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1.
Chemosphere ; 282: 130897, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34470145

RESUMO

Mussel shell (MS) and biochar (BC) are commonly used for the remediation of metal contaminated soil. However, less research has been focused to examine the efficacy of their combinations to reduce metal toxicity in crop plants. This study was therefore conducted to investigate the effects of BC, MS and their activated concoctions on the soil properties, enzyme activities and nickel (Ni) immobilization in aged Ni contaminated soil. Moreover, the growth, photosynthetic pigments and anti-oxidative machnery of Brassica napus plants has also been investigated in order to determine amendments efficiency in reducing soil Ni toxicity for plants. The results showed that the application of Ni adversely affected soil health and trigged stress responses by inducing oxidative stress in B. napus. However, the incorporation of amendments reduced the bioavailability of Ni, and the concoctions of BC and MS showed promising results in the immobilization of Ni. Among various combinations of BC and MS, treatment with BC + MS (3:1) significantly reduced Ni uptake, decreased reactive oxygen species (ROS) and enhanced antioxidant defense of B. napus plants. Results showed that amendment's combinations stimulated the transcriptional levels of ROS scavenging enzymes and suppressed the expression level of Ni transporters. The morphological and physical characterization techniques (i.e. SEM, BET, EDS, FTIR and X-ray diffraction analyses) showed that amendment's combinations had relatively higher Ni adsorption capacity, indicating that BC and MS concoctions are efficient immobilizing agents for minimizing Ni availability, preventing oxidative toxicity and promoting growth and biomass production in rapeseed plants under metal stress conditions.


Assuntos
Bivalves , Brassica napus , Poluentes do Solo , Animais , Carvão Vegetal , Níquel/análise , Níquel/toxicidade , Solo , Poluentes do Solo/análise , Poluentes do Solo/toxicidade
2.
Ecotoxicol Environ Saf ; 225: 112695, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34478972

RESUMO

The advent of the nanotechnology era offers a unique opportunity for sustainable agriculture, and the contribution of nanoparticles (NPs) to ameliorate abiotic stresses became the new area of interest for researchers due to their special physiochemical characteristics in the biological system. Salinity is a key devastating abiotic factor that hinders the development and yield of rapeseed. On the flip side, the impact of nanoparticles on plant hormones upon salt stress during seed imbibition and germination has been poorly understood. Hence, we aimed to study the influence of nanopriming on plant hormones and germination processes using selenium and zinc oxide nanoparticles (SeNPs and ZnONPs) during seed imbibition and the early seedling stage upon salinity stress. Nanopriming showed a positive effect on final germination percentage, germination rate, seed microstructure, and antioxidant enzyme activity of two rapeseed cultivars under salt stress. Moreover, nano-treatment decreased the expression of abscisic acid related genes BnCYP707A1, 3, and 4 during the priming time and after sowing, where the levels of BnCYP707A1, and 3 genes showed a slightly significant difference between the nanopriming and hydropriming, which gave an evidence that the nanopriming influenced the ABA levels then elevated the seed germination with SeNPs and ZnONPs. Likewise, nanoparticles significantly elevated the expression levels of BnGA20ox, BnGA3ox and BnCPS genes during the germination stage, especially at 24 h after being sown in salt stress. That confirms the positive role of SeNPs and ZnONPs in regulating gibberellic acid level, which increases the germination in primed seeds as compared to unprimed seeds and hydroprimed seeds. Additionally, our results demonstrated that nanopriming regulated the expression level of BnCAM and BnPER during priming time and after sowing, along with the various levels of expression remarkably in BnEXP4 and BnRAB28, especially at 24 h of being sown under salt stress, which promoted seed germination and early seedling growth. Overall, this work provides new insights into mechanisms underlying the interactions of SeNPs and ZnONPs with plant hormones during the seed imbibition and early seedling stage, consequently enhanced plant growth and development. Additionally, these findings portrayed that the application of SeNPs and ZnONPs could be a new strategy and useful approach to enhance tolerance against salinity in rapeseed plants.


Assuntos
Brassica napus , Nanopartículas , Selênio , Óxido de Zinco , Brassica napus/genética , Germinação , Nanopartículas/toxicidade , Estresse Salino , Plântula , Sementes , Óxido de Zinco/toxicidade
3.
J Hazard Mater ; 416: 125921, 2021 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-34492853

RESUMO

In-situ stabilization has been considered an effective way to remediate metal contaminated soil. Thus, pot experiments were undertaken to investigate the effectiveness of multiple stabilization agents such as biochar (BC), mussel shell (MS), zeolite (ZE) and limestone (LS) on the immobilization of Ni, physicochemical features and enzyme activities in polluted soil. Results showed that the sole application of Ni adversely affected the rapeseed growth, photosynthetic pigments, and antioxidative defense. However, the addition of amendments to the contaminated soil significantly reduced Ni bioavailability. The XRD analysis confirmed the formation of Ni related ligands and FTIR showed the presence of hydroxyl, carboxyl and sulfur functional groups, as well as complexation and adsorption of Ni on amendments. Among multiple amendments, biochar significantly enhanced plant biomass attributes and total chlorophyll content. Moreover, addition of amendments also strengthened the antioxidant defense by decreasing Ni induced oxidative stress (H2O2 and O2.-), increased macronutrient availability, reduced Ni uptake and improved soil health. The qPCR analysis showed that the Ni transporters were significantly suppressed by amendments, which is correlated with the lower accumulation of Ni in rapeseed. The present study showed that immobilizing agents, especially biochar, is an effective amendment to immobilize Ni in soil, which restricts its entry into the food chain.


Assuntos
Brassica napus , Poluentes do Solo , Carvão Vegetal , Peróxido de Hidrogênio , Níquel/análise , Estresse Oxidativo , Solo , Poluentes do Solo/análise
4.
J Plant Physiol ; 265: 153505, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34481359

RESUMO

Brassicaceae seeds consist of three genetically distinct structures: the embryo, endosperm and seed coat, all of which are involved in assimilate allocation during seed development. The complexity of their metabolic interrelations remains unresolved to date. In the present study, we apply state-of-the-art imaging and analytical approaches to assess the metabolic environment of the Brassica napus embryo. Nuclear magnetic resonance imaging (MRI) provided volumetric data on the living embryo and endosperm, revealing how the endosperm envelops the embryo, determining endosperm's priority in assimilate uptake from the seed coat during early development. MRI analysis showed higher levels of sugars in the peripheral endosperm facing the seed coat, but a lower sugar content within the central vacuole and the region surrounding the embryo. Feeding intact siliques with 13C-labeled sucrose allowed tracing of the post-phloem route of sucrose transfer within the seed at the heart stage of embryogenesis, by means of mass spectrometry imaging. Quantification of over 70 organic and inorganic compounds in the endosperm revealed shifts in their abundance over different stages of development, while sugars and potassium were the main determinants of osmolality throughout these stages. Our multidisciplinary approach allows access to the hidden aspects of endosperm metabolism, a task which remains unattainable for the small-seeded model plant Arabidopsis thaliana.


Assuntos
Brassica napus/crescimento & desenvolvimento , Brassica napus/metabolismo , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Brassica napus/genética , Brassica napus/ultraestrutura , Endosperma/genética , Endosperma/ultraestrutura , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Sementes/genética , Sementes/ultraestrutura
5.
Ecotoxicol Environ Saf ; 225: 112765, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34530260

RESUMO

We aimed to investigate how sulfur (S) application prior to oilseed rape cultivation influences the uptake of cadmium (Cd) by rice grown in low- and high-Cd soils. A pot experiment involving four S levels (0, 30, 60, 120 mg S kg-1) combined with two Cd rates (low and high-0.35 and 10.35 mg Cd kg-1, respectively) was conducted. Soil pore water during rice growth and plant tissues at maturity were analyzed. The soil pore water results indicated that S application decreased Cd solubility under submergence due to the S-induced increase of soil pH and the enhancement of sulfide formation in soil micropores. When S was applied at rates of 30, 60 and 120 mg S kg-1, brown rice Cd concentrations decreased by 18%, 18%, and 55% (p < 0.05) in the low-Cd soil but increased by 20%, 40%, and 40% in the high-Cd soil compared with those in the non-S treatment. The different effects of S on Cd accumulation in brown rice were related to Cd-induced oxidative stress in the rice plants. In low-Cd soils, a S-induced increase in phytochelatins in rice roots restricted and inhibited Cd translocation in brown rice. In high-Cd soils, the Cd-induced oxidative stress in rice plants weakened the protective effects of S, while highlighted the promotion of Cd uptake by S. Overall, S fertilizer is recommended for oilseed rape-rice rotations in low-Cd paddy fields. In high Cd-contaminated fields, oilseed rape-rice rotations are suitable for the simultaneous remediation by oilseed rape and production of rice without S fertilization.


Assuntos
Brassica napus , Oryza , Poluentes do Solo , Cádmio/toxicidade , Solo , Poluentes do Solo/toxicidade , Enxofre
6.
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
7.
BMC Plant Biol ; 21(1): 359, 2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-34353289

RESUMO

BACKGROUND: Plant height is an important architecture trait which is a fundamental yield-determining trait in crops. Variety with dwarf or semi-dwarf phenotype is a major objective in the breeding because dwarfing architecture can help to increase harvest index, increase planting density, enhance lodging resistance, and thus be suitable for mechanization harvest. Although some germplasm or genes associated with dwarfing plant type have been carried out. The molecular mechanisms underlying dwarfism in oilseed rape (Brassica napus L.) are poorly understood, restricting the progress of breeding dwarf varieties in this species. Here, we report a new dwarf mutant Bndwarf2 from our B. napus germplasm. We studied its inheritance and mapped the dwarf locus BnDWARF2. RESULTS: The inheritance analysis showed that the dwarfism phenotype was controlled by one semi-dominant gene, which was mapped in an interval of 787.88 kb on the C04 chromosome of B. napus by Illumina Brassica 60 K Bead Chip Array. To fine-map BnDWARF2, 318 simple sequence repeat (SSR) primers were designed to uniformly cover the mapping interval. Among them, 15 polymorphic primers that narrowed down the BnDWARF2 locus to 34.62 kb were detected using a F2:3 family population with 889 individuals. Protein sequence analysis showed that only BnaC04.BIL1 (BnaC04g41660D) had two amino acid residues substitutions (Thr187Ser and Gln399His) between ZS11 and Bndwarf2, which encoding a GLYCOGEN SYNTHASE KINASE 3 (GSK3-like). The quantitative real-time PCR (qRT-PCR) analysis showed that the BnaC04.BIL1 gene expressed in all tissues of oilseed rape. Subcellular localization experiment showed that BnaC04.BIL1 was localized in the nucleus in tobacco leaf cells. Genetic transformation experiments confirmed that the BnaC04.BIL1 is responsible for the plant dwarf phenotype in the Bndwarf2 mutants. Overexpression of BnaC04.BIL1 reduced plant height, but also resulted in compact plant architecture. CONCLUSIONS: A dominant dwarfing gene, BnaC04.BIL1, encodes an GSK3-like that negatively regulates plant height, was mapped and isolated. Our identification of a distinct gene locus may help to improve lodging resistance in oilseed rape.


Assuntos
Brassica napus/crescimento & desenvolvimento , Brassica napus/genética , Proteínas de Plantas/genética , Mapeamento Cromossômico , Clonagem Molecular , Regulação da Expressão Gênica de Plantas , Quinase 3 da Glicogênio Sintase/genética , Mutação , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Tabaco/genética
8.
BMC Plant Biol ; 21(1): 366, 2021 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-34380425

RESUMO

BACKGROUND: Small RNAs are short non-coding RNAs that are key gene regulators controlling various biological processes in eukaryotes. Plants may regulate discrete sets of sRNAs in response to pathogen attack. Sclerotinia sclerotiorum is an economically important pathogen affecting hundreds of plant species, including the economically important oilseed B. napus. However, there are limited studies on how regulation of sRNAs occurs in the S. sclerotiorum and B. napus pathosystem. RESULTS: We identified different classes of sRNAs from B. napus using high throughput sequencing of replicated mock and infected samples at 24 h post-inoculation (HPI). Overall, 3999 sRNA loci were highly expressed, of which 730 were significantly upregulated during infection. These 730 up-regulated sRNAs targeted 64 genes, including disease resistance proteins and transcriptional regulators. A total of 73 conserved miRNA families were identified in our dataset. Degradome sequencing identified 2124 cleaved mRNA products from these miRNAs from combined mock and infected samples. Among these, 50 genes were specific to infection. Altogether, 20 conserved miRNAs were differentially expressed and 8 transcripts were cleaved by the differentially expressed miRNAs miR159, miR5139, and miR390, suggesting they may have a role in the S. sclerotiorum response. A miR1885-triggered disease resistance gene-derived secondary sRNA locus was also identified and verified with degradome sequencing. We also found further evidence for silencing of a plant immunity related ethylene response factor gene by a novel sRNA using 5'-RACE and RT-qPCR. CONCLUSIONS: The findings in this study expand the framework for understanding the molecular mechanisms of the S. sclerotiorum and B. napus pathosystem at the sRNA level.


Assuntos
Ascomicetos/fisiologia , Brassica napus/genética , Brassica napus/microbiologia , Doenças das Plantas/microbiologia , RNA de Plantas , Pequeno RNA não Traduzido , Sequência Conservada , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/genética , Análise de Sequência de RNA , Regulação para Cima
9.
Biomolecules ; 11(8)2021 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-34439819

RESUMO

With the introduction of the new auxinic herbicide halauxifen-methyl into the oilseed rape (Brassica napus) market, there is a need to understand how this new molecule interacts with indigenous plant hormones (e.g., IAA) in terms of crop response. The aim of this study was to investigate the molecular background by using different growth conditions under which three different auxinic herbicides were administered. These were halauxifen-methyl (Hal), alone and together with aminopyralid (AP) as well as picloram (Pic). Three different hormone classes were determined, free and conjugated indole-3-acetic acid (IAA), aminocyclopropane carboxylic acid (ACC) as a precursor for ethylene, and abscisic acid (ABA) at two different temperatures and growth stages as well as over time (2-168 h after treatment). At 15 °C growth temperature, the effect was more pronounced than at 9 °C, and generally, the younger leaves independent of the developmental stage showed a larger effect on the alterations of hormones. IAA and ACC showed reproducible alterations after auxinic herbicide treatments over time, while ABA did not. Finally, a transcriptome analysis after treatment with two auxinic herbicides, Hal and Pic, showed different expression patterns. Hal treatment leads to the upregulation of auxin and hormone responses at 48 h and 96 h. Pic treatment induced the hormone/auxin response already after 2 h, and this continued for the other time points. The more detailed analysis of the auxin response in the datasets indicate a role for GH3 genes and genes encoding auxin efflux proteins. The upregulation of the GH3 genes correlates with the increase in conjugated IAA at the same time points and treatments. Also, genes for were found that confirm the upregulation of the ethylene pathway.


Assuntos
Ácido Abscísico/farmacologia , Aminoácidos Cíclicos/farmacologia , Brassica napus/efeitos dos fármacos , Herbicidas/farmacologia , Ácidos Indolacéticos/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Ácido Abscísico/metabolismo , Aminoácidos Cíclicos/metabolismo , Brassica napus/genética , Brassica napus/metabolismo , Ácidos Carboxílicos/farmacologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Ontologia Genética , Ácidos Indolacéticos/metabolismo , Anotação de Sequência Molecular , Picloram/farmacologia , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Piridinas/farmacologia , Temperatura , Transcriptoma
10.
Int J Mol Sci ; 22(16)2021 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-34445724

RESUMO

Plant height is one of the most important agronomic traits of rapeseeds. In this study, we characterized a dwarf Brassica napus mutant, named ndf-2, obtained from fast neutrons and DES mutagenesis. Based on BSA-Seq and genetic properties, we identified causal mutations with a time-saving approach. The ndf-2 mutation was identified on chromosome A03 and can result in an amino acid substitution in the conserved degron motif (GWPPV to EWPPV) of the Auxin/indole-3-acetic acid protein 7 (BnaA03.IAA7) encoded by the causative gene. Aux/IAA protein is one of the core components of the auxin signaling pathway, which regulates many growth and development processes. However, the molecular mechanism of auxin signal regulating plant height is still not well understood. In the following work, we identified that BnaARF6 and BnaARF8 as interactors of BnaA03.IAA7 and BnaEXPA5 as a target of BnaARF6 and BnaARF8. The three genes BnaA03.IAA7, BnaARF6/8 and BnaEXPA5 were highly expressed in stem, suggesting that these genes were involved in stem development. The overexpression of BnaEXPA5 results in larger rosettes leaves and longer inflorescence stems in Arabidopsis thaliana. Our results indicate that BnaA03.IAA7- and BnaARF6/8-dependent auxin signal control stem elongation and plant height by regulating the transcription of BnaEXPA5 gene, which is one of the targets of this signal.


Assuntos
Brassica napus/genética , Parede Celular/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Caules de Planta/crescimento & desenvolvimento , Arabidopsis , Brassica napus/crescimento & desenvolvimento , Brassica napus/metabolismo , Mutação com Ganho de Função , Ácidos Indolacéticos/metabolismo , Proteínas de Plantas/metabolismo
11.
Plant Physiol Biochem ; 167: 529-540, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34425397

RESUMO

Proline biosynthesis and accumulation is a common response to unfavorable environment in many plants. This work aimed to elucidate the effects of boron (B)-deficiency and toxicity on proline metabolism and biosynthesis in Brassica napus in a hydroponic experiment. The results showed that B-deficiency and toxicity exert injurious impact on plant growth, accumulated high malondialdehyde (MDA) content, and caused the destruction of subcellular structure. Proline accumulated in both B deprivation and B toxicity plants, except B toxicity-treated root. In roots, B-deficiency increased ornithine content and pyrroline-5-carboxylate reductase (P5CR) activity, with the higher expression of BnaC03.P5CR, whilst decreased glutamate, glutamate-1-semialdehyde (GSA), pyrroline-5-carboxylate (P5C) contents and ornithine-δ-aminotransferase (δ-OAT), pyrroline-5-carboxylate synthetase (P5CS), proline dehydrogenase (ProDH) activities in terms of down-regulated the BnaC04.P5CS2, BnaA04.P5CS2, and BnaAnn.ProDH expression. The glutamate and GSA contents were decreased while P5C, arginine, and ornithine contents were enhanced in leaves under B-deficient and toxicity conditions. Lower glutamate pathway-related substance contents, P5CR, and δ-OAT activities while higher ProDH activity along with the same trend of related-gene expression were observed in B-toxicity-treated roots. Importantly, principal component analysis (PCA) in conjunction with correlation analysis indicated that ornithine pathway-related substances and enzymes contributed more to proline accumulation in B-deficient plant and B toxicity-treated leaves. Collectively, proline accumulation is caused by increased synthesis and decreased decomposition, and positively contributed, at least partly, by regulated ornithine pathway.


Assuntos
Brassica napus , Boro/toxicidade , Ácido Glutâmico , Prolina , Prolina Oxidase
12.
Theor Appl Genet ; 134(10): 3287-3303, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34410456

RESUMO

KEY MESSAGE: Two CO paralogs in Brassica napus were confirmed and shown distinct expression pattern and function in promoting flowering and allelic variation s within BnaCO.A10 were found closely associated with ecotype divergence. CONSTANS (CO) is a key gene that responds to photoperiod and in Arabidopsis can promote flowering under long-day (LD) conditions. Brassica napus L. is a major oil crop and close relative of Arabidopsis, and arose via allopolyploidization from the diploids B. rapa (A genome) and B. oleracea (C genome). In this study, we confirmed that B. napus has two CO genes located on the A10 (BnaCO.A10) and C9 (BnaCO.C9) chromosomes. Significant differences in level and temporal pattern of transcription, as well as in protein function, of these homoeologous may have resulted from sequence variation in the promoter as well as in the coding region. Apart from two insertions of 527 bp and 2002 bp in the promoter of BnaCO.C9 that function as transcriptional enhancers, this gene is otherwise highly conserved in both promoter and coding region. However, BnaCO.A10 was classified into two haplotypes and transgene analysis in Arabidopsis and backcross analysis in rapeseed indicated that the winter-type haplotype had a greater effect in promoting flowering than the spring type. We discuss the contribution of CO alleles to species evolution, and for eco-geographic radiation following crop domestication, alongside scope for managing this locus in future breeding.


Assuntos
Brassica napus/crescimento & desenvolvimento , Cromossomos de Plantas/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Fenótipo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Alelos , Brassica napus/genética , Mapeamento Cromossômico/métodos , Ecótipo , Evolução Molecular , Flores/genética , Fotoperíodo , Filogenia , Melhoramento Vegetal , Proteínas de Plantas/genética , Locos de Características Quantitativas , Estações do Ano , Fatores de Transcrição/genética
13.
Nat Plants ; 7(7): 899-905, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34211132

RESUMO

Plant organelles including mitochondria and chloroplasts contain their own genomes, which encode many genes essential for respiration and photosynthesis, respectively. Gene editing in plant organelles, an unmet need for plant genetics and biotechnology, has been hampered by the lack of appropriate tools for targeting DNA in these organelles. In this study, we developed a Golden Gate cloning system1, composed of 16 expression plasmids (8 for the delivery of the resulting protein to mitochondria and the other 8 for delivery to chloroplasts) and 424 transcription activator-like effector subarray plasmids, to assemble DddA-derived cytosine base editor (DdCBE)2 plasmids and used the resulting DdCBEs to efficiently promote point mutagenesis in mitochondria and chloroplasts. Our DdCBEs induced base editing in lettuce or rapeseed calli at frequencies of up to 25% (mitochondria) and 38% (chloroplasts). We also showed DNA-free base editing in chloroplasts by delivering DdCBE mRNA to lettuce protoplasts to avoid off-target mutations caused by DdCBE-encoding plasmids. Furthermore, we generated lettuce calli and plantlets with edit frequencies of up to 99%, which were resistant to streptomycin or spectinomycin, by introducing a point mutation in the chloroplast 16S rRNA gene.


Assuntos
Brassica napus/genética , Clonagem de Organismos/métodos , DNA de Cloroplastos , DNA Mitocondrial , Edição de Genes/métodos , Alface/genética , Melhoramento Vegetal/métodos , Produtos Agrícolas/genética
14.
Int J Mol Sci ; 22(14)2021 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-34299178

RESUMO

Timely flowering is important for seed formation and maximization of rapeseed (Brassica napus) yield. Here, we performed flowering-time quantitative trait loci (QTL) mapping using a double haploid (DH) population grown in three environments to study the genetic architecture. Brassica 60 K Illumina Infinium™ single nucleotide polymorphism (SNP) array and simple sequence repeat (SSR) markers were used for genotyping of the DH population, and a high-density genetic linkage map was constructed. QTL analysis of flowering time from the three environments revealed five consensus QTLs, including two major QTLs. A major QTL located on chromosome A03 was detected specifically in the semi-winter rapeseed growing region, and the one on chromosome C08 was detected in all environments. Ribonucleic acid sequencing (RNA-seq) was performed on the parents' leaves at seven time-points in a day to determine differentially expressed genes (DEGs). The biological processes and pathways with significant enrichment of DEGs were obtained. The DEGs in the QTL intervals were analyzed, and four flowering time-related candidate genes were found. These results lay a foundation for the genetic regulation of rapeseed flowering time and create a rapeseed gene expression library for seven time-points in a day.


Assuntos
Brassica napus/crescimento & desenvolvimento , Brassica napus/genética , Perfilação da Expressão Gênica/métodos , Locos de Características Quantitativas , Mapeamento Cromossômico/métodos , Flores/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Haploidia , Polimorfismo de Nucleotídeo Único
15.
Sensors (Basel) ; 21(14)2021 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-34300368

RESUMO

The three-dimensional reconstruction method using RGB-D camera has a good balance in hardware cost and point cloud quality. However, due to the limitation of inherent structure and imaging principle, the acquired point cloud has problems such as a lot of noise and difficult registration. This paper proposes a 3D reconstruction method using Azure Kinect to solve these inherent problems. Shoot color images, depth images and near-infrared images of the target from six perspectives by Azure Kinect sensor with black background. Multiply the binarization result of the 8-bit infrared image with the RGB-D image alignment result provided by Microsoft corporation, which can remove ghosting and most of the background noise. A neighborhood extreme filtering method is proposed to filter out the abrupt points in the depth image, by which the floating noise point and most of the outlier noise will be removed before generating the point cloud, and then using the pass-through filter eliminate rest of the outlier noise. An improved method based on the classic iterative closest point (ICP) algorithm is presented to merge multiple-views point clouds. By continuously reducing both the size of the down-sampling grid and the distance threshold between the corresponding points, the point clouds of each view are continuously registered three times, until get the integral color point cloud. Many experiments on rapeseed plants show that the success rate of cloud registration is 92.5% and the point cloud accuracy obtained by this method is 0.789 mm, the time consuming of a integral scanning is 302 s, and with a good color restoration. Compared with a laser scanner, the proposed method has considerable reconstruction accuracy and a significantly ahead of the reconstruction speed, but the hardware cost is much lower when building a automatic scanning system. This research shows a low-cost, high-precision 3D reconstruction technology, which has the potential to be widely used for non-destructive measurement of rapeseed and other crops phenotype.


Assuntos
Brassica napus , Imageamento Tridimensional , Algoritmos , Produtos Agrícolas
16.
J Agric Food Chem ; 69(30): 8536-8545, 2021 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-34296617

RESUMO

pH shift is an effective technique for modifying functional properties of food proteins. However, it can increase lysinoalanine (LAL) content under alkali conditions, thus limiting the use of proteins. This study investigated the inhibition effect of ultrasonic parameters on LAL formation in rapeseed protein isolates (RPI) during pH shift treatment (pH-ST). Results showed that the content of LAL decreased by 49.5% and 74.1%, following the use of ultrasound (28 kHz, 40 W/L, 40 °C, and 30 min) under alkali and acidic treatment, respectively. Structural analysis showed that after ultrasonic irradiation, increased sulfhydryl groups and amino acids reduced the dehydroalanine and, thus, decreased LAL content. Particle size, secondary structure, and microstructure (SEM, AFM) analyses showed relative dispersion in protein distribution, reducing intermolecular or intramolecular cross-linking, thereby lowering the LAL content. Thus, ultrasonic-aided pH-ST may be an operational technique toward minimizing LAL formation in RPI.


Assuntos
Brassica napus , Lisinoalanina , Aminoácidos , Concentração de Íons de Hidrogênio , Proteínas
17.
Plant Sci ; 310: 110963, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34315588

RESUMO

Ammonium (NH4+) toxicity has become a serious ecological and agricultural issue owing to increasing soil nitrogen inputs and atmospheric nitrogen deposition. There is accumulating evidence for the mechanisms underlying NH4+-tolerance in rice and Arabidopsis, but similar knowledge for dryland crops is currently limited. We investigated the responses of a natural population of allotetraploid rapeseed to NH4+ and nitrate (NO3-) and screened one NH4+-tolerant genotype (T5) and one NH4+-sensitive genotype (S211). Determination of the shoot and root NH4+ concentrations showed that levels were higher in S211 than in T5. 15NH4+ uptake assays, glutamine synthetase (GS) activity quantification, and relative gene transcriptional analysis indicated that the significantly higher GS activity observed in T5 roots than that in S211 was the main reason for its NH4+-tolerance. In-depth metabolomic analysis verified that Gln metabolism plays an important role in rapeseed NH4+-tolerance. Furthermore, adaptive changes in carbon metabolism were much more active in T5 shoots than in S211. Interestingly, we found that N-glycosylation pathway was significantly induced by NH4+, especially the mannose metabolism, which concentration was 2.75-fold higher in T5 shoots than in S211 with NH4+ treatment, indicating that mannose may be a metabolomic marker which also confers physiological adaptations for NH4+ tolerance in rapeseed. The corresponding amino acid and soluble sugar concentrations and gene expression in T5 and S211 were consistent with these results. Genomic sequencing identified variations in the GLN (encoding GS) and GMP1 (encoding the enzyme that provides GDP-mannose) gene families between the T5 and S211 lines. These genes will be utilized as candidate genes for future investigations of the molecular mechanisms underlying NH4+ tolerance in rapeseed.


Assuntos
Compostos de Amônio/toxicidade , Glutamato-Amônia Ligase/metabolismo , Glutamina/metabolismo , Manose/metabolismo , Brassica napus/metabolismo , Regulação da Expressão Gênica de Plantas , Genótipo , Glutamato-Amônia Ligase/genética , Raízes de Plantas/genética , Raízes de Plantas/metabolismo
18.
Plant Sci ; 310: 110980, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34315596

RESUMO

Flowering is an important turning point from vegetative growth to reproductive growth, and vernalization is an essential condition for the flowering of annual winter plants. To investigate the genetic architecture of flowering time in rapeseed, we used the 60 K Brassica Infinium SNP array to perform a genome-wide analysis of haplotype blocks associated with flowering time in 203 Chinese semi-winter rapeseed inbred lines. Twenty-one haplotype regions carrying one or more candidate genes showed a significant association with flowering time. Interestingly, we detected a SNP (Bn-scaff_22728_1-p285715) located in exon 3 of the BnVIN3-C03 gene that showed a significant association with flowering time on chromosome C03. Based on the SNP alleles A and G, two groups of accessions with early and late flowering time phenotypes were selected, respectively, and PCR amplification and gene expression analysis were combined to reveal the structural variation of the BnVIN3-C03 gene that affected flowering time. Moreover, we found that BnVIN3-C03 inhibited the expression of BnFLC-A02, BnFLC-A03.1, BnFLC-A10 and BnFLC-C03.1, thus modulating the flowering time of Brassica napus. This result provides insight into the genetic improvement of flowering time in B. napus.


Assuntos
Brassica napus/genética , Estudo de Associação Genômica Ampla/métodos , Transcriptoma/genética , Alelos , Mapeamento Cromossômico , Flores/genética , Haplótipos/genética , Proteínas de Plantas/genética , Locos de Características Quantitativas/genética
19.
BMC Genomics ; 22(1): 548, 2021 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-34273948

RESUMO

BACKGROUND: Lipid phosphate phosphatases (LPP) are critical for regulating the production and degradation of phosphatidic acid (PA), an essential signaling molecule under stress conditions. Thus far, the LPP family genes have not been reported in rapeseed (Brassica napus L.). RESULTS: In this study, a genome-wide analysis was carried out to identify LPP family genes in rapeseed that respond to different stress conditions. Eleven BnLPPs genes were identified in the rapeseed genome. Based on phylogenetic and synteny analysis, BnLPPs were classified into four groups (Group I-Group IV). Gene structure and conserved motif analysis showed that similar intron/exon and motifs patterns occur in the same group. By evaluating cis-elements in the promoters, we recognized six hormone- and seven stress-responsive elements. Further, six putative miRNAs were identified targeting three BnLPP genes. Gene ontology analysis disclosed that BnLPP genes were closely associated with phosphatase/hydrolase activity, membrane parts, phosphorus metabolic process, and dephosphorylation. The qRT-PCR based expression profiles of BnLPP genes varied in different tissues/organs. Likewise, several gene expression were significantly up-regulated under NaCl, PEG, cold, ABA, GA, IAA, and KT treatments. CONCLUSIONS: This is the first report to describe the comprehensive genome-wide analysis of the rapeseed LPP gene family. We identified different phytohormones and abiotic stress-associated genes that could help in enlightening the plant tolerance against phytohormones and abiotic stresses. The findings unlocked new gaps for the functional verification of the BnLPP gene family during stresses, leading to rapeseed improvement.


Assuntos
Brassica napus , Brassica napus/genética , Brassica napus/metabolismo , Regulação da Expressão Gênica de Plantas , Família Multigênica , Fosfolipases , Fosfolipídeos , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genética
20.
BMC Genomics ; 22(1): 539, 2021 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-34256693

RESUMO

BACKGROUND: Associative transcriptomics has been used extensively in Brassica napus to enable the rapid identification of markers correlated with traits of interest. However, within the important vegetable crop species, Brassica oleracea, the use of associative transcriptomics has been limited due to a lack of fixed genetic resources and the difficulties in generating material due to self-incompatibility. Within Brassica vegetables, the harvestable product can be vegetative or floral tissues and therefore synchronisation of the floral transition is an important goal for growers and breeders. Vernalisation is known to be a key determinant of the floral transition, yet how different vernalisation treatments influence flowering in B. oleracea is not well understood. RESULTS: Here, we present results from phenotyping a diverse set of 69 B. oleracea accessions for heading and flowering traits under different environmental conditions. We developed a new associative transcriptomics pipeline, and inferred and validated a population structure, for the phenotyped accessions. A genome-wide association study identified miR172D as a candidate for the vernalisation response. Gene expression marker association identified variation in expression of BoFLC.C2 as a further candidate for vernalisation response. CONCLUSIONS: This study describes a new pipeline for performing associative transcriptomics studies in B. oleracea. Using flowering time as an example trait, it provides insights into the genetic basis of vernalisation response in B. oleracea through associative transcriptomics and confirms its characterisation as a complex G x E trait. Candidate leads were identified in miR172D and BoFLC.C2. These results could facilitate marker-based breeding efforts to produce B. oleracea lines with more synchronous heading dates, potentially leading to improved yields.


Assuntos
Brassica napus , Brassica , Brassica/genética , Brassica napus/genética , Estudo de Associação Genômica Ampla , Melhoramento Vegetal , Transcriptoma
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