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italic>Dendrobium officinale Kimura et Migo is a rare Chinese herbal medicine, while Dendrobium crepidatum Lindl is a local medicine in Yunnan, both of which have the function of nourishing yin and stomach. To reveal the differences in chemical composition between the two species, ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was used to analyze the chemical composition of stems and leaves of D. officinale and D. crepidatum. Principal component analysis (PCA) and partial least squares discriminant analysis (OPLS-DA) were used to determine the differences in metabolites between species and parts of Dendrobium. Fifty-eight chemical compounds were identified in the two species. Analysis indicated that the side ring of alkaloids connected with nitrogen was readily cleaved during analysis. The results of PCA analysis showed that the stems and leaves of D. officinale and D. crepidatum could be easily differentiated, and the chemical constituents of D. officinale and D. crepidatum were significantly different. OPLS-DA analysis showed that there were 16 metabolite differences between the stems and 22 differences in metabolites between the leaves of D. officinale and D. crepidatum. The main metabolite differences in components between the two Dendrobium species were dendrocrepidine B, dendrocrepidine C and dendrocrepine. There were 14 differences in metabolites between the stems and leaves of D. crepidatum. In conclusion, the chemical compositions of D. officinale and D. crepidatum are quite different; the small molecular compounds of D. officinale are mainly terpenoids and flavonoids, and the content of alkaloids is low. There is no significant difference between stem and leaf. In contrast, D. crepidatum is mainly composed of alkaloids and terpenoids, with crepidamine and dendrocrepine as its unique components, and there are great differences in the components between stems and leaves. This study provides a theoretical basis for the development and utilization of Dendrobium resources.
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italic>Dendrobium officinale Kimura et Migo (D. officinale) has been used as a valuable traditional Chinese medicine for more than 2 000 years in China. Modern research has confirmed a wide range of pharmacological activities, such as regulating blood sugar, improving gastrointestinal inflammation, and regulating immunity. Polysaccharides are the main active ingredients of D. officinale. With the intensive studies of the pharmacological activities of D. officinale, evidence for the pharmacological effects and potential mechanisms of D. officinale polysaccharides has increased dramatically. In this review, we summarized the latest progress in the pharmacological and mechanical studies of D. officinale polysaccharides, and based on the pharmacological efficacy and oral absorption and utilization characteristics of D. officinale polysaccharides, it is proposed that regulating the gut microbiota may be one of the key mechanisms for D. officinale to exert its beneficial effects. Research on the mechanism of D. officinale polysaccharides puts forward new research directions and prospects.
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Objective: To clone an F-box protein namely S phase kinase-associated protein gene DoSKP2A with full length cDNA, in a rare endangered medicinal orchid species Dendrobium officinale, followed by bioinformatics analysis and expression pattern analysis. Methods: RACE technology were used for gene identification. Characteristics of physiochemical properties, conserved domains and subcellular localization of the protein were determined using a series of bioinformatics tools. The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR 7.0 and MEGA 7.0, respectively. Real time quantitative PCR was used for gene expression analysis. Results: DoSKP2A gene was cloned (GenBank accession KU160472). The full length cDNA of DoSKP2A was 1 507 bp in length, and ORF was 1 101 bp, encoding a 366-aa protein with a molecular weight of 39 590 and an isoelectric point of 7.9. The deduced DoSKP2A protein, without transmembrane or signal peptide residues, contained an F-box core domain (26-88), a leucine-rich repeat (202-226), and multiple conserved motifs. DoSKP2A had high identities (64.6%-72.4%) with SKP2As proteins from various plants. DoSKP2A belonged to the monocotyledons subgroup of the SKP2As evolutionary tree. DoSKP2A gene was differentially expressed in the three included organs. The transcripts were more abundant in the roots, with 6.16 fold, then the stems and the lowest in the leaves. Conclusion: The novel full-length F-box protein gene DoSKP2A was obtained, along with bioinformatics and expression characteristics, which provided molecular basis for the growth and development, signal transduction, and stress resistance of D. officinale.
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Dendrobium officinale is one of the most precious medicinal plants in the family of Orchidaceae, and rich in polysaccharides, astragalus, bibenzyls and alkaloids. It has effects such as antioxidant, anti-cancer, immuno-enhancing, lowering blood sugar, and alleviating liver injury. Its huge medicinal, scientific and commercial value has raised a research upsurge, especially in the past five years, nucleic acid molecular biology of Dendrobium officinale has made more and more exciting results. In order to provide scientific guidance for further research on functional genes, this paper reviews the recent progress in research on genomics, transcriptomics and functional genes of D. officinale.
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Based on the previous studies in recent years, this advances made a summary of immunoregulatory effects from Dendrobium officinale. And the research advances on related bioactive compounds of polysaccharides in extraction, separation and purification, structural composition and content determination were systemically reviewed. This paper intended to provide a theoretical basis for reasonable research and exploitation of related health foods of D. Officinale.
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Objective: To identifiy a receptor-like kinase (RLK) encoding gene named as DoRLK (GenBank accession No. ANC68272.1) from Dendrobium officinale. Methods: RT-PCR and RACE technologies were used to isolate the full length cDNA of DoRLK. Characteristics of physiochemical properties, conserved domains, and subcellular localization of the deduced DoRLK protein were determined by a series of bioinformatics tools. The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR 6.0 and MEGA 7.0 softwares, respectively. Quantitative PCR was used for gene expression analysis. Results: The results showed that the full length cDNA of DoRLK was 1 715 bp in length and encoded a 423 aa protein with a molecular weight of 47 800.88 and an isoelectric point (pI) of 9.47. The deduced DoRLK protein, like other RLK proteins, constituted one conserved domains (85—370) and one transmembrane motifs (250—270). Multiple sequence alignment and phylogenetic analyses demonstrated that DoRLK had high identity (43.62%—63.35%) to a number of RLK genes from various plants and was closely related to Phalaenopsis equestris, Phoenix dactylifera and Asparagus officinalis. Real time quantitative PCR (qPCR) analysis revealed that DoRLK was expressed in the three included organs. The transcripts were the most abundant in the roots with 2.22 fold over that in the leaves,followed by that in the stems with 2.75 fold. Conclusion: Molecular characterization of DoRLK will be useful for further functional elucidation of the gene involving in D. officinale responses to environmental factors.
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Objective: To analyze the effect of endophytic fungi GXRz2, GXRz3 and GXRz10 on content of poly saccharide and alkaloid, and the expression of key enzyme genes UGPase, HMGR, and FPS in Dendrobium officinale. Methods: The endophytic fungi liquid were added to D. officinale seedlings. Polysaccharide and alkaloid content were measured by spectrophotometry method. With 18S rRNA as internal control gene, the expression of key enzyme genes was detected by real-time quantitative PCR method. Results: It was found that the content of polysaccharide in D. officinale was higher, mainly concentrated in stem, but the lowest in root. And the content of alkaloid in D. officinale was lower, mainly accumulated in leaf, but the lowest in root. In addition, the three endophytic fungi strains could promote accumulation of polysaccharide and alkaloid in D. officinale to a certain extent. The expression of UGPase, HMGR and FPS genes in D. officinale induced by different strains was detected by real-time quantitative PCR. The results showed that endophytic fungi GXRz3 and GXRz10 could significantly increase the expression of UGPase, HMGR and FPS genes in D. officinale. In the polysaccharide synthesis pathway, the UGPase gene had the highest relative expression in the stem, followed by the leaf, and the least in the root. In the alkaloid synthesis pathway, HMGR gene had the highest relative expression in the stem, followed by the leaf, and the least in the root. However, FPS gene had the highest relative expression in the leaf, followed by the stem, and the least in the root. Conclusion: Endophytic fungi may affect the synthesis of polysaccharide by regulating the expression of UGPase. Considering the accumulation of polysaccharide, it is speculated that UGPase may be a key enzyme in the polysaccharide synthesis pathway of D. officinale. Endophytic fungi may affect the synthesis of alkaloid by regulating the expression of HMGR and FPS. Considering the accumulation of alkaloid, it is speculated that FPS may be a key enzyme in the alkaloid synthesis pathway of D. officinale.
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Objective To investigate the metabolites related to the course of anthracnosis of Dendrobium officinale and its possible mechanism based on the technique of metabonomics, so as to provide theoretical support for the creation or breeding of disease resistant varieties of D. officinale. Methods The normal and infected by Colletotrichum gloeosporioides leaves of D. officinale were collected respectively, and to find out the differential metabolites through the sample pretreatment, GC-MS analysis, and bioinformatics analysis. Then the differential metabolites and analysis of metabolic pathways involved in the course of disease were carried on the preliminary discussion. Results The multidimensional statistical models of each analysis group were successfully established. The dispersion points of VIP > 1.0 were selected as the potential differential materials, combined with P < 0.05 in the analysis of single dimensional Statistics (test) as the standard to verify. A total of 84 differential metabolites screened from 305 identified metabolites were considered to be pathogenesis related metabolites. A total of 34 differential metabolites were found out to be involved in the metabolic pathways through the pathway enrichment analysis, and the obtained ZC-GB metabolic pathways were of significance. Conclusion Based on GC-MS technology, the metabolomics analysis of D. officinale samples (normal/infected) was carried out and the metabolites related to the course of anthracnosis of D. officinale were found out. It could lay the foundation for studying the disease of D. officinale and cultivating resistant varieties on molecular level.
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Objective: To isolate and characterize a protein phosphatase (PP) encoding gene DoPP2C1 in a rare endangered medicinal orchid species Dendrobium officinale, followed by bioinformatics analysis and expression pattern detection. Methods: qRT-PCR and RACE technologies were used to isolate the full length cDNA of DoPP2C1. Characteristics of physiochemical properties, conserved domains, and subcellular localization of DoPP2C1 protein were determined using a series of bioinformatics tools. The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR 7.0 and MEGA 6.0 softwares, respectively. Quantitative PCR was used for gene expression analysis. Results: The full length cDNA of DoPP2C1 (GenBank accession KJ995533) was 1 221 bp in length, and encoded a 285-aa protein with a molecular weight of 31 080 and an isoelectric point of 6.18; The deduced DoPP2C1 protein had one PP2C domain (27-285), which are all conserved among the PP2C proteins. DoPP2C1 protein did not contain a signal peptide or a transmembrane region, and was predicted to locate in cytoplasm; DoPP2C1 had high identities (56.3%-73.7%) with various PP2C proteins in plants; DoPP2C1 was closely related to Oryza sativa OsPP2C62, Sorghum bicolor XP_002462907, Hordeum vulgare BAK00362 and Triticum urartu EMS47641 proteins, and belonged to the Group F1 of the PP2C evolutionary tree; DoPP2C1 gene was differentially expressed in the three included organs. The transcripts were more abundant in the roots and stems, with 7.57 and 1.79 fold, respectively, over that in the leaves. Conclusion: Molecular characterization of DoPP2C1 gene was obtained, which will be useful for further functional determination of the gene involving in the growth and development, physiological stress adaptations, and secondary metabolic regulations of D. officinale.
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Objective: To clone novel member of alkaline/neutral invertase (NI) gene in a rare and endangered medicinal plant of Dendrobium officinale, conduct bioinformatic analysis and detect the quantitative expression in different organs. Methods: Primers were designed according to NI gene segment which was selected from leaf transcriptome sequencing results of D. officinale. The full-length cDNA of NI gene was cloned via homology-based cloning and rapid amplification of cDNA ends (RACE) approach. The physical and chemical properties, secondary structure and tertiary structure of NI protein were forecasted and analyzed using related software. The expression levels of NI gene in roots, stems, and leaves of D. officinale were detected using real-time PCR. Results: A novel gene encoding a NI protein was cloned from D. officinale. This gene (named as DoNI2, GenBank accession number: KY794404) had a total length of 2 397 bp with an open reading frame of 1 836 bp, and encoded a predicted polypeptide of 611 amino acids with a molecular weight of 69 050. Bioinformatics predicted that the isoelectric point of DoNI2 gene encoding protein was 6.38, the instability coefficient was 44.95, and the hydrophobic coefficient was −0.232. RT-PCR showed that DoNI2 gene expressed in all organs with highest expression level in stems and the lowest in roots. DoNI2 gene expression was significantly positively correlation with NI enzymatic activities at different growth years of D. officinale. Conclusion: The full length cDNA sequence in a mitochondrial DoNI2 gene was identified, facilitating future functional analysis of the gene involving in the regulation of sugar metabolism in D. officinale.
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Objective To clone the SnRK2 gene in Dendrobium officinale and investigate its characteristics and expression pattern. Methods RT-PCR and RACE techniques were used to clone the full-length cDNA of DoSRK2E, with the aids of a series of online bioinformatic software, characteristics including molecular mass, isoelectric point, conserved domain, transmembrane structure, signal peptide, and subcellular localization of the deduced protein were analyzed. Besides, the sequence of the deduced protein was aligned with those of other plant SnRK2 by DNASTAR, and phylogenetic relationships were reconstructed utilizing MEGA. Finally, tissue specific expression pattern of DoSRK2E was tested by quantitative real-time PCR (qRT-PCR). Results The full-length cDNA of DoSRK2E (GenBank accession API65110) is 1 795 bp with a 1 086 bp complete open reading frame (ORF). The predicted molecular mass and isoelectric point of the deduced protein DoSRK2E were 40 850 and 4.80, respectively. No signal peptide nor transmembrane region were detected, this protein contains one protein kinase domain, one ATP binding site, and one Ser/Thr active site, which was predicted most likely to be located in the endoplasmic reticulum membrane. DoSRK2E protein showed high similarity with those from other plant SnRK2, and its phylogenetic location was in Group III of SnRK2 subfamily, and phylogenetically closest to AtSnRK2.6 from Arabidopsis. In addition, qPCR analysis revealed that DoSRK2E showed the highest expression level in root, followed by stem and leaf. Conclusion A SnRK2 family gene DoSRK2E was cloned from the rare and endangered medicinal plant D. officinale for the first time. The Characteristics and expression pattern of this gene were analyzed. This study will provide a basis for further exploration of the regulation mechanisms of DoSRK2E in D. officinale under stress conditions.
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To explore the effects of crude toxins from Colletotrichum gloeosporioides(C. gloeosporioides)on the growth of Dendrobium officinale Kimura et Migo(D. officinale),and to provide early basis for thescreening and cultivation of the resistant variants of C. gloeosporioides. Methods Seedlings of D. officinal werecultivated in MS medium added with different concentrations of the crude toxins from C. gloeosporioides. Theeffects of the crude toxin on the growth of seedlings were observed, and the optimum resistance -selectionthreshold was preliminarily screened. Results In the concentration range of 5％ - 15％(volume fraction),crudetoxins from C. gloeosporioides increased the plant height,stem diameter,number of new bud,root number,and fresh mass of D. officinale,among which the effect of 5％(volume fraction)of crude toxins was the moststrongest. In the concentration range of 35％ - 40％(volume fraction),crude toxins suppressed the plant height,leaf number,number of new bud,root number,and the fresh mass of D. officinale. When cultured with 20％(volume fraction)crude toxins, D. officinale was blooming, and the flowers appeared variation phenomenon.Conclusion The crude toxins from C. gloeosporioides have biological activity and certain toxicity,which can beconsidered as a selection agent instead of pathogenin in vitro to screen the resistant variant of D. officinale,butthe optimum resistance-selection threshold still needs further research.
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Objective: To study the distribution of endophytic fungi from Dendrobium officinale and screen the distinctive fungi with antimicrobial and antitumor activities. Methods: Endophytic fungi were isolated by the conventional method and cylinder-plate method with Staphylococcus aureus, Escherichia coli, and Bacillus subtilis as subjects. The metabolites of ethyl acetate extracts with antitumor activity were identified based on MTT assay. Results: A total of 28 strains of the endophytic fungi were isolated from D. officinale. By analyzing the data of molecular identity, there were 10 species in D. officinale, in which Fusarium Link is a dominant. The ethyl acetate extracts of the fermentation broth had different antibiotic activities on the indicator microorganisms, and had antitumor effect on HepG2 cells. Conclusion: Endophytic fungi isolated from D. officinale have rich diversity and have antimicrobial and antitumor activities, which could be potential for developing antimicrobial and antitumor reagents.
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Objective To isolate and characterize a WRKY transcription factor encoding gene DoWRKY3 in a rare endangered medicinal orchid species Dendrobium officinale, followed by bioinformatics analysis and expression pattern detection. Methods RT-PCR and RACE technologies were used to isolate the full length cDNA of DoWRKY3. Characteristics of physiochemical properties, conserved domains, and subcellular localization of the deduced DoWRKY3 protein were determined by a series of bioinformatics tools. The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR 6.0 and MEGA 6.0 softwares, respectively. Quantitative PCR was used for gene expression analysis. Results The full length cDNA of DoWRKY3 (GenBank accession KT957549) was 2 065 bp in length, and encoded a 509-aa protein with a molecular weight of 55 580 and an isoelectric point of 6.58; The deduced DoWRKY3 protein sequence had two WRKYGQK motifs, two WRKY domains (217-279, 381-449), and two C2H2-type zinc-finger signatures (C-X4-C-X22-23-H-X1-H), which are all conserved among the WRKY proteins.DoWRKY3 protein did not contain a signal peptide or a transmembrane region, and was predicted to locate in nucleus; DoWRKY3 had high identities (46.3%-57.4%) with various WRKY proteins from several plants; DoWRKY3 was closely related to Arabidopsis AtWRKY3, AtWRKY4, and Salvia miltiorrhiza SmWRKY54 proteins, and belonged to the Group 1 of the WRKY evolutionary tree; DoWRKY3 gene was differentially expressed in the three included organs. The transcripts were more abundant in the roots and leaves, with 2.32 and 1.69 fold, respectively, over that in the stems. Conclusion Molecular cloning and characterization of the full length DoWRKY3 gene will be useful for further functional determination of the gene involving in the growth and development, physiological stress adaptations, and secondary metabolic regulations of D. officinale.
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Objective: To clone and characterize the F family ATP-binding cassette (ABC) transporter genes in Dendrobium officinale. Methods: RACE was used to isolate ABC transporter genes from the leaf cDNA of D. officinale. Characteristics including molecular weight, theoretical pI (isoelectric point), conserved domain, transmembrane structure, signal peptide, and subcellular localization of the deduced protein were analyzed by serials of bioinformatics algorithms. The analyses of multiple alignment and phylogenetic tree were respectively performed by DNASTAR and MEGA6. Tissue specific expression patterns were determined by real-time quantitative PCR (qPCR) analyses. Results: Two full length genes DoABCF1 and DoABCF2 (GenBank accessions KU160474 and KU160475), 2 104 and 2 193 bp in length, respectively, were obtained. DoABCF1 was deduced to a 600 aa (amino acid) protein with a molecular weight of 67 030 and pI of 6.20, while DoABCF2 encoded a 659 aa protein with a molecular weight of 74 140 and pI of 5.71. The two deduced DoABCF1 and DoABCF2 proteins both had two conserved ABC domains (74-314 and 385-600 for the former, 65-323 and 392-607 for the latter) and several functional motifs. The proteins did not contain any signal peptide or transmembrane domain, and were predicted to locate in the chloroplast. The two genes were highly identical to the plant F family ABC transporter genes with more than 80% similarity, and were mostly close to monocots ABC F family members from maize and rice. DoABCF1 and DoABCF2 showed different expression among the three organs and both had relatively highest expression levels in the leaves, whereas no significant difference could be observed in the roots and stems. Taken the stem as the calibrator sample, DoABCF1 transcripts were 1.74 fold over that in the stems and DoABCF1 transcripts were 3.44 fold, respectively. Conclusion: Two F family DoABCF1 and DoABCF2 genes with full length cDNAs are successfully cloned in this study. The highest expression levels of the two ABC genes in the leaves of D. officinale suggest that they should play an important role during the growth and development of D. officinale.
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Objective To establish an efficient tissue culture and rapid propagation system, and study the protocorm proliferation and regeneration conditions using seeds as explants in Dendrobium officinale. Methods Seeds of D. officinale were used as explants, protocorm was induced on inducement medium. After proliferation, the protocorm were transferred to the regeneration medium. Then the regenerated shoots were transferred into rooting medium to induce rooting of plantlets, and developing complete plant. Results The basic culture medium for D. officinale growth was 1/2 MS; The best culture medium formula for inducing protocorms was 1/2 MS +1.0 mg/L 6-BA + 0.2 mg/L NAA + 50 g/L mashed potato; The optimal proliferation medium for protocorm was 1/2 MS + 1.0 mg/L 6-BA + 0.5 mg/L NAA, and the maximum multiplication factor could reach 23. The optimal regeneration medium was 1/2 MS + 2.0 mg/L 6-BA + 0.2 mg/L NAA, regeneration rate can reach 95%; The best culture medium for seedling rooting was 1/2 MS + 0.3 mg/L NAA + 50 g/L potato juice, and rooting rate reached 100%. Conclusion This research provides an effective way for keeping good varieties of features and rapid propagation of D. officinale, at the same time helps to solve some theoretical problems in factory production of D. officinale.
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Objective Conserved DNA-Derived Polymorphism (CDDP) markers were used in the study of the genetic diversity of 43 Dendrobium officinale and preliminary resistance screening, in order to provide a theoretical basis for the screening of D. officinale and the selection of fine varieties. Methods A total of 21 CDDP primers were used to amplify the genomic DNA of the test material with clear and polymorphic primers. Results Sixteen primers generated 151 bands, of which 144 (95.7%) were polymorphic. The results of data analysis on three population showed that the percentage of polymorphic locus (PPL) were between 65.56% and 82.12%, coefficient of genetic differentiation among natural populations (Gst) was 0.110 2, and the total gene flow (Nm) was 4.035 4. Indicating that anthropogenic factors may also promote the genetic diversity of D. officinale. Conclusion The genetic diversity of D. officinale was rich, and the results showed that there were eight plants with potentially good resistance among 43 materials.
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Objective To clone the full-length cDNA encoding sesquiterpene synthase (SES) gene from Dendrobium officinale (DoSES), then to analyze the expression difference in different tissues and expression patterns of DoSES induced by methyl jasmonate (MeJA). Methods RACE was used to clone the full length cDNA of DoSES. Bioinformatics analysis was carried out by using related software and online website. Then, the expression patterns of DoSES were studied by quantitative real-time PCR (qRT-PCR). Results The DoSES gene was obtained (GenBank accession number KX278311), and the full-length cDNA was 1 890 bp, encoding the protein of 520 amino acids. DoSES had highest homology ( ≈ 60%) with SES proteins from other plants. Tissue expression analysis showed that DoSES had the highest expression in the stems, followed by leaves, flowers, protocorm and roots. The results of qRT-PCR analysis suggested that DoSES could be induced by MeJA. Conclusion The cDNA encoding DoSES was cloned. The research results provide basic materials for further function alanalysis of this gene in the future.
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Objective: In order to isolate and analysize the bioinformatics and expression pattern of DoWRKY5 gene from Dendrobium officinale. Methods: A WRKY gene was first obtained by transcriptome sequencing and reverse transcription-polymerase chain reaction (RT-PCR) from D. officinale and analyzed by bioinformatics tools. The tissue expression pattern and the low temperature stress, abscisic acid (ABA) stress, and sucrose stress responses were analyzed by qRT-PCR. Results: The cDNA sequence of DoWRKY5 gene was isolated, which was 1 336 bp in length, with an open reading frame (ORF) of 834 bp and an encoded polypeptide of 277 amino acid. The amino acid sequence contained a conserved WRKY domains and a zinc finger structures (C2H2), belonging to Group II of WRKY family. Expression analysis by qRT-PCR showed that DoWRKY5 was expressed in the roots, stems, and leaves of D. officinale, and the most abundant in leaves. The amount of DoWRKY5 expression were significantly increased under low temperature of 4℃ and different time. Moreover, the expression of DoWRKY5 could be induced by ABA and source. Conclusion: DoWRKY5 may be an important transcription factor to response cold stress and other abiotic stresses in D. officinale, which provides a foundation for further study of cold tolerance mechanism and cold-resistant breeding of D. officinale.
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Objective: To clone the 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase (HDS) gene from Dendrobium officinale, induce fusion protein in Escherichia coli, and explore the regular pattern about HDS gene in different tissues of D. officinale. Methods: RT-PCR and RACE technologies were used to clone the full-length cDNA of DoHDS. Using relevant softwares and online sites to analyze bioinformatics. Then the expression patterns of DoHDS were studied by real-time PCR. Constructing prokaryotic expression vector pET-28a (+)-DoHDS to induce the expression protein in E. coli BL21 (DE3). Results: The DoHDS gene was successfully obtained (GenBank accession number KJ161312), the full-length cDNA was 2666 bp and ORF was 2238 bp, coding the protein containing 745 amino acids. Relative real-time PCR analysis indicated that DoHDS showed the higher transcript abundance in the stems, 5 fold higher than protocorm-like bodies. The SDS-PAGE results showed that a relative molecular weight of 82700 recombinant protein was produced. Conclusion: The cDNA encoding HDS from D. officinale is cloned. The prokaryotic expression vector and different tissues expression patterns of DoHDS are constructed. It is helpful for the future research on the mechanism of terpenoid biosynthesis in medicinal plants D. officinale.