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Research Article
Xiasheng Zheng
Guangzhou University of Chinese Medicine
Corresponding Author
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Helicteres angustifolia has long been used in Chinese traditional medicine. It has multiple pharmacological benefits, including anti-inflammatory, anti-viral and anti-tumor effects. Its main active chemicals include betulinic acid, oleanolic acid, helicteric acid, helicterilic acid, and other triterpenoid sapions. It is worth noting that the acylation of triterpenoids, such as helicteric acid and helicterilic acid, are characteristic components of Helicteres and are relatively rare among other plants. However, reliance on natural plants as the only sources of these restricts the potential use of H. angustifolia. Therefore, engineering of its metabolic pathway is of high research value for enhancing the production of secondary metabolites. Unfortunately, little is known of the biosynthesis of acylated triterpenoids, hindering its further investigation. Here, the RNAs of different groups treated by metabolic stimulation were sequenced with an Illumina high-throughput sequencing platform, resulting in 121 gigabases of data. A total of 424,824 unigenes were obtained after the trimming and assembly of the raw data, and 22,430 unigenes were determined to be differentially expressed. In addition, three oxidized squalene cyclases (OSCs) and four Cytochrome P450 (CYP450s) were screened, of which one OSC (HaOSC1) and one CYP450 (HaCYPi3) achieved functional verification, suggesting that they could catalyze the production of lupeol and oleanolic acid, respectively. At the same time, we also screened two triterpenoid acetyl transferases (TATs) and one triterpenoid benzoyl transferase (TBT), as subsequent structural modificatory genes, their preliminary study laid a foundation for the acylation of triterpenoids. In a nutshell, these results shed light on the regulation of acylated triterpenoids biosynthesis.
Keywords
Helicteres Angustifolia, Acylated triterpenoids, Transcriptome, , Functional expressions， Triterpenoids acetyl transferases, Triterpenoids benzoyl transferases.
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No competing interests reported.
This is a list of supplementary files associated with this preprint. Click to download.
- FigureS1.png
Figure S1 The electrophoresis diagrams of RNA extracted from different treatment groups of H. angustifolia tree leaves.
- FigureS2.png
Figure S2 Investigation of hormone concentration.
- FigureS3.png
Figure S3. Comparisons of the total triterpenoid content among different treatment groups.
- FigureS4.png
Figure S4 Functional annotation of the transcripts and unigenes against six databases.
- FigureS5.png
Figure S5 Venn diagram indicating the numbers of common and specific expressed unigenes among six databases.
- FigureS6.png
Figure S6 KEGG enrichment results of the DEGs among negative control group and different treatment groups.
- FigureS7.jpg
Figure S7 The expression pattern of key related genes involved with triterpenoids biosynthesis were shown using a heatmap.
- FigureS8.png
Figure S8 Three dimention structure of target gene coding proteins.
- FigureS9.png
Figure S9 Analysis of amino acid sequences of TATs and TBT.
- FigureS10.png
Figure S10 SDS-PAGE analysis of TATs and TBT.
- TableS1.doc
Table S1 Primers for qRT-PCR
- TableS2.doc
Table S2 Primers for amplification of ten putative genes
- TableS3.doc
Table S3 Gene-specific forward and reverse primers
- TableS4.doc
Table S4 Transcriptome sequencing data of the H. angustifolia
- TableS5.doc
Table S5 The overview statistics of annotation
- TableS7.doc
Table S7 KEGG enrichment of DEGs from the NC vs SA comparison
- TableS8.doc
Table S8 KEGG enrichment of DEGs from the NC vs MeJA comparison
- TableS12.doc
Table S12 Analysis of physicochemical properties of target gene coding proteins.
- TableS13.doc
Table S13 Two-dimensional structure information of of target gene coding proteins.
- TableS14.doc
Table S14 Summary table of OSC genes
- TableS16.doc
Table S16 Summary table of BAHD genes
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This preprint is under consideration at BMC Plant Biology. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research Article
Xiasheng Zheng
Guangzhou University of Chinese Medicine
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 08 Apr, 2021
No community comments so far
Reviewers invited
Invitations sent on 08 Apr, 2021
Editor assigned
On 07 Apr, 2021
Editor invited
On 07 Apr, 2021
Submission checks complete
On 07 Apr, 2021
First submitted
On 05 Apr, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Helicteres angustifolia has long been used in Chinese traditional medicine. It has multiple pharmacological benefits, including anti-inflammatory, anti-viral and anti-tumor effects. Its main active chemicals include betulinic acid, oleanolic acid, helicteric acid, helicterilic acid, and other triterpenoid sapions. It is worth noting that the acylation of triterpenoids, such as helicteric acid and helicterilic acid, are characteristic components of Helicteres and are relatively rare among other plants. However, reliance on natural plants as the only sources of these restricts the potential use of H. angustifolia. Therefore, engineering of its metabolic pathway is of high research value for enhancing the production of secondary metabolites. Unfortunately, little is known of the biosynthesis of acylated triterpenoids, hindering its further investigation. Here, the RNAs of different groups treated by metabolic stimulation were sequenced with an Illumina high-throughput sequencing platform, resulting in 121 gigabases of data. A total of 424,824 unigenes were obtained after the trimming and assembly of the raw data, and 22,430 unigenes were determined to be differentially expressed. In addition, three oxidized squalene cyclases (OSCs) and four Cytochrome P450 (CYP450s) were screened, of which one OSC (HaOSC1) and one CYP450 (HaCYPi3) achieved functional verification, suggesting that they could catalyze the production of lupeol and oleanolic acid, respectively. At the same time, we also screened two triterpenoid acetyl transferases (TATs) and one triterpenoid benzoyl transferase (TBT), as subsequent structural modificatory genes, their preliminary study laid a foundation for the acylation of triterpenoids. In a nutshell, these results shed light on the regulation of acylated triterpenoids biosynthesis.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
No competing interests reported.
This is a list of supplementary files associated with this preprint. Click to download.
- FigureS1.png
Figure S1 The electrophoresis diagrams of RNA extracted from different treatment groups of H. angustifolia tree leaves.
- FigureS2.png
Figure S2 Investigation of hormone concentration.
- FigureS3.png
Figure S3. Comparisons of the total triterpenoid content among different treatment groups.
- FigureS4.png
Figure S4 Functional annotation of the transcripts and unigenes against six databases.
- FigureS5.png
Figure S5 Venn diagram indicating the numbers of common and specific expressed unigenes among six databases.
- FigureS6.png
Figure S6 KEGG enrichment results of the DEGs among negative control group and different treatment groups.
- FigureS7.jpg
Figure S7 The expression pattern of key related genes involved with triterpenoids biosynthesis were shown using a heatmap.
- FigureS8.png
Figure S8 Three dimention structure of target gene coding proteins.
- FigureS9.png
Figure S9 Analysis of amino acid sequences of TATs and TBT.
- FigureS10.png
Figure S10 SDS-PAGE analysis of TATs and TBT.
- TableS1.doc
Table S1 Primers for qRT-PCR
- TableS2.doc
Table S2 Primers for amplification of ten putative genes
- TableS3.doc
Table S3 Gene-specific forward and reverse primers
- TableS4.doc
Table S4 Transcriptome sequencing data of the H. angustifolia
- TableS5.doc
Table S5 The overview statistics of annotation
- TableS7.doc
Table S7 KEGG enrichment of DEGs from the NC vs SA comparison
- TableS8.doc
Table S8 KEGG enrichment of DEGs from the NC vs MeJA comparison
- TableS12.doc
Table S12 Analysis of physicochemical properties of target gene coding proteins.
- TableS13.doc
Table S13 Two-dimensional structure information of of target gene coding proteins.
- TableS14.doc
Table S14 Summary table of OSC genes
- TableS16.doc
Table S16 Summary table of BAHD genes
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