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1.
Tree Physiol ; 2019 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-31860707

RESUMO

WRKY transcription factors, one of the largest transcription factor families, play important roles in regulating the synthesis of secondary metabolites. In sweet osmanthus (Osmanthus fragrans), the monoterpenes have been demonstrated as the most important volatile compounds, and the W-box, which is the cognate binding site of WRKY transcription factors could be identified in most of terpene synthesis related genes' promoters. However, the role of the WRKY family in terpene synthesis in sweet osmanthus has rarely been examined. In this study, 154 WRKY genes with conserved WRKY domain were identified and classified into three groups. The group II was further divided into five subgroups and almost all members of IId contained a plant zinc cluster domain. Eight OfWRKYs (OfWRKY7/19/36/38/42/84/95/139) were screened from 20 OfWRKYs for their flower-specific expression patterns in different tissues. Simultaneously, the expression patterns of OfWRKYs and emission patterns of volatile compounds during the flowering process were determined and GC-MS results showed that monoterpenes, such as linalool and ocimene, accounted for the highest proportion, contributing to the floral scent of sweet osmanthus in two cultivars. In addition, correlation analysis revealed the expression patterns of OfWRKYs (OfWRKY7/19/36/139) were each correlated with distinct monoterpenes (linalool, linalool derivatives, ocimene, and ocimene derivatives). Subcellular localization analysis showed that p35S::GFP-OfWRKY7/38/95/139 were localized in the nucleus and OfWRKY139 had very strong transactivation activity. Collectively, the results indicated potential roles of OfWRKY139 and OfWRKYs with plant zinc cluster domain in regulating synthesis of aromatic compounds in sweet osmanthus, laying the foundation for use of OfWRKYs to improve aroma of ornamental plants.

2.
PLoS One ; 14(11): e0225451, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31747430

RESUMO

Clerodendrum trichotomum, a member of the Lamiaceae (Verbenaceae) family, is an ornamental plant widely distributed in South Asia. Previous studies have focused primarily on its growth characteristics, stress resistance, and pharmacological applications; however, molecular investigations remain limited. Considering germplasm conservation and the extensive applications of this plant, it is necessary to explore transcriptome resources and SSR makers for C. trichotomum. In the present study, RNA sequencing was used to determine the transcriptome of C. trichotomum. Subsequently, unigene annotations and classifications were obtained, and SSRs were mined with MIcroSAtellite. Finally, primer pairs designed with Oligo 6.0 were selected for polymorphism validation. In total, 127,325,666 high-quality reads were obtained, and 58,345 non-redundant unigenes were generated, of which 36,900 (63.24%) were annotated. Among the annotated unigenes, 35,980 (97.51%) had significant similarity to 607 species in Nr databases. In addition, a total of 6,444 SSRs were identified in 5,530 unigenes, and 200 random primer pairs were designed for polymorphism validation. Furthermore, after primary polymorphism identification, 30 polymorphic primer pairs were selected for the further polymorphism screening, and 200 alleles were identified, 197 of which showed polymorphism. In this work, a large number of unigenes were generated, and numerous SSRs were detected. These findings should be beneficial for further investigations into germplasm conservation and various applications of C. trichotomum. These results should also provide a solid foundation for future molecular biology studies in C. trichotomum.

3.
Plants (Basel) ; 8(10)2019 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-31627411

RESUMO

Lycoris, which is known as the 'Chinese tulip,' has diverse flower colors and shapes, and some species have a delicate fragrance. However, limited studies have reported the volatile organic compounds (VOCs) of Lycoris. In this study, headspace solid-phase microextraction combined with gas chromatography-mass spectrometry was used to analyze the floral VOCs of six typical Lycoris taxa. Thirty-two VOCs were identified, including terpenoids, alcohols, esters, aldehydes, ketones, and phenols. The aldehyde and terpenoid contents in Lycoris aurea were higher than in the other taxa, and the ester and alcohol contents in L. sprengeri were the highest compared to all taxa tested. Compared with other species and cultivars, L. longituba and L. longituba var. flava were the two most scented taxa and the VOCs were dominated by terpenoids and esters. L. radiate and L. chinensis were two unscented taxa and, accordingly, the VOC content was weak. A partial least squares discriminate analysis of the floral VOCs among the six Lycoris taxa showed that the six taxa could be successfully separated. Moreover, the VOCs of L. longituba and L. longituba var. flava clustered together. ß-Ocimene was verified as the most important aroma compound, as determined via the calculation of the variable importance in projection values and significance analysis. ß-Ocimene and its trans isomer, trans-ß-ocimene, had a high relative content in L. longituba, L. longituba var. flava, L. aurea, and L. chinensis but were not detected in L. sprengeri and L. radiata. These results indicate that floral VOCs might be selected during the evolutional processes of Lycoris, and ß-ocimene could be the most typical VOC among the different Lycoris taxa.

5.
Plants (Basel) ; 8(3)2019 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-30823447

RESUMO

Lycoris longituba, belonging to the Amaryllidaceae family, is a perennial bulb bearing flowers with diverse colors and fragrance. Selection of cultivars with excellent colored and scented flowers has always been the breeding aim for ornamental plants. However, the molecular mechanisms underlying color fading and aroma production during flower expansion in L. longituba remain unclear. Therefore, to systematically investigate these important biological phenomena, the tepals of L. longituba from different developmental stages were used to screen and analyze the metabolic components and relevant genes. Utilizing the Illumina platform, a total of 144,922 unigenes were obtained from the RNA-Seq libraries. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that the phenylpropanoid biosynthesis and flavonoid biosynthesis pathways might play important roles during color and aroma changes. Metabolomic analysis identified 29 volatile organic components (VOCs) from different developmental stages of L. longituba tepals, and orthogonal partial least-squares discriminate analysis (OPLS-DA) revealed that trans-ß-ocimene-a terpene-was the most important aroma compound. Meanwhile, we found the content of anthocyanin was significantly reduced during the tepal color fading process. Then, we identified two dihydroflavonol-4-reductase (DFR) and three terpene synthase (TPS) genes, for which expression changes coincided with the production patterns of anthocyanins and trans-ß-ocimene, respectively. Furthermore, a number of MYB and bHLH transcription factors (TFs) which might be involved in color- and aroma-formation were also identified in L. longituba tepal transcriptomes. Taken together, this is the first comprehensive report of the color and fragrance in tepals of L. longituba and these results could be helpful in understanding these characteristics and their regulation networks.

6.
Hortic Res ; 5: 72, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30479779

RESUMO

Sweet osmanthus (Osmanthus fragrans) is a very popular ornamental tree species throughout Southeast Asia and USA particularly for its extremely fragrant aroma. We constructed a chromosome-level reference genome of O. fragrans to assist in studies of the evolution, genetic diversity, and molecular mechanism of aroma development. A total of over 118 Gb of polished reads was produced from HiSeq (45.1 Gb) and PacBio Sequel (73.35 Gb), giving 100× depth coverage for long reads. The combination of Illumina-short reads, PacBio-long reads, and Hi-C data produced the final chromosome quality genome of O. fragrans with a genome size of 727 Mb and a heterozygosity of 1.45 %. The genome was annotated using de novo and homology comparison and further refined with transcriptome data. The genome of O. fragrans was predicted to have 45,542 genes, of which 95.68 % were functionally annotated. Genome annotation found 49.35 % as the repetitive sequences, with long terminal repeats (LTR) being the richest (28.94 %). Genome evolution analysis indicated the evidence of whole-genome duplication 15 million years ago, which contributed to the current content of 45,242 genes. Metabolic analysis revealed that linalool, a monoterpene is the main aroma compound. Based on the genome and transcriptome, we further demonstrated the direct connection between terpene synthases (TPSs) and the rich aromatic molecules in O. fragrans. We identified three new flower-specific TPS genes, of which the expression coincided with the production of linalool. Our results suggest that the high number of TPS genes and the flower tissue- and stage-specific TPS genes expressions might drive the strong unique aroma production of O. fragrans.

7.
Molecules ; 23(7)2018 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-30004428

RESUMO

Osmanthus fragrans, or "RiXiangGui", is an ornamental, woody, evergreen plant that is cultivated widely because it blooms recurrently and emits a strong fragrance. Recently, the germplasm resources, classification, and aroma compositions of O. fragrans have been investigated. However, the molecular mechanisms of the floral scent formation and regulation have remained largely unknown. To obtain a global perspective on the molecular mechanism of the aroma formation during blooming, nine RNA Sequencing (RNA-Seq) libraries were constructed from three flowering stages: The initial, full, and final flowering stage. In short, a total of 523,961,310 high-quality clean reads were assembled into 136,611unigenes, with an average sequence length of 792 bp. About 47.43% of the unigenes (64,795) could be annotated in the NCBI non-redundant protein database. A number of candidate genes were identified in the terpenoid metabolic pathways and 1327 transcription factors (TFs), which showed differential expression patterns among the floral scent formation stages, were also identified, especially OfMYB1, OfMYB6, OfWRKY1, and OfWRKY3, which could play critical roles in the floral scent formation. These results indicated that the floral scent formation of O. fragrans was a very complex process which involved a large number of TFs. This study provides reliable resources for further studies of the O.fragrans floral scent formation.


Assuntos
Oleaceae/genética , Transcriptoma/genética , Flores/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Anotação de Sequência Molecular/métodos , Proteínas de Plantas/genética , Análise de Sequência de RNA/métodos , Fatores de Transcrição/genética
8.
J Genet ; 96(2): 273-281, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28674226

RESUMO

Accurate normalized data is a primary requisite for quantifying gene expression using RT-qPCR technology. Despite this importance, however, suitable reference genes in Osmanthus fragrans are not available. In this study, seven potential candidate reference genes (OfL25-1, OfL25-10, OfRP2, OfTUA, OfTUB3, OfUBQ2 and Of18S) were evaluated to determine which one would be the most reliable reference genes. The expression levels of the candidate reference genes were analysed by RT-qPCR in flower, leaf, pedicel, blossom bud tissues, as well as in floral organs at different developmental stages.GeNormand NormFinderwere used to statistically analyse transcript variation.Results indicated that OfRP2 and OfL25-10 were the optimal reference genes for use in RT-qPCR when analysing different stages of floral development; while OfTUB3 and OfL25-1 were optimal across tissues. The selected reference genes were used to examineOfMYB1 expression. The results appeared to be useful for future gene expression analyses aiming to characterize developmental stages and tissues of O. fragrans.


Assuntos
Flores/genética , Oleaceae/genética , Transcriptoma/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas , Oleaceae/crescimento & desenvolvimento , Proteínas de Plantas/genética , Padrões de Referência
9.
Genes (Basel) ; 8(2)2017 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-28157171

RESUMO

The authors wish to make the following correction to their paper [...].

10.
Genes (Basel) ; 7(10)2016 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-27690108

RESUMO

The 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway is responsible for the biosynthesis of many crucial secondary metabolites, such as carotenoids, monoterpenes, plastoquinone, and tocopherols. In this study, we isolated and identified 10 MEP pathway genes in the important aromatic plant sweet osmanthus (Osmanthus fragrans). Multiple sequence alignments revealed that 10 MEP pathway genes shared high identities with other reported proteins. The genes showed distinctive expression profiles in various tissues, or at different flower stages and diel time points. The qRT-PCR results demonstrated that these genes were highly expressed in inflorescences, which suggested a tissue-specific transcript pattern. Our results also showed that OfDXS1, OfDXS2, and OfHDR1 had a clear diurnal oscillation pattern. The isolation and expression analysis provides a strong foundation for further research on the MEP pathway involved in gene function and molecular evolution, and improves our understanding of the molecular mechanism underlying this pathway in plants.

11.
PLoS One ; 11(8): e0158228, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27508410

RESUMO

Different mulches have variable effects on soil physical properties and plant growth. This study aimed to compare the effects of mulching with inorganic (round gravel, RG), organic (wood chips, WC), and living (manila turf grass, MG) materials on soil properties at 0-5-cm and 5-10-cm depths, as well as on the growth and physiological features of Osmanthus fragrans L. 'Rixianggui' plants. Soil samples were collected at three different time points from field plots of O. fragrans plants treated with the different mulching treatments. Moisture at both soil depths was significantly higher after mulching with RG and WC than that in the unmulched control (CK) treatment. Mulching did not affect soil bulk density, pH, or total nitrogen content, but consistently improved soil organic matter. The available nitrogen in the soil increased after RG and WC treatments, but decreased after MG treatment during the experimental period. Mulching improved plant growth by increasing root activity, soluble sugar, and chlorophyll a content, as well as by providing suitable moisture conditions and nutrients in the root zone. Plant height and trunk diameter were remarkably increased after mulching, especially with RG and WC. However, while MG improved plant growth at the beginning of the treatment, the 'Rixianggui' plants later showed no improvement in growth. This was probably because MG competed with the plants for water and available nitrogen in the soil. Thus, our findings suggest that RG and WC, but not MG, improved the soil environment and the growth of 'Rixianggui' plants. Considering the effect of mulching on soil properties and plant growth and physiology, round gravel and wood chips appear to be a better choice than manila turf grass in 'Rixianggui' nurseries. Further studies are required to determine the effects of mulch quality and mulch-layer thickness on shoot and root growths.


Assuntos
Agricultura/métodos , Oleaceae/crescimento & desenvolvimento , Solo/química , Fertilizantes/análise , Concentração de Íons de Hidrogênio , Nitrogênio/análise , Compostos Orgânicos/química , Raízes de Plantas/crescimento & desenvolvimento
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