Metabolic Engineering of Glycyrrhizin Pathway by Over-Expression of Beta-amyrin 11-Oxidase in Transgenic Roots of Glycyrrhiza glabra

Shirazi, Zahra; Aalami, Ali; Tohidfar, Masoud; Sohani, Mohammad Mehdi

doi:10.1007/s12033-018-0082-7

Cited by 15 publications

(5 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These data showed the great impact of these two genes on the glycyrrhizin biosynthetic pathway. In the case of a research on metabolic engineering of glycyrrhizin biosynthetic pathways including SQS1 and CYP88D6, an increased amount of glycyrrhizin was observed in hairy root cultures of Glycyrrhiza plants that confirmed the results of this study (Lu et al 2008;Shirazi et al 2018). Production pattern of glycyrrhizin was more similar to the expression pattern of SQS1 gene so that the stepwise regression analysis also confirmed that 92% of the change in glycyrrhizin content is associated with SQS1.…”

Section: Effect Of Salt Stress On Glycyrrhizin Content In Rootssupporting

confidence: 84%

Triterpenoid gene expression and phytochemical content in Iranian licorice under salinity stress

et al. 2019

Self Cite

View full text Add to dashboard Cite

Licorice is a well-known medicinal plant, containing various secondary metabolites of triterpenoid and phenolic families. The aim of this study is to evaluate the effect of salinity stress on the expression of key genes involved in the biosynthetic pathway of triterpenoids such as glycyrrhizin, betulinic acid, soyasaponins, and phytosterols in licorice root, as well as providing a phonemic platform to characterize antioxidant properties, glycyrrhizin, and total phenolic content. This study also includes measuring the gene expression level and glycyrrhizin content in leaves and roots of control plants. The studied genes included squalene synthase (SQS1 and SQS2), β-amyrin synthase (bAS), lupeol synthase (LUS), cycloartenol synthase (CAS), β-amyrin 11-oxidase (CYP88D6), and β-amyrin 24-hydroxylase (CYP93E6). Our results revealed that all of the mentioned genes were upregulated following the stress condition with different transcription rates. The highest increase (12-fold) was observed for the expression of the LUS gene, which is related to the betulinic acid pathway. Also, the highest content of glycyrrhizin was observed at 72 h posttreatment, which was consistent with the upregulated transcription levels of the glycyrrhizin pathway genes especially SQS1 and CYP88D6 at the same time. Correlation and stepwise regression analysis proved the key role of SQS1 gene in the biosynthetic pathway of glycyrrhizin. Antioxidant activity and phenolic content also were increased following stress condition. A comparison between the expression levels of SQS1 and other genes involved in the production of glycyrrhizin, phytosterols, and soyasaponins revealed a similar transcription trend, which shows the gene expression in the roots was significantly higher than the leaves. In contrast, SQS2 and LUS genes displayed a higher expression in leaf tissues. The genes related to betulinic acid biosynthetic pathway exhibited an expression rate different from other triterpenoid pathway genes, which could be observed in the leaves and roots of control plants and the roots of salt-treated plants. Furthermore, results showed that these two SQS genes have different expression rates due to different plant tissues (roots and leaves) and stress conditions. Importantly, in contrast to previous reports, we detected the glycyrrhizin in leaf tissues. This result may indicate the presence of a different genetic background in native Iranian licorice germplasm. Keywords Gene expression . Glycyrrhiza glabra . Glycyrrhizin . Quantitative real-time PCR . Secondary metabolites Abbreviations bAS β-Amyrin synthase CAS Cycloartenol synthase CYP88D6 β-Amyrin 11-oxidase CYP93E6 β-Amyrin 24-hydroxylase FDP Farnesyl diphosphate GA3 Gibberellic acid HPLC High-performance liquid chromatography LUS Lupeol synthase MeJA Methyl jasmonate OSCs Oxidosqualene cyclases QRT-PCR Quantitative reverse transcription PCR SQS Squalene synthase

show abstract

Section: Effect Of Salt Stress On Glycyrrhizin Content In Rootssupporting

confidence: 84%

Triterpenoid gene expression and phytochemical content in Iranian licorice under salinity stress

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…GA shows a wide range of effects such as antibacterial, hepatoprotective, antiproliferative, antiallergic, and antiviral [98].…”

Section: Glycyrrhizin (Ga)mentioning

confidence: 99%

“…This molecule inhibits the chemotactic and pathogenic functions of HMGB1 by binding directly the A and B boxes [ 97 ]. GA shows a wide range of effects such as antibacterial, hepatoprotective, antiproliferative, antiallergic, and antiviral [ 98 ].…”

Section: Future Therapeutic Approachesmentioning

confidence: 99%

The Complex Relationship between Diabetic Retinopathy and High-Mobility Group Box: A Review of Molecular Pathways and Therapeutic Strategies

et al. 2020

View full text Add to dashboard Cite

High-mobility group box 1 (HMGB1) is a protein that is part of a larger family of non-histone nuclear proteins. HMGB1 is a ubiquitary protein with different isoforms, linked to numerous physiological and pathological pathways. HMGB1 is involved in cytokine and chemokine release, leukocyte activation and migration, tumorigenesis, neoangiogenesis, and the activation of several inflammatory pathways. HMGB1 is, in fact, responsible for the trigger, among others, of nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), toll-like receptor-4 (TLR-4), and vascular endothelial growth factor (VEGF) pathways. Diabetic retinopathy (DR) is a common complication of diabetes mellitus (DM) that is rapidly growing in number. DR is an inflammatory disease caused by hyperglycemia, which determines the accumulation of oxidative stress and cell damage, which ultimately leads to hypoxia and neovascularization. Recent evidence has shown that hyperglycemia is responsible for the hyperexpression of HMGB1. This protein activates numerous pathways that cause the development of DR, and HMGB1 levels are constantly increased in diabetic retinas in both proliferative and non-proliferative stages of the disease. Several molecules, such as glycyrrhizin (GA), have proven effective in reducing diabetic damage to the retina through the inhibition of HMGB1. The main focus of this review is the growing amount of evidence linking HMGB1 and DR as well as the new therapeutic strategies involving this protein.

show abstract

“…Considering these, the aim of this research was to study the metabolic pathway and biosynthetic genes functioning in diosgenin formation. The results could be applied to engineer the metabolic flux toward diosgenin biosynthesis in order to improve the accumulation of diosgenin in either the whole plant or the hairy root cultures of fenugreek, which have been considered as promising alternative platforms as compared to the wild plant . In the present study, the full‐length CDS of 14 genes involved in the diosgenin biosynthesis pathway were identified (Figures S5B–S15C).…”

Section: Discussionmentioning

confidence: 99%

New insights into diosgenin biosynthesis pathway and its regulation in Trigonella foenum‐graecum L.

Mohammadi

Mashayekh

Rashidi-Monfared

et al. 2019

Phytochemical Analysis

View full text Add to dashboard Cite

Introduction Throughout history, thousands of medicinal and aromatic plants have been widely utilised by people worldwide. Owing to them possessing of valuable compounds with little side effects in comparison with chemical drugs, herbs have been of interest to humans for a number of purposes. Diosgenin, driven from fenugreek, Trigonella foenum‐graecum L., has extensively drawn scientist's attention owing to having curable properties and being a precursor of steroid hormones synthesis. Nonetheless, complete knowledge about the biosynthesis pathway of this metabolite is still elusive. Objective In the present research, we isolated the full‐length CDS of 14 genes involving in diosgenin formation and measured their expression rate in various genotypes, which had illustrated different amount of diosgenin. Methodology The genes were successfully isolated, and functional motifs were also assessed using in silico approaches. Results Moreover, combining transcript and metabolite analysis revealed that there are many genes playing the role in diosgenin formation, some of which are highly influential. Among them, ∆24‐reductase, which converts cycloartenol to cycloartanol, is the first‐committed and rate‐limiting enzyme in this pathway. Additionally, no transcripts indicating to the presence or expression of lanosterol synthase were detected, contradicting the previous hypothesis about the biosynthetic pathway of diosgenin in fenugreek. Conclusion Considering all these, therefore, we propose the most possible pathway of diosgenin. This knowledge will then pave the way toward cloning the genes as well as engineering the diosgenin biosynthesis pathway.

show abstract

Metabolic Engineering of Glycyrrhizin Pathway by Over-Expression of Beta-amyrin 11-Oxidase in Transgenic Roots of Glycyrrhiza glabra

Cited by 15 publications

References 47 publications

Triterpenoid gene expression and phytochemical content in Iranian licorice under salinity stress

Triterpenoid gene expression and phytochemical content in Iranian licorice under salinity stress

The Complex Relationship between Diabetic Retinopathy and High-Mobility Group Box: A Review of Molecular Pathways and Therapeutic Strategies

New insights into diosgenin biosynthesis pathway and its regulation in Trigonella foenum‐graecum L.

Contact Info

Product

Resources

About