Saponin determination, expression analysis and functional characterization of saponin biosynthetic genes in Chenopodium quinoa leaves

Fiallos-Jurado, Jennifer; Pollier, Jacob; Moses, Tessa; Arendt, Philipp; Barriga-Medina, Noelia; Morillo, Eduardo; Arahana, Venancio; Torres, María de Lourdes; Goossens, Alain; León-Reyes, Antonio

doi:10.1016/j.plantsci.2016.05.015

Cited by 97 publications

(71 citation statements)

References 59 publications

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“…The LR reaction was performed between pENTR207[OsONS1] vector (previously created for N. benthamiana transient transformation) and pESC-URA-tHMG1-DEST (Fiallos-Jurado et al, 2016) to obtain pESC-URA (GAL10/tHMG1; GAL1/OsONS1) plasmid for expression in yeast. The yeast strain GIL77 (gal2,erg7, was transformed with this plasmid and precultured at 30°C in synthetic defined medium containing Glc (Clontech) and the uracil dropout (-U) supplement (Clontech) and supplemented with 20 mg mL 21 ergosterol and 13 mg mL 21 hemin (all from Sigma-Aldrich).…”

Section: Expression Of Osons1 In Yeastmentioning

confidence: 99%

A Single Oxidosqualene Cyclase Produces the Seco-Triterpenoid α-Onocerin

et al. 2017

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Section: Expression Of Osons1 In Yeastmentioning

confidence: 99%

A Single Oxidosqualene Cyclase Produces the Seco-Triterpenoid α-Onocerin

et al. 2017

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“…In this 291 study, we aimed to select suitable reference genes for diurnal expression studies to normalize expression data from two different quinoa accessions. We selected eight candidate reference genes (ACT, GAPDH, RAN-3, IDH-A, IDH-B, PTB, UBQ, TUB) based on C. quinoa transcriptome data [17] and literature survey [16,23,24] and tested them for stability during the course of a day.…”

Section: Discussionmentioning

confidence: 99%

“…quinoa to normalize the expression of saponin biosynthesis genes after methyl jasmonate treatment [23]. In beet, BBX19 acts together with BOLTING TIME CONTROL 1 (BTC1) in the circadian regulation of the flowering pathway [33] and shows diurnal regulation with an expression peak at dawn [20] .…”

Section: Discussionmentioning

confidence: 99%

“…First, we performed a selection of candidate genes based on a literature survey. We preferentially selected reference genes that have been validated in species of the Amaranthacea family [16,23,24]. Afterwards, we validated these genes with C. quinoa transcriptome data, where plants had been grown under abiotic stress conditions [17].…”

Section: Selection Of Candidate Genesmentioning

confidence: 99%

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Validation of suitable genes for normalization of diurnal gene expression studies inChenopodium quinoa

Maldonado-Taipe

Sarange

Schmöckel

et al. 2020

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Quinoa depicts high nutritional quality and abiotic stress resistance attracting strong interest in the last years. To unravel the function of candidate genes for agronomically relevant traits, studying their transcriptional activities by RT-qPCR is an important experimental approach. The accuracy of such experiments strongly depends on precise data normalization. To date, validation of potential candidate genes for normalization of diurnal expression studies has not been performed in C. quinoa. We selected eight candidate genes based on transcriptome data and literature survey, including conventionally used reference genes. We used three statistical algorithms (BestKeeper, geNorm and NormFinder) to test their stability and added further validation by a simulation-based strategy. We demonstrated that using different reference genes, including those top ranked by 1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20stability, causes significant differences among the resulting diurnal expression patterns, and that our novel approach overcomes failures related to stability-based selection of reference genes. Our results show that isocitrate dehydrogenase enzyme (IDH-A) and polypyrimidine tract-binding protein (PTB) are suitable genes to normalize diurnal expression data of two different quinoa accessions. The validated reference genes obtained in this study will improve the accuracy of RT-qPCR data normalization and facilitate gene expression studies in quinoa.

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“…dendroidea cDNA using the primers LdCAS _Fw and LdCAS _RV (Table 1). The obtained PCR fragment was Gateway TM recombined into the Gateway TM vector pDONR207 and the resulting entry clone was sequence verified and further recombined into the in-house generated destination vector pESC-URA-tHMG1-DEST [41] to yield pESC-URA-tHMG1-DEST[ GAL1/LdCAS ].…”

Section: Methodsmentioning

confidence: 99%

Cloning and Functional Characterization of Cycloartenol Synthase from the Red Seaweed Laurencia dendroidea

et al. 2016

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The red seaweed Laurencia dendroidea belongs to the Rhodophyta, a phylum of eukaryotic algae that is widely distributed across the oceans and that constitute an important source of bioactive specialized metabolites. Laurencia species have been studied since 1950 and were found to contain a plethora of specialized metabolites, mainly halogenated sesquiterpenes, diterpenes and triterpenes that possess a broad spectrum of pharmacological and ecological activities. The first committed step in the biosynthesis of triterpenes is the cyclization of 2,3-oxidosqualene, an enzymatic reaction carried out by oxidosqualene cyclases (OSCs), giving rise to a broad range of different compounds, such as the sterol precursors cycloartenol and lanosterol, or triterpene precursors such as cucurbitadienol and β-amyrin. Here, we cloned and characterized the first OSC from a red seaweed. The OSC gene was identified through mining of a L. dendroidea transcriptome dataset and subsequently cloned and heterologously expressed in yeast for functional characterization, which indicated that the corresponding enzyme cyclizes 2,3-oxidosqualene to the sterol precursor cycloartenol. Accordingly, the gene was named L. dendroidea cycloartenol synthase (LdCAS). A phylogenetic analysis using OSCs genes from plants, fungi and algae revealed that LdCAS grouped together with OSCs from other red algae, suggesting that cycloartenol could be the common product of the OSC in red seaweeds. Furthermore, profiling of L. dendroidea revealed cholesterol as the major sterol accumulating in this species, implicating red seaweeds contain a ‘hybrid’ sterol synthesis pathway in which the phytosterol precursor cycloartenol is converted into the major animal sterol cholesterol.

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Saponin determination, expression analysis and functional characterization of saponin biosynthetic genes in Chenopodium quinoa leaves

Cited by 97 publications

References 59 publications

A Single Oxidosqualene Cyclase Produces the Seco-Triterpenoid α-Onocerin

A Single Oxidosqualene Cyclase Produces the Seco-Triterpenoid α-Onocerin

Validation of suitable genes for normalization of diurnal gene expression studies inChenopodium quinoa

Cloning and Functional Characterization of Cycloartenol Synthase from the Red Seaweed Laurencia dendroidea

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