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1.
Theor Appl Genet ; 134(1): 159-169, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33011819

RESUMO

KEY MESSAGE: Overexpression and virus-induced gene silencing verified BoDFR1 conferred the anthocyanin accumulation in pink-leaved ornamental kale. Leaf color is an essential trait in the important horticultural biennial plant ornamental kale (Brassica oleracea var. acephala). The identity of the gene conferring this striking trait and its mode of inheritance are topics of debate. Based on an analysis of F1, F2, BC1P1, and BC1P2 ornamental kale populations derived from a cross between a pink-leaved P28 and white-leaved D10 line, we determined that the pink leaf trait is controlled by a semi-dominant gene. We cloned two genes potentially involved in anthocyanin biosynthesis in ornamental kale: Bo9g058630 and Bo6g100940. Based on their variation in sequence, we speculated that Bo9g058630, encoding the kale dihydroflavonol-4 reductase (BoDFR1) enzyme, plays a critical role in the development of the pink leaf trait. Indeed, an InDel marker specific for BoDFR1 completely co-segregated with the pink leaf trait in our F2 population. We then generated the 35Spro: DFR-GUS overexpression vector, which we transformed into D10. Overexpression of BoDFR1 indeed restored some anthocyanin accumulation in this white-leaved parental line. In addition, we targeted BoDFR1 in P28 using virus-induced gene silencing. Again, silencing of BoDFR1 resulted in a substantial decrease in anthocyanin accumulation. This work lays the foundation for further exploration of the mechanism underlying anthocyanin accumulation in pink-leaved ornamental kale.


Assuntos
Oxirredutases do Álcool/metabolismo , Antocianinas/biossíntese , Brassica/enzimologia , Proteínas de Plantas/metabolismo , Oxirredutases do Álcool/genética , Brassica/genética , Cruzamentos Genéticos , Inativação Gênica , Genes de Plantas , Marcadores Genéticos , Mutação INDEL , Pigmentação/genética , Folhas de Planta , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas
2.
J Chem Ecol ; 47(1): 112-122, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33180275

RESUMO

Plants evolved in close contact with a myriad of microorganisms, some of which formed associations with their roots, benefitting from carbohydrates and other plant resources. In exchange, they evolved to influence important plant functions, e.g. defense against insect herbivores and other antagonists. Here, we test whether a fungus, Metarhizium brunneum, which is mostly known as an insect pathogen, can also associate with plant roots and contribute to above-ground plant defense. Cauliflower (Brassica oleracea var. botrytis) seeds were sown together with M. brunneum-inoculated rice grains, and the resulting plants subjected to leaf herbivory by the specialist Plutella xylostella. Activity of myrosinases, the enzymes activating glucosinolates, was measured before and after herbivory; larval consumption and plant weight at the end of experiments. Metarhizium brunneum clearly established in the plant roots, and after herbivory myrosinase activity was substantially higher in M. brunneum-treated plants than in controls; before herbivory, M. brunneum-treated and control plants did not differ. Leaf consumption was slightly lower in the M. brunneum-treated plants whereas total biomass and allocation to above- or below-ground parts was not affected by the Metarhizium treatment. Thus, M. brunneum associates with roots and primes the plant for a stronger or faster increase in myrosinase activity upon herbivory. Consistent with this, myrosinase function has been suggested to be rate-limiting for induction of the glucosinolate-myrosinase defense system. Our results show that M. brunneum, in addition to being an insect pathogen, can associate with plant roots and prime plant defense.


Assuntos
Brassica/enzimologia , Glicosídeo Hidrolases/metabolismo , Metarhizium/fisiologia , Mariposas/fisiologia , Defesa das Plantas contra Herbivoria , Raízes de Plantas/enzimologia , Animais , Brassica/crescimento & desenvolvimento , Brassica/microbiologia , Herbivoria , Larva/fisiologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia
3.
J Sci Food Agric ; 101(4): 1629-1635, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32893880

RESUMO

BACKGROUND: Chlorophyll is the most abundant pigment on Earth, essential for the capture of light energy during photosynthesis. During senescence, chlorophyll degradation is highly regulated in order to diminish toxicity of the free chlorophyll molecule due to its photoactivity. The first step in the chlorophyll degradation pathway is the conversion of chlorophyll b to chlorophyll a by means of two consecutive reactions catalyzed by enzymes coded by NYC1 (NON-YELLOW COLORING 1), NOL (NYC1-LIKE) and HCAR. RESULTS: In this work, we studied the expression of NOL and HCAR genes during postharvest senescence of broccoli. We found that the expression of BoNOL increase during the first days of storage and then decrease. In the case of BoHCAR, its expression is maintained during the first days and then it also diminishes. Additionally, the effect of different postharvest treatments on the expression of these genes was also analyzed. It was observed that the expression of BoNOL is lower in the treatments performed with 1-methylcyclopropene (1-MCP), 6-benzylaminopurine (6-BAP) and modified atmospheres, while BoHCAR expression showed an increase in these same treatments, and a decrease in the treatment with ethylene. There were no variations in the expression of both genes in heat treatment, UV-C treatment and visible light treatment. CONCLUSIONS: These results suggest that both BoHCAR and BoNOL show a lower regulation of their expression than other genes involved in chlorophyll degradation during senescence. © 2020 Society of Chemical Industry.


Assuntos
Brassica/enzimologia , Brassica/genética , Proteínas de Plantas/genética , Brassica/crescimento & desenvolvimento , Brassica/metabolismo , Clorofila/metabolismo , Clorofila A/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo
4.
Int J Mol Sci ; 21(16)2020 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-32784897

RESUMO

Plant polygalacturonases (PGs) are closely related to cell-separation events during plant growth and development by degrading pectin. Identifying and investigating their diversification of evolution and expression could shed light on research on their function. We conducted sequence, molecular evolution, and gene expression analyses of PG genes in Brassica oleracea. Ninety-nine B. oleracea PGs (BoPGs) were identified and divided into seven clades through phylogenetic analysis. The exon/intron structures and motifs were conserved within, but divergent between, clades. The second conserved domain (GDDC) may be more closely related to the identification of PGs. There were at least 79 common ancestor PGs between Arabidopsis thaliana and B. oleracea. The event of whole genome triplication and tandem duplication played important roles in the rapid expansion of the BoPG gene family, and gene loss may be an important mechanism in the generation of the diversity of BoPGs. By evaluating the expression in five tissues, we found that most of the expressed BoPGs in clades A, B, and E showed ubiquitous expression characteristics, and the expressed BoPGs in clades C, D, and F were mainly responsible for reproduction development. Most of the paralogous gene pairs (76.2%) exhibited divergent expression patterns, indicating that they may have experienced neofunctionalization or subfunctionalization. The cis-elements analysis showed that up to 96 BoPGs contained the hormone response elements in their promoters. In conclusion, our comparative analysis may provide a valuable data foundation for the further functional analysis of BoPGs during the development of B. oleracea.


Assuntos
Brassica/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Família Multigênica , Proteínas de Plantas/genética , Poligalacturonase/genética , Arabidopsis/enzimologia , Arabidopsis/genética , Sequência de Bases , Brassica/enzimologia , Sequência Conservada/genética , Evolução Molecular , Duplicação Gênica/genética , Genoma de Planta/genética , Filogenia , Proteínas de Plantas/classificação , Poligalacturonase/classificação , Homologia de Sequência do Ácido Nucleico
5.
J Agric Food Chem ; 68(10): 3017-3025, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32059105

RESUMO

Chlorsulfuron has been applied in wheat fields as a recognized herbicide worldwide, yet it was officially banned in China since 2014 for its soil persistence problem. On the basis of our previous research that 5-dimethylamino distinctively accelerated degradation rate in soils, a modified amino moiety (Ia-c) and monosubstituted amino group (Id-e) were introduced onto the fifth position of the benzene ring in sulfonylurea structures, as well as heterocyclic amino substituents (If-g) to seek a suitable soil degradation rate during such an in situ crop rotation system. Referring to the biological data and ScAHAS inhibition and ScAHAS docking results, they turned out to be AHAS inhibitors with high potent herbicidal activities. The various influence on soil degradation rate along with crop safety indicated that different substituents on the fifth position have exerted an apparent impact. Their united study of structure-activity-safety-degradation relationship has great potential to provide valuable information for further development of eco-friendly agrochemicals.


Assuntos
Acetolactato Sintase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Herbicidas/farmacologia , Proteínas de Plantas/antagonistas & inibidores , Poluentes do Solo/química , Compostos de Sulfonilureia/farmacologia , Acetolactato Sintase/metabolismo , Amaranthus/efeitos dos fármacos , Amaranthus/enzimologia , Brassica/efeitos dos fármacos , Brassica/enzimologia , Inibidores Enzimáticos/química , Herbicidas/química , Cinética , Modelos Moleculares , Proteínas de Plantas/metabolismo , Poluentes do Solo/farmacologia , Relação Estrutura-Atividade , Compostos de Sulfonilureia/química
6.
Physiol Plant ; 168(1): 174-187, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30706476

RESUMO

Tryptophan is one of the amino acids that cannot be produced in humans and has to be acquired primarily from plants. In Arabidopsis thaliana (Arabidopsis), the tryptophan synthase beta subunit (TSB) genes have been found to catalyze the biosynthesis of tryptophan. Here, we report the isolation and characterization of two TSB genes from Brassica oleracea (broccoli), designated BoTSB1 and BoTSB2. Overexpressing BoTSB1 or BoTSB2 in Arabidopsis resulted in higher tryptophan content and the accumulation of indole-3-acetic acid (IAA) and indole glucosinolates in rosette leaves. Therefore, the transgenic plants showed a series of high auxin phenotypes, including long hypocotyls, large plants and a high number of lateral roots. The spatial expression of BoTSB1 and BoTSB2 was detected by quantitative real-time PCR in broccoli and by expressing the ß-glucuronidase reporter gene (GUS) controlled by the promoters of the two genes in Arabidopsis. BoTSB1 was abundantly expressed in vascular tissue of shoots and inflorescences. Compared to BoTSB1, BoTSB2 was expressed at a very low level in shoots but at a higher level in roots. We further investigated the expression response of the two genes to several hormone and stress treatments. Both genes were induced by methyl jasmonate (MeJA), salicylic acid (SA), gibberellic acid (GA), Flg22 (a conserved 22-amino acid peptide derived from bacterial flagellin), wounding, low temperature and NaCl and were repressed by IAA. Our study enhances the understanding of tryptophan biosynthesis and its regulation in broccoli and Arabidopsis. In addition, we provide evidence that TSB genes can potentially be a good tool to breed plants with high biomass and high nutrition.


Assuntos
Brassica/genética , Glucosinolatos/biossíntese , Ácidos Indolacéticos/metabolismo , Triptofano/biossíntese , Arabidopsis , Brassica/enzimologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Plantas Geneticamente Modificadas
7.
Genes (Basel) ; 10(12)2019 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-31810369

RESUMO

The plant U-box (PUB) protein family plays an important role in plant growth and development. The U-box gene family has been well studied in Arabidopsis thaliana, Brassica rapa, rice, etc., but there have been no systematic studies in Brassica oleracea. In this study, we performed genome-wide identification and evolutionary analysis of the U-box protein family of B. oleracea. Firstly, based on the Brassica database (BRAD) and the Bolbase database, 99 Brassica oleracea PUB genes were identified and divided into seven groups (I-VII). The BoPUB genes are unevenly distributed on the nine chromosomes of B. oleracea, and there are tandem repeat genes, leading to family expansion from the A. thaliana genome to the B. oleracea genome. The protein interaction network, GO annotation, and KEGG pathway enrichment analysis indicated that the biological processes and specific functions of the BoPUB genes may mainly involve abiotic stress. RNA-seq transcriptome data of different pollination times revealed spatiotemporal expression specificity of the BoPUB genes. The differential expression profile was consistent with the results of RT-qPCR analysis. Additionally, a large number of pollen-specific cis-acting elements were found in promoters of differentially expressed genes (DEG), which verified that these significantly differentially expressed genes after self-pollination (SP) were likely to participate in the self-incompatibility (SI) process, including gene encoding ARC1, a well-known downstream protein of SI in B. oleracea. Our study provides valuable information indicating that the BoPUB genes participates not only in the abiotic stress response, but are also involved in pollination.


Assuntos
Brassica , Bases de Dados Genéticas , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Complexos Ubiquitina-Proteína Ligase , Brassica/enzimologia , Brassica/genética , Evolução Molecular , Genoma de Planta , Estudo de Associação Genômica Ampla , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Pólen , Polinização , Complexos Ubiquitina-Proteína Ligase/biossíntese , Complexos Ubiquitina-Proteína Ligase/genética
8.
J Biochem ; 166(5): 441-448, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31504617

RESUMO

Glycosylinositol phosphoceramide (GIPC) is the most abundant sphingolipid in plants and fungi. Recently, we detected GIPC-specific phospholipase D (GIPC-PLD) activity in plants. Here, we found that GIPC-PLD activity in young cabbage leaves catalyzes transphosphatidylation. The available alcohol for this reaction is a primary alcohol with a chain length below C4. Neither secondary alcohol, tertiary alcohol, choline, serine nor glycerol serves as an acceptor for transphosphatidylation of GIPC-PLD. We also found that cabbage GIPC-PLD prefers GIPC containing two sugars. Neither inositol phosphoceramide, mannosylinositol phosphoceramide nor GIPC with three sugar chains served as substrate. GIPC-PLD will become a useful catalyst for modification of polar head group of sphingophospholipid.


Assuntos
Biocatálise , Brassica/enzimologia , Ceramidas/metabolismo , Inositol/metabolismo , Fosfatidilcolinas/metabolismo , Fosfolipase D/metabolismo , Folhas de Planta/enzimologia , Brassica/química , Ceramidas/química , Inositol/análogos & derivados , Inositol/química , Estrutura Molecular , Fosfatidilcolinas/química , Fosfolipase D/química , Folhas de Planta/química
9.
Environ Pollut ; 251: 45-55, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31071632

RESUMO

Hydrogen gas (H2) has been shown as an important factor in plant tolerance to abiotic stresses, but the underlying mechanisms remain unclear. In the present study, the effects of H2 and its interaction with nitric oxide (NO) on alleviating cadmium (Cd) stress in Brassica campestris seedlings were investigated. NO donor (SNP) or hydrogen-rich water (HRW) treatment showed a significant improvement in growth of Cd-stressed seedlings. Cd treatment upregulated both endogenous NO and H2 (36% and 66%, respectively), and the increase of H2 was prior to NO increase. When treated with NO scavenger (PTIO) or NO biosynthesis enzyme inhibitors (L-NAME and Gln), HRW-induced alleviation under Cd stress was prevented. Under Cd stress, HRW pretreatment significantly enhanced the NO accumulation, and together up-regulated the activity of NR (nitrate reductase) and expression of NR. HRW induced lower reactive oxygen species (ROS), higher AsA content, enhanced activity of POD (peroxidase) and SOD (superoxide dismutase) in seedling roots were inhibited by PTIO, L-NAME and Gln. Through proteomic analysis, the level of 29 proteins were changed in response to H2 and NO-induced amelioration of Cd stress. Nearly half of them were involved in oxidation-reduction processes (about 20%) or antioxidant enzymes (approximately 20%). These results strongly indicate that in Cd-stressed seedlings, pretreatment with HRW induces the accumulation of H2 (biosynthesized or permeated), which further stimulates the biosynthesis of NO through the NR pathway. Finally, H2 and NO together enhance the antioxidant capabilities of seedlings in response to Cd toxicity.


Assuntos
Antioxidantes/metabolismo , Brassica/efeitos dos fármacos , Cádmio/toxicidade , Hidrogênio/farmacologia , Óxido Nítrico/biossíntese , Poluentes do Solo/toxicidade , Brassica/enzimologia , Brassica/metabolismo , Cádmio/metabolismo , Doadores de Óxido Nítrico/farmacologia , Oxirredução , Proteômica , Plântula/efeitos dos fármacos , Plântula/enzimologia , Plântula/metabolismo , Poluentes do Solo/metabolismo , Regulação para Cima/efeitos dos fármacos
10.
Ecotoxicol Environ Saf ; 180: 179-184, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31082582

RESUMO

Selenium (Se) and zinc (Zn) are necessary mineral nutrients for human body but millions of people have an inadequate intake of them, and eat food enriched with Se and Zn may minimize these problems. Chinese cabbage is an important food in people's daily life. The aim of this study was to evaluate the effects of single Se, Zn and their combination treatment in soil on their accumulation, antioxidant system and lipid peroxidation in roots and leaves of Chinese cabbage using soil pot culture experiment. When 0.5 mg kg-1 Se +30 mg kg-1 Zn and 1.0 mg kg-1 Se +30 mg kg-1 Zn were spiked in soils, Zn concentrations in roots and leaves of Chinese cabbage were significantly increased (p < 0.05) by 20.2%, 37.8% and 17.9%, 34.1% respectively compared to the treatment of 30 mg kg-1 Zn added, and the latter was significantly higher (p < 0.05) than that of former, indicating Se significantly promoted Zn accumulation. Almost all physiological indexes including POD, SOD, CAT, APX, GR, Chlorophyll a, Chlorophyll b, Carotenoids, MDA and Free proline in the treatments of Se or Zn spiked were significantly improved (p < 0.05) or basically unaffected compared to the control without Se or Zn added. The biomass change trends were similar with these indexes either. These results showed that the addition in soil of Se and Zn significantly increased their accumulation in Chinese cabbage without affected its formal growth. Particularly, the addition of Se promoted Zn accumulation. The conclusions were more important reference for the production practice of cash crop enriched of Se and Zn either.


Assuntos
Brassica/efeitos dos fármacos , Selênio/farmacologia , Solo , Zinco/metabolismo , Antioxidantes/metabolismo , Brassica/enzimologia , Brassica/metabolismo , Carotenoides/metabolismo , Clorofila/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/enzimologia , Folhas de Planta/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/enzimologia , Raízes de Plantas/metabolismo , Selênio/metabolismo
11.
Int J Mol Sci ; 20(9)2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31083282

RESUMO

Petal color, size, and morphology play important roles in protecting other floral organs, attracting pollinators, and facilitating sexual reproduction in plants. In a previous study, we obtained a petal degeneration mutant (pdm) from the 'FT' doubled haploid line of Chinese cabbage and found that the candidate gene for pdm, Bra040093, encodes the enzyme acyl-CoA oxidase1. In this study, we sought to examine the gene networks regulating petal development in pdm plants. We show that the mRNA and protein expression of Bra040093, which is involved in the jasmonic acid (JA) biosynthetic pathway, were significantly lower in the petals of pdm plants than in those of 'FT' plants. Similarly, the JA and methyl jasmonate (MeJA) contents of petals were significantly lower in pdm plants than in 'FT' plants and we found that exogenous application of these hormones to the inflorescences of pdm plants restored the 'FT' phenotype. Comparative analyses of the transcriptomes of 'FT', pdm and pdm + JA (pJA) plants revealed 10,160 differentially expressed genes (DEGs) with consistent expression tendencies in 'FT' vs. pdm and pJA vs. pdm comparisons. Among these DEGs, we identified 69 DEGs related to floral organ development, 11 of which are involved in petal development regulated by JA. On the basis of qRT-PCR verification, we propose regulatory pathways whereby JA may mediate petal development in the pdm mutant. We demonstrate that mutation of Bra040093 in pdm plants leads to reduced JA levels and that this in turn promotes changes in the expression of genes that are expressed in response to JA, ultimately resulting in petal degeneration. These findings thus indicate that JA is associated with petal development in Chinese cabbage. These results enhance our knowledge on the molecular mechanisms underlying petal development and lay the foundations for further elucidation of the mechanisms associated with floral organ development in Chinese cabbage.


Assuntos
Brassica/enzimologia , Brassica/genética , Ciclopentanos/metabolismo , Flores/enzimologia , Mutação/genética , Oxilipinas/metabolismo , Proteínas de Plantas/genética , Acetatos/metabolismo , Acetatos/farmacologia , Brassica/efeitos dos fármacos , Brassica/crescimento & desenvolvimento , Ciclopentanos/farmacologia , Flores/anatomia & histologia , Flores/efeitos dos fármacos , Flores/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Oxilipinas/farmacologia , Fenótipo , Proteínas de Plantas/metabolismo
12.
Environ Pollut ; 249: 716-727, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30933769

RESUMO

Understanding the chemical response and characteristics of bacterial communities in soil is critical to evaluate the effects of selenium (Se) supplement on plant growth and chromium (Cr)/Se uptake in Cr contaminated soil. The rhizosphere soil characteristics of pak choi (Brassica campestris L. ssp. Chinensis Makino) were investigated in soil contaminated with different levels and forms of Cr when supplemented with Se. Although inhibition of plant growth caused by Cr stress was not completely alleviated by Se, Cr content in plant tissues decreased in Cr(VI)120Se5 treatment (Cr(VI): 120 mg kg-1 soil; Se: 5 mg kg-1 soil) and its bioavailability in soil decreased in Cr(III)200Se5 (Cr(III): 200 mg kg-1 soil; Se: 5 mg kg-1 soil) treatment. Moreover, antagonism of Cr and Se on soil enzyme activities and bacterial communities were revealed. Notably, results of Cr(VI) reduction and Se metabolism functional profiles confirmed that bacterial communities play a critical role in regulating Cr/Se bioavailability. Additionally, the increases of Se bioavailability in Cr contaminated soil were ascribed to oxidation of Cr(VI) and reduction of Se reductases proportions, as well as the enhancing of pH in soil. These findings reveal that Se has the potential capacity to sustain the stability of microdomain in Cr contaminated soil.


Assuntos
Brassica/enzimologia , Cromo/análise , Rizosfera , Selênio/farmacologia , Microbiologia do Solo , Poluentes do Solo/análise , Solo/química , Disponibilidade Biológica , Transporte Biológico , Brassica/metabolismo , Cromo/metabolismo , Selênio/metabolismo , Poluentes do Solo/metabolismo
13.
Plant Cell Physiol ; 60(2): 421-435, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30462304

RESUMO

Long non-coding RNAs (lncRNAs) are non-protein-coding transcripts longer than 200 nt that are distributed widely in organisms and play many physiological roles. The BoNR8 lncRNA is a 272 nt long transcript yielded by RNA polymerase III in cabbage that was identified as the closest homolog of the AtR8 lncRNA in Arabidopsis. The BoNR8 lncRNA was expressed extensively in the epidermal tissue in the root elongation zone of germinated seeds, and its accumulation was induced by abiotic stresses, auxins and ABA. To investigate the correlation between the BoNR8 lncRNA and germination, BoNR8-overexpressing Arabidopsis plants (BoNR8-AtOX) were prepared. Three independent BoNR8-AtOX lines showed less primary root elongation, incomplete silique development and decreased germination rates. The germination efficiencies were affected strongly by ABA and slightly by salt stress, and ABA-related gene expression was changed in the BoNR8-AtOX lines.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Brassica/genética , Germinação , Proteínas de Plantas/fisiologia , RNA Polimerase III/fisiologia , RNA Longo não Codificante/fisiologia , Sementes/genética , Arabidopsis/genética , Brassica/enzimologia , Brassica/crescimento & desenvolvimento , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , RNA Polimerase III/metabolismo , RNA Longo não Codificante/genética
14.
Int J Mol Sci ; 19(11)2018 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-30373125

RESUMO

The activities of pectin methylesterases (PMEs) are regulated by pectin methylesterase inhibitors (PMEIs), which consequently control the pectin methylesterification status. However, the role of PMEI genes in Brassica oleracea, an economically important vegetable crop, is poorly understood. In this study, 95 B. oleracea PMEI (BoPMEI) genes were identified. A total of 77 syntenic ortholog pairs and 10 tandemly duplicated clusters were detected, suggesting that the expansion of BoPMEI genes was mainly attributed to whole-genome triplication (WGT) and tandem duplication (TD). During diploidization after WGT, BoPMEI genes were preferentially retained in accordance with the gene balance hypothesis. Most homologous gene pairs experienced purifying selection with ω (Ka/Ks) ratios lower than 1 in evolution. Five stamen-specific BoPMEI genes were identified by expression pattern analysis. By combining the analyses of expression and evolution, we speculated that nonfunctionalization, subfunctionalization, neofunctionalization, and functional conservation can occur in the long evolutionary process. This work provides insights into the characterization of PMEI genes in B. oleracea and contributes to the further functional studies of BoPMEI genes.


Assuntos
Brassica/genética , Hidrolases de Éster Carboxílico/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Proteínas de Plantas/genética , Brassica/enzimologia , Diploide , Evolução Molecular , Duplicação Gênica , Família Multigênica , Transcriptoma
15.
Ecotoxicol Environ Saf ; 166: 157-164, 2018 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-30267988

RESUMO

Silicon (Si) and selenium (Se) are beneficial for many higher plants when grown on stress conditions. However, the mechanisms underlying the differential effects between foliar Si and Se in alleviation of plant toxicity exposed to cadmium (Cd) stress are remained unclear. In this study, we investigated the discrepant mechanisms of foliar Si and Se on Cd absorption and compartmentation by roots, its translocation in xylem, and the antioxidant system within Chinese flowering cabbage (Brassica campestris L. ssp. chinensis var. utilis) under low and high Cd stress. Results showed that plant growth was significantly enhanced by foliar additions of Si or/and Se according to an increased plant tissue biomass at high Cd exposure. In addition, the foliar coupled addition of Si and Se showed little effects on the concentrations of Si or Se in plant tissues in comparison with the single addition of foliar Si or Se respectively. The foliar Si alone or combined with Se markedly reduced the Cd concentrations in plant shoots under two Cd treatments. This might be explained by the lower Cd concentrations in symplast and apoplast and the higher Cd concentrations in cell walls of plant roots, and the lower Cd concentrations in xylem sap. However, no great changes in these values were observed under the treatments of foliar Se alone. Moreover, the foliar additions of Si or/and Se all increased the antioxidant enzyme activities of SOD, CAT and APX in plant tissues, especially at high Cd dosage. No significant differences in the increasing degrees of these three antioxidant enzymes were found between the foliar Si and Se treatments. However, only the foliar Se alone or combined with Si markedly promoted the antioxidant enzyme activities of GR and DHAR in plant tissues. Our findings demonstrate that the alleviation of Cd toxicity by foliar Si maybe mainly responsible for inhibition of Cd absorption and its translocation to plant shoots, reinforcing its compartmentation into root cell walls, whilst enhancing the antioxidant enzyme system may be employed by foliar Se.


Assuntos
Brassica/metabolismo , Cádmio/farmacocinética , Selênio/farmacologia , Silício/farmacologia , Absorção Fisiológica , Antioxidantes/metabolismo , Transporte Biológico , Biomassa , Brassica/enzimologia , Brassica/crescimento & desenvolvimento , Parede Celular/metabolismo , Brotos de Planta/metabolismo , Xilema/metabolismo
16.
Molecules ; 23(9)2018 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-30200303

RESUMO

Myrosinase is an enzyme present in many functional foods and spices, particularly in Cruciferous vegetables. It hydrolyses glucosinolates which thereafter rearrange into bioactive volatile constituents (isothiocyanates, nitriles). We aimed to develop a simple reversible method for on-gel detection of myrosinase. Reagent composition and application parameters for native PAGE and SDS-PAGE gels were optimized. The proposed method was successfully applied to detect myrosinase (or sulfatase) on-gel: the detection solution contains methyl red which gives intensive red bands where the HSO4- is enzymatically released from the glucosinolates. Subsequently, myrosinase was successfully distinguished from sulfatase by incubating gel bands in a derivatization solution and examination by LC-ESI-MS: myrosinase produced allyl isothiocyanate (detected in conjugate form) while desulfo-sinigrin was released by sulfatase, as expected. After separation of 80 µg protein of crude extracts of Cruciferous vegetables, intensive color develops within 10 min. On-gel detection was found to be linear between 0.031⁻0.25 U (pure Sinapis alba myrosinase, R² = 0.997). The method was successfully applied to detection of myrosinase isoenzymes from horseradish, Cruciferous vegetables and endophytic fungi of horseradish as well. The method was shown to be very simple, rapid and efficient. It enables detection and partial characterization of glucosinolate decomposing enzymes without protein purification.


Assuntos
Bioquímica/métodos , Glicosídeo Hidrolases/análise , Brassica/enzimologia , Misturas Complexas , Glucosinolatos/química , Glucosinolatos/metabolismo , Padrões de Referência , Espectrometria de Massas por Ionização por Electrospray , Sulfatases/metabolismo
17.
Food Chem ; 269: 96-102, 2018 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-30100490

RESUMO

Five Brassicaceae sprouts (white cabbage, kale, broccoli, Chinese cabbage, arugula) were comparatively analyzed based on phytochemicals (polyphenols, glucosinolates, carotenoids, chlorophylls, ascorbic acid) content and accompanying enzymes associated with phytochemical stability and bioavailability (peroxidases, myrosinase, and polyphenol-oxidase) that consequently impact food quality. Significantly high content of polyphenols and glucosinolates, as well as a high antioxidant activity were found in white cabbage, followed by kale sprouts. In addition, white cabbage contained higher amount of fibers and lower polyphenol-oxidase activity which potentially indicates prevention of browning and consequently better sprout quality. Arugula and broccoli showed higher activity of myrosinase that may result in higher bioavailability of active glucosinolates forms. According to our data, sprouts are cheap, easy- and fast-growing source of phytochemicals but also they are characterized by different endogenous enzymes activity. Consequently, this parameter should also be taken into consideration in the studies related to the health benefits of the plant-based food.


Assuntos
Brassica/química , Brassica/enzimologia , Qualidade dos Alimentos , Compostos Fitoquímicos/análise , Disponibilidade Biológica , Catecol Oxidase/metabolismo , Análise de Alimentos , Glucosinolatos , Glicosídeo Hidrolases/metabolismo , Humanos , Peroxidases/metabolismo , Compostos Fitoquímicos/farmacocinética
18.
Plant Signal Behav ; 13(5): e1467698, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29944433

RESUMO

Phosphoglucomutases (PGM) (5.4.2.2.) belong to the Phosphohexomutases superfamily and are highly specific in catalyzing the interconversion of Glc-1-P to Glc-6-P. In this study, we characterize the expression and activity of two cytosolic PGMs (cPGM2 and cPGM3) stigmas of ornamental kale during flower development. In stigmas, cPGM expression and activity showed a gradual increase during stigma development with the highest activity around the time of anthesis. Blocking of cPGM activity in the stigmas using a known inhibitor, resulted in breakdown of self-incompatibility in immature S3 and S4 stigmas, but had no effect on the fully mature S5 stigmas. It is likely that cPGMs are required for accumulation of factors necessary for SI response in mature stigmas.


Assuntos
Brassica/enzimologia , Brassica/metabolismo , Flores/enzimologia , Flores/metabolismo , Fosfoglucomutase/metabolismo , Proteínas de Plantas/metabolismo , Brassica/genética , Flores/genética , Fosfoglucomutase/genética , Proteínas de Plantas/genética
19.
Nutrients ; 10(6)2018 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-29890681

RESUMO

Dietary broccoli is anti-inflammatory. Past studies have typically investigated raw broccoli, even though most consumers prefer cooked broccoli, where the plant myrosinase is inactivated by heat, resulting in failure of formation of the anti-inflammatory bioactive compound sulforaphane (SF). This study compareed efficacy of lightly cooked broccoli (CB) containing greatly diminished myrosinase activity, with raw broccoli (RB), in mitigating colitis in dextran sulfate sodium (DSS)-treated mice. Male C57BL/6 mice were fed for two weeks on a 10% RB, 10% CB or control diet, all based on the AIN-93M diet. Half (n = 9) of each group received drinking water, half received 2.5% DSS in water for one week, starting from Day 7 of the diet. Even with far less plant myrosinase activity, CB was essentially as effective as RB in lessening damage by DSS, evidenced by decreased disease activity index, attenuated colon length shrinkage, less endotoxin (lipopolysaccharide) leakage into blood, and less severe colon lesions as assessed by histopathology. mRNA expression of pro-inflammatory cytokines indicated that broccoli anti-inflammatory action may be through inhibition of the IL-6 trans-signaling pathway, as evidenced by reversal of the DSS-increased expression of IL-6, CCR2 and vascular cell adhesion molecule 1 (VCAM-1).


Assuntos
Brassica , Colite/prevenção & controle , Colo , Culinária , Sulfato de Dextrana , Animais , Brassica/enzimologia , Colite/induzido quimicamente , Colite/metabolismo , Colite/patologia , Colo/metabolismo , Colo/patologia , Modelos Animais de Doenças , Estabilidade Enzimática , Glicosídeo Hidrolases/metabolismo , Temperatura Alta , Hidrólise , Interleucina-6/metabolismo , Isotiocianatos/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/metabolismo , Permeabilidade , Desnaturação Proteica , Receptores CCR2/metabolismo , Transdução de Sinais , Molécula 1 de Adesão de Célula Vascular/metabolismo
20.
Int J Mol Sci ; 19(5)2018 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-29724020

RESUMO

Pectin methylesterase inhibitor genes (PMEIs) are a large multigene family and play crucial roles in cell wall modifications in plant growth and development. Here, a comprehensive analysis of the PMEI gene family in Brassicacampestris, an important leaf vegetable, was performed. We identified 100 BrassicacampestrisPMEI genes (BcPMEIs), among which 96 BcPMEIs were unevenly distributed on 10 chromosomes and nine tandem arrays containing 20 BcPMEIs were found. We also detected 80 pairs of syntenic PMEI orthologs. These findings indicated that whole-genome triplication (WGT) and tandem duplication (TD) were the main mechanisms accounting for the current number of BcPMEIs. In evolution, BcPMEIs were retained preferentially and biasedly, consistent with the gene balance hypothesis and two-step theory, respectively. The molecular evolution analysis of BcPMEIs manifested that they evolved through purifying selection and the divergence time is in accordance with the WGT data of B. campestris. To obtain the functional information of BcPMEIs, the expression patterns in five tissues and the cis-elements distributed in promoter regions were investigated. This work can provide a better understanding of the molecular evolution and biological function of PMEIs in B. campestris.


Assuntos
Brassica/enzimologia , Hidrolases de Éster Carboxílico/metabolismo , Evolução Molecular , Perfilação da Expressão Gênica , Estudo de Associação Genômica Ampla , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Brassica/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Genoma de Planta/genética , Filogenia , Proteínas de Plantas/classificação , Análise de Sequência de DNA , Análise de Sequência de Proteína , Sintenia , Sequências de Repetição em Tandem/genética
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