Abstract
Erysimum cheiranthoides L (Brassicaceae; wormseed wallflower) accumulates not only glucosinolates, which are characteristic of the Brassicaceae, but also abundant and diverse cardenolides. These steroid toxins, primarily glycosylated forms of digitoxigenin, cannogenol, and strophanthidin, inhibit the function of essential Na+/K+-ATPases in animal cells. We screened a population of 659 ethylmethanesulfonate-mutagenized E. cheiranthoides plants to identify isolates with altered cardenolide profiles. One mutant line exhibited 66% lower cardenolide content, resulting from greatly decreased cannogenol and strophanthidin glycosides, partially compensated for by increases in digitoxigenin glycosides. This phenotype was likely caused by a single-locus recessive mutation, as evidenced by a wildtype phenotype of F1 plants from a backcross, a 3:1 wildtype:mutant segregation in the F2 generation, and genetic mapping of the altered cardenolide phenotype to one position in the genome. The mutation created a more even cardenolide distribution, decreased the average cardenolide polarity, but did not impact most glucosinolates. Growth of generalist herbivores from two feeding guilds, Myzus persicae Sulzer (Hemiptera: Aphididae; green peach aphid) and Trichoplusia ni Hübner (Lepidoptera: Noctuidae; cabbage looper), was decreased on the mutant line compared to wildtype. Both herbivores accumulated cardenolides in proportion to the plant content, with T. ni accumulating higher total concentrations than M. persicae. Helveticoside, a relatively abundant cardenolide in E. cheiranthoides, was not detected in M. persicae feeding on these plants. Our results support the hypothesis that increased digitoxigenin glycosides provide improved protection against M. persicae and T. ni, despite an overall decrease in cardenolide content of the mutant line.
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References
Agrawal AA, Petschenka G, Bingham RA, Weber MG, Rasmann S (2012) Toxic cardenolides: chemical ecology and coevolution of specialized plant-herbivore interactions. New Phytol 194:28–45. https://doi.org/10.1111/j.1469-8137.2011.04049.x
Berenbaum MR, Nitao JK, Zangerl AR (1991) Adaptive significance of furanocoumarin diversity in Pastinaca sativa (Apiaceae). J Chem Ecol 17:207–215. https://doi.org/10.1007/BF00994434
Christmann J, Kreis W, Reinhard E (1993) Uptake, transport and storage of cardenolides in foxglove - cardenolide sinks and occurrence of cardenolides in the sieve tubes of Digitalis lanata. Botanica Acta 106:419–427. https://doi.org/10.1111/j.1438-8677.1993.tb00769.x
Dimock MB, Renwick JA, Radke CD, Sachdev-Gupta K (1991) Chemical constituents of an unacceptable crucifer, Erysimum cheiranthoides, deter feeding by Pieris rapae. J Chem Ecol 17:525–533. https://doi.org/10.1007/BF00982123
Douglas AE, Minto LB, Wilkinson TL (2001) Quantifying nutrient production by the microbial symbionts in an aphid. J Exp Biol 204:349–358
Dzimiri N, Fricke U, Klaus W (1987) Influence of derivation on the lipophilicity and inhibitory actions of cardiac glycosides on myocardial Na+-K+-ATPase. Br J Pharmacol 91:31–38. https://doi.org/10.1111/j.1476-5381.1987.tb08980.x
Fahey JW, Zalcmann AT, Talalay P (2001) The chemical diversity and distribution of glucosinolates and isothiocyanates among plants. Phytochemistry 56:5–51
Farr CD, Burd C, Tabet MR, Wang X, Welsh WJ, Ball WJ Jr (2002) Three-dimensional quantitative structure-activity relationship study of the inhibition of Na(+),K(+)-ATPase by cardiotonic steroids using comparative molecular field analysis. Biochemistry 41:1137–1148. https://doi.org/10.1021/bi011511g
Frerigmann H, Gigolashvili T (2014) MYB34, MYB51, and MYB122 distinctly regulate indolic glucosinolate biosynthesis in Arabidopsis thaliana. Mol plant 7:814–828. https://doi.org/10.1093/mp/ssu004
Groen SC, LaPlante ER, Alexandre NM, Agrawal AA, Dobler S, Whiteman NK (2017) Multidrug transporters and organic anion transporting polypeptides protect insects against the toxic effects of cardenolides Insect. Biochem Mol Biol 81:51–61. https://doi.org/10.1016/j.ibmb.2016.12.008
Jander G, Baerson SR, Hudak JA, Gonzalez KA, Gruys KJ, Last RL (2003) Ethylmethanesulfonate saturation mutagenesis in Arabidopsis to determine frequency of herbicide resistance. Plant Physiol 131:139–146
Karageorgi M et al (2019) Genome editing retraces the evolution of toxin resistance in the monarch butterfly. Nature 574:409–412. https://doi.org/10.1038/s41586-019-1610-8
Kim JH, Jander G (2007) Myzus persicae (green peach aphid) feeding on Arabidopsis induces the formation of a deterrent indole glucosinolate. Plant J 49:1008–1019
Kim JH, Lee BW, Schroeder FC, Jander G (2008) Identification of indole glucosinolate breakdown products with antifeedant effects on Myzus persicae (green peach aphid). Plant J 54:1015–1026
Li H (2011) A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data. Bioinformatics 27:2987–2993. https://doi.org/10.1093/bioinformatics/btr509
Li H, Durbin R (2009) Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 25:1754–1760. https://doi.org/10.1093/bioinformatics/btp324
Li H et al (2009) The sequence alignment/map format and SAMtools. Bioinformatics 25:2078–2079. https://doi.org/10.1093/bioinformatics/btp352
Müller R, De Vos M, Sun JY, Sønderby IE, Halkier BA, Wittstock U, Jander G (2010) Differential effects of indole and aliphatic glucosinolates on lepidopteran herbivores. J Chem Ecol 36:905–913
Paula S, Tabet MR, Ball WJ Jr (2005) Interactions between cardiac glycosides and sodium/potassium-ATPase: three-dimensional structure-activity relationship models for ligand binding to the E2-pi form of the enzyme versus activity inhibition. Biochemistry 44:498–510. https://doi.org/10.1021/bi048680w
Petschenka G, Fei CS, Araya JJ, Schroder S, Timmermann BN, Agrawal AA (2018) Relative selectivity of plant cardenolides for Na(+)/K(+)-ATPases from the monarch butterfly and non-resistant insects. Front Plant Sci 9:1424. https://doi.org/10.3389/fpls.2018.01424
Petschenka G, Pick C, Wagschal V, Dobler S (2013) Functional evidence for physiological mechanisms to circumvent neurotoxicity of cardenolides in an adapted and a non-adapted hawk-moth species. Proc Biol Sci 280:20123089. https://doi.org/10.1098/rspb.2012.3089
R Core Team (2020) R: A language and environment for statistical computing. Foundation for Statistical Computing. https://www.R-project.org/. 2020
Ramsey JS, Elzinga DA, Sarkar P, Xin Y-R, Ghanim M, Jander G (2014) Adaptation to nicotine feeding in Myzus persicae. J Chem Ecol 40:869–877. https://doi.org/10.1007/s10886-014-0482-5
Ramsey JS et al (2007) Genomic resources for Myzus persicae: EST sequencing, SNP identification, and microarray design. BMC Genomics 8:423
Rasmann S, Agrawal AA (2011) Latitudinal patterns in plant defense: evolution of cardenolides, their toxicity and induction following herbivory. Ecol Lett 14:476–483. https://doi.org/10.1111/j.1461-0248.2011.01609.x
Renwick JA, Radke CD, Sachdev-Gupta K (1989) Chemical constituents of Erysimum cheiranthoides deterring oviposition by the cabbage butterfly Pieris rapae. J Chem Ecol 15:2161–2169. https://doi.org/10.1007/BF01014106
Sachdev-Gupta K, Radke C, Renwick JA, Dimock MB (1993) Cardenolides from Erysimum cheiranthoides: Feeding deterrents to Pieris rapae larvae. J Chem Ecol 19:1355–1369. https://doi.org/10.1007/BF00984881
Sachdev-Gupta K, Renwick JA, Radke CD (1990) Isolation and identification of oviposition deterrents to cabbage butterfly, Pieris rapae, from Erysimum cheiranthoides. J Chem Ecol 16:1059–1067. https://doi.org/10.1007/BF01021010
Salazar D, Lokvam J, Mesones I, Vasquez Pilco M, Ayarza Zuniga JM, de Valpine P, PVA F (2018) Origin and maintenance of chemical diversity in a species-rich tropical tree lineage. Nat Ecol Evol 2:983–990. https://doi.org/10.1038/s41559-018-0552-0
Schonfeld W et al (1985) The lead structure in cardiac glycosides is 5 beta, 14 beta-androstane-3 beta 14-diol Naunyn Schmiedebergs. Arch Pharmacol 329:414–426. https://doi.org/10.1007/bf00496377
Smith B, Wilson JB (1996) A consumer's guide to evenness indices Oikos 76:70-82 doi:Doi. https://doi.org/10.2307/3545749
Wickham H, Francois R, Henry L, Müller K (2020) Dplyr: a grammar of data manipulation. https://CRAN.R-project.org/package=dplyr
Wittstock U, Agerbirk N, Stauber EJ, Olsen CE, Hippler M, Mitchell-Olds T, Gershenzon J, Vogel H (2004) Successful herbivore attack due to metabolic diversion of a plant chemical defense. Proc Natl Acad Sci U S A 101:4859–4864
Zhao Y et al (2002) Trp-dependent auxin biosynthesis in Arabidopsis: involvement of cytochrome P450s CYP79B2 and CYP79B3. Genes Dev 16:3100–3112
Zhu Y, Mang HG, Sun Q, Qian J, Hipps A, Hua J (2012) Gene discovery using mutagen-induced polymorphisms and deep sequencing: application to plant disease resistance. Genetics 192:139–146. https://doi.org/10.1534/genetics.112.141986
Züst T, Agrawal AA (2015) Population growth and sequestration of plant toxins along a gradient of specialization in four aphid species on the common milkweed Asclepias syriaca. Functional Ecology 186:E1–E15
Züst T, Mirzaei M, Jander G (2018) Erysimum cheiranthoides, an ecological research system with potential as a genetic and genomic model for studying cardiac glycoside biosynthesis. Phytochem Rev 17:1239–1251
Züst T, Petschenka G, Hastings AP, Agrawal AA (2019) Toxicity of milkweed leaves and latex: Chromatographic quantification versus biological activity of cardenolides in 16 Asclepias species. J Chem Ecol 45:50–60. https://doi.org/10.1007/s10886-018-1040-3
Züst T et al (2020) Rapid and independent evolution of ancestral and novel defenses in a genus of toxic plants (Erysimum, Brassicaceae). eLife 9:e51712. https://doi.org/10.7554/eLife.51712
Acknowledgements
We thank the Biotechnology Resource Center and Bioinformatics Facility at Cornell University for assistance in developing data analysis scripts.
Funding
This research was funded by US National Science Foundation awards 1907491 to AAA and 1645256 to GJ and AAA, Swiss National Science Foundation grant PZ00P3–161472 to TZ, and a Triad Foundation grant to GJ.
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Mirzaei, M., Züst, T., Younkin, G.C. et al. Less Is More: a Mutation in the Chemical Defense Pathway of Erysimum cheiranthoides (Brassicaceae) Reduces Total Cardenolide Abundance but Increases Resistance to Insect Herbivores. J Chem Ecol 46, 1131–1143 (2020). https://doi.org/10.1007/s10886-020-01225-y
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DOI: https://doi.org/10.1007/s10886-020-01225-y