Production of plant bioactive triterpenoid saponins: from metabolites to genes and back

Magedans, Yve Verônica da Silva; Phillips, Michael A.; Fett-Neto, Arthur Germano

doi:10.1007/s11101-020-09722-4

Cited by 31 publications

(39 citation statements)

References 180 publications

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“…The results of this study have widened the knowledge on the effect of biotic elicitors, particularly selected phytohormones, on the biosynthesis of triterpenoid saponins in HRCs. Saponin production in plant in vitro cultures is currently a subject of intensive research [ 40 , 41 ]. However, to avoid any generalizations, it should be emphasized that the final effect of any elicitation strongly depends on various factors as the plant species, the type of culture, elicitor specificity, elicitor concentration, duration of exposure, and timing of elicitor addition.…”

Section: Discussionmentioning

confidence: 99%

Effect of Ethylene and Abscisic Acid on Steroid and Triterpenoid Synthesis in Calendula officinalis Hairy Roots and Saponin Release to the Culture Medium

Markowski

Alsoufi

Szakiel

et al. 2022

Plants

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Phytohormones (plant growth regulators) can be applied as efficient elicitors to enhance the productivity of plant in vitro cultures, due to their significance in regulating the plant metabolism and strong influence on plant defense responses. In the present study, the effects of exogenous ethylene (ETY, applied in the form of ethephon as an ethylene-generating agent) and abscisic acid (ABA) on the synthesis of triterpenoids and steroids in Calendula officinalis hairy roots were investigated. ABA appeared to be an efficient elicitor of the biosynthesis of triterpenoid oleanolic acid (almost two-fold) and the release of its glycosides (saponins) to the culture medium (up to 6.6-fold). ETY had only a slight effect on triterpenoid metabolism; instead, it strongly influenced steroid metabolism, leading to profound modifications of the quantitative profiles of these compounds, particularly the ratio of stigmasterol to sitosterol. Both the applied phytohormones influenced the interplay between steroid and triterpenoid biosynthetic pathways, revealing the symptoms of their competition.

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Section: Discussionmentioning

confidence: 99%

Effect of Ethylene and Abscisic Acid on Steroid and Triterpenoid Synthesis in Calendula officinalis Hairy Roots and Saponin Release to the Culture Medium

Markowski

Alsoufi

Szakiel

et al. 2022

Plants

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show abstract

“…Nevertheless, studies on the competition between sterols and triterpenoids were crucial for revealing the pathway of their biosynthesis, understanding the role of particular enzymes, and discovering the possibility of gene regulation. Currently, adventitious and hairy roots of many plant species can be considered not only as an important and efficient tool for controlled production of valuable natural products, but also as suitable experimental models for investigations on regulatory mechanisms of plant development or various aspects of the specific interplay between general and specialized metabolism [ 88 , 89 ].…”

Section: Discussionmentioning

confidence: 99%

Enhancement of Phytosterol and Triterpenoid Production in Plant Hairy Root Cultures—Simultaneous Stimulation or Competition?

Rogowska

Szakiel

2021

Plants

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Plant in vitro cultures, including hairy roots, can be applied for controlled production of valuable natural products, such as triterpenoids and sterols. These compounds originate from the common precursor squalene. Sterols and triterpenoids distinctly differ in their functions, and the 2,3-oxidosqualene cyclization step is often regarded as a branch point between primary and secondary (more aptly: general and specialized) metabolism. Considering the crucial role of phytosterols as membrane constituents, it has been postulated that unconstrained biosynthesis of triterpenoids can occur when sterol formation is already satisfied, and these compounds are no longer needed for cell growth and division. This hypothesis seems to follow directly the growth-defense trade-off plant dilemma. In this review, we present some examples illustrating the specific interplay between the two divergent pathways for sterol and triterpenoid biosynthesis appearing in root cultures. These studies were significant for revealing the steps of the biosynthetic pathway, understanding the role of particular enzymes, and discovering the possibility of gene regulation. Currently, hairy roots of many plant species can be considered not only as an efficient tool for production of phytochemicals, but also as suitable experimental models for investigations on regulatory mechanisms of plant metabolism.

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“…Also, the development of plants with modified saponin profiles could yield further understanding of their biosynthetic route and regulation mechanisms. The metabolic engineering of both the early and late stages of the saponin biosynthetic pathway has been the topic of several reviews (Yendo et al, 2010 ; Lambert et al, 2011 ; da Silva Magedans et al, 2020 ). The major requirement for the successful engineering of specific saponins is the knowledge of the genes/enzymes involved in their biosynthesis.…”

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9-Mediated Targeted Mutagenesis of CYP93E2 Modulates the Triterpene Saponin Biosynthesis in Medicago truncatula

et al. 2021

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In the Medicago genus, triterpene saponins are a group of bioactive compounds extensively studied for their different biological and pharmaceutical properties. In this work, the CRISPR/Cas9-based approach with two single-site guide RNAs was used in Medicago truncatula (barrel medic) to knock-out the CYP93E2 and CYP72A61 genes, which are responsible for the biosynthesis of soyasapogenol B, the most abundant soyasapogenol in Medicago spp. No transgenic plants carrying mutations in the target CYP72A61 gene were recovered while fifty-two putative CYP93E2 mutant plant lines were obtained following Agrobacterium tumefaciens-mediated transformation. Among these, the fifty-one sequenced plant lines give an editing efficiency of 84%. Sequencing revealed that these lines had various mutation patterns at the target sites. Four T0 mutant plant lines were further selected and examined for their sapogenin content and plant growth performance under greenhouse conditions. The results showed that all tested CYP93E2 knock-out mutants did not produce soyasapogenols in the leaves, stems and roots, and diverted the metabolic flux toward the production of valuable hemolytic sapogenins. No adverse influence was observed on the plant morphological features of CYP93E2 mutants under greenhouse conditions. In addition, differential expression of saponin pathway genes was observed in CYP93E2 mutants in comparison to the control. Our results provide new and interesting insights into the application of CRISPR/Cas9 for metabolic engineering of high-value compounds of plant origin and will be useful to investigate the physiological functions of saponins in planta.

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Production of plant bioactive triterpenoid saponins: from metabolites to genes and back

Cited by 31 publications

References 180 publications

Effect of Ethylene and Abscisic Acid on Steroid and Triterpenoid Synthesis in Calendula officinalis Hairy Roots and Saponin Release to the Culture Medium

Effect of Ethylene and Abscisic Acid on Steroid and Triterpenoid Synthesis in Calendula officinalis Hairy Roots and Saponin Release to the Culture Medium

Enhancement of Phytosterol and Triterpenoid Production in Plant Hairy Root Cultures—Simultaneous Stimulation or Competition?

CRISPR/Cas9-Mediated Targeted Mutagenesis of CYP93E2 Modulates the Triterpene Saponin Biosynthesis in Medicago truncatula

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