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1.
Plants (Basel) ; 12(7)2023 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-37050100

RESUMO

Sugars Will Eventually be Exported Transporter (SWEET) gene family plays indispensable roles in plant physiological activities, development processes, and responses to biotic and abiotic stresses, but no information is known for roses. In this study, a total of 25 RcSWEET genes were identified in Rosa chinensis 'Old Blush' by genome-wide analysis and clustered into four subgroups based on their phylogenetic relationships. The genomic features, including gene structures, conserved motifs, and gene duplication among the chromosomes of RcSWEET genes, were characterized. Seventeen types of cis-acting elements among the RcSWEET genes were predicted to exhibit their potential regulatory roles during biotic and abiotic stress and hormone responses. Tissue-specific and cold-response expression profiles based on transcriptome data showed that SWEETs play widely varying roles in development and stress tolerance in two rose species. Moreover, the different expression patterns of cold-response SWEET genes were verified by qRT-PCR between the moderately cold-resistant species R. chinensis 'Old Blush' and the extremely cold-resistant species R. beggeriana. Especially, SWEET2a and SWEET10c exhibited species differences after cold treatment and were sharply upregulated in the leaves of R. beggeriana but not R. chinensis 'Old Blush', indicating that these two genes may be the crucial candidates that participate in cold tolerance in R. beggeriana. Our results provide the foundation for function analysis of the SWEET gene family in roses, and will contribute to the breeding of cold-tolerant varieties of roses.

2.
Front Plant Sci ; 13: 919294, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35979072

RESUMO

Essential oils have attracted wide attention in recent years due to their extensive applications in natural functional ingredients, pharmaceutical preparations, biomedical products, and the cosmetics industry. In this study, the chemical compositions and biological activities of essential oils extracted from six Lamiaceae herbs, including Pogostemon cablin (Blanco) Benth. (PCEO), Perilla frutescens (L.) Britton (PFEO), Salvia japonica Thunb. (SJEO), Rosmarinus officinalis L. (ROEO), Lavandula angustifolia Mill. (LAEO), and Agastache rugosa (Fisch. & C. A. Mey.) Kuntze (AREO), were determined and analyzed. A total of 167 components were identified from the six essential oils by GC-MS analysis, with 35, 24, 47, 46, 54, and 37 components in PCEO, PFEO, SJEO, ROEO, LAEO, and AREO, respectively. Hierarchical cluster analysis of chemical compositions showed that the composition of the six essential oils was significantly different in content, and they were clearly divided into six classes. However, all of these six essential oils exhibited promising anti-inflammatory activity by inhibiting the expression of interleukin-1, interleukin-6, tumor necrosis factor-α, and cyclooxygenase-2 in rats with adjuvant arthritis, among which PFEO had the best performance. In addition, the six essential oils displayed significant cytotoxicity on B16 (IC50 = 86.91-228.91 µg/mL) and LNCaP cell lines (IC50 = 116.4-189.63 µg/mL). Meanwhile, all of them presented satisfactory antioxidant activity (IC50 = 4.88-13.89 µg/mL) compared with Trolox C (IC50 = 13.83 µg/mL), and SJEO (IC50 = 7.93 µg/mL) served as an optimal candidate natural antioxidant by DPPH assay. Taken together, these results indicate that the six Lamiaceae essential oils manifest excellent and diverse biological activities, enabling them to be used as perfect natural functional ingredients in antioxidant, antitumor, or anti-arthritic drugs. This study provides more references for pharmaphylogeny research and drug discovery from folk medicinal plants.

3.
Front Plant Sci ; 13: 845662, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35401615

RESUMO

Rosa beggeriana 'Aurea' is a yellow-green leaf (yl) mutant and originated from Rosa beggeriana Schrenk by 60Co-γ irradiation, which is an important ornamental woody species. However, the molecular mechanism of the yl mutant remains unknown. Herein, comparative transcriptome profiling was performed between the yl type and normal green color type (WT) by RNA sequencing. A total of 3,372 significantly differentially expressed genes (DEGs) were identified, consisting of 1,585 upregulated genes and 1,787 downregulated genes. Genes that took part in metabolic of biological process (1,090), membrane of cellular component (728), catalytic (1,114), and binding of molecular function (840) were significantly different in transcription level. DEGs involved in chlorophyll biosynthesis, carotenoids biosynthesis, cutin, suberine, wax biosynthesis, photosynthesis, chloroplast development, photosynthesis-antenna proteins, photosystem I (PSI) and photosystem II (PSII) components, CO2 fixation, ribosomal structure, and biogenesis related genes were downregulated. Meanwhile, linoleic acid metabolism, siroheme biosynthesis, and carbon source of pigments biosynthesis through methylerythritol 4-phosphate (MEP) pathways were upregulated. Moreover, a total of 147 putative transcription factors were signification different expression, involving NAC, WRKY, bHLH, MYB and AP2/ERF, C2H2, GRAS, and bZIP family gene. Our results showed that the disturbed pigments biosynthesis result in yl color by altering the ratio of chlorophylls and carotenoids in yl mutants. The yl mutants may evoke other metabolic pathways to compensate for the photodamage caused by the insufficient structure and function of chloroplasts, such as enhanced MEP pathways and linoleic acid metabolism against oxidative stress. This research can provide a reference for the application of leaf color mutants in the future.

4.
Breed Sci ; 72(4): 275-284, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36699823

RESUMO

The induction of 2n pollen is an important technique for breeding polyploid plants. Here, we observed meiosis in the pollen mother cells (PMCs) of six Phalaenopsis cultivars and attempted to induce 2n pollen. The meiotic stage was related to flower bud length. During meiosis, Phalaenopsis cultivars with flower widths of approximately 20-40 mm and 50-60 mm had bud lengths of approximately 3-8 mm and 5-13 mm, respectively. The duration of meiosis ranged from 4.2 to 14 d. This was the first study to characterize meiosis of the PMCs of Phalaenopsis. The natural generation frequency of 2n pollen varied from 0.68% to 1.78%. Meiotic stage and colchicine concentration significantly affected the induction of 2n pollen. The most effective treatment for obtaining 2n pollen was 0.05% colchicine in the leptotene to zygotene stage for 3 d, which achieved a 2n pollen frequency of 10.04%.

5.
J Agric Food Chem ; 68(15): 4473-4484, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32208653

RESUMO

Long-chain acyl-coenzyme A (CoA) synthetase (LACS) catalyzes the formation of acyl-CoAs from free fatty acids, which is pivotal for lipid metabolism. Here, we confirmed the presence of six CzLACS genes in Chromochloris zofingiensis. Functional complementation and in vitro enzymatic assay indicated that CzLACS2 through CzLACS5 rather than CzLACS1 or CzLACS6 are bona fide LACS enzymes and they have overlapping yet distinct substrate preference. The results of the subcellular colocalization experiment and different expression patterns under three triacylglycerol (TAG)-inducing conditions showed that CzLACS2 through CzLACS4 reside at endoplasmic reticulum (ER) and are involved in TAG biosynthesis, while CzLACS5 resides in peroxisome and participates in fatty acid ß-oxidation. The yeast one-hybrid assay using a library of 50 transcription factors (TFs) constructed in our study identified 12 TFs potentially involved in regulating the expression of CzLACSs. Moreover, heterologous expression of CzLACSs demonstrated their engineering potential for modulating TAG synthesis in yeast and algal cells.


Assuntos
Clorofíceas/enzimologia , Coenzima A Ligases/metabolismo , Família Multigênica , Sequência de Aminoácidos , Clorofíceas/química , Clorofíceas/classificação , Clorofíceas/genética , Coenzima A Ligases/genética , Retículo Endoplasmático/enzimologia , Retículo Endoplasmático/metabolismo , Filogenia , Transporte Proteico , Alinhamento de Sequência , Especificidade por Substrato , Triglicerídeos/metabolismo
6.
J Agric Food Chem ; 67(34): 9569-9578, 2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31385495

RESUMO

Acetyl-CoA synthetase (ACS) plays a key role in microalgal lipid biosynthesis and acetyl-CoA industrial production. In the present study, two ACSs were cloned and characterized from the oleaginous microalga Chromochloris zofingiensis. In vitro kinetic analysis showed that the Km values of CzACS1 and CzACS2 for potassium acetate were 0.99 and 0.81 mM, respectively. Moreover, CzACS1 and CzACS2 had outstanding catalytic efficiencies (kcat/Km), which were 70.67 and 79.98 s-1 mM-1, respectively, and these values were higher than that of other reported ACSs. CzACS1 and CzACS2 exhibited differential expression patterns at the transcriptional level under various conditions. Screening a recombinant library of 52 transcription factors (TFs) constructed in the present study via yeast one-hybrid assay pointed to seven TFs with potential involvement in the regulation of the two ACS genes. Expression correlation analysis implied that GATA20 was likely an important regulator of CzACS2 and that ERF9 could regulate two CzACSs simultaneously.


Assuntos
Acetato-CoA Ligase/metabolismo , Clorófitas/enzimologia , Regulação Enzimológica da Expressão Gênica , Microalgas/enzimologia , Acetato-CoA Ligase/química , Acetato-CoA Ligase/genética , Biocatálise , Clorófitas/química , Clorófitas/genética , Cinética , Metabolismo dos Lipídeos , Microalgas/química , Microalgas/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
7.
Biotechnol Biofuels ; 12: 28, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30792816

RESUMO

BACKGROUND: The green alga Chlorella zofingiensis has been recognized as an industrially relevant strain because of its robust growth under multiple trophic conditions and the potential for simultaneous production of triacylglycerol (TAG) and the high-value keto-carotenoid astaxanthin. Nevertheless, the mechanism of TAG synthesis remains poorly understood in C. zofingiensis. Diacylglycerol acyltransferase (DGAT) is thought to catalyze the committed step of TAG assembly in the Kennedy pathway. C. zofingiensis genome is predicted to possess eleven putative DGAT-encoding genes, the greatest number ever found in green algae, pointing to the complexity of TAG assembly in the alga. RESULTS: The transcription start site of C. zofingiensis DGATs was determined by 5'-rapid amplification of cDNA ends (RACE), and their coding sequences were cloned and verified by sequencing, which identified ten DGAT genes (two type I DGATs designated as CzDGAT1A and CzDGAT1B, and eight type II DGATs designated as CzDGTT1 through CzDGTT8) and revealed that the previous gene models of seven DGATs were incorrect. Function complementation in the TAG-deficient yeast strain confirmed the functionality of most DGATs, with CzDGAT1A and CzDGTT5 having the highest activity. In vitro DGAT assay revealed that CzDGAT1A and CzDGTT5 preferred eukaryotic and prokaryotic diacylglycerols (DAGs), respectively, and had overlapping yet distinctive substrate specificity for acyl-CoAs. Subcellular co-localization experiment in tobacco leaves indicated that both CzDGAT1A and CzDGTT5 were localized at endoplasmic reticulum (ER). Upon nitrogen deprivation, TAG was drastically induced in C. zofingiensis, accompanied by a considerable up-regulation of CzDGAT1A and CzDGTT5. These two genes were probably regulated by the transcription factors (TFs) bZIP3 and MYB1, as suggested by the yeast one-hybrid assay and expression correlation. Moreover, heterologous expression of CzDGAT1A and CzDGTT5 promoted TAG accumulation and TAG yield in different hosts including yeast and oleaginous alga. CONCLUSIONS: Our study represents a pioneering work on the characterization of both type I and type II C. zofingiensis DGATs by systematically integrating functional complementation, in vitro enzymatic assay, subcellular localization, yeast one-hybrid assay and overexpression in yeast and oleaginous alga. These results (1) update the gene models of C. zofingiensis DGATs, (2) shed light on the mechanism of oleaginousness in which CzDGAT1A and CzDGTT5, have functional complementarity and probably work in collaboration at ER contributing to the abundance and complexity of TAG, and (3) provide engineering targets for future trait improvement via rational manipulation of this alga as well as other industrially relevant ones.

8.
Biotechnol Biofuels ; 12: 287, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31890015

RESUMO

BACKGROUND: Chromochloris zofingiensis is emerging as an industrially relevant alga given its robust growth for the production of lipids and astaxanthin, a value-added carotenoid with broad applications. Nevertheless, poor understanding of astaxanthin synthesis has limited engineering of this alga for rational improvements. RESULTS: To reveal the molecular mechanism underlying astaxanthin accumulation in C. zofingiensis, here we conducted an integrated analysis by combining the time-resolved transcriptomes and carotenoid profiling in response to nitrogen deprivation (ND). A global response was triggered for C. zofingiensis to cope with the ND stress. Albeit the little variation in total carotenoid content, individual carotenoids responded differentially to ND: the primary carotenoids particularly lutein and ß-carotene decreased, while the secondary carotenoids increased considerably, with astaxanthin and canthaxanthin being the most increased ones. The carotenogenesis pathways were reconstructed: ND had little effect on the carbon flux to carotenoid precursors, but stimulated astaxanthin biosynthesis while repressing lutein biosynthesis, thereby diverting the carotenoid flux from primary carotenoids to secondary carotenoids particularly astaxanthin. Comparison between C. zofingiensis and Haematococcus pluvialis revealed the distinctive mechanism of astaxanthin synthesis in C. zofingiensis. Furthermore, potential bottlenecks in astaxanthin synthesis were identified and possible engineering strategies were proposed for the alga. CONCLUSIONS: Collectively, these findings shed light on distinctive mechanism of carotenogenesis for astaxanthin biosynthesis in C. zofingiensis, identify key functional enzymes and regulators with engineering potential and will benefit rational manipulation of this alga for improving nutritional traits.

9.
Plant Cell Rep ; 37(7): 1049-1060, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29687169

RESUMO

KEY MESSAGE: We find that the DREB subfamily transcription factor, CmERF053, has a novel function to regulate the development of shoot branching and lateral root in addition to affecting abiotic stress. Dehydration-responsive element binding proteins (DREBs) are important plant transcription factors that regulate various abiotic stresses. Here, we isolated an APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor from chrysanthemum (Chrysanthemum morifolium 'Jinba'), CmERF053, the expression of which was rapidly up-regulated by main stem decapitation. Phylogenetic analysis indicated that it belongs to the A-6 group of the DREB subfamily, and the subcellular localization assay confirmed that CmERF053 was a nuclear protein. Overexpression of CmERF053 in Arabidopsis exhibited positive effects of plant lateral organs, which had more shoot branching and lateral roots than did the wild type. We also found that the expression of CmERF053 in axillary buds was induced by exogenous cytokinins. These results suggested that CmERF053 may be involved in cytokinins-related shoot branching pathway. In this study, an altered auxin distribution was observed during root elongation in the seedlings of the overexpression plants. Furthermore, overexpress CmERF053 gene could enhance drought tolerance. Together, these findings indicated that CmERF053 plays crucial roles in regulating shoot branching, lateral root, and drought stress in plant. Moreover, our study provides potential application value for improving plant productivity, ornamental traits, and drought tolerance.


Assuntos
Chrysanthemum/fisiologia , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/fisiologia , Chrysanthemum/genética , Citocininas/metabolismo , Secas , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Plantas Geneticamente Modificadas , Plântula , Fatores de Transcrição/metabolismo
10.
Front Plant Sci ; 7: 557, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27200031

RESUMO

Thidiazuron (N-phenyl-N'-1,2,3-thiadiazol-5-ylurea; TDZ) is an artificial plant growth regulator that is widely used in plant tissue culture. Protocorm-like bodies (PLBs) induced by TDZ serve as an efficient and rapid in vitro regeneration system in Rosa species. Despite this, the mechanism of PLB induction remains relatively unclear. TDZ, which can affect the level of endogenous auxins and cytokinins, converts the cell fate of rhizoid tips and triggers PLB formation and plantlet regeneration in Rosa canina L. In callus-rhizoids, which are rhizoids that co-develop from callus, auxin and a Z-type cytokinin accumulated after applying TDZ, and transcription of the auxin transporter gene RcPIN1 was repressed. The expression of RcARF4, RcRR1, RcCKX2, RcCKX3, and RcLOG1 increased in callus-rhizoids and rhizoid tips while the transcription of an auxin response factor (RcARF1) and auxin transport proteins (RcPIN2, RcPIN3) decreased in callus-rhizoids but increased in rhizoid tips. In situ hybridization of rhizoids showed that RcWUS and RcSERK1 were highly expressed in columella cells and root stem cells resulting in the conversion of cell fate into shoot apical meristems or embryogenic callus. In addition, transgenic XVE::RcWUS lines showed repressed RcWUS overexpression while RcWUS had no effect on PLB morphogenesis. Furthermore, higher expression of the root stem cell marker RcWOX5 and root stem cell maintenance regulator genes RcPLT1 and RcPLT2 indicated the presence of a dedifferentiation developmental pathway in the stem cell niche of rhizoids. Viewed together, our results indicate that different cells in rhizoid tips acquired regeneration competence after induction by TDZ. A novel developmental pathway containing different cell types during PLB formation was identified by analyzing the endogenous auxin and cytokinin content. This study also provides a deeper understanding of the mechanisms underlying in vitro regeneration in Rosa.

11.
Plant Physiol Biochem ; 96: 241-53, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26310142

RESUMO

Shoot branching plays an important role in determining plant architecture. Strigolactones (SLs) negatively regulate shoot branching, and can respond to conditions of low or absent phosphate or nitrogen. The D14 gene is a probable candidate as an SL receptor in rice, petunia, and Arabidopsis. To investigate the roles of D14 in shoot branching of chrysanthemum, we isolated the D14 homolog DgD14. Functional analysis showed that DgD14 was a nuclear-localized protein, and restored the phenotype of Arabidopsis d14-1. Exogenous SL (GR24) could down-regulate DgD14 expression, but this effect could be overridden by apical auxin application. Decapitation could down-regulate DgD14 expression, but this effect could be restored by exogenous auxin. In addition, DgD14 transcripts produced rapid responses in shoot and root under conditions of phosphate absence, but only a mild variation in bud and stem with low nitrogen treatment. Indistinct reductions of P levels in shoot were observed in plants grown under low nitrogen conditions. The absence of phosphate and low levels of nitrogen negatively affected plant growth. These results demonstrate that P levels in shoot had a close relationship with phosphate, whereas nitrogen did not directly regulate DgD14 expression in shoot. Taken together, these results demonstrated that DgD14 was the functional strigolactone signaling component in chrysanthemum.


Assuntos
Chrysanthemum/crescimento & desenvolvimento , Genes de Plantas , Brotos de Planta/crescimento & desenvolvimento , Sequência de Aminoácidos , Chrysanthemum/genética , Dados de Sequência Molecular , Proteínas de Plantas/química , Proteínas de Plantas/genética , Brotos de Planta/genética , Homologia de Sequência de Aminoácidos
12.
Plant Mol Biol ; 86(6): 671-9, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25301174

RESUMO

Homeobox (HB) proteins are important transcription factors that regulate the developmental decisions of eukaryotes. WUSCHEL-related homeobox (WOX) transcription factors, known as a plant-specific HB family, play a key role in plant developmental processes. Our previous work has indicated that rhizoids are induced by auxin in rose (Rosa spp.), which acts as critical part of an efficient plant regeneration system. However, the function of WOX genes in auxin-induced rhizoid formation remains unclear. Here, we isolated and characterized a WUSCHEL-related homeobox gene from Rosa canina, RcWOX1, containing a typical homeodomain with 65 amino acid residues. Real-time reverse transcription PCR (qRT-PCR) analysis revealed that RcWOX1 was expressed in the whole process of callus formation and in the early stage of rhizoid formation. Moreover, its expression was induced by auxin treatment. In Arabidopsis transgenic lines expressing the RcWOX1pro::GUS and 35S::GFP-RcWOX1, RcWOX1 was specifically expressed in roots and localized to the nucleus. Overexpression of RcWOX1 in Arabidopsis increased lateral root density and induced upregulation of PIN1 and PIN7 genes. Therefore, we postulated that RcWOX1 is a functional transcription factor that plays an essential role in auxin-induced rhizoid formation.


Assuntos
Regulação da Expressão Gênica de Plantas/genética , Ácidos Indolacéticos/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/genética , Rosa/genética , Sequência de Aminoácidos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Núcleo Celular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Genes Reporter , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Dados de Sequência Molecular , Especificidade de Órgãos , Filogenia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas/genética , Rosa/citologia , Rosa/crescimento & desenvolvimento , Plântula/genética , Plântula/crescimento & desenvolvimento , Alinhamento de Sequência , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
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