Simple and robust determination of the activity signature of key carbohydrate metabolism enzymes for physiological phenotyping in model and crop plants

Jammer, Alexandra; Gasperl, Anna; Luschin-Ebengreuth, Nora; Heyneke, Elmien; Chu, Hyosub; Cantero-Navarro, Elena; Großkinsky, Dominik K.; Albacete, Alfonso; Stabentheiner, Edith; Franzaring, Jürgen; Fangmeier, Andreas; Graaff, Eric van der; Roitsch, Thomas

doi:10.1093/jxb/erv228

Cited by 87 publications

(117 citation statements)

References 59 publications

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“…Although the method we established to determine an activity signature for key enzymes of carbohydrate metabolism performed well with various monocot and dicot model and crop plants 11 and proved to be useful for physiological phenotyping, 8 we encountered a number of difficulties in applying the "universal" extraction protocol to grapevine, probably due to interfering phenolic and anthocyanin compounds in the extracts. 12,13…”

Section: Resultsmentioning

confidence: 99%

“…The workflow for measuring the enzyme activities of AGPase, SuSy, cytINV, cwINV and vacINV is depicted in the assays' scheme ( Figure 1). Major changes to the original protocols 11 were required, and these are detailed in the Results and Discussion section.…”

Section: Methodsmentioning

confidence: 99%

“…Four previously reported extraction buffers, including Jammer's extraction buffer B 11 and three buffers used in grapevine enzyme studies, 22,24,26 were tested in parallel for highest activities (see recipes in Table 2). Because activity of maize SuSy requires magnesium sulfate in extraction buffer, 24 buffers were tested with and without added Mg if it was not included in the published recipe.…”

Section: Adaptation Of the Extraction Protocol For Grapevinementioning

confidence: 99%

“…8 We have recently developed a universal protein extraction and fractionation method for determining activities of key carbohydrate metabolism enzymes from a wide variety of plant species. 11 These published semi-highthroughput spectrophotometric assays for determination of primary carbohydrate metabolism enzymes from a single extraction 11 failed when applied to grapevine leaves.…”

Section: Introductionmentioning

confidence: 99%

“…15 In the present work we have adapted our general assays 11 for application in uninfected grapevine samples and those infected with the most widespread grapevine phytoplasma in Europe, šCandidatus Phytoplasma solani'. 16 …”

Section: Introductionmentioning

confidence: 99%

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Determination of the Activity Signature of Key Carbohydrate Metabolism Enzymes in Phenolic-rich Grapevine Tissues

Covington

Roitsch

Dermastia

2016

ACSi

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Physiological studies in plants often require enzyme extraction from tissues containing high concentrations of phenols and polyphenols. Unless removed or neutralized, such compounds may hinder extraction, inactivate enzymes, and interfere with enzyme detection. The following protocol for activity assays for enzymes of primary carbohydrate metabolism, while based on our recently published one for quantitative measurement of activities using coupled spectrophotometric assays in a 96-well format, is tailored to the complexities of phenolic-and anthocyanin-rich extracts from grapevine leaf. As a case study we applied the protocol to grapevine leaf samples infected with plant pathogenic bacteria šCandidatus Phytoplasma solani', known to alter carbohydrate metabolism in grapevine. The described adaptations may be useful for determination of metabolic fingerprints for physiological phenotyping of other plant species with inherently high levels of phenolic compounds.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Adaptation Of the Extraction Protocol For Grapevinementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Determination of the Activity Signature of Key Carbohydrate Metabolism Enzymes in Phenolic-rich Grapevine Tissues

Covington

Roitsch

Dermastia

2016

ACSi

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Measurement of Enzyme Activities and Optimization of Continuous and Discontinuous Assays

Bénard

Gibon

2016

CP Plant Biology

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The measurement of enzyme activities represents an important step towards the understanding of biological networks. Continuous or discontinuous assays can be used, as well as highly sensitive assays, depending on the abundance of the enzymes under study. To exemplify such methods, two protocols for phosphoenolpyruvate carboxylase activity (EC 4.1.1.31) in plant extracts are given. For this, an extraction protocol is also described. Then, an optimization protocol for enzyme assays using enzymatic, chemical, or biological standards is proposed. This protocol evaluates in one run the optimal extract dilution, the recovery of a standard, and the technical error in a given matrix. The interest of using biological standard in routine measurements is highlighted. © 2016 by John Wiley & Sons, Inc.

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Carbohydrate metabolism enzymes and phenotypic characterization of diverse lines of the climate‐resilient food, feed, and bioenergy crop Camelina sativa

Stasnik

Vollmann

Großkinsky

et al. 2023

Food and Energy Security

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Climate change increasingly challenges agricultural production systems and thus food and bioenergy security. To meet this challenge, the climate-resilient, low-input, versatile Brassicaceae oilseed crop camelina (Camelina sativa [L.] Crantz) is a promising alternative species. Camelina, also known as gold-of-pleasure, false-flax, Siberian oilseed, German sesame and linseed dodder is native to Southeast Europe and Southwest Asia and

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Simple and robust determination of the activity signature of key carbohydrate metabolism enzymes for physiological phenotyping in model and crop plants

Cited by 87 publications

References 59 publications

Determination of the Activity Signature of Key Carbohydrate Metabolism Enzymes in Phenolic-rich Grapevine Tissues

Determination of the Activity Signature of Key Carbohydrate Metabolism Enzymes in Phenolic-rich Grapevine Tissues

Measurement of Enzyme Activities and Optimization of Continuous and Discontinuous Assays

Carbohydrate metabolism enzymes and phenotypic characterization of diverse lines of the climate‐resilient food, feed, and bioenergy crop Camelina sativa

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