Skip to main content
SpringerLink
Log in

The Role of the Glucosinolate-Myrosinase System in Mediating Greater Resistance of Barbarea verna than B. vulgaris to Mamestra brassicae Larvae

  • Published:
Journal of Chemical Ecology Aims and scope Submit manuscript

Abstract

We investigated the influences of two structurally similar glucosinolates, phenethylglucosinolate (gluconasturtiin, NAS) and its (S)-2-hydroxyl derivative glucobarbarin (BAR), as well as their hydrolysis products on larvae of the generalist Mamestra brassicae (Lepidoptera: Noctuidae). Previous results suggested a higher defensive activity of BAR than NAS based on resistance toward M. brassicae larvae of natural plant genotypes of Barbarea vulgaris R. Br. (Brassicaceae) dominated by BAR. In the present study, the hypothesis of a higher defensive activity of BAR than NAS was tested by comparing two Barbarea species similarly dominated either by BAR or by NAS and by testing effects of isolated BAR and NAS on larval survival and feeding preferences. Larvae reared on leaf disks of B. verna (Mill.) Asch. had a lower survival than those reared on B. vulgaris P- and G-chemotypes. Leaves of B. verna were dominated by NAS, whereas B. vulgaris chemotypes were dominated by BAR or its epimer. In addition, B. verna leaves showed a threefold higher activity of the glucosinolate-activating myrosinase enzymes. The main product of NAS from breakdown by endogenous enzymes including myrosinases (“autolysis”) in B. verna leaves was phenethyl isothiocyanate, while the main products of BAR in autolyzed B. vulgaris leaves were a cyclized isothiocyanate product, namely an oxazolidine-2-thione, and a downstream metabolite, an oxazolidin-2-one. The glucosinolates BAR and NAS were isolated and offered to larvae on disks of cabbage. Both glucosinolates exerted similar negative effects on larval survival but effects of NAS tended to be more detrimental. Low concentrations of BAR, but not of NAS, stimulated larval feeding, whereas high BAR concentrations acted deterrent. NAS only tended to be deterrent at the highest concentration, but the difference was not significant. Recoveries of NAS and BAR on cabbage leaf disks were similar, and when hydrolyzed by mechanical leaf damage, the same isothiocyanate-type products as in Barbarea plants were formed with further conversion of BAR to cyclic products, (R)-5-phenyloxazolidine-2-thione [(R)-barbarin] and (R)-5-phenyloxazolidin-2-one [(R)-resedine]. We conclude that a previously proposed generally higher defensive activity of BAR than NAS to M. brassicae larvae could not be confirmed. Indeed, the higher resistance of NAS-containing B. verna plants may be due to a combined effect of rather high concentrations of NAS and a relatively high myrosinase activity or other plant traits not investigated yet.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  • Abdalsamee MK, Müller C (2012) Effects of indole glucosinolates on performance and sequestration by the sawfly Athalia rosae and consequences of feeding on the plant defence system. J Chem Ecol 38:1366–1375

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE (2011) Isoferuloyl derivatives of five seed glucosinolates in the crucifer genus Barbarea. Phytochemistry 72:610–623

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE (2012) Glucosinolate structures in evolution. Phytochemistry 77:16–45

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE (2015) Glucosinolate hydrolysis products in the crucifer Barbarea vulgaris include a thiazolidine-2-one from a specific phenolic isomer as well as oxazolidine-2-thiones. Phytochemistry 115:143–151

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE, Nielsen JK (2001) Seasonal variation in leaf glucosinolates and insect resistance in two types of Barbarea vulgaris ssp. arcuata. Phytochemistry 58:91–100

    Article  CAS  Google Scholar 

  • Agerbirk N, Ørgaard M, Nielsen JK (2003a) Glucosinolates, flea beetle resistance, and leaf pubescence as taxonomic markers in the genus Barbarea (Brassicaceae). Phytochemistry 63:69–80

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE, Bibby BM, Frandsen HO, Brown LD, Nielsen JK, Renwick JAA (2003b) A saponin correlated with variable resistance of Barbarea vulgaris to the diamondback moth Plutella xylostella. J. Chem Ecol 29:1417–1433

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE, Topbjerg HB, Sørensen JC (2007) Host plant dependent metabolism of 4-hydroxybenzylglucosinolate in Pieris rapae: substrate specificity and effects of genetic modification and plant nitrile hydratase. Insect Biochem Mol Biol 37:1119–1130

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE, Cipollini D, Ørgaard M, Linde-Laursen I, Chew FC (2014) Specific glucosinolate analysis reveals variable levels of glucobarbarins, dietary precursors of 5-phenyloxazolidine-2-thiones, in watercress types with contrasting chromosome number. J Agric. Food Chem 62:9586–9596

    Article  CAS  Google Scholar 

  • Agerbirk N, Olsen CE, Heimes C, Christensen S, Bak S, Hauser T (2015a) Multiple hydroxyphenethyl glucosinolate isomers and their tandem mass spectrometric distinction in a geographically structured polymorphism in the crucifer Barbarea vulgaris. Phytochemistry 115:130–142

    Article  CAS  Google Scholar 

  • Agerbirk N, De Nicola GR, Olsen CE, Müller C, Iori R (2015b) Derivatization of isothiocyanates and their reactive adducts for chromatographic analysis. Phytochemistry 118:109–115

    Article  CAS  Google Scholar 

  • Agerbirk N, Matthes A, Erthmann PØ, Ugolini L, Cinti S, Lazaridi E, Nuzillard J-M, Müller C, Bak S, Rollin P, Lazzeri L (2018) Glucosinolate turnover in Brassicales species to an oxazolidin-2-one, formed via the 2-thione and without formation of thioamide. Phytochemistry 153:79–93

    Article  CAS  Google Scholar 

  • Ahuja I, van Dam NM, Winge P, Traelnes M, Heydarova A, Rohloff J, Langaas M, Bones AM (2015) Plant defence responses in oilseed rape MINELESS plants after attack by the cabbage moth Mamestra brassicae. J Exp Bot 66(2):579–592

    Article  CAS  Google Scholar 

  • Badenes-Pérez FR, López-Pérez JA (2018) Resistance and susceptibility to powdery mildew, root-knot nematode, and western flower thrips in two types of winter cress (Brassicaceae). Crop Protect 110:41–47

    Article  Google Scholar 

  • Badenes-Pérez FR, Reichelt M, Gershenzon J, Heckel DG (2011) Phylloplane location of glucosinolates in Barbarea spp. (Brassicaceae) and misleading assessment of host suitability by a specialist herbivore. New Phytol 189:549–556

    Article  Google Scholar 

  • Badenes-Pérez FR, Reichelt M, Gershenzon J, Heckel DG (2013) Interaction of glucosinolate content of Arabidopsis thaliana mutant lines and feeding and oviposition by generalist and specialist lepidopterans. Phytochemistry 86:36–43

    Article  Google Scholar 

  • Badenes-Pérez FR, Gershenzon J, Heckel DG (2014a) Insect attraction versus plant defense: young leaves high in glucosinolates stimulate oviposition by a specialist herbivore despite poor larval survival due to high saponin content. PLOS One 9:e95766

    Article  Google Scholar 

  • Badenes-Pérez FR, Reichelt M, Gershenzon J, Heckel DG (2014b) Using plant chemistry and insect preference to study the potential of Barbarea (Brassicaceae) as a dead-end trap crop for diamondback moth (Lepidoptera: Plutellidae). Phytochemistry 98:137–144

    Article  Google Scholar 

  • Barillari J, Gueyrard D, Rollin P, Iori R (2001) Barbarea verna as a source of 2-phenylethyl glucosinolate, precursor of cancer chemopreventive phenylethyl isothiocyanate. Fitoterapia 72:760–764

    Article  CAS  Google Scholar 

  • Beekwilder J, van Leeuwen W, van Dam NM, Bertossi M, Grandi V, Mizzi L, Soliviev M, Szabados L, Molthoff JW, Schipper B, Verbocht H, de Vos RCH, Morandini P, Aarts MGM, Bovy A (2008) The impact of the absence of aliphatic glucosinolates on insect herbivory in Arabidopsis. PLoS One 3:e2068

    Article  Google Scholar 

  • Burow M, Halkier BA (2017) How does a plant orchestrate defense in time and space? Using glucosinolates in Arabidopsis as a case study. Curr Opin Plant Biol 38:142–147

    Article  CAS  Google Scholar 

  • Byrne S, Erthmann PØ, Agerbirk N, Bak S, Hauser TP, Nagy I, Paina C, Asp T (2017) The genome sequence of Barbarea vulgaris facilitates the study of ecological biochemistry. Sci Rep 7:40728

    Article  CAS  Google Scholar 

  • Christensen S, Heimes C, Agerbirk N, Kuzina V, Olsen CE, Hauser TP (2014) Different geographical distributions of two chemotypes of Barbarea vulgaris that differ in resistance to insects and a pathogen. J Chem Ecol 40:491–501

    Article  CAS  Google Scholar 

  • Christensen S, Enge S, Munk KR, Müller C, Kiær LP, Agerbirk N, Heimes C, Hauser TP (2018) Different herbivore responses to two co-occuring chemotypes of the wild crucifer Barbarea vulgaris. Arthropod Plant Interact. https://doi.org/10.1007/s11829-018-9633-x

  • ElSayed G, Louveaux A, Mavratzotis M, Rollin P, Quinsac A (1996) Effects of glucobrassicin, epiprogoitrin and related breakdown products on locusts feeding: Schouwia purpurea and desert locust relationships. Entomol Exp Appl 78:231–236

    Article  CAS  Google Scholar 

  • Felkner P, Bunch R, Leung AM (2016) Concentrations of thiocyanate and goitrin in human plasma, their precursor concentrations in brassica vegetables, and associated potential risk for hypothyroidism. Nutrition Rev 74:248–258

    Article  Google Scholar 

  • Frisch T, Agerbirk N, Davis S, Cipollini D, Olsen CE, Motavia MS, Bjarnholt N, Møller BL (2014) Glucosinolate-related glucosides in Alliaria petiolata: sources of variation in the plant and different metabolism in an adapted specialist herbivore, Pieris rapae. J Chem Ecol 40:1063–1079

    Article  CAS  Google Scholar 

  • Gmelin R, Kjær A, Schuster A (1970) Glucosinolates in seeds of Sibara virginica (L.) Rollins: Two new glucosinolates. Acta Chem. Scand. 24:3031–3037

    Article  Google Scholar 

  • Gols R, Wagenaar R, Bukovinszky T, van Dam NM, Dicke M, Bullock JM, Harvey JA (2008) Genetic variation in defense chemistry in wild cabbages affects herbivores and their endoparasitoids. Ecology 89:1616–1626

    Article  Google Scholar 

  • Hanschen FS, Lamy E, Schreiner M, Rohn S (2014) Reactivity and stability of glucosinolates and their breakdown products in foods. Angew Chem Int Ed 53:2–23

    Article  Google Scholar 

  • Hansen BG, Kerwin RE, Ober J, Lambrix VM, Mitchell-Olds T, Gershenzon J, Halkier BA, Kliebenstein DJ (2008) A novel 2-oxoacid-dependent dioxygenase involved in the formation of goiterogenic 2-hydroxybut-3-enyl glucosinolate and generalist insect resistance in Arabidopsis. Plant Phys 148:2096–2108

    Article  CAS  Google Scholar 

  • Heimes C, Agerbirk N, Sørensen H, van Mölken T, Hauser TP (2016) Ecotopic differentiation of two sympatric chemotypes of Barbarea vulgaris (Brassicaceae) with different biotic resistances. Plant Ecol 217:1055–1068

    Article  Google Scholar 

  • Jeschke V, Gershenzon J, Vassão DG (2016a) Insect detoxification of glucosinolates and their hydrolysis products. Adv Bot Res 80:199–245

    Article  Google Scholar 

  • Jeschke V, Gershenzon J, Vassão DG (2016b) A mode of action of glucosinolate-derived isothiocyanates: Detoxification depletes glutathione and cysteine levels with ramifications on protein metabolism in Spodoptera littoralis. Insect Biochem Mol Biol 71:37–48

    Article  CAS  Google Scholar 

  • Jeschke V, Kearney EE, Schramm K, Kunert G, Shekhov A, Gershenzon J, Vassão D (2017) How glucosinolates affect generalist lepidopteran larvae: growth, development and glucosinolate content. Front. Plant Sc. 8:1995

    Article  Google Scholar 

  • Kerwin RE, Feusier J, Muok A, Lin C, Larson B, Copeland D, Corwin JA, Rubin MJ, Francisco M, Li B, Joseph B, Weinig C, Kliebenstein DJ (2017) Epistasis x environment interactions among Arabidopsis thaliana glucosinolate genes impact complex traits and fitness in the field. New Phytol 215:1249–1263

    Article  CAS  Google Scholar 

  • Kissen R, Pope TW, Grant M, Pickett JA, Rossiter JT, Powell G (2009) Modifying the alkylglucosinolate profile in Arabidopsis thaliana alters the tritrophic interaction with the herbivore Brevicoryne brassicae and parasitoid Diaeretiella rapae. J Chem Ecol 35:958–969

    Article  CAS  Google Scholar 

  • Klopsch R, Witzel K, Artemyeva A, Ruppel S, Hanschen F (2018) Genotypic variation of glucosinolates and their breakdown products in leaves of Brassica rapa. J Agric Food Chem 66:5481–5490

    Article  CAS  Google Scholar 

  • Kumar P, Augustine R, Singh AK, Bisht NC (2017) Feeding behaviour of generalist pests on Brassica juncea: implication for manipulation of glucosinolate biosynthesis pathway for enhanced resistance. Plant Cell Environ 40:2109–2120

    Article  CAS  Google Scholar 

  • Lazzeri L, Malaguti L, Cinti S, Ugolini L, De Nicola GR, Bagatta M, Matteo R, Casadei N, Patalano G, D’Avino L (2013) The Brassicaceae biofumigation system for plant cultivation and defence. An Italian twenty-year experience of study and application. Acta Hortic 1005:375–382

    Article  Google Scholar 

  • Li Q, Eigenbrode SD, Stringham GR, Thiagarajah MR (2000) Feeding and growth of Plutella xylostella and Spodoptera eridania on Brassica juncea with varying glucosinolate concentrations and myrosinase activities. J Chem Ecol 26:2401–2419

    Article  CAS  Google Scholar 

  • Liu T, Zhang X, Yang H, Agerbirk N, Qiu Y, Wang H, Shen D, Song J, Li X (2016) Aromatic glucosinolate biosynthesis pathway in Barbarea vulgaris and its response to Plutella xylostella infestation. Front Plant Sci 7:83

    PubMed  PubMed Central  Google Scholar 

  • Müller C, Sieling N (2006) Effects of glucosinolate and myrosinase levels in Brassica juncea on a glucosinolate-sequestering herbivore - and vice versa. Chemoecology 16:191–201

    Article  Google Scholar 

  • Müller C, van Loon J, Ruschioni S, De Nicola GR, Olsen CE, Iori R, Agerbirk N (2015) Taste detection of the non-volatile isothiocyanate moringin results in deterrence to glucosinolate-adapted insect larvae. Phytochemistry 118:139–148

    Article  Google Scholar 

  • Nielsen JK (1999) Specificity of a Y-linked gene in the flea beetle Phylotreta nemorum for defences in Barbarea vulgaris. Entomol. Exp. Appl. 91:359–368

    Article  Google Scholar 

  • Olsen CE, Huang X-C, Hansen CIC, Cipollini D, Ørgaard M, Matthes A, Geu-Flores F, Koch MA, Agerbirk N (2016) Glucosinolate diversity within a phylogenetic framework of the tribe Cardamineae (Brassicaceae) unravelled with HPLC-MS/MS and NMR-based analytical distinction of 70 desulfoglucosinolates. Phytochemistry 132:33–56

    Article  CAS  Google Scholar 

  • Pagnotta E, Agerbirk N, Olsen CE, Ugolini L, Cinti S, Lazzeri L (2017) Hydroxyl and methoxyl derivatives of benzylglucosinolate in Lepidium densiflorum with hydrolysis to isothiocyanates and non-isothiocyanate products: Substitution governs product type and mass spectral fragmentation. J Agric Food Chem 65:3167–3178

    Article  CAS  Google Scholar 

  • Radulovic NS, Todorovska MM, Zlatkovic DB, Stojanovic NM, Randjelovic PJ (2017) Two goitrogenic 1,3-oxazolidine-2-thione derivatives from Brassicales taxa: challenging identification, occurrence and immunomodulatory effects. Food Chem Toxicol 110:94–108

    Article  CAS  Google Scholar 

  • Rojas JC, Wyatt TD, Birch MC (2000) Flight and oviposition behavior toward different host plant species by the cabbage moth, Mamestra brassicae (L.) (Lepidoptera : Noctuidae). J Insect Behav 13:247–254

    Article  Google Scholar 

  • Siemens DH, Mitchell-Olds T (1996) Glucosinolates and herbivory by specialists (Coleoptera: Chrysomelidae, Lepidoptera: Plutellidae): Consequences of concentration and induced resistance. Environ Entomol 25:1344–1353

    Article  CAS  Google Scholar 

  • Sønderby IE, Geu-Flores F, Halkier B (2010) Biosynthesis of glucosinolates – gene discovery and beyond. Trends Pl Sci 15:283–290

    Article  Google Scholar 

  • Tanton MT (1965) Agar and chemostimulant concentrations and their effect on intake of synthetic food by larvae of the mustard beetle, Phaedon cochleariae. Entomol Exp Appl 8:74–82

    Article  CAS  Google Scholar 

  • Travers-Martin N, Kuhlmann F, Müller C (2008) Revised determination of free and complexed myrosinase activities in plant extracts. Plant Phys Biochem 46:506–516

    Article  CAS  Google Scholar 

  • van Leur H (2008) Genetics, chemistry and ecology of a qualitative glucosinolate polymorphism in Barbarea vulgaris. PhD thesis, Wageningen University, The Netherlands

    Google Scholar 

  • van Leur H, Raaijmakers CE, van Dam NM (2006) A heritable glucosinolate polymorphism within natural populations of Barbarea vulgaris. Phytochemistry 67:1214–1223

    Article  Google Scholar 

  • van Leur H, Vet LEM, van der Putten W, van Dam NM (2008) Barbarea vulgaris glucosinolate phenotypes differentially affect performance and preference of two different species of lepidopteran hervivores. J Chem Ecol 34:121–131

    Article  Google Scholar 

  • van Mölken T, Kuzina V, Munk KR, Olsen CE, Sundelin T, van Dam N, Hauser TP (2014) Consequences of combined herbivore feeding and pathogen infection for fitness of Barbarea vulgaris plants. Oecologia 175:589–600

    Article  Google Scholar 

  • Visentin M, Tava A, Iori R, Palmieri S (1992) Isolation and Identification for trans-4-(methylthio)-3-butenyl glucosinolate from radish roots (Raphanus sativus L.). J Agric Food Chem 40:1687–1691

    Article  CAS  Google Scholar 

  • Wittstock U, Kurzbach E, Herfurth A-M, Stauber EJ (2016) Glucosinolate breakdown. Adv Bot Res 80:125–169

    Article  Google Scholar 

  • Zalucki MP, Zalucki JM, Perkins LE, Schramm K, Vassão DG, Gershenzon J, Heckel DG (2017) A generalist herbivore copes with specialized plant defence: the effects of induction and feeding by Helicoverpa armigera (Lepidoptera: Noctuidae) larvae on intact Arabidopsis thaliana (Brassicales) plants. J Chem Ecol 43:608–616

    Article  CAS  Google Scholar 

  • Zheng SJ, Zhang PJ, van Loon JJA, Dicke M (2011) Silencing defense pathways in Arabidopsis by heterologous gene sequences from Brassica oleracea enhances the performance of a specialist and a generalist herbivorous insect. J Chem Ecol 37(8):818–829

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank two anonymous reviewers for constructive comments that improved this paper, Dr. Carl Erik Olsen for NMR analysis of glucosinolate preparations and Dr. David Ian Pattison and Eleni Lazaridi for optimizing the chiral separation method. Ting Yang acknowledges support by the Novo Nordisk Foundation through a Postdoctoral Fellowship (Biotechnology-based Synthesis and Production Research, NNF16OC0019608), while Niels Agerbirk thanks Torben og Alice Frimodts Fond for financial support.

Author information

Authors and Affiliations

  1. Department of Chemical Ecology, Bielefeld University, Universitätsstr. 25, 33615, Bielefeld, Germany

    Caroline Müller & Monique Schulz

  2. Council for Agricultural Research and Economics, Research Centre for Cereal and Industrial Crops, Via di Corticella 133, 40128, Bologna, Italy

    Eleonora Pagnotta, Luisa Ugolini & Luca Lazzeri

  3. Copenhagen Plant Science Center and Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark

    Ting Yang, Annemarie Matthes & Niels Agerbirk

Authors
  1. Caroline Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Monique Schulz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eleonora Pagnotta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Luisa Ugolini
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ting Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Annemarie Matthes
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Luca Lazzeri
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Niels Agerbirk
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceived overall experiment: NA, CM. Input to experimental design: all. Provided glucosinolates: LL, EP, LU. Analyzed glucosinolate purity: LL, EP, LU, NA. Performed insect experiments and analysis of glucosinolates, myrosinases and C/N: MS, CM. Analyzed products of NAS by GC: EP, LU supervised by LL. Analyzed glucosinolate products by HPLC: NA, TY, AM. Wrote draft: CM, NA. Wrote paper: All.

Corresponding authors

Correspondence to Caroline Müller or Niels Agerbirk.

Electronic Supplementary Material

ESM 1

(PDF 1944 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Müller, C., Schulz, M., Pagnotta, E. et al. The Role of the Glucosinolate-Myrosinase System in Mediating Greater Resistance of Barbarea verna than B. vulgaris to Mamestra brassicae Larvae. J Chem Ecol 44, 1190–1205 (2018). https://doi.org/10.1007/s10886-018-1016-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10886-018-1016-3

Keywords

Search

Navigation