High-Throughput Screening and Characterization of a High-Density Soybean Mutant Library Elucidate the Biosynthesis Pathway of Triterpenoid Saponins

Krishnamurthy, Panneerselvam; Fujisawa, Yukiko; Takahashi, Yuya; Abe, Hanako; Yamane, Kentaro; Mukaiyama, Kyosuke; Son, Hae Reon; Hiraga, Susumu; Kaga, Akito; Anai, Toyoaki; Tsukamoto, Chigen; Ishimoto, Masao

doi:10.1093/pcp/pcz025

Cited by 16 publications

(26 citation statements)

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“…The in vivo activity of UGT73P2 was never characterized before. We thus identified missense mutations causing various amino acid changes in PSPG box or its proximal region of UGT73P2 but none of them affected the soyasaponin composition [22]. We reckon that the in vivo characterization of UGT73P2 is essential to validate its function.…”

Section: Duplication History and Functional Divergence Of Triterpene mentioning

confidence: 97%

Reconstruction of the Evolutionary Histories of UGT Gene Superfamily in Legumes Clarifies the Functional Divergence of Duplicates in Specialized Metabolism

Krishnamurthy

Tsukamoto

Ishimoto

2020

IJMS

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Plant uridine 5′-diphosphate glycosyltransferases (UGTs) influence the physiochemical properties of several classes of specialized metabolites including triterpenoids via glycosylation. To uncover the evolutionary past of UGTs of soyasaponins (a group of beneficial triterpene glycosides widespread among Leguminosae), the UGT gene superfamily in Medicago truncatula, Glycine max, Phaseolus vulgaris, Lotus japonicus, and Trifolium pratense genomes were systematically mined. A total of 834 nonredundant UGTs were identified and categorized into 98 putative orthologous loci (POLs) using tree-based and graph-based methods. Major key findings in this study were of, (i) 17 POLs represent potential catalysts for triterpene glycosylation in legumes, (ii) UGTs responsible for the addition of second (UGT73P2: galactosyltransferase and UGT73P10: arabinosyltransferase) and third (UGT91H4: rhamnosyltransferase and UGT91H9: glucosyltransferase) sugars of the C-3 sugar chain of soyasaponins were resulted from duplication events occurred before and after the hologalegina–millettoid split, respectively, and followed neofunctionalization in species-/ lineage-specific manner, and (iii) UGTs responsible for the C-22-O glycosylation of group A (arabinosyltransferase) and DDMP saponins (DDMPtransferase) and the second sugar of C-22 sugar chain of group A saponins (UGT73F2: glucosyltransferase) may all share a common ancestor. Our findings showed a way to trace the evolutionary history of UGTs involved in specialized metabolism.

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Section: Duplication History and Functional Divergence Of Triterpene mentioning

confidence: 97%

Reconstruction of the Evolutionary Histories of UGT Gene Superfamily in Legumes Clarifies the Functional Divergence of Duplicates in Specialized Metabolism

Krishnamurthy

Tsukamoto

Ishimoto

2020

IJMS

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“…Gene IDs were obtained from NCBI (https://www.ncbi.nlm.nih.gov/). (B) Soyasaponin biosynthetic genes listed in Krishnamurthy et al (2019) and Sundaramoorthy et al (2019). The shaded areas show nighttime.…”

Section: Resultsmentioning

confidence: 99%

“…The expression levels of genes encoding β-Amyrin synthase ( BAS ), cytochrome P450 ( CYP ) for triterpenes, and UDP-glucuronosyltransferase ( UGT ) involved in soyasaponin biosynthesis (Sundaramoothy et al, 2019; Krishnamurthy et al, 2019) increased from ZT18 (0:00 am) to ZT0 (6:00 am) and decreased at ZT6, which was an inverse pattern to that of isoflavone biosynthesis.…”

Section: Resultsmentioning

confidence: 99%

“…Gene Ontology (GO) enrichment analysis of gene set was performed using the SoyBase GO Term Enrichment Tool (http://www.soybase.org) in accordance with a diurnal pattern of interests. Gene IDs for isoflavone and soyasaponin biosynthesis, ABC and MATE transporters and ICHG were collected based on the procedures described in previous literatures (Yoo et al, 2013; Liu et al, 2016; Ahmad et al, 2017; Krishnamurthy et al, 2019; Mishra et al, 2019; Sundaramoothy et al, 2019). The networks were constructed using the network visualization software Cytoscape (v. 3.7.2; Shannon et al, 2003).…”

Section: Methodsmentioning

confidence: 99%

“…Biochemical analysis has suggested the involvement of ABC-type transporters and apoplast-localized isoflavone conjugate-hydrolyzing beta-glucosidase (ICHG) in isoflavone aglycone secretion to the rhizosphere (Suzuki et al, 2006; Sugiyama et al, 2007), although these processes have not been genetically characterized (Figure 1A). In contrast to isoflavones, the secretory mechanism involving the transporters that are responsible for the vacuolar accumulation and secretion to the rhizosphere are still unknown for soyasaponins, whereas the soyasaponin biosynthetic pathway in soybeans has been intensively studied and characterized (Sundaramoorthy et al, 2019; Krishnamurthy et al, 2019) (Figure 1B).…”

Section: Introductionmentioning

confidence: 99%

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