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Transcriptomic responses to cold stress in Dendrobium huoshanense C.Z. Tang et S.J. Cheng

ABSTRACT: Dendrobium huoshanense C.Z. Tang et S.J. Cheng is a perennial epiphytic herb of family Orchidaceae, which main metabolites are polysaccharides, flavonoids, etc. Low temperature is the main environmental factor that limits the growth and development of plants, and even threatens the survival of plants. However, Changes that occur at the molecular level in response to low temperatures are poorly understood in D. huoshanense. To understand the molecular mechanism of cold tolerance, we performed transcriptomic analysis on two time points of 0 d (control group) and 7 d (cold stress group). A total of 37.63 Gb transcriptomic data were generated using the MGI 2000 platform. These reads were assembled into 170,754 transcripts, and 23,724 differentially expressed genes (DEGs) were obtained. Pathway analysis indicated that “flavonoid biosynthesis”, “anthocyanin biosynthesis”, “flavone and flavonol biosynthesis”, and “plant hormone signal transduction” might play a vital role in D. huoshanense responses to cold stress. Several important pathways genes such as genes encoding polysaccharides, flavonoid, and plant hormone-signaling transduction kinase were identified under cold stress. In addition, the contents of mannose and total flavonoids increased under cold stress. Twelve DEGs in polysaccharides, flavonoid, and hormone pathways were selected from transcriptome analysis for quantitative real-time PCR (qRT-PCR) validation. Our results provide a transcriptome database and candidate genes for further study of D. huoshanense cold stress.

ORGANISM(S): Dendrobium huoshanense

PROVIDER: GSE217439 | GEO | 2022/11/10

REPOSITORIES: GEO

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Project description:Background: Dendrobium officinale, an endangered Chinese herb, has extensive therapeutic effects and contains bioactive ingredients including a large number of polysaccharides and alkaloids, and minimal flavonoids. Firstly, this study attempts to obtain the protocorm-like bodies of this plant through tissue culture to produce the main secondary metabolites whose distribution in each organelle and protocorm like bodies is analyzed. Then, analysis of the correlation between comparative transcriptome sequence and the metabolite content in different organs enables the discovery of putative genes encoding enzymes involved in the biosynthesis of polysaccharides and alkaloids, and flavonoids. Results: The optimum condition for protocorm-like bodies (PLBs) induction and propagation of D. officinale is established. For protocorm induction, we use the seed as the explant, and the optimum medium formula for PLBs propagation is 1/2 MS + α-NAA 0.5 mg·L-1 +6-BA 1.0 mg·L-1 + 2, 4-D 1.5-2.0 mg·L-1 + potato juice 100 g·L-1. The distribution of polysaccharides, alkaloids and flavonoids in D. officinale organs was clarified. Stems, PLBs and leaves have the highest content of polysaccharides, alkaloids and flavonoids, respectively. PLBs replace organs to produce alkaloids in D. officinale, and naringenin was only produced in stem. Hot water extraction (HWE) method was found outperforming the ultrasound-assisted extraction (UAE) method for polysaccharides from D. officinale. A comparative transcriptome analysis of the protocorm-like bodies and leaves of D. officinale showed genes encoding enzymes involved in polysaccharides, alkaloids and flavonoids biosynthetic pathway were differentially expressed. Putative genes encoding enzymes involved in polysaccharides, alkaloids and flavonoids synthetic pathway were identified. Notably, genes encoding enzymes of strictosidine beta-glucosidase, geissoschizine synthase and vinorine synthase in alkaloids biosynthesis of D. officinale are first reported. Conclusions: Our works, especially the identification of candidate genes encoding enzymes involved in metabolites biosynthesis will help to explore and protect the endangered genetic resources and will also facilitate further analysis of the molecular mechanism of secondary metabolites’ biosynthesis in D. officinale.

Project description:The aerial parts of Glycyrrhiza uralensis supply substantial raw material for the extraction of active pharmaceutical ingredients comprehensively utilized in many industries. Liquiritin is the unique and most abundant flavonoid active component in G. uralensis. It has a wide range of pharmacological activities and high economic value. Our previous study indicated that moderate salt stress increased the content of active ingredients. However, the regulatory mechanism remains unclear. In this study, RNA-sequencing (RNA-seq) of the aerial parts of G. uralensis treated with 150 mM NaCl for 0, 2, 6, and 12 h was performed to identify the key genes and metabolic pathways regulating pharmacological active component accumulation. The main active component detection showed that liquiritin was the major ingredient and exhibited more than a 10-fold significant increase in the 6-h NaCl treatment. Temporal expression analysis of the obtained 4245 differentially expressed genes (DEGs) obtained by RNA-seq revealed two screened profiles that included the significant up-regulated DEGs (UDEGs) at different treatment points. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of these UDEGs identified phenylpropanoid metabolism and flavonoid biosynthesis as the most significantly enriched pathways in 2-h treated materials. Interestingly, the carotenoid biosynthesis pathway that is related to ABA synthesis was also discovered, and the ABA content was significantly promoted after 6-h NaCl treatment. Following ABA stimulation, the content of liquiritin demonstrated a significant and immediate increase after 2-h treatment, with the corresponding consistent expression of genes involved in the pathways of ABA signal transduction and flavonoid biosynthesis, but not in the pathway of glycyrrhizic acid biosynthesis. Therefore, salt stress might promote liquiritin accumulation through the ABA-mediated signaling pathway in G. uralensis. Our study provides candidate genes and pathways controlling liquiritin synthesis, and would drive progress in genetic improvement and promote the comprehensive utilization of the aerial part of G. uralensis.

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Transcriptomic responses to cold stress in Dendrobium huoshanense C.Z. Tang et S.J. Cheng

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