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1.
Int J Mol Sci ; 23(2)2022 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-35055168

RESUMO

Dendrobium catenatum Lindl is a valuable medicinal herb and gardening plant due to its ornamental value and special medical value. Low temperature is a major bottleneck restricting D. catenatum expansion towards the north, which influences the quality and yield of D. catenatum. In this study, we analysed the cold response of D. catenatum by RNA-Seq. A total of 4302 differentially expressed genes were detected under cold stress, which were mainly linked to protein kinase activity, membrane transport and the glycan biosynthesis and metabolism pathway. We also identified 4005 differential alternative events in 2319 genes significantly regulated by cold stress. Exon skipping and intron retention were the most common alternative splicing isoforms. Numerous genes were identified that differentially modulated under cold stress, including cold-induced transcription factors and splicing factors mediated by AS (alternative splicing). GO enrichment analysis found that differentially alternatively spliced genes without differential expression levels were related to RNA/mRNA processing and spliceosomes. DAS (differentially alternative splicing) genes with different expression levels were mainly enriched in protein kinase activity, plasma membrane and cellular response to stimulus. We further identified and cloned DcCBP20 in D. catenatum; we found that DcCBP20 promotes the generation of alternative splicing variants in cold-induced genes under cold stress via genetic experiments and RT-PCR. Taken together, our results identify the main cold-response pathways and alternative splicing events in D. catenatum responding to cold treatment and that DcCBP20 of D. catenatum get involved in regulating the AS and gene expression of cold-induced genes during this process. Our study will contribute to understanding the role of AS genes in regulating the cold stress response in D. catenatum.


Assuntos
Processamento Alternativo , Dendrobium/crescimento & desenvolvimento , Perfilação da Expressão Gênica/métodos , Proteínas de Plantas/genética , Resposta ao Choque Frio , Dendrobium/genética , Regulação da Expressão Gênica de Plantas , Ontologia Genética , RNA-Seq , Fatores de Transcrição/genética
2.
Int J Mol Sci ; 22(19)2021 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-34638610

RESUMO

Teosinte branched1/cycloidea/proliferating cell factor (TCP) gene family members are plant-specific transcription factors that regulate plant growth and development by controlling cell proliferation and differentiation. However, there are no reported studies on the TCP gene family in Dendrobium catenatum Lindl. Here, a genome-wide analysis of TCP genes was performed in D. catenatum, and 25 TCP genes were identified. A phylogenetic analysis classified the family into two clades: Class I and Class II. Genes in the same clade share similar conserved motifs. The GFP signals of the DcaTCP-GFPs were detected in the nuclei of tobacco leaf epidermal cells. The activity of DcaTCP4, which contains the miR319a-binding sequence, was reduced when combined with miR319a. A transient activity assay revealed antagonistic functions of Class I and Class II of the TCP proteins in controlling leaf development through the jasmonate-signaling pathway. After different phytohormone treatments, the DcaTCP genes showed varied expression patterns. In particular, DcaTCP4 and DcaTCP9 showed opposite trends after 3 h treatment with jasmonate. This comprehensive analysis provides a foundation for further studies on the roles of TCP genes in D. catenatum.


Assuntos
Dendrobium/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Motivos de Aminoácidos , Sequência Conservada , Ciclopentanos/farmacologia , Dendrobium/efeitos dos fármacos , Dendrobium/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genoma de Planta , Estudo de Associação Genômica Ampla , Levanogestrel , Lipoxigenase/genética , Família Multigênica , Oxilipinas/farmacologia , Filogenia , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/química , Plantas Geneticamente Modificadas , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Fatores de Transcrição/química
3.
Biomolecules ; 11(5)2021 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-34063498

RESUMO

Dendrobium officinale Kimura et Migo is a precious traditional Chinese medicine. Despite D. officinale displaying a good salt-tolerance level, the yield and growth of D. officinale were impaired drastically by the increasing soil secondary salinization. The molecular mechanisms of D. officinale plants' adaptation to salt stress are not well documented. Therefore, in the present study, D. officinale plants were treated with 250 mM NaCl. Transcriptome analysis showed that salt stress significantly altered various metabolic pathways, including phenylalanine metabolism, flavonoid biosynthesis, and α-linolenic acid metabolism, and significantly upregulated the mRNA expression levels of DoAOC, DoAOS, DoLOX2S, DoMFP, and DoOPR involved in the jasmonic acid (JA) biosynthesis pathway, as well as rutin synthesis genes involved in the flavonoid synthesis pathway. In addition, metabolomics analysis showed that salt stress induced the accumulation of some compounds in D. officinale leaves, especially flavonoids, sugars, and alkaloids, which may play an important role in salt-stress responses of leaf tissues from D. officinale. Moreover, salt stress could trigger JA biosynthesis, and JA may act as a signal molecule that promotes flavonoid biosynthesis in D. officinale leaves. To sum up, D. officinale plants adapted to salt stress by enhancing the biosynthesis of secondary metabolites.


Assuntos
Ciclopentanos/metabolismo , Dendrobium/fisiologia , Flavonoides/metabolismo , Oxilipinas/metabolismo , Vias Biossintéticas , Dendrobium/genética , Dendrobium/crescimento & desenvolvimento , Dendrobium/metabolismo , Metaboloma , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , Estresse Salino , Transcriptoma
4.
Int J Mol Sci ; 22(10)2021 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-34069261

RESUMO

The APETALA2 (AP2) transcription factors (TFs) play crucial roles in regulating development in plants. However, a comprehensive analysis of the AP2 family members in a valuable Chinese herbal orchid, Dendrobium officinale, or in other orchids, is limited. In this study, the 14 DoAP2 TFs that were identified from the D. officinale genome and named DoAP2-1 to DoAP2-14 were divided into three clades: euAP2, euANT, and basalANT. The promoters of all DoAP2 genes contained cis-regulatory elements related to plant development and also responsive to plant hormones and stress. qRT-PCR analysis showed the abundant expression of DoAP2-2, DoAP2-5, DoAP2-7, DoAP2-8 and DoAP2-12 genes in protocorm-like bodies (PLBs), while DoAP2-3, DoAP2-4, DoAP2-6, DoAP2-9, DoAP2-10 and DoAP2-11 expression was strong in plantlets. In addition, the expression of some DoAP2 genes was down-regulated during flower development. These results suggest that DoAP2 genes may play roles in plant regeneration and flower development in D. officinale. Four DoAP2 genes (DoAP2-1 from euAP2, DoAP2-2 from euANT, and DoAP2-6 and DoAP2-11 from basal ANT) were selected for further analyses. The transcriptional activation of DoAP2-1, DoAP2-2, DoAP2-6 and DoAP2-11 proteins, which were localized in the nucleus of Arabidopsis thaliana mesophyll protoplasts, was further analyzed by a dual-luciferase reporter gene system in Nicotiana benthamiana leaves. Our data showed that pBD-DoAP2-1, pBD-DoAP2-2, pBD-DoAP2-6 and pBD-DoAP2-11 significantly repressed the expression of the LUC reporter compared with the negative control (pBD), suggesting that these DoAP2 proteins may act as transcriptional repressors in the nucleus of plant cells. Our findings on AP2 genes in D. officinale shed light on the function of AP2 genes in this orchid and other plant species.


Assuntos
Dendrobium/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Dendrobium/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Estudo de Associação Genômica Ampla , Família Multigênica , Filogenia , Folhas de Planta/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Mapas de Interação de Proteínas , Sequências Reguladoras de Ácido Nucleico , Estresse Fisiológico/genética , Tabaco/genética , Fatores de Transcrição/metabolismo
5.
Plant Physiol ; 186(4): 2021-2036, 2021 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-33930147

RESUMO

A major obstacle for orchid (Orchidaceae) breeding and production is a long juvenile phase before orchid reproductive development. The molecular basis for prolonged vegetative growth in orchids remains largely unclear despite many efforts to clarify the relevant mechanisms. In this study, we report functional characterization of Dendrobium Orchid TERMINAL FLOWER1 (DOTFL1), an ortholog of TFL1 in Arabidopsis (Arabidopsis thaliana), from the orchid Dendrobium Chao Praya Smile. DOTFL1 is highly expressed in pseudobulbs and the shoot apical meristem (SAM) before and during the floral transition, but is downregulated in inflorescence apices and open flowers. Ectopic expression of DOTFL1 rescues the early-flowering and terminal-flower phenotypes of tfl1-20 in Arabidopsis. Overexpression of DOTFL1 in Dendrobium orchids delays flowering and produces defective inflorescence meristems and flowers with vegetative traits, whereas knockdown of DOTFL1 accelerates flowering and perturbs the maintenance of the inflorescence meristem. Notably, DOTFL1 suppresses orchid flowering and associated pseudobulb formation during the floral transition. We further reveal that two orchid MADS-box transcription factors, Dendrobium Orchid SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (DOSOC1) and AGAMOUS-LIKE 24 (DOAGL24), could interact with each other and bind to the CArG-box motif at DOTFL1, implying a regulatory hierarchy similar to their counterparts in Arabidopsis. Taken together, our findings suggest that DOTFL1 promotes vegetative growth, modulates successive developmental events required for reproductive success in Dendrobium orchids, and may have evolved with a previously unknown role in controlling pseudobulb formation in the Orchidaceae family.


Assuntos
Dendrobium/genética , Flores/crescimento & desenvolvimento , Proteínas de Plantas/genética , Dendrobium/crescimento & desenvolvimento , Dendrobium/metabolismo , Flores/genética , Proteínas de Plantas/metabolismo
6.
Genes (Basel) ; 12(3)2021 03 17.
Artigo em Inglês | MEDLINE | ID: mdl-33802927

RESUMO

Dendrobium officinale is a rare and traditional medicinal plant with high pharmacological and nutritional value. The self-incompatibility mechanism of D. officinale reproductive isolation was formed in the long-term evolution process, but intraspecific hybridization of different germplasm resources leads to a large gap in the yield, quality, and medicinal value of D. officinale. To investigate the biological mechanism of self-incompatibility in D. officinale, cytological observation and the transcriptome analysis was carried out on the samples of self-pollination and cross-pollination in D. officinale. Results for self-pollination showed that the pollen tubes could grow in the style at 2 h, but most of pollen tubes stopped growing at 4 h, while a large number of cross-pollinated pollen tubes grew along the placental space to the base of ovary, indicating that the self-incompatibility of D. officinale may be gametophyte self-incompatibility. A total of 63.41 G basesum of D. officinale style samples from non-pollinated, self-pollination, and cross-pollination by RNA-seq were obtained, and a total of 1944, 1758, and 475 differentially expressed genes (DEGs) in the comparison of CK (non-pollinated) vs. HF (cross-pollination sample), CK vs. SF (self-pollination sample) and SF vs. HF were identified, respectively. Forty-one candidate genes related to self-incompatibility were found by function annotation of DEGs, including 6 Ca2+ signal genes, 4 armed repeat containing (ARC) related genes, 11 S-locus receptor kinase (SRK) related genes, 2 Exo70 family genes, 9 ubiquitin related genes, 1 fatty acid related gene, 6 amino acid-related genes, 1 pollen-specific leucine-rich repeat extensin-like protein (LRX) related gene and 1 lectin receptor-like kinases (RLKs) related gene, showed that self-incompatibility mechanism of D. officinale involves the interaction of multiple genes and pathways. The results can provide a basis for the study of the self-incompatibility mechanism of D. officinale, and provide ideas for the preservation and utilization of high-quality resources of D. officinale.


Assuntos
Dendrobium/fisiologia , Perfilação da Expressão Gênica/métodos , Proteínas de Plantas/genética , Dendrobium/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Anotação de Sequência Molecular , Pólen/genética , Pólen/crescimento & desenvolvimento , Polinização , Análise de Sequência de RNA
8.
Molecules ; 25(21)2020 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-33143136

RESUMO

Dendrobium officinale Kimura et Migo, a rare and traditional medicinal plant, contains many nutrients such as polysaccharides, alkaloids, amino acids and so on. Different growth environment and intraspecific hybridization of different germplasm resources lead to large differences in the yield, quality and medicinal value of D. officinale. Here, the volatile compounds of D. officinale from four producing regions (Zhejiang, Fujian, Yunnan and Jiangxi) were analyzed to provide a certain reference value for the selection of a specific medicinal component in D. officinale breeding. Fresh stems of D. officinale germplasm resources were collected, and the chemical constituents were determined by gas chromatography-mass spectrometry. A total of 101 volatile compounds were identified, of which esters and alcohols accounted for 23 and 22. Hexacosane is the highest relative content of all volatile components. The highest content of hexacosane was observed in YA1 from Yunnan was 34.41%, and the lowest (23.41%) in JA1 from Jiangxi. Moreover, 5-10 unique substances were determined in different regions. A total of 17 medicinal components were detected, and three unique medicinal components were detected only in YA1, revealing that YA1 can provide raw materials for the application of specific medicinal substances extraction. A total of four toxic components were detected, but no toxic components were detected in JA1 from Jiangxi, suggested that the germplasm resources from Jiangxi could be exploited efficiently for breeding superior D. officinale specimens. The results provide a theoretical basis for the collection, protection and utilization of D. officinale germplasm resources in different regions.


Assuntos
Dendrobium , Plantas Medicinais , Compostos Orgânicos Voláteis , China , Dendrobium/química , Dendrobium/crescimento & desenvolvimento , Plantas Medicinais/química , Plantas Medicinais/crescimento & desenvolvimento , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/metabolismo
9.
Int J Mol Sci ; 21(17)2020 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-32872385

RESUMO

The acetylation or deacetylation of polysaccharides can influence their physical properties and biological activities. One main constituent of the edible medicinal orchid, Dendrobium officinale, is water-soluble polysaccharides (WSPs) with substituted O-acetyl groups. Both O-acetyl groups and WSPs show a similar trend in different organs, but the genes coding for enzymes that transfer acetyl groups to WSPs have not been identified. In this study, we report that REDUCED WALL ACETYLATION (RWA) proteins may act as acetyltransferases. Three DoRWA genes were identified, cloned, and sequenced. They were sensitive to abscisic acid (ABA), but there were no differences in germination rate and root length between wild type and 35S::DoRWA3 transgenic lines under ABA stress. Three DoRWA proteins were localized in the endoplasmic reticulum. DoRWA3 had relatively stronger transcript levels in organs where acetyl groups accumulated than DoRWA1 and DoRWA2, was co-expressed with polysaccharides synthetic genes, so it was considered as a candidate acetyltransferase gene. The level of acetylation of polysaccharides increased significantly in the seeds, leaves and stems of three 35S::DoRWA3 transgenic lines compared to wild type plants. These results indicate that DoRWA3 can transfer acetyl groups to polysaccharides and is a candidate protein to improve the biological activity of other edible and medicinal plants.


Assuntos
Dendrobium/crescimento & desenvolvimento , Proteínas de Plantas/genética , Polissacarídeos/metabolismo , Ácido Abscísico/farmacologia , Acetilação , Clonagem Molecular , Dendrobium/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Germinação/efeitos dos fármacos , Filogenia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/fisiologia , Análise de Sequência de DNA
10.
Plant Mol Biol ; 104(4-5): 529-548, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32876816

RESUMO

KEY MESSAGE: This research provides comprehensive insight into the molecular networks and molecular mechanisms underlying D. officinale flower development. Flowers are complex reproductive organs and play a crucial role in plant propagation, while also providing sustenance for insects and natural bioactive metabolites for humans. However, knowledge about gene regulation and floral metabolomes in flowers is limited. In this study, we used an important orchid species (Dendrobium officinale), whose flowers can be used to make herbal tea, to perform transcriptome sequencing and metabolic profiling of early- and medium-stage flower buds, as well as opened flowers, to provide comprehensive insight into the molecular mechanisms underlying flower development. A total of 8019 differentially expressed genes (DEGs) and 239 differentiated metabolites were found. The transcription factors that were identified and analyzed belong exclusively to the MIKC-type MADS-box proteins and auxin responsive factors that are known to be involved in flower development. The expression of genes involved in chlorophyll and carotenoid biosynthesis strongly matched the metabolite accumulation patterns. The genes related to flavonoid and polysaccharide biosynthesis were active during flower development. Interestingly, indole-3-acetic acid and abscisic acid, whose trend of accumulation was inverse during flower development, may play an important role in this process. Collectively, the identification of DEGs and differentiated metabolites could help to illustrate the regulatory networks and molecular mechanisms important for flower development in this orchid.


Assuntos
Dendrobium/crescimento & desenvolvimento , Dendrobium/genética , Dendrobium/metabolismo , Flores/crescimento & desenvolvimento , Ácido Abscísico/metabolismo , Carotenoides/metabolismo , Clorofila/metabolismo , Flavonoides/metabolismo , Flores/genética , Flores/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Proteínas de Domínio MADS/genética , Filogenia , Proteínas de Plantas/genética , Polissacarídeos/metabolismo
11.
Plant Physiol ; 184(3): 1469-1481, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32900977

RESUMO

Orchids (members of the Orchidaceae family) possess unique flower morphology and adaptive reproduction strategies. Although the mechanisms underlying their perianth development have been intensively studied, the molecular basis of reproductive organ development in orchids remains largely unknown. Here, we report the identification and functional characterization of two AGAMOUS (AG)-like MADS-box genes, Dendrobium 'Orchid' AG1 (DOAG1) and DOAG2, which are putative C- and D-class genes, respectively, from the orchid Dendrobium 'Chao Praya Smile'. Both DOAG1 and DOAG2 are highly expressed in the reproductive organ, known as the column, compared to perianth organs, while DOAG2 expression gradually increases in pace with pollination-induced ovule development and is localized in ovule primordia. Ectopic expression of DOAG1, but not DOAG2, rescues floral defects in the Arabidopsis (Arabidopsis thaliana) ag-4 mutant, including reiteration of stamenoid perianth organs in inner whorls and complete loss of carpels. Downregulation of DOAG1 and DOAG2 in orchids by artificial microRNA interference using l-Met sulfoximine selection-based gene transformation systems shows that both genes are essential for specifying reproductive organ identity, yet they, exert different roles in mediating floral meristem determinacy and ovule development, respectively, in Dendrobium spp. orchids. Notably, knockdown of DOAG1 and DOAG2 also affects perianth organ development in orchids. Our findings suggest that DOAG1 and DOAG2 not only act as evolutionarily conserved C- and D-class genes, respectively, in determining reproductive organ identity, but also play hitherto unknown roles in mediating perianth organ development in orchids.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Dendrobium/crescimento & desenvolvimento , Dendrobium/genética , Flores/crescimento & desenvolvimento , Flores/genética , Proteínas de Domínio MADS/metabolismo , Desenvolvimento Vegetal/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Proteínas de Domínio MADS/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento
12.
Transgenic Res ; 29(4): 429-442, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32691287

RESUMO

Four Dendrobium Sonia 'Earsakul' lines were generated by insertion of one, two or three antisense copies of a Carica papaya gene encoding 1-aminocyclopropane-1-carboxylic acid oxidase (CpACO). Whole vegetative plants of the transgenic lines showed about 50% of the basal ethylene production rate, while the increase in ethylene production in floral buds during opening and open flowers prior to visible senescence was delayed. Detailed analysis of more than 100 parameters in flowering plants showed no effect of antisense ACO on plant morphology and coloration, except for shorter length and width of some of the sepals and petals. In intact plants the water-soaking of floral buds as well as bud abscission were delayed by ACO antisense, as was the time to senescence of open flowers. Pollen viability and pollen tube growth were not affected in the transgenic lines. In cut inflorescences placed in water, bud yellowing, bud water soaking, and bud abscission were considerably delayed by the antisense construct, while the life span of open flowers were increased and abscission of open flowers were delayed. It is concluded that the reduction of ACO activity affected the shape of some petals/sepals and delayed the abortion in floral buds, and the senescence and abscission of open flowers.


Assuntos
Aminoácido Oxirredutases/antagonistas & inibidores , Elementos Antissenso (Genética) , Senescência Celular , Dendrobium/enzimologia , Flores/anatomia & histologia , Flores/fisiologia , Regulação Enzimológica da Expressão Gênica , Aminoácido Oxirredutases/genética , DNA de Plantas/genética , Dendrobium/genética , Dendrobium/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento
13.
Bioengineered ; 11(1): 386-396, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32172675

RESUMO

Dendrobium cariniferum is a valuable ornamental and medicinal plant rich with polysaccharides, alkaloid, and other bioactive compounds, which are potential raw materials for pharmacological utilization. In this study, an efficient protocol for the rapid propagation of D. cariniferum was developed. By using the tissue culture protocol, the effects of pH, hormone combinations, temperatures, light intensity, culture time protocorm proliferation, seedlings rooting, and accumulation of biomass with bioactive compounds were investigated. The experiments showed that the medium [1/2 MS + activated carbon1.0 g/L+ agar strip 7.5 g/L + sucrose 25 g/L] effectively promoted the germination of D. cariniferum seeds. The optimal culture conditions were found at pH 5.7, temperature 23 ± 2°C, and light intensity of 1000 Lx in the protocorm proliferation stage. Adding 1.5 g/L peptone in the medium effectively promoted the seedling rooting. The optimal culture conditions for accumulation of bioactive compounds (polysaccharides and alkaloids) of seedlings were found at temperature of 25 ± 2°C, light intensity of 1500-2000 Lx after the 60-day (d). Our study constructed a rapid propagation system in vitro for D. cariniferum, as well as the methods for efficient accumulation of active substances in seedling culture, which will serve as guidance for industrial production of D. cariniferum seedlings for both medicinal raw materials and ornamental plants. In addition, our study provided a new idea that we can directly use the high bioactive compound seedlings to extract medicinal components in industry conditions without transferring to the field.


Assuntos
Dendrobium/metabolismo , Dendrobium/crescimento & desenvolvimento , Germinação/fisiologia , Plantas Medicinais/crescimento & desenvolvimento , Plantas Medicinais/metabolismo , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Sementes/crescimento & desenvolvimento , Sementes/metabolismo
14.
Sci Rep ; 10(1): 2857, 2020 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-32071345

RESUMO

The stem of Dendrobium huoshanense C.Z. Tang and S.J. Cheng was widely used as a medicinal herb in health care products due to its broad pharmacological activities. However, the molecular regulation mechanism of stem development and biosynthetic pathways of important bioactive substances are still unclear in D. huoshanense. In this study, the bioactive compounds in leaves, stems and roots, and the identification of candidate genes involved in stem formation and biosynthesis of active compounds via transcriptome sequence were analyzed. The accumulation of total polysaccharides and flavonoids were varied significantly in different tissues. A comparative transcriptomic analysis revealed several differentially expressed genes (DEGs) involved in polysaccharides biosynthesis (103 genes), including fructose and mannose related genes (29 genes) and glycosyltransferase genes (74 genes), and flavonoids biosynthesis (15 genes). Some candidate genes that participated in photoperiod regulation (27 genes), starch and sucrose metabolism (46 genes), and hormone-induced activation of signaling pathways (38 genes) may be involved in stem formation. In sum, this study provides a foundation for investigating the molecular processes in the biosynthesis of active compounds and stem development. The transcriptome data presented here provides an important resource for the future studies of the molecular genetics and functional genomics in D. huoshanense and optimized control of the active compounds produced by D. huoshanense.


Assuntos
Dendrobium/genética , Flavonoides/genética , Caules de Planta/genética , Transcriptoma/genética , Vias Biossintéticas/genética , Dendrobium/crescimento & desenvolvimento , Flavonoides/biossíntese , Perfilação da Expressão Gênica , Folhas de Planta , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Caules de Planta/crescimento & desenvolvimento , Plantas Medicinais , Polissacarídeos/genética
15.
Bioengineered ; 10(1): 668-678, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31722607

RESUMO

Auxin/Indole-3-Acetic Acid (Aux/IAA) genes are involved in auxin signaling pathway and play an important role in plant growth and development. However, many studies focus on Aux/IAA gene families and much less known in Bletilla striata. In this study, a total of 27 Aux/IAA genes (BsIAA1-27) were cloned from the transcriptome of Bletilla striata. Based on a phylogenetic analysis of the Aux/IAA protein sequences from B. striata, Arabidopsis thaliana and Dendrobium officinale, the Aux/IAA genes of B. striata (BsIAAs) were categorized into 2 subfamilies and 9 groups. While BsIAAs were more closer to those of D. officinale compared to A. thaliana. EST-SSR marker mining test showed that 4 markers could be stably amplified with obvious polymorphisms among 4 landraces. Our results suggested that BsIAAs were involved in the process of tuber development and provided insights into functional roles of Aux/IAA genes in B. striata and other plants.


Assuntos
Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Orchidaceae/genética , Reguladores de Crescimento de Plantas/genética , Proteínas de Plantas/genética , Transcriptoma , Sequência de Aminoácidos , Arabidopsis/classificação , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Evolução Biológica , Biologia Computacional/métodos , Dendrobium/classificação , Dendrobium/genética , Dendrobium/crescimento & desenvolvimento , Dendrobium/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Marcadores Genéticos , Orchidaceae/classificação , Orchidaceae/crescimento & desenvolvimento , Orchidaceae/metabolismo , Filogenia , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Tubérculos/genética , Tubérculos/crescimento & desenvolvimento , Tubérculos/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos
16.
PLoS One ; 14(9): e0222666, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31539401

RESUMO

Dendrobium, an important medicinal plant, is a source of widely used herbal medicine to nourish the stomach and treat throat inflammation. The present study is aimed at distinguishing and evaluating three major Dendrobium species by comparing physiochemical characteristics and understanding differences between different growth years in the Ta-pieh Mountains. Polysaccharides and total alkaloids of Dendrobium were determined, and the amino acids and trace elements were determined by UPLC (Ultra High-Performance Liquid Chromatography) and ICP-MS (Inductively coupled plasma mass spectrometry). It can be seen from the results that the polysaccharide content of these three kinds of Dendrobium in different growth years ranges from 249.31 mg·g-1 to 547.66 mg·g-1, and the highest content is in the 3-year-old Dendrobium huoshanense. The total alkaloid content ranges from 0.21 mg·g-1 to 0.54 mg·g-1, and the highest content is also the 3-year-old Dendrobium huoshanense. We determined the amino acid content of these three Dendrobium in different growth years, and we can see that each of the three kinds of Dendrobium contain seven kinds of amino acids required by the human body. We conducted a safety evaluation of the essential trace elements of Dendrobium, and the results showed that the dosage of 12g·d-1 Dendrobium prescribed in China Pharmacopoeia is in accordance with the recommended daily intake of trace elements recommended by the Food and Drug Administration of the United States, and will not cause trace element poisoning. Linear discriminant analysis was carried out on the basis of amino acids and trace elements and confirmed the applicability of multi-elemental analysis for identifying different Dendrobium species.


Assuntos
Dendrobium/crescimento & desenvolvimento , Alcaloides/análise , Aminoácidos/análise , Cromatografia Líquida de Alta Pressão , Dendrobium/química , Dendrobium/fisiologia , Plantas Medicinais/química , Plantas Medicinais/crescimento & desenvolvimento , Plantas Medicinais/fisiologia , Polissacarídeos/análise , Oligoelementos/análise
17.
Zhongguo Zhong Yao Za Zhi ; 44(3): 614-618, 2019 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-30989931

RESUMO

The Dendrobium species are rare and endangered medicinal plants, and it is difficult to investigate their wild resources with conventional methods because of typical epiphytic herbaceous. We explored Dendrobium resources(include culture resource) of Qinba Mountains and the boundary Mountain area in Hubei, Chongqing using the methods of literatures and field investigation, and found that the cultural base of Dendrobium were profound in Qinba Mountains region. Furthermore, its germplasm resources of Dendrobium were established for the first time in Wanzhou Luotian town. In case the advantages of local rock resources and poverty alleviation demand, we have actively carried out the cultivating mode of Dendrobium which grow on rock, and the poverty alleviation model of local characteristic Dendrobium industry were established preliminarily. Our application case can provide reference for the mining and transformation of traditional Chinese medicine resources census results.


Assuntos
Agricultura/economia , Dendrobium/crescimento & desenvolvimento , Plantas Medicinais/crescimento & desenvolvimento , Pobreza , China , Medicina Tradicional Chinesa
18.
Plant Sci ; 277: 43-54, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30466600

RESUMO

Dendrobium officinale is a precious traditional Chinese medicinal herb because it is abundant in mannose-containing polysaccharides (MCPs). GDP-mannose transporter (GMT), which translocates GDP-mannose into the Golgi lumen, is indispensable for the biosynthesis of MCPs. In this study, we found that the dominant polysaccharides in D. officinale were MCPs in a range of varieties and different physiological phases. After a positive correlation between the accumulation of mannose and the transcript levels of candidate GMT genes was found, three GMT genes (DoGMT1-3) were identified in D. officinale. DoGMT1, DoGMT2 and DoGMT3 exhibited the highest transcript level in stem that an organ for MCPs storage. All three DoGMT proteins were targeted to Golgi apparatus, and had a GDP binding domain (GXL/VNK) that was homologous to a specially characterized GMT protein GONST1 in Arabidopsis thaliana. Moreover, DoGMT1, DoGMT2 and DoGMT3 complemented a GDP-mannose transport-defective yeast mutant (vrg4-2), meanwhile they also demonstrated a higher GDP-mannose uptake activity. Therefore, we conclude that DoGMT1, DoGMT2 and DoGMT3 are able to transport GDP-mannose while the expression patterns of these genes correspond to the accumulation of MCPs in D. officinale. These findings support the importance of GMT genes from D. officinale in the biosynthesis of MCPs.


Assuntos
Proteínas de Transporte/metabolismo , Dendrobium/crescimento & desenvolvimento , Dendrobium/metabolismo , Mananas/metabolismo , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Transporte/genética , Dendrobium/genética , Complexo de Golgi/metabolismo , Manose/metabolismo , Proteínas de Plantas/genética , Polissacarídeos/metabolismo
19.
Molecules ; 23(10)2018 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-30262777

RESUMO

Dendrobium officinale is a widely used medicinal plant in China with numerous bio-activities. However, the main structure and anti-tumor activity of the polysaccharides from this plant have not been investigated. In this study, we elucidated the main structure of polysaccharides purified with DEAE and Sephadex G-25 from Dendrobium officinale grown under different planting conditions. In addition, the anti-tumor activity was tested via MTT assays. The results showed that the polysaccharides of Dendrobium officinale grown under different conditions were almost the same, with slight differences in the branched chain; both polysaccharide fractions consisted of (1→4)-linked mannose and (1→4)-linked glucose, with an O-acetyl group in the mannose. After degradation, the polysaccharide fractions from wild plants showed significant anti-proliferation activity in HeLa cells. The fractions F1 and F3 induced apoptosis by up-regulating the expression of ERK, JNK, and p38. We concluded that polysaccharides from Dendrobium officinale planted in the wild exhibit significant anti-tumor effects only after being degraded to smaller molecular weight species. The planting mode is a significant factor in the pharmacological activity of Dendrobium officinale. We advise that the planting conditions for Dendrobium officinale should be changed.


Assuntos
Antineoplásicos Fitogênicos , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Dendrobium/química , Polissacarídeos , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Dendrobium/crescimento & desenvolvimento , Células HeLa , Humanos , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia
20.
Sci Rep ; 8(1): 6373, 2018 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-29686299

RESUMO

In this study, we applied transcriptome and UHPLC-MS technologies to investigate the flavonoids and their biosynthesis- and accumulation-related genes in Dendrobium catenatum from three different locations. Eight flavonoid glycosides were identified using standard references or previously isolated substances with MS data analysis. The total flavonoid contents were determined by reagents, and all the data were analyzed. In total, 23139 unigenes were obtained using the Dendrobium catenatum genome data. Of these, 10398 were annotated in the Gene Ontology (GO) database, 4203 were annotated in the KEGG database, and 10917 were annotated in the EuKaryotic Orthologous Groups (KOG) database. Thirty-one of the unigenes annotated by the KEGG database were involved in flavonoid pathways. The genes involved in bio-modification, accumulation, transportation and the regulation of the flavonoid bio-synthesis process were investigated. In conclusion, the flavonoids in Dendrobium catenatum from three different locations were different in quantitative and qualitative which may contribute to the establishment of quality control method for this herbal plant. These differences were determined by flavonoids biosynthesis process and they were concluded by sorting out the expression level of certain biosynthesis related genes.


Assuntos
Dendrobium/genética , Dendrobium/metabolismo , Flavonoides/biossíntese , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Transcriptoma , Dendrobium/crescimento & desenvolvimento , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Anotação de Sequência Molecular
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