysis suggested that BnaAAPs were subjected to strong purifying selection pressure. Cis-element analysis showed potential differential transcriptional regulation of AAPs between the model Arabidopsis and B. napus. Differential expression of BnaAAPs under nitrate limitation, ammonium excess, phosphate shortage, boron deficiency, cadmium toxicity, and salt stress conditions indicated their potential involvement in diverse nutrient stress responses. CONCLUSIONS: The genome-wide identification of BnaAAPs will provide a comprehensive insight into their family evolution and AAP-mediated AA transport under diverse abiotic stresses. The molecular characterization of core AAPs can provide elite gene resources and contribute to the genetic improvement of crop stress resistance through the modulation of AA transport.


Assuntos
Sistemas de Transporte de Aminoácidos/genética , Brassica napus/enzimologia , Regulação da Expressão Gênica de Plantas , Estresse Fisiológico , Sistemas de Transporte de Aminoácidos/metabolismo , Brassica napus/genética , Estudo de Associação Genômica Ampla , Filogenia
9.
J Exp Bot ; 70(19): 5437-5455, 2019 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-31232451

RESUMO

Oilseed rape (Brassica napus) has great potential for phytoremediation of cadmium (Cd)-polluted soils due to its large plant biomass production and strong metal accumulation. Enhanced plant Cd resistance (PCR) is a crucial prerequisite for phytoremediation through hyper-accumulation of excess Cd. However, the complexity of the allotetraploid genome of rapeseed hinders our understanding of PCR. To explore rapeseed Cd-resistance mechanisms, we examined two genotypes, 'ZS11' (Cd-resistant) and 'W10' (Cd-sensitive), that exhibit contrasting PCR while having similar tissue Cd concentrations, and characterized their different fingerprints in terms of plant morphophysiology (electron microscopy), ion abundance (inductively coupled plasma mass spectrometry), DNA variation (whole-genome resequencing), transcriptomics (high-throughput mRNA sequencing), and metabolomics (ultra-high performance liquid chromatography-mass spectrometry). Fine isolation of cell components combined with ionomics revealed that more Cd accumulated in the shoot vacuoles and root pectins of the resistant genotype than in the sensitive one. Genome and transcriptome sequencing identified numerous DNA variants and differentially expressed genes involved in pectin modification, ion binding, and compartmentalization. Transcriptomics-assisted gene co-expression networks characterized BnaCn.ABCC3 and BnaA8.PME3 as the central members involved in the determination of rapeseed PCR. High-resolution metabolic profiles revealed greater accumulation of shoot Cd chelates, and stronger biosynthesis and higher demethylation of root pectins in the resistant genotype than in the sensitive one. Our comprehensive examination using a multiomics approach has greatly improved our understanding of the role of subcellular reallocation of Cd in the determination of PCR.


Assuntos
Brassica napus/genética , Brassica napus/metabolismo , Cádmio/metabolismo , Genoma de Planta , Poluentes do Solo/metabolismo , Biodegradação Ambiental , Cádmio/toxicidade , Metaboloma , Poluentes do Solo/toxicidade , Transcriptoma
10.
Mol Biol Rep ; 39(5): 5851-7, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22207172

RESUMO

Sinapis alba has many desirable agronomic traits including tolerance to drought. In this investigation, we performed the genome-wide transcriptional profiling of S. alba leaves under drought stress and rewatering growth conditions in an attempt to identify candidate genes involved in drought tolerance, using the Illumina deep sequencing technology. The comparative analysis revealed numerous changes in gene expression level attributable to the drought stress, which resulted in the down-regulation of 309 genes and the up-regulation of 248 genes. Gene ontology analysis revealed that the differentially expressed genes were mainly involved in cell division and catalytic and metabolic processes. Our results provide useful information for further analyses of the drought stress tolerance in Sinapis, and will facilitate molecular breeding for Brassica crop plants.


Assuntos
Secas , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Folhas de Planta/genética , Sinapis/crescimento & desenvolvimento , Sinapis/genética , Estresse Fisiológico/genética , Análise por Conglomerados , Regulação para Baixo/genética , Dosagem de Genes/genética , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Genes de Plantas/genética , Anotação de Sequência Molecular , Folhas de Planta/crescimento & desenvolvimento , Regulação para Cima/genética , Água
11.
Mol Biol Rep ; 38(2): 1055-61, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20571911

RESUMO

To screen the genes related to Brassica napus seed development at pattern formation and maturation stages, the suppression subtractive cDNA libraries of B. napus cultivar Zhongshuang 6 were constructed with its embryos at 10 days after flowering (10 DAF) and 30 days after flowering (30 DAF) through suppression subtractive hybridization (SSH) technology. The positive clones were screened by PCR and dot blot hybridization, and then sequenced. High quality expressed sequence tags (ESTs) were used for COG functional classification with COGNITOR software, as well as analysis and annotation with BLAST software. Tissue-specific detection of five genes screened was performed by RT-PCR in root, stem, leaf, flower, bud, pod, and embryo tissues. The insert size ranged from 100 to 900 bp, with an average size of about 500 bp. According to COG functional classification database, the differentially expressed genes mainly involved in lipid and amino acid metabolism, signal transduction, post-transcriptional modification, and etc. The results from RT-PCR detection of the five differentially expressed genes indicated that genes 2-96 and 2-352 presented embryo-specific expression, gene 1-385 expressed in parts of tissues, and genes 1-71 and 1-682 expressed in all tissues. Two genes were found to be involved in seed development, lipid and protein metabolisms, two genes may be involved in signal transduction, one gene could not match with the homologous sequences known to date and was likely a new gene. These results are helpful for future gene cloning and their functional analysis.


Assuntos
Brassica/genética , Brassica/metabolismo , Regulação da Expressão Gênica de Plantas , Sementes/metabolismo , Clonagem Molecular , DNA Complementar/metabolismo , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Biblioteca Gênica , Genes de Plantas , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Software
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