A Straightforward Method for Glucosinolate Extraction and Analysis with High-pressure Liquid Chromatography (HPLC)

Grosser, Katharina; Dam, Nicole M. van

doi:10.3791/55425

Cited by 51 publications

(59 citation statements)

References 22 publications

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“…Glucosinolate extraction and analysis was performed as described in detail in Grosser and van Dam (). In brief, after plant material collection the samples were freeze‐dried, ground and weight to c .…”

Section: Methodsmentioning

confidence: 99%

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Locally and systemically induced glucosinolates follow optimal defence allocation theory upon root herbivory

2018

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Herbivore‐induced defences in plants are considered a strategy to manage multiple interactions while saving resources. The optimal defence theory (ODT) is one of the most prominent theoretical frameworks to explain the defence allocation patterns within plants. It was recently shown that the ODT generally applies to constitutive glucosinolate (GSL) allocation in shoot and root organs. Previous studies showed that both root and shoot herbivore feeding may alter defence allocation over plant organs. For shoots, the effect depends on where the herbivores feed. It is as yet unknown whether similar principles apply to root‐herbivore‐induced GSLs. To analyse the effects of root localized herbivore feeding on GSL allocation, we conducted a pot experiment using Anomala cuprea grubs and four Brassicaceae; Brassica rapa, B. nigra, B. oleracea and Sinapis alba. Individuals of these four plant species were grown in dedicated mesocosms. The grubs were confined either to the bottom soil, the middle section or the topsoil. Plants grown in the same set‐ups but without root herbivores served as controls. Glucosinolate levels of the leaf lamina, petiole and stem as well as of the taproot, lateral roots and fine roots were measured after 8 days of herbivory. Plant biomass reduction due to herbivory was the largest when herbivores were confined to the topsoil. In the three Brassica species, taproot GSL levels increased upon herbivory independent of where the root herbivores were feeding. Glucosinolate levels in fine roots and shoots, on the other hand, hardly responded to root herbivory. Indole GSLs, which are more effective to pathogens than to herbivores, were more strongly induced than aliphatic and aromatic GSLs, especially in the taproots. Sinapis alba did not show remarkable increments in any GSL level upon herbivory. These results show that locally and systemically induced defences in roots are consistent with the ODT: The taproot which is the most vulnerable and valuable to plant performance shows the highest increase in defence induction. The induced GSL profiles suggest that the response may not only target herbivores, but may also help to prevent secondary infection by microbial pathogens. A http://onlinelibrary.wiley.com/doi/10.1111/1365-2435.13147/suppinfo is available for this article.

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Section: Methodsmentioning

confidence: 99%

“…Glucosinolate extraction and analysis was performed as described in detail in Grosser and van Dam (2017 Table S1). GSL concentrations were calculated using an external sinigrin reference curve (EC 1990) and…”

Section: Glucosinolate Analysismentioning

confidence: 99%

Locally and systemically induced glucosinolates follow optimal defence allocation theory upon root herbivory

2018

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“…All plants but S. alba and D. erucoides were in the vegetative stage when harvested. Harvested plant organs were flash‐frozen in liquid nitrogen and stored at −20 °C until GSL analysis, which was performed by high performance liquid chromatography–UV detecter following van Dam, Witjes & Svatoš () (see Grosser & van Dam and Appendix S1).…”

Section: Methodsmentioning

confidence: 99%

Root and shoot glucosinolate allocation patterns follow optimal defence allocation theory

2017

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Summary Optimal defence allocation theory (ODT) is one of the most prominent theoretical frameworks to explain the allocation of defence compounds within plants. It predicts that the most valuable and vulnerable plant organs have the highest levels of chemical defence. The ODT has been well worked out and experimentally tested for shoot defences, but not for root defences. To assess if ODT principles apply similarly to roots and shoots, we examined glucosinolates in above‐ground and below‐ground organs of nine plant species belonging to two families. In order to evaluate whether ODT equally applies to shoot and root organs, we designed a conceptual model in which above‐ground and below‐ground organs were assigned to orders of importance to plant performance. We hypothesized that organs constituting the plant's core structure are better protected than more distal organs. The nine plant species were cultivated, and their roots and shoots were harvested and divided into three orders for glucosinolate analysis. Using a specialist (Delia radicum) and a generalist (Amphimallon solstitiale) root herbivore, we also experimentally tested the hypothesis that the generalist herbivore prefers to feed on fine roots (FRs) with a low glucosinolate concentration, while the specialist prefers taproots (TRs) with a high glucosinolate concentration. We found that both in roots and shoots, the higher ordered core structural organs (TRs and stems) had the highest levels of glucosinolates. Below‐ground, TRs and lateral roots were better protected than the more distal, and less costly, FRs in seven out of nine species tested. The specialist root herbivore preferred feeding on the highly defended TRs, which is in line with what has been found for above‐ground specialist herbivores. Moreover, the glucosinolate concentration in roots overall was significantly higher than that in shoots. Synthesis. These results support the hypothesis that Optimal defence allocation theory (ODT) generally applies to glucosinolate allocation in above‐ground and below‐ground organs and may mainly serve to maintain the integrity of the main plant structure. Moreover, it suggests that above‐ground and below‐ground insect herbivores independently exert similar selection pressures on defence allocation patterns in roots and shoots.

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“…Detection was performed at 229 nm using a photodiode array. Glucoerucin UV spectra acquisition and analysis were carried following methods described by Grosser and Dam (2017) and Madsen, Kunert, Reichelt, Gershenzon, and Halkier (2015).…”

Section: Methionine-derived Aliphatic Glucosinolate Hplc Analysismentioning

confidence: 99%

Mechanisms underlying the enhanced biomass and abiotic stress tolerance phenotype of an Arabidopsis MIOX over‐expresser

et al. 2019

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Myo ‐inositol oxygenase (MIOX) is the first enzyme in the inositol route to ascorbate (L‐ascorbic acid, AsA, vitamin C). We have previously shown that Arabidopsis plants constitutively expressing MIOX have elevated foliar AsA content and displayed enhanced growth rate, biomass accumulation, and increased tolerance to multiple abiotic stresses. In this work, we used a combination of transcriptomics, chromatography, microscopy, and physiological measurements to gain a deeper understanding of the underlying mechanisms mediating the phenotype of the At MIOX4 line. Transcriptomic analysis revealed increased expression of genes involved in auxin synthesis, hydrolysis, transport, and metabolism, which are supported by elevated auxin levels both in vitro and in vivo, and confirmed by assays demonstrating their effect on epidermal cell elongation in the At MIOX4 over‐expressers. Additionally, we detected up‐regulation of transcripts involved in photosynthesis and this was validated by increased efficiency of the photosystem II and proton motive force. We also found increased expression of amylase leading to higher intracellular glucose levels. Multiple gene families conferring plants tolerance/expressed in response to cold, water limitation, and heat stresses were found to be elevated in the At MIOX4 line. Interestingly, the high AsA plants also displayed up‐regulation of transcripts and hormones involved in defense including jasmonates, defensin, glucosinolates, and transcription factors that are known to be important for biotic stress tolerance. These results overall indicate that elevated levels of auxin and glucose, and enhanced photosynthetic efficiency in combination with up‐regulation of abiotic stresses response genes underly the higher growth rate and abiotic stresses tolerance phenotype of the At MIOX4 over‐expressers.

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A Straightforward Method for Glucosinolate Extraction and Analysis with High-pressure Liquid Chromatography (HPLC)

Cited by 51 publications

References 22 publications

Locally and systemically induced glucosinolates follow optimal defence allocation theory upon root herbivory

Locally and systemically induced glucosinolates follow optimal defence allocation theory upon root herbivory

Root and shoot glucosinolate allocation patterns follow optimal defence allocation theory

Mechanisms underlying the enhanced biomass and abiotic stress tolerance phenotype of an Arabidopsis MIOX over‐expresser

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