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1.
Planta ; 253(4): 80, 2021 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-33742226

RESUMO

MAIN CONCLUSION: Chitinase family genes were involved in the response of Brassica oleracea to Fusarium wilt, powdery mildew, black spot and downy mildew. Abstract Chitinase, a category of pathogenesis-related proteins, is believed to play an important role in defending against external stress in plants. However, a comprehensive analysis of the chitin-binding gene family has not been reported to date in cabbage (Brassica oleracea L.), especially regarding the roles that chitinases play in response to various diseases. In this study, a total of 20 chitinase genes were identified using a genome-wide search method. Phylogenetic analysis was employed to classify these genes into two groups. The genes were distributed unevenly across six chromosomes in cabbage, and all of them contained few introns (≤ 2). The results of collinear analysis showed that the cabbage genome contained 1-5 copies of each chitinase gene (excluding Bol035470) identified in Arabidopsis. The heatmap of the chitinase gene family showed that these genes were expressed in various tissues and organs. Two genes (Bol023322 and Bol041024) were relatively highly expressed in all of the investigated tissues under normal conditions, exhibiting the expression characteristics of housekeeping genes. In addition, under four different stresses, namely, Fusarium wilt, powdery mildew, black spot and downy mildew, we detected 9, 5, 8 and 8 genes with different expression levels in different treatments, respectively. Our results may help to elucidate the roles played by chitinases in the responses of host plants to various diseases.


Assuntos
Brassica , Quitina/metabolismo , Genoma de Planta , Imunidade Vegetal , Proteínas de Plantas/genética , Brassica/genética , Brassica/microbiologia , Quitinases/genética , Regulação da Expressão Gênica de Plantas , Filogenia
2.
Plant Mol Biol ; 106(1-2): 193-206, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33742369

RESUMO

KEY MESSAGE: We provided a study on homeologous gene evolution of homeologous genes by comparing Brassica genomes. Polyploidy has played fundamental roles during the evolution of plants. Following polyploidization, many duplicated genes are diversified or lost in a process termed diploidization. Understanding the retention and diversification of homeologs after polyploidization will help elucidate the process of diploidization. Here, we investigated the evolution of homeologous genes in Brassica genomes and observed similarly asymmetrical gene retention among different functional groups and consistent retention after recurrent polyploidizations. In the comparative analysis of Brassica diploid genomes, we found that preferentially retained genes show different patterns on sequence and expression divergence: genes with the function of 'biosynthetic process' and 'transport' were under much stronger purifying selection, while transcriptional regulatory genes diverged much faster than other genes. Duplicate pairs of the former two functional groups show conserved high expression patterns, while most of transcriptional regulatory genes are simultaneously lowly expressed. Furthermore, homeologs in diploids and allotetraploids showed similar loss and retention patterns: duplicates in progenitor genomes were more likely to be retained and accumulated fewer substitutions. However, transcriptional regulation is also enriched in the genes that do not have any non-synonymous mutations in the Brassica allotetraploids, indicating that some of these genes were under strong purifying selection. Overall, our study provided insight into the evolution of homeologs genes during diploidization process.


Assuntos
Brassica/genética , Genes de Plantas , Poliploidia , Arabidopsis/genética , Sequência de Bases , Diploide , Evolução Molecular , Dosagem de Genes , Regulação da Expressão Gênica de Plantas , Genes Duplicados , Filogenia , Seleção Genética , Sintenia/genética , Transcrição Genética
3.
Arch Microbiol ; 203(4): 1641-1647, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33432379

RESUMO

To investigate the molecular mechanism of Trichoderma L-amino acid oxidase (Th-LAAO) in protecting and in promoting growth of cabbage infected with Botrytis cinerea, a three-way interaction system was established. Cabbage leaves treated with purified Th-LAAO significantly constrained damaged leaf area caused by B. cinerea infection. In response to Th-LAAO treatment, the expression levels of genes involved in photosynthesis, such as ribulose-1,5-bisphosphate carboxylase oxygenase, Rubisco activase, and ATP synthase increased 2.54, 2.18, and 1.41 folds, respectively. The transcription levels of sucrose transport protein 1 increased 7.6 fold. As to the expression of defense-related genes, the transcription level of ascorbate peroxidase increased 1.46 fold. On the contrary, pathogenesis-related protein 1, chitinase, ß-1,3 glucanase, and glutathione S-transferase decreased significantly. Overall, the results indicated that Th-LAAO may stimulate CO2 fixation and sucrose transport and elicit host defense responses in cabbage against B. cinerea, and this elicitation of defense response is likely to contribute to induced systemic resistance of host plant.


Assuntos
Brassica , Resistência à Doença , L-Aminoácido Oxidase , Trichoderma , Botrytis/fisiologia , Brassica/efeitos dos fármacos , Brassica/genética , Brassica/microbiologia , Resistência à Doença/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas/genética , L-Aminoácido Oxidase/isolamento & purificação , L-Aminoácido Oxidase/farmacologia , Fotossíntese/efeitos dos fármacos , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/microbiologia , Trichoderma/química , Trichoderma/enzimologia
4.
Int J Mol Sci ; 22(2)2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33467698

RESUMO

The genus Brassica comprises a highly diverse range of vegetable crops varying in morphology, harvestable crop product, and postharvest shelf-life that has arisen through domestication, artificial selection and plant breeding. Previous postharvest studies on the shelf-life of Brassica species has mainly focused on the variable rates of physiological changes including respiration and transpiration. Therefore, further understanding of the molecular basis of postharvest senescence in Brassica vegetables is needed to understand its progression in improving their postharvest shelf-life. The aim of this study was to better understand the trajectory of molecular responses in senescence-associated genes but not induced by ethylene and ethylene-induced genes towards altered postharvest storage conditions. After storage at different temperatures, the expression levels of the key senescence-associated genes (SAGs) and the ethylene biosynthesis, perception, and signaling genes were quantitatively analyzed in cabbage, broccoli and kale. The expression levels of these genes were tightly linked to storage temperature and phase of senescence. Expression of ORE15, SAG12, and NAC29 were continuously increased during the twelve days of postharvest storage at room temperature. Prolonged exposure of these three vegetables to cold temperature reduced the variation in the expression levels of ORE15 and SAG12, observed as mostly decreased which resulted in limiting senescence. The transcript levels of the ethylene receptor were also decreased at lower temperature, further suggesting that decreased ethylene biosynthesis and signaling in cabbage during postharvest storage would delay the senescence mechanism. These results enhanced our understanding of the transcriptional changes in ethylene-independent SAGs and ethylene-related genes in postharvest senescence, as well as the timing and temperature sensitive molecular events associated with senescence in cabbage, broccoli and kale and this knowledge can potentially be used for the improvement of postharvest storage in Brassica vegetables.


Assuntos
Brassica/genética , Senescência Celular , Etilenos/química , Armazenamento de Alimentos , Regulação da Expressão Gênica de Plantas , Verduras/genética , Brassica/metabolismo , Produtos Agrícolas/genética , Produtos Agrícolas/metabolismo , Etilenos/metabolismo , Melhoramento Vegetal , Proteínas de Plantas/metabolismo , Transdução de Sinais , Temperatura , Verduras/metabolismo
5.
Planta ; 253(1): 21, 2021 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-33399991

RESUMO

MAIN CONCLUSION: Starch and sucrose metabolism and plant-pathogen interaction pathways play a dominate role in recessive genic male sterility (RGMS) of cabbage (Brassica oleracea L. var. capitata). RGMS is common in plants and has been widely applied as an effective and economic system for hybrid seed production in many crops. However, little is known regarding the molecular mechanisms of RGMS in cabbage. Hence, full-length transcriptomic and physiological analysis were performed in the spontaneous RGMS mutant RMS3185A and its near-isogenic fertile line (NIL) RMS3185B of small (< 1.6 mm in diameter), medium (~ 2.5 mm in diameter), and large floral buds (~ 3.4 mm in diameter) to identify the differentially expressed genes (DEGs) associated with RMGS. The pollen abnormalities between RMS3185B and RMS3185A appeared at the large floral bud stage. In contrast with RMS3185B, the mature anthers and stamens of RMS3185A were shorter than those of RMS3185B, and the anthers did not dehiscent. The concentrations of glucose, fructose, trehalose, starch, and cellulose in RMS3185A were all significantly lower than those in large floral buds of RMS3185B. PacBio sequencing results showed that DEGs were mainly concentrated in large floral bud stage. In combination with the KEGG enrichment analysis of DEGs in GO terms related to cell wall, pollen and anther, pentose and glucuronate interconversions (ko00040), starch and sucrose metabolism (ko00500), and plant-pathogen interaction (ko04626) were significantly enriched. Among which, cell-wall/pectin-related genes of eighteen PEI, twenty-two PEL, three PG, and fifteen PGL involved in ko00040, and one UGDH, one SPS, four CWINV, four TPP/TPS, and four EGL involved in ko00500, as well as plant-pathogen interaction genes, including sixteen calcium-dependent protein kinase (CDPK), one cyclic nucleotide-gated ion channel (CNGC), and twenty-three calcium-binding protein CML (CML), were significantly down-regulated in RMS3185A relative to that in RMS3185B. Besides, genes involved in ko04626, including two CML and one transcription factor WRKY33, were up-regulated in RMS3185A relative to that in RMS3185B. In conclusion, we hypothesized that the expression alterations of these genes were responsible for calcium signaling and sugar metabolism, thus affecting the occurrence of RGMS in cabbage.


Assuntos
Brassica , Flores , Infertilidade das Plantas , Proteínas de Plantas , Transcriptoma , Brassica/genética , Flores/genética , Regulação da Expressão Gênica de Plantas , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Transcriptoma/genética
6.
Ecotoxicol Environ Saf ; 208: 111616, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396136

RESUMO

Our previous studies showed that exogenous glutathione (GSH) decreased cadmium (Cd) concentration in shoots and alleviated the growth inhibition in pakchoi (Brassica chinensis L.) under Cd stress. Nevertheless, it is largely unknown how GSH decreases Cd accumulation in edible parts of pakchoi. This experiment mainly explored the mechanisms of GSH-induced reduction of Cd accumulation in shoot of pakchoi. The results showed that compared with sole Cd treatment, Cd + GSH treatment remarkably increased the expression of BcIRT1 and BcIRT2, and further enhanced the concentrations of Cd and Fe in root. By contrast, GSH application declined the concentration of Cd in the xylem sap. However, these results were not caused by xylem loading process because the expression of BcHMA2 and BcHMA4 had not significant difference between sole Cd treatment and Cd + GSH treatment. In addition, exogenous GSH significantly enhanced the expression of BcPCS1 and promoted the synthesis of PC2, PC3 and PC4 under Cd stress. At the same time, exogenous GSH also significantly improved the expression of BcABCC1 and BcABCC2 in the roots of seedling under Cd stress, suggesting that more PCs-Cd complexes may be sequestrated into vacuoles by ABCC1 and ABCC2 transporters. The results showed that exogenous GSH could up-regulate the expression of BcIRT1/2 to increase the Cd accumulation in root, and the improvement of PCs contents and the expression of BcABCC1/2 enhanced the compartmentalization of Cd in root vacuole of pakchoi under Cd stress. To sum up, exogenous GSH reduce the concentration of free Cd2+ in the cytoplast of root cells and then dropped the loading of Cd into the xylem, which eventually given rise to the reduction of Cd accumulation in edible portion of pakchoi.


Assuntos
Brassica/metabolismo , Cádmio/metabolismo , Glutationa/metabolismo , Raízes de Plantas/metabolismo , Plântula/metabolismo , Poluentes do Solo/metabolismo , Vacúolos/metabolismo , Transporte Biológico , Brassica/efeitos dos fármacos , Brassica/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Plântula/efeitos dos fármacos
7.
Food Chem ; 334: 127519, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32721832

RESUMO

We aimed to characterize and quantify glucosinolate compounds and contents in broccoli, and a total of 80 genotypes and eight developmental organs were analyzed with UHPLC-Triple-TOF-MS. The method was validated in terms of performance, and the coefficients of determination (R2) were 0.97 and 0.99 for glucoraphanin and gluconapin, respectively. In 80 genotypes, twelve glucosinolates were found in broccoli florets ranging from 0.467 to 57.156 µmol/g DW, with the highest glucosinolate content being approximately 122-fold higher than the lowest value. The principal component of glucobrassicin, neoglucobrassicin and glucoraphanin explained 60.53% of the total variance. There were positive correlations among hydroxyglucobrassicin, methoxyglucobrassicin, glucobrassicin, glucoerucin, gluconasturtiin, glucoraphanin, and glucotropaeolin (P < 0.05). The root contained 43% of total glucosinolates in 80 genotypes, and glucoraphanin represented 29% of the total glucosinolate content in different organs. The mutant broccoli genotypes were found by analysis of gluconapin contents in different organs.


Assuntos
Brassica/metabolismo , Glucosinolatos/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Brassica/genética , Cromatografia Líquida de Alta Pressão , Genótipo , Glucosinolatos/análise , Imidoésteres/análise , Indóis/análise
8.
Food Chem ; 338: 127997, 2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-33091988

RESUMO

Flavonols are gaining increasing interests due to their diverse health benefits for humans. Broccoli is a main flavonol source in our diet, but the genetic variation of flavonols and their correlation with antioxidant capacity remain to be understood. Here, we examined variations of the two major flavonols kaempferol and quercetin in florets and leaves of 15 diverse broccoli accessions by ultra-performance liquid chromatography. Broccoli accumulated more kaempferol than quercetin in most of the accessions tested, with the ratios varying from 4.4 to 27.9 in leaves and 0.4 to 4.4 in florets. Total flavonoids showed 2.5-fold and 3.3-fold differences in leaves and florets of these accessions, respectively. Principle component analysis revealed that flavonols, along with the key biosynthetic pathway genes, correlated with antioxidant capacity related indicators. This study provides important information for broccoli flavonol genotypic variations and correlation with antioxidant capacity, and will facilitate the development of flavonol enriched cultivars in broccoli.


Assuntos
Antioxidantes/química , Brassica/genética , Flavonóis/análise , Variação Genética , Antioxidantes/metabolismo , Brassica/química , Brassica/metabolismo , Catalase/metabolismo , Cromatografia Líquida de Alta Pressão , Flavonóis/metabolismo , Genótipo , Humanos , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Análise de Componente Principal , RNA de Plantas/química , RNA de Plantas/metabolismo , Superóxido Dismutase/metabolismo
9.
Food Chem ; 340: 127901, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32889204

RESUMO

Cauliflowers are generally associated with healthy diets due to their positive impact on health. This research aims to evaluate the effects of cooking processes (boiling, steaming and microwaving) and different preparation times, on the content of carotenoids and provitamin A and tocopherols, in cauliflowers and to verify the effect of the cooking process on maintaining the coloring. The results revealed that the thermal processinfluencedthe antioxidant compounds releaseindependent of genotype. The highest content of zeaxanthin and lutein was found in 'Verde di Macerata' after boiling for 20 min. 'Cheddar' presented the highest content of all carotenoids and when steamed for 20 min, the highest levels ofprovitamin Awere observed.Microwaved and bolied 'Grafitti' for longer times showed the highest retention of tocopherol. The cooking did not negatively affect the visual aspect. 'Verde di Macerata' and 'Cheddar' may be good sources of carotenoids and tocopherols.


Assuntos
Brassica/química , Carotenoides/análise , Culinária/métodos , Tocoferóis/análise , Antioxidantes/análise , Antioxidantes/química , Brassica/genética , Carotenoides/química , Genótipo , Pigmentação , Fatores de Tempo , Tocoferóis/química , Vitamina E/análise , Vitamina E/química
10.
Nat Commun ; 11(1): 3847, 2020 07 31.
Artigo em Inglês | MEDLINE | ID: mdl-32737299

RESUMO

Reporter systems are routinely used in plant genetic engineering and functional genomics research. Most such plant reporter systems cause accumulation of foreign proteins. Here, we demonstrate a protein-independent reporter system, 3WJ-4 × Bro, based on a fluorescent RNA aptamer. Via transient expression assays in both Escherichia coli and Nicotiana benthamiana, we show that 3WJ-4 × Bro is suitable for transgene identification and as an mRNA reporter for expression pattern analysis. Following stable transformation in Arabidopsis thaliana, 3WJ-4 × Bro co-segregates and co-expresses with target transcripts and is stably inherited through multiple generations. Further, 3WJ-4 × Bro can be used to visualize virus-mediated RNA delivery in plants. This study demonstrates a protein-independent reporter system that can be used for transgene identification and in vivo dynamic analysis of mRNA.


Assuntos
Aptâmeros de Nucleotídeos/genética , Arabidopsis/genética , Brassica/genética , Engenharia Genética/métodos , RNA Mensageiro/genética , Tabaco/genética , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/metabolismo , Arabidopsis/metabolismo , Compostos de Benzil/química , Brassica/metabolismo , Fluorescência , Corantes Fluorescentes/química , Regulação da Expressão Gênica , Genes Reporter , Imidazolinas/química , Folhas de Planta/genética , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas , RNA Mensageiro/metabolismo , Tabaco/metabolismo , Transformação Genética
11.
PLoS One ; 15(6): e0234411, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32511257

RESUMO

The AINTEGUMENTA-like (AIL) proteins, which belong to the AP2 family, play important roles in regulating the growth and development of plant organs. The AIL family has not yet been comprehensively studied in rapeseed (Brassica napus), an allotetraploid and model organism for the study of polyploid evolution. In the present study, 99 AIL family genes were identified and characterized from B. rapa, B. oleracea, B. napus, B. juncea, and B. nigra using a comprehensive genome-wide study, including analyses of phylogeny, gene structure, chromosomal localization, and expression pattern. Using a phylogenetic analysis, the AIL genes were divided into eight groups, which were closely related to the eight AtAIL genes, and which shared highly conserved structural features within the same subfamily. The non-synonymous/synonymous substitution ratios of the paralogs and orthologs were less than 1, suggesting that the AIL genes mainly experienced purifying selection during evolution. In addition, the RNA sequencing data and qRT-PCR analysis revealed that the B. napus AIL genes exhibited organ- and developmental stage-specific expression patterns. Certain genes were highly expressed in the developing seeds (BnaAIL1, BnaAIL2, BnaAIL5, and BnaAIL6), the roots (BnaANT, BnaAIL5, and BnaAIL6), and the stem (BnaAIL7B). Our results provide valuable information for further functional analysis of the AIL family in B. napus and related Brassica species.


Assuntos
Brassica napus/genética , Brassica/genética , Genes de Plantas , Sequência de Aminoácidos , Proteínas de Arabidopsis/genética , Brassica/crescimento & desenvolvimento , Brassica napus/crescimento & desenvolvimento , Mapeamento Cromossômico , Sequência Conservada , Duplicação Gênica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Proteínas de Homeodomínio/genética , Família Multigênica , Filogenia , Proteínas de Plantas/genética , Sintenia
12.
Proc Natl Acad Sci U S A ; 117(25): 14561-14571, 2020 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-32518116

RESUMO

Recombination between homeologous chromosomes, also known as homeologous exchange (HE), plays a significant role in shaping genome structure and gene expression in interspecific hybrids and allopolyploids of several plant species. However, the molecular mechanisms that govern HEs are not well understood. Here, we studied HE events in the progeny of a nascent allotetraploid (genome AADD) derived from two diploid progenitors of hexaploid bread wheat using cytological and whole-genome sequence analyses. In total, 37 HEs were identified and HE junctions were mapped precisely. HEs exhibit typical patterns of homologous recombination hotspots, being biased toward low-copy, subtelomeric regions of chromosome arms and showing association with known recombination hotspot motifs. But, strikingly, while homologous recombination preferentially takes place upstream and downstream of coding regions, HEs are highly enriched within gene bodies, giving rise to novel recombinant transcripts, which in turn are predicted to generate new protein fusion variants. To test whether this is a widespread phenomenon, a dataset of high-resolution HE junctions was analyzed for allopolyploid Brassica, rice, Arabidopsis suecica, banana, and peanut. Intragenic recombination and formation of chimeric genes was detected in HEs of all species and was prominent in most of them. HE thus provides a mechanism for evolutionary novelty in transcript and protein sequences in nascent allopolyploids.


Assuntos
Cromossomos de Plantas/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Poliploidia , Recombinação Genética , Arabidopsis/genética , Arachis/genética , Brassica/genética , Biologia Computacional , Evolução Molecular , Fusão Gênica , Cariotipagem , Musa/genética , Oryza/genética , Transcrição Genética , Triticum/genética
13.
RNA ; 26(7): 784-793, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32241834

RESUMO

Long noncoding RNAs (lncRNAs) have recently emerged as prominent regulators of gene expression in eukaryotes. LncRNAs often drive the modification and maintenance of gene activation or gene silencing states via chromatin conformation rearrangements. In plants, lncRNAs have been shown to participate in gene regulation, and are essential to processes such as vernalization and photomorphogenesis. Despite their prominent functions, only over a dozen lncRNAs have been experimentally and functionally characterized. Similar to its animal counterparts, the rates of sequence divergence are much higher in plant lncRNAs than in protein coding mRNAs, making it difficult to identify lncRNA conservation using traditional sequence comparison methods. Beyond this, little is known about the evolutionary patterns of lncRNAs in plants. Here, we characterized the splicing conservation of lncRNAs in Brassicaceae. We generated a whole-genome alignment of 16 Brassica species and used it to identify synthenic lncRNA orthologs. Using a scoring system trained on transcriptomes from A. thaliana and B. oleracea, we identified splice sites across the whole alignment and measured their conservation. Our analysis revealed that 17.9% (112/627) of all intergenic lncRNAs display splicing conservation in at least one exon, an estimate that is substantially higher than previous estimates of lncRNA conservation in this group. Our findings agree with similar studies in vertebrates, demonstrating that splicing conservation can be evidence of stabilizing selection. We provide conclusive evidence for the existence of evolutionary deeply conserved lncRNAs in plants and describe a generally applicable computational workflow to identify functional lncRNAs in plants.


Assuntos
Sequência Conservada/genética , Processamento de RNA/genética , RNA Longo não Codificante/genética , RNA de Plantas/genética , Arabidopsis/genética , Brassica/genética , Evolução Molecular , Genoma de Planta/genética , RNA Mensageiro/genética
14.
Sci Rep ; 10(1): 4295, 2020 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-32152363

RESUMO

Annexins (ANN) are a multigene, evolutionarily conserved family of calcium-dependent and phospholipid-binding proteins that play important roles in plant development and stress resistance. However, a systematic comprehensive analysis of ANN genes of Brassicaceae species (Brassica rapa, Brassica oleracea, and Brassica napus) has not yet been reported. In this study, we identified 13, 12, and 26 ANN genes in B. rapa, B. oleracea, and B. napus, respectively. About half of these genes were clustered on various chromosomes. Molecular evolutionary analysis showed that the ANN genes were highly conserved in Brassicaceae species. Transcriptome analysis showed that different group ANN members exhibited varied expression patterns in different tissues and under different (abiotic stress and hormones) treatments. Meanwhile, same group members from Arabidopsis thaliana, B. rapa, B. oleracea, and B. napus demonstrated conserved expression patterns in different tissues. The weighted gene coexpression network analysis (WGCNA) showed that BnaANN genes were induced by methyl jasmonate (MeJA) treatment and played important roles in jasmonate (JA) signaling and multiple stress response in B. napus.


Assuntos
Anexinas/metabolismo , Brassica/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Família Multigênica , Proteínas de Plantas/metabolismo , Estresse Fisiológico , Anexinas/genética , Brassica/classificação , Brassica/genética , Brassica/metabolismo , Brassica napus/genética , Brassica napus/crescimento & desenvolvimento , Brassica napus/metabolismo , Brassica rapa/genética , Brassica rapa/crescimento & desenvolvimento , Brassica rapa/metabolismo , Filogenia , Proteínas de Plantas/genética
15.
Mol Phylogenet Evol ; 147: 106777, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32126279

RESUMO

Evolution of Brassica genome post-polyploidization reveals asymmetrical genome fractionation and copy number variation. Herein, we describe the impact of promoter divergence among SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) homeologs on expression and function in Brassica spp. SOC1, a regulated floral pathway integrator, is conserved as 3 redundant homeologs in diploid Brassicas. Even with high sequence identity within coding regions (92.8-100%), the spatio-temporal expression patterns of 9 SOC1 homologs in B. juncea and B. nigra indicates regulatory divergence. While LF and MF2 SOC1 homeologs are upregulated during floral transition, MF1 is barely expressed. Also, MF2 homeolog levels do not decline post-flowering, unlike LF. To investigate the underlying source of divergence, we analyzed the sequence and phylogeny of all reported (22) and isolated (21) upstream regions of Brassica SOC1. Full length upstream regions (4712-19189 bp) reveal 5 ubiquitously conserved ancestral Blocks, harboring binding sites of 18 TFs (TFBSs) characterized in Arabidopsis thaliana. The orthologs of these TFBSs are differentially conserved among Brassica SOC1 homeologs, imparting expression divergence. No crucial TFBSs are exclusively lost from LF_SOC1 promoter, while MF1_SOC1 has lost NF-Y binding site crucial for SOC1 activation by CONSTANS. MF2_SOC1 homeologs have lost important TFBSs (SEP3, AP1 and SMZ), responsible for SOC1 repression post-flowering. BjuAALF_SOC1 promoter (proximal 2 kb) shows ubiquitous reporter expression in B. juncea cv. Varuna transgenics, while BjuAAMF1_SOC1 promoter shows absence of reporter expression, validating the impact of TFBS divergence. Conservation of the original primary protein sequence is discovered in B. rapa homeologs (46) of 18 TFs. Co-regulation pattern of these TFs appeared similar for B. rapa LF and MF2 SOC1 homeologs; MF1 shows significant variation. Strong regulatory association is recorded for AP1, AP2, SEP3, FLC and CONSTANS/NF-Y, highlighting their importance in homeolog-specific SOC1 regulation. Correlation of B. juncea AP1, AP2 and FLC expression with SOC1 homeologs also complies with the TFBS differences. We thus conclude that redundant SOC1 loci contribute differentially to cumulative expression of SOC1 due to divergent selection of ancestral TFBSs.


Assuntos
Brassica/genética , Sequência Conservada , Evolução Molecular , Flores/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Homologia de Sequência do Ácido Nucleico , Fatores de Transcrição/metabolismo , Sequência de Bases , Sítios de Ligação , Brassica/metabolismo , Sequência Conservada/genética , Variações do Número de Cópias de DNA/genética , Genes Reporter , Filogenia , Poliploidia , Regiões Promotoras Genéticas , Ligação Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Tempo
16.
Int J Mol Sci ; 21(6)2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32183438

RESUMO

"Riceyness" refers to the precocious development of flower bud initials over the curd surface of cauliflower, and it is regarded as undesirable for the market. The present study aimed to identify the candidate loci and genes responsible for the morphological difference in riceyness between a pair of cauliflower sister lines. Genetic analysis revealed that riceyness is controlled by a single dominant locus. An F2 population derived from the cross between these sister lines was used to construct "riceyness" and "non-riceyness" bulks, and then it was subjected to BSA-seq. On the basis of the results of Δ(SNP-index) analysis, a 4.0 Mb candidate region including 22 putative SNPs was mapped on chromosome C04. Combining the RNA-seq, gene function annotation, and target sequence analysis among two parents and other breeding lines, an orthologous gene of the Arabidopsis gene SOC1, Bo4g024850 was presumed as the candidate gene, and an upstream SNP likely resulted in riceyness phenotype via influencing the expression levels of Bo4g024850. These results are helpful to understand the genetic mechanism regulating riceyness, and to facilitate the molecular improvement on cauliflower curds.


Assuntos
Brassica , Cromossomos de Plantas/genética , Flores , Locos de Características Quantitativas , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Brassica/genética , Brassica/crescimento & desenvolvimento , Mapeamento Cromossômico , Flores/genética , Flores/crescimento & desenvolvimento
17.
Nat Commun ; 11(1): 1264, 2020 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-32152287

RESUMO

Wildfires can encourage the establishment of invasive plants by releasing potent germination stimulants, such as karrikins. Seed germination of Brassica tournefortii, a noxious weed of Mediterranean climates, is strongly stimulated by KAR1, the archetypal karrikin produced from burning vegetation. In contrast, the closely-related yet non-fire-associated ephemeral Arabidopsis thaliana is unusual because it responds preferentially to KAR2. The α/ß-hydrolase KARRIKIN INSENSITIVE 2 (KAI2) is the putative karrikin receptor identified in Arabidopsis. Here we show that B. tournefortii expresses three KAI2 homologues, and the most highly-expressed homologue is sufficient to confer enhanced responses to KAR1 relative to KAR2 when expressed in Arabidopsis. We identify two amino acid residues near the KAI2 active site that explain the ligand selectivity, and show that this combination has arisen independently multiple times within dicots. Our results suggest that duplication and diversification of KAI2 proteins could confer differential responses to chemical cues produced by environmental disturbance, including fire.


Assuntos
Arabidopsis/metabolismo , Brassica/metabolismo , Proteínas de Transporte/efeitos dos fármacos , Proteínas de Transporte/metabolismo , Furanos/farmacologia , Plantas Daninhas/efeitos dos fármacos , Plantas Daninhas/metabolismo , Piranos/farmacologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Proteínas de Arabidopsis , Brassica/efeitos dos fármacos , Brassica/genética , Domínio Catalítico , Fogo , Regulação da Expressão Gênica de Plantas , Germinação/efeitos dos fármacos , Germinação/fisiologia , Hidrolases/genética , Hidrolases/metabolismo , Magnoliopsida , Proteínas de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plântula , Sementes/efeitos dos fármacos , Sementes/fisiologia , Análise de Sequência de Proteína , Transcriptoma , Incêndios Florestais
18.
Plant Reprod ; 33(1): 43-58, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32080762

RESUMO

Polyploidy or whole genome duplication is a frequent and recurrent phenomenon in flowering plants that has played a major role in their diversification, adaptation and speciation. The adaptive success of polyploids relates to the different evolutionary fates of duplicated genes. In this study, we explored the impact of the whole genome triplication (WGT) event in the Brassiceae tribe on the genes involved in the self-incompatibility (SI) signalling pathway, a mechanism allowing recognition and rejection of self-pollen in hermaphrodite plants. By taking advantage of the knowledge acquired on this pathway as well as of several reference genomes in Brassicaceae species, we determined copy number of the different genes involved in this pathway and investigated their structural and functional evolutionary dynamics. We could infer that whereas most genes involved in the SI signalling returned to single copies after the WGT event (i.e. ARC1, JDP1, THL1, THL2, Exo70A01) in diploid Brassica species, a few were retained in duplicated (GLO1 and PLDα) or triplicated copies (MLPK). We also carefully studied the gene structure of these latter duplicated genes (including the conservation of functional domains and active sites) and tested their transcription in the stigma to identify which copies seem to be involved in the SI signalling pathway. By taking advantage of these analyses, we then explored the putative origin of a contrasted SI phenotype between two Brassica rapa varieties that have been fully sequenced and shared the same S-allele (S60).


Assuntos
Evolução Biológica , Brassica , Genoma de Planta , Transdução de Sinais , Brassica/genética , Genoma de Planta/genética , Pólen , Transdução de Sinais/genética
19.
BMC Genomics ; 21(1): 178, 2020 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-32093614

RESUMO

BACKGROUND: The basic helix-loop-helix (bHLH) is the second largest gene family in the plant, some members play important roles in pistil development and response to drought, waterlogging, cold stress and salt stress. The bHLH gene family has been identified in many species, except for Brassica oleracea and B. napus thus far. This study aims to identify the bHLH family members in B. oleracea, B. rapa and B. napus, and elucidate the expression, duplication, phylogeny and evolution characters of them. RESULT: A total of 268 bHLH genes in B. oleracea, 440 genes in B. napus, and 251 genes in B. rapa, including 21 new bHLH members, have been identified. Subsequently, the analyses of the phylogenetic trees, conserved motifs and gene structures showed that the members in the same subfamily were highly conserved. Most Ka/Ks values of homologous gene were < 1, which indicated that these genes suffered from strong purifying selection for retention. The retention rates of BrabHLH and BolbHLH genes were 51.6 and 55.1%, respectively. The comparative expression patterns between B. rapa and B. napus showed that they had similar expression patterns in the root and contrasting patterns in the stems, leaves, and reproductive tissues. In addition, there were 41 and 30 differential expression bHLH genes under the treatments of ABA and JA, respectively, and the number of down regulation genes was significantly more than up regulation genes. CONCLUSION: In the present study, we identified and performed the comparative genomics analysis of bHLH gene family among B. oleracea, B. rapa and B. napus, and also investigated their diversity. The expression patterns between B. rapa and B. napus shows that they have the similar expression pattern in the root and opposite patterns in the stems, leaves, and reproduction tissues. Further analysis demonstrated that some bHLH gene members may play crucial roles under the abiotic and biotic stress conditions. This is the first to report on the bHLH gene family analysis in B. oleracea and B. napus, which can offer useful information on the functional analysis of the bHLH gene in plants.


Assuntos
Brassica/genética , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Sequências Hélice-Alça-Hélice , Brassica napus/genética , Brassica rapa/genética , Duplicação Gênica , Perfilação da Expressão Gênica , Genoma de Planta , Genômica , Filogenia
20.
Theor Appl Genet ; 133(4): 1303-1312, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31996972

RESUMO

KEY MESSAGE: BoFL, a candidate gene conferring the feathered-leaved trait in ornamental kale, is located in a 374.532-kb region on chromosome C9. Leaf shape is an important economic trait in ornamental kale (Brassica oleracea var. acephala). Identifying the genes that influence leaf shape would provide insight into the mechanism underlying leaf development. In this study, we constructed F1, F2, BC1P1, BC1P2, and F2:3 populations from a cross between a feathered-leaved inbred line, F0819, and a smooth-leaved inbred line, S0835, of ornamental kale. Genetic analysis showed that the feathered-leaved trait was controlled by a semi-dominant gene, BoFL. Using bulked segregant analysis sequencing, we mapped the BoFL gene to a 4.17-Mb interval on chromosome C9. Next, we narrowed down the candidate region to 374.532-kb by fine-scale mapping between the two flanking markers INDEL940 and INDEL5443. We identified 38 genes in the candidate region, among which only Bo9g184610 showed significant variation in expression level between the two parental lines. Sequencing of the gene in the parental lines identified three single-nucleotide polymorphism (SNP) differences, containing two nonsynonymous and one synonymous SNPs, which resulted in coding variations of an aspartate and a phenylalanine residue in F0819, but an alanine and a leucine residue in S0835. A cleaved amplified polymorphic sequence (CAPS) marker, CAPS4610, corresponding to the first nonsynonymous SNP co-segregated with the leaf shape trait. We thus speculated that the gene conferring the feathered-leaved trait is BoALG10, a homolog of ALG10, which encodes an alpha-1,2-glucosyltransferase in Arabidopsis thaliana. This work will be useful for understanding the mechanism of leaf development and provides important information for the breeding of kale with novel leaf shapes.


Assuntos
Brassica/genética , Genes de Plantas , Mapeamento Físico do Cromossomo , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Sequência de Bases , Segregação de Cromossomos/genética , Cromossomos de Plantas/genética , Cruzamentos Genéticos , Regulação da Expressão Gênica de Plantas , Estudos de Associação Genética , Marcadores Genéticos , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
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