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Fig. 2 in Development of an efficient transient expression system for Siraitia grosvenorii fruit and functional characterization of two NADPH-cytochrome P450 reductases

Fig. 2. (A) Cloning of full-length and specific fragments of the SgCPR1 and SgCPR2 genes from S. grosvenorii cDNA. M, marker (2000 bp). The fulllength SgCPR1 and SgCPR2 genes are shown in red brackets, whereas the blue arrow indicates the specific fragments of the SgCPR1 and SgCPR2 genes. (B) Vector map of PBI121. (C) Construction of PBI12- SgCPR recombinant plasmids with the GUS marker and CaMV35S promoter. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 5 in Development of an efficient transient expression system for Siraitia grosvenorii fruit and functional characterization of two NADPH-cytochrome P450 reductases

Fig. 5. qRT-PCR analysis of SgCPR1 and SgCPR2 in fruit after infiltration (at t = 0 h). Relative expression levels were normalized to t = 0 h. (A) Relative transcript levels of SgCPR1 in fruit harbouring PBI121- SgCPR1 and PBI121. (B) SgCPR2 gene expression levels in fruit harbouring PBI121 and PBI121-SgCPR2. (C) Relative expression levels of SgCPR1 in fruit harbouring pTRV-SgCPR1 and pTRV-0. (D) Expression levels of SgCPR2 in fruit harbouring pTRV-SgCPR2 and pTRV-0. The relative expression levels normalized to the level of the SgUBQ gene. All the data are shown as means±SDs. * indicates significant differences at p <0.05 (LSD test).

Fig. 3 in Development of an efficient transient expression system for Siraitia grosvenorii fruit and functional characterization of two NADPH-cytochrome P450 reductases

Fig. 3. (A) Selective nucleotide sequences for VIGS in S. grosvenorii, which were inserted into EcoRI and BamHI. (B) pTRV2-SgCPR recombinant plasmid construction.

Fig. 1 in Development of an efficient transient expression system for Siraitia grosvenorii fruit and functional characterization of two NADPH-cytochrome P450 reductases

Fig. 1. The mogroside biosynthetic pathway in S. grosvenorii. The blue box on the left shows the generation of mogrol, which is a unique nonglycosylated tetra-hydroxycucurbitane in S. grosvenorii. The red box on the right represents a series of glycosylation reactions. The enzyme names are abbreviated as follows: SgSQE, squalene epoxidase; SgCS, cucurbitadienol synthase; SgEPH, epoxide hydrolase; SgCYP450, cytochrome P450 monooxygenase; SgCPR, NADPHcytochrome P450 reductase; SgUGT, UDP-glucosyltransferase. Among these, the candidate SgCPRs are highlighted in red. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 6 in Development of an efficient transient expression system for Siraitia grosvenorii fruit and functional characterization of two NADPH-cytochrome P450 reductases

Fig. 6. Accumulation of MIII and MIIE. (A) Total ion chromatograms of ten standard mixtures determined by LC-MS/MS. Among these, the retention times of MIII and MIIE were 10.01 and 12.09 min, respectively. (B) Extracted ion chromatograms of the sample. The red arrow indicates the Q1/Q3 mass chromatograms of MIII, which are in the red box. The Q1/Q3 mass chromatograms of MIIE are shown in the blue box, as indicated by the blue arrow. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4 in Development of an efficient transient expression system for Siraitia grosvenorii fruit and functional characterization of two NADPH-cytochrome P450 reductases

Fig. 4. Histochemical GUS assay of transient expression in S. grosvenorii. GUS expression was detected in fruit but not in CK or leaves.

Fig. 6 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 6. Heatmap representing organ specific significantly enriched genes corresponding to (A) Isosteroidal alkaloid biosynthesis, (B) Sucrose and starch metabolism, (C) UGTs and CYPs, (D) aquaporins, (E) ABC transporters, (F) Transcription factor and Transposable elements. The red-blue scale represents positive enrichment (red) and negative enrichment (Blue) of transcripts. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 5 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 5. Significantly enriched transcripts (nodes) in predicted interactome network. (A) Steroidal alkaloid biosynthesis pathways (B) Isopentenyl diphosphate biosynthesis pathway, (C) Sucrose and starch metabolic pathways and (D) Aquaporins. The nodes encircled in red color represents higher enrichment in the bulb while the nodes encircled in grey and blue represents higher enrichment in arial organs (leaf and stem). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 7 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 7. qRT-PCR expression-based validation of RNA-seq data using potential 20 genes involved in isosteroidal alkaloid biosynthetic pathway. (A) Bulb vs. Leaf, (B) Leaf vs. Stem and (C) Bulb vs. Stem.

Fig. 3 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 3. Significant KEGG pathway enrichment analysis in tissues from aerial organs (leaf & Stem) and bulb. The green colour enrichment indicates higher expression in aerial tissue while pink represents higher enrichment in bulb. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 4. Transcriptional protein-protein interactome (PPIN) prediction in F. roylei along the network statistics. (A) Overall prediction of PPI network based on significantly differential expressed transcripts. Spatial PPI network prediction of significantly enriched transcripts in (B) Bulb, (C) Leaf and (D) Stem.

Fig. 1 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 1. Summary of organ specific spatial transcriptome sequencing in F. roylei. (A): Overall quality filtering of sequenced reads; (B): Tissue specific high quality filtered reads obtained after removing low quality/adaptor contaminated sequences; (C): Assembly statistics details; (D): Venn diagram representing functional annotation with six different public protein databases.

Fig. 8 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 8. Representation of isosteroidal alkaloid biosynthesis pathway in F. roylei and heat map representing expression of genes in Stem, Leaf and Bulb tissue using red-blue scale (red: positive enrichment and Blue: negative enrichment of transcripts). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 2 in Genome-wide transcriptional analysis unveils the molecular basis of organ-specific expression of isosteroidal alkaloids biosynthesis in critically endangered Fritillaria roylei Hook

Fig. 2. Clustering of 2488 significant differentially expressed transcripts in leaf, stem and bulb in F. roylei (A) sub-cluster 1 represent transcripts with significant higher expression in bulb; (B): Sub-cluster 2 in Stem and (C): Sub-cluster 3 in leaf; (D): Pearson's correlation of organ specific significant differentially expressed clustered transcripts in leaf, stem and bulb tissues.

Fig. 6 in Impact of potential stimulants on asiaticoside and madecassoside levels and expression of triterpenoid-related genes in axenic shoot cultures of Centella asiatica (L.) Urb

Fig. 6. Relative fold change of genes involved in the production of asiaticoside and madecassoside in C. asiatica multiple shoot cultures, after three and four weeks of a treatment with AgNO3. Abbreviation: HMGR: hydroxymethylglutaryl-CoA reductase; DXR: deoxylulosephosphate reductoisomerase; FPS: farnesyl diphosphate synthase; βAS: beta amyrin synthase; SE: squalene epoxidase; SS: squalene synthase; GT: glycosyltransferase.

Fig. 7 in Impact of potential stimulants on asiaticoside and madecassoside levels and expression of triterpenoid-related genes in axenic shoot cultures of Centella asiatica (L.) Urb

Fig. 7. Proposed biosynthetic pathway of triterpenoids in C.asiatica (Prasad et al., 2019). Abbreviation: HMGS: hydroxy methylglutaryl-CoA synthase; HMGR: hydroxymethylglutaryl-CoA reductase; MVK: mevalonate kinase; DXS: deoxylulosephosphate synthase; DXR: deoxylulosephosphate reductoisomerase; DMAPP: dimethylallyl pyrophosphate; IPP: isopentenyl pyrophosphate; GPP: geranyl pyrophosphate; FPP: farnesyl pyrophosphate; FPPS: farnesyl diphosphate synthase; βAS: beta amyrin synthase; αAS: alpha amyrin synthase; SE: squalene epoxidase; SS: squalene synthase; CYP450: cytochrome P450; GT'S: glycosyltransferase; MVA: mevalonate; MEP: methylerythritol 4-phosphate.

Fig. 5 in Impact of potential stimulants on asiaticoside and madecassoside levels and expression of triterpenoid-related genes in axenic shoot cultures of Centella asiatica (L.) Urb

Fig. 5. Madecassoside content (mg/g DW) after three and four weeks, in response to two different concentrations of each elicitor. Mean values followed by different letters under different treatments and time within similarly colored bars are significantly different from each other at p <0.05 (Duncan's multiple range tests).

Fig. 4 in Impact of potential stimulants on asiaticoside and madecassoside levels and expression of triterpenoid-related genes in axenic shoot cultures of Centella asiatica (L.) Urb

Fig. 4. Asiaticoside content (mg/g DW) after three and four weeks, in response to two different concentrations of each elicitor. Mean values followed by different letters under different treatments and time within similarly colored bars are significantly different from each other at p <0.05 (Duncan's multiple range tests).

Fig. 2. A in Impact of potential stimulants on asiaticoside and madecassoside levels and expression of triterpenoid-related genes in axenic shoot cultures of Centella asiatica (L.) Urb

Fig. 2. A) Nodal segment of C. asiatica used for culture initiation; B) Shoot induction in MS media with 1 mg/L BAP and 0.2 mg/L NAA; C) Multiple shoot cultures observed after three weeks; D) Established cultures; E) and F) Biomass cultivation and harvest.

Fig. 1 in Impact of potential stimulants on asiaticoside and madecassoside levels and expression of triterpenoid-related genes in axenic shoot cultures of Centella asiatica (L.) Urb

Fig. 1. Different growth stages showing an increase in the number of shoots in C. asiatica cultures at two weeks (A), three weeks (B), four weeks (C), and five weeks (D).