De novo leaf and root transcriptome analysis to identify putative genes involved in triterpenoid saponins biosynthesis in Hedera helix L.

Sun, Hua; Li, Fang; Xu, Zijian; Sun, Mengli; Cong, Hanqing; Qiao, Fei; Zhong, Xiaohong

doi:10.1371/journal.pone.0182243

Cited by 22 publications

(20 citation statements)

References 47 publications

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“…After the assembly and annotation of the A. elata transcriptome in the present study, we were able to begin examining the terpenoid biosynthesis backbone and sesquiterpenoid and triterpenoid biosynthesis pathways in these samples, leading to the identification of 19 functional genes. While these two pathways are well-known to be important for the biosynthesis of terpenoids and sterols in P. notoginseng [22] and Hedera helix L. [23], the results of the present analysis are the first such comprehensive analysis of these pathways in A. elata. Triterpenoids and sesquiterpenoids are biosynthesized via the MVA pathway, that takes place primarily in the cytoplasm, whereas monoterpenoids, diterpenoid, and tetraterpenoids are biosynthesized via the MEP pathway, mostly in the plastid [11].…”

Section: Discussionmentioning

confidence: 80%

“…Additionally, 28 unigenes encoding 4 putative enzymes (FPS, SS, SE, and bAS) were found to be associated with carbocyclic biosynthesis (Additional file 3: Table S1). For the majority of these genes, we were able to map >1 unigene to a given gene or gene family (Additional file 3: Table S1), suggesting that these sequences may correspond to multiple copies or partial fragments of a given gene [23]. In each of enzymatic steps, those unigenes that were showed a high identity with functionally characterized genes and differentially expressed in different tissues were next selected and arranged into a heat map.…”

Section: Enrichment Of Terpenoid Backbone and Triterpenoid Biosynthetmentioning

confidence: 99%

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Identification and analysis of CYP450 and UGT supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponin biosynthesis

Cheng

Liu

Tong

et al. 2020

Preprint

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Background: Members of the cytochrome P450 (CYP450) and UDP-glycosyltransferases (UGT) gene superfamily have been shown to play essential roles in regulating secondary metabolites biosynthesis. However, the systematic identification of CYP450s and UGTs have not been reported in Aralia elata (Miq.) Seem , a highly valued medicinal plant. Results: In the present study we conducted the RNA-sequencing (RNA-seq) analysis of the leaves, stems, and roots of A. elata, yielding 66,713 total unigenes. Following the annotation and KEGG pathway analysis, we were able to identify 64 unigenes related to triterpenoid skeleton biosynthesis, 254 CYP450s and 122 UGTs, respectively. 150 CYP450s and 92 UGTs encoding >300 amino acid proteins were utilized for phylogenetic and tissue-specific expression analyses. This allowed us to cluster 150 CYP450s into 9 clans and 40 families, and then these CYP450 proteins were further grouped into two primary branches: A-type (53%) and non-A type (47%). A phylogenetic analysis of 92 UGTs and other plant UGTs led to clustering into 16 groups (A-P). We further assessed the expression patterns of these CYP450 and UGT genes across A. elata tissues, with 23 CYP450 and 16 UGT members being selected for qRT-PCR validation, respectively. From these data, we identified CYP716A295 and CYP716A296 as the candidate genes most likely to be associated with oleanolic acid synthesis, while CYP72A763 and CYP72A776 was identified as being the most likely to play a role in hederagenin biosynthesis. We also selected five unigenes as the best candidates for oleanolic acid 3-O-glucosyltransferase. Finally, we assessed the subcellular localization of three CYP450 proteins within Arabidopsis protoplasts, highlighting the fact that they localize to the endoplasmic reticulum. Conclusions: This study presents a systematic analysis of the CYP450 and UGT gene family in A. elata and provided a foundation for further functional characterization of these two multigene family.

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Section: Discussionmentioning

confidence: 80%

Section: Enrichment Of Terpenoid Backbone and Triterpenoid Biosynthetmentioning

confidence: 99%

Identification and analysis of CYP450 and UGT supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponin biosynthesis

Cheng

Liu

Tong

et al. 2020

Preprint

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“…It was found that the content of triterpenes accumulated in the leaves of P. ginseng are higher in the early growth stage, while the content of triterpenes in the roots of old plants are higher [27]. Comparative transcriptome analysis including root and leaf tissues to excavate transcripts related to saponin biosynthesis is already reported for many plants, such as Hedera helix [28], Panax notoginseng [29] and Asparagus racemosus [26]. The maximum triterpenoid saponin content was identi ed in stem.…”

Section: Introductionmentioning

confidence: 85%

Comparative transcriptome analysis of root, stem, and leaf tissues of Entada phaseoloides reveals potential genes involved in triterpenoid saponin biosynthesis

Liao

Mei

Miao

et al. 2020

Preprint

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Background Entada phaseoloides (L.) Merr. is an important traditional medicinal plant. The stem of Entada phaseoloides is popularly used as traditional medicine because of its significance in dispelling wind and dampness and remarkable anti-inflammatory activities. Triterpenoid saponins are the major bioactive compounds of Entada phaseoloides. However, genomic or transcriptomic technologies have not been used to study the triterpenoid saponin biosynthetic pathway in this plant. Results We performed comparative transcriptome analysis of the root, stem, and leaf tissues of Entada phaseoloides with three independent biological replicates and obtained a total of 53.26 Gb clean data and 116,910 unigenes, with an average N50 length of 1218 bp. Putative functions could be annotated to 42,191 unigenes (36.1%) based on BLASTx searches against the Non-redundant, Uniprot, KEGG, Pfam, GO, KEGG and COG databases. Most of the unigenes related to triterpenoid saponin backbone biosynthesis were specifically upregulated in the stem. A total of 26 cytochrome P450 and 17 uridine diphosphate glycosyltransferase candidate genes related to triterpenoid saponin biosynthesis were identified. The differential expressions of selected genes were further verified by qPT-PCR. Conclusions The dataset reported here will facilitate the research about the functional genomics of triterpenoid saponin biosynthesis and genetic engineering of Entada phaseoloides.

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“…Total RNA from the mixed sample (three individual plants) was isolated using the RNA EasySpin Isolation System (Aidlab Biotech, Beijing, China) according to the manufacturer's protocol. Following treatment with RNase-free DNase I (New England BioLabs) for 30 min at 37 • C to remove residual DNA, sequencing was performed by Shanghai Biotechnology Co. Ltd., where a cDNA library was constructed and subjected to Illumina HiSeq2500 sequencing, as described by Sun et al [35].…”

Section: Cdna Library Preparation and Illumina Transcriptomic Sequencingmentioning

confidence: 99%

Heterologous Expression of the DREB Transcription Factor AhDREB in Populus tomentosa Carrière Confers Tolerance to Salt without Growth Reduction under Greenhouse Conditions

Guo

Sun

et al. 2019

Forests

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The DREB transcription factors regulate multiple stress response genes, and are therefore useful for molecular plant breeding. AhDREB, a stress-inducible gene, was isolated from Atriplex hortensis L. and introduced into Populus tomentosa Carrière under the control of the CaMV35S promoter. Under salt stress, the chlorophyll content and net photosynthetic rate were higher in transgenic lines than in the wild type (WT). Moreover, the rate of electrolyte penetration (REC) was lower in the transgenic lines. Additional analyses revealed that the AhDREB transgenic plants generally displayed lower malondialdehyde (MDA) activity but higher superoxide dismutase (SOD) and peroxidase (POD) activities and proline content than the WT under salt stress. RNA sequencing indicated that AhDREB could enhance tolerance to salt by activating various downstream genes in the transgenic plants. Furthermore, no growth inhibition was detected in transgenic plants expressing AhDREB driven by the constitutive CaMV35S promoter. The transcriptome showed 165 and 52 differentially expressed genes in transgenic plants under stress and non-stress conditions, respectively, among which no significant metabolic pathway was enriched and no unintended effects have yet been identified. Together, these results suggest that AhDREB may be a good candidate gene for increasing salt tolerance in transgenic poplar breeding.

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De novo leaf and root transcriptome analysis to identify putative genes involved in triterpenoid saponins biosynthesis in Hedera helix L.

Cited by 22 publications

References 47 publications

Identification and analysis of CYP450 and UGT supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponin biosynthesis

Identification and analysis of CYP450 and UGT supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponin biosynthesis

Comparative transcriptome analysis of root, stem, and leaf tissues of Entada phaseoloides reveals potential genes involved in triterpenoid saponin biosynthesis

Heterologous Expression of the DREB Transcription Factor AhDREB in Populus tomentosa Carrière Confers Tolerance to Salt without Growth Reduction under Greenhouse Conditions

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