Illuminating the lineage-specific diversification of resin glycoside acylsugars in the morning glory (Convolvulaceae) family using computational metabolomics

Abstract

Acylsugars are a class of plant defense compounds produced across many distantly related families. Members of the horticulturally important morning glory (Convolvulaceae) family produce a diverse subclass of acylsugars called resin glycosides (RGs), which comprise oligosaccharide cores, hydroxyacyl chain(s), and decorating aliphatic and aromatic acyl chains. While many RG structures are characterized, the extent of structural diversity of this class in different genera and species is not known. In this study, we asked whether there has been lineage-specific diversification of RG structures in different Convolvulaceae species that may suggest diversification of the underlying biosynthetic pathways. Liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) was performed from root and leaf extracts of 26 species sampled in a phylogeny-guided manner. LC–MS/MS revealed thousands of peaks with signature RG fragmentation patterns with one species producing over 300 signals, mirroring the diversity in Solanaceae-type acylsugars. A novel RG from Dichondra argentea was characterized using nuclear magnetic resonance spectroscopy, supporting previous observations of RGs with open hydroxyacyl chains instead of closed macrolactone ring structures. Substantial lineage-specific differentiation in utilization of sugars, hydroxyacyl chains, and decorating acyl chains was discovered, especially among Ipomoea and Convolvulus-the two largest genera in Convolvulaceae. Adopting a computational, knowledge-based strategy, we further developed a high-recall workflow that successfully explained ∼72% of the MS/MS fragments, predicted the structural components of 11/13 previously characterized RGs, and partially annotated ∼45% of the RGs. Overall, this study improves our understanding of phytochemical diversity and lays a foundation for characterizing the evolutionary mechanisms underlying RG diversification.

References

[1] Fan P, Leong BJ, Last RL . Tip of the trichome: evolution of acylsugar metabolic diversity in Solanaceae. Curr Opin Plant Biol. 2019; 49: 8-16.

[2] Landis JB, Miller CM, Broz AK et al. Migration through a major Andean ecogeographic disruption as a driver of genetic and phenotypic diversity in a wild tomato species. Mol Biol Evol. 2021; 8: 3202-19.

[3] Leong BJ, Hurney SM, Fiesel PD et al. Specialized metabolism in a nonmodel nightshade: trichome acylinositol biosynthesis. Plant Physiol. 2020; 183: 915-24.

[4] Leong BJ, Lybrand DB, Lou YR et al. Evolution of metabolic novelty: a trichome-expressed invertase creates specialized metabolic diversity in wild tomato. Sci Adv. 2019; 5: eaaw3754.

[5] Moghe GD, Leong BJ, Hurney S et al. Evolutionary routes to biochemical innovation revealed by integrative analysis of a plant-defense related specialized metabolic pathway. eLife. 2017; 136937. https://doi.org/10.7554/eLife.28468.

[6] Asai T, Hara N, Fujimoto Y . Fatty acid derivatives and dammarane triterpenes from the glandular trichome exudates of Ibicella lutea and Proboscidea louisiana. Phytochemistry. 2010; 71: 877-94.

[7] Asai T, Fujimoto Y . Cyclic fatty acyl glycosides in the glandular trichome exudate of Silene gallica. Phytochemistry. 2010; 71: 1410-7.

[8] Asai T, Nakamura Y, Hirayama Y et al. Cyclic glycolipids from glandular trichome exudates of Cerastium glomeratum. Phytochemistry. 2012; 82: 149-57.

[9] Asai T, Sakai T, Ohyama K et al. N-Octyl α-L-Rhamnopyranosyl-(1→2)-β-D-glucopyranoside derivatives from the glandular trichome exudate of Geranium carolinianum . Chem Pharm Bull (Tokyo). 2011; 59: 747-52.

[10] Anaya AL, Calera MR, Mata R et al. Allelopathic potential of compounds isolated from Ipomoea tricolor cav. (Convolvulaceae). J Chem Ecol. 1990; 16: 2145-52.

[11] Pereda-Miranda R, Mata R, Anaya AL et al. Tricolorin A, major phytogrowth inhibitor from Ipomoea tricolor . J Nat Prod. 1993; 56: 571-82.

[12] Eich, E. Solanaceae and Convolvulaceae: Secondary Metabolites. Biosynthesis, Chemotaxonomy, Biological and Economic Significance (A Handbook) . Berlin: Springer, 2008, 525-82.

[13] Pereda-Miranda R, Rosas-Ramírez D, Castañeda-Gómez J . Resin glycosides from the morning glory family. Prog Chem Org Nat Prod. 2010; 92: 77-153.

[14] Harrison H, Peterson J, Jackson D et al. Periderm resin glycoside contents of sweetpotato clones and their biological activity. Allelopathy J. 2003; 12: 53-60.

[15] Jackson DM, Peterson JK. Sublethal effects of resin glycosides from the periderm of sweetpotato storage roots on Plutella xylostella (Lepidoptera: Plutellidae). J Econ Entomol. 2000;93:388-93.

[16] Stefanović S, Austin DF, Olmstead RG . Classification of Convolvulaceae: a phylogenetic approach. Syst Bot. 2003; 28: 791-806.

[17] Stefanović S, Krueger L, Olmstead RG . Monophyly of the Convolvulaceae and circumscription of their major lineages based on DNA sequences of multiple chloroplast loci. Am J Bot. 2002; 89: 1510-22.

[18] Song W, Wang W, Zhang S et al. Multidrug resistance-reversal effects of resin glycosides from Dichondra repens. Bioorg Med Chem Lett. 2015; 25: 795-8.

[19] Du X-M, Kohinata K, Kawasaki T et al. Components of the ether-insoluble resin glycoside-like fraction from Cuscuta chinensis. Phytochemistry. 1998; 48: 843-50.

[20] Stefanović S, Olmstead RG . Testing the phylogenetic position of a parasitic plant (Cuscuta, Convolvulaceae, Asteridae): Bayesian inference and the parametric bootstrap on data drawn from three genomes. Syst Biol. 2004; 53: 384-99.

[21] Kim J, Kang K, Gonzales-Vigil E et al. Striking natural diversity in glandular trichome acylsugar composition is shaped by variation at the Acyltransferase2 locus in the wild tomato Solanum habrochaites . Plant Physiol. 2012; 160: 1854-70.

[22] Schilmiller AL, Moghe GD, Fan P et al. Functionally divergent alleles and duplicated loci encoding an acyltransferase contribute to acylsugar metabolite diversity in Solanum trichomes. Plant Cell. 2015; 27: 1002-17.

[23] Ning J, Moghe GD, Leong B et al. A feedback insensitive isopropylmalate synthase affects acylsugar composition in cultivated and wild tomato. Plant Physiol. 2015; 169: 1821-35.

[24] Gaspar EMM . New pentasaccharide macrolactone from the European Convolvulaceae Calystegia soldanella. Tetrahedron Lett. 1999; 40: 6861-4.

[25] Takigawa A, Muto H, Kabata K et al. Calysolins I-IV, resin glycosides from Calystegia soldanella . J Nat Prod. 2011; 74: 2414-9.

[26] Ding W, Jiang Z-H, Wu P et al. Resin glycosides from the aerial parts of Operculina turpethum. Phytochemistry. 2012; 81: 165-74.

[27] Lira-Ricárdez J, Pereda-Miranda R, Castañeda-Gomez J et al. Resin glycosides from the roots of Operculina macrocarpa (Brazilian jalap) with purgative activity. J Nat Prod. 2019; 82: 1664-77.

[28] Degtyarenko K, de Matos P, Ennis M et al. ChEBI: a database and ontology for chemical entities of biological interest. Nucleic Acids Res. 2008; 36: D344-50.

[29] Noda N, Yoda S, Kawasaki T et al. Resin glycosides. XV. Simonins I-V, ether-soluble resin glycosides (Jalapins) from the roots of Ipomoea batatas (cv. Simon). Chem Pharm Bull (Tokyo). 1992; 40: 3163-8.

[30] Barnes CC, Smalley MK, Manfredi KP et al. Characterization of an anti-tuberculosis resin glycoside from the prairie medicinal plant Ipomoea leptophylla. J Nat Prod. 2003; 66: 1457-62.

[31] Castañeda-Gómez J, Pereda-Miranda R . Resin glycosides from the herbal drug jalap (Ipomoea purga). J Nat Prod. 2011; 74: 1148-53.

[32] Ono M, Yuhura N, Shimohora T et al. Calyhedins I-VI: resin glycosides from the rhizomes of Calystegia hederacea . Phytochemistry. 2021; 190: 112888.

[33] Fan B-Y, Lu Y, Yin H et al. Arvensic acids A-D, novel heptasaccharide glycosidic acids as the alkaline hydrolysis products of crude resin glycosides from Convolvulus arvensis. Fitoterapia. 2018; 131: 209-14.

[34] Lu Y, He Y, Yang M et al. Arvensic acids K and L, components of resin glycoside fraction from Convolvulus arvensis. Nat Prod Res. 2021; 35: 2303-7.

[35] Cruz-Morales S, Castañeda-Gomez J, Rosas-Ramirez D et al. Resin glycosides from Ipomoea alba seeds as potential chemosensitizers in breast carcinoma cells. J Nat Prod. 2016; 79: 3093-104.

[36] Sumner LW, Amberg A, Barret D et al. Proposed minimum reporting standards for chemical analysis Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI). Metabolomics. 2007; 3: 211-21.

[37] Demarque D, Crotti AE, Vessecchi R et al. Fragmentation reactions using electrospray ionization mass spectrometry: an important tool for the structural elucidation and characterization of synthetic and natural products. Nat Prod Rep. 2016; 33: 432-55.

[38] Park Y-K, Nicaud J-M . Metabolic engineering for unusual lipid production in Yarrowia lipolytica. Microorganisms. 2020; 8: 1937.

[39] Kroumova AB, Wagner GJ . Different elongation pathways in the biosynthesis of acyl groups of trichome exudate sugar esters from various solanaceous plants. Planta. 2003; 216: 1013-21.

[40] Jansen GA, Wanders RJA . Alpha-oxidation. Biochim Biophys Acta. 2006; 1763: 1403-12.

[41] Kruse LH, Weigle A, Martinez-Gomez J et al. Ancestral class-promiscuity as a driver of functional diversity in the BAHD acyltransferase family in plants. bioRxiv 2020.11.18.385815. 2020. https://doi.org/10.1101/2020.11.18.385815.

[42] Murayama K, Kato-Murayama M, Sato T et al. Anthocyanin 5,3^′-aromatic acyltransferase from Gentiana triflora, a structural insight into biosynthesis of a blue anthocyanin . Phytochemistry. 2021; 186: 112727.

[43] Afendi FM, Okada T, Yamazaki M et al. KNApSAcK family databases: integrated metabolite-plant species databases for multifaceted plant research. Plant Cell Physiol. 2012; 53: e1-1.

[44] Kind T, Liu KH, Lee DY et al. LipidBlast - in-silico tandem mass spectrometry database for lipid identification. Nat Methods. 2013; 10: 755-8.

[45] Dührkop K, Nothais LF, Fleischauer M et al. Systematic classification of unknown metabolites using high-resolution fragmentation mass spectra. Nat Biotechnol. 2021; 39: 462-71.

[46] Bennett AA, Mahood EH, Fan K et al. Untargeted metabolomics of purple and orange-fleshed sweet potatoes reveals a large structural diversity of anthocyanins and flavonoids. Sci Rep. 2021; 11: 1-13.

[47] Moghe GD, Kruse LH . The study of plant specialized metabolism: challenges and prospects in the genomics era. Am J Bot. 2018; 105: 959-62.

[48] Camacho C, Coulouris G, Avagyan V et al. BLAST+: architecture and applications. BMC Bioinformatics. 2009; 10: 421.

[49] Katoh K, Misawa K, Kuma K et al. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res. 2002; 30: 3059-66.

[50] Nguyen L-T, Schmidt HA, von Haeseler A et al. IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Mol Biol Evol. 2015; 32: 268-74.

[51] Kalyaanamoorthy S, Minh BQ, Wong TKF et al. ModelFinder: fast model selection for accurate phylogenetic estimates. Nat Methods. 2017; 14: 587-9.

[52] Guindon S, Dufayard JF, Lefort V et al. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol. 2010; 59: 307-21.

[53] Tsugawa H, Cajka T, Kind T et al. MS-DIAL: data-independent MS/MS deconvolution for comprehensive metabolome analysis. Nat Methods. 2015; 12: 523-6.

[54] Wang M, Carver JJ, Phelan VV et al. Sharing and community curation of mass spectrometry data with global natural products social molecular networking. Nat Biotechnol. 2016; 34: 828-37.

[55] Shannon P, Markeil A, Ozier O et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003; 13: 2498-504.