Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Dec;4(4):e20102.
doi: 10.1002/cppb.20102.

Cassava Metabolomics and Starch Quality

Laise Rosado-Souza  1 , Laure C David  2 , Margit Drapal  3 , Paul D Fraser  3 , Jörg Hofmann  4 , Patrick A W Klemens  5 , Frank Ludewig  4 , H Ekkehard Neuhaus  5 , Toshihiro Obata  1   6 , Laura Perez-Fons  3 , Armin Schlereth  1 , Uwe Sonnewald  4 , Mark Stitt  1 , Samuel C Zeeman  2 , Wolfgang Zierer  4 , Alisdair R Fernie  1
Affiliations

Cassava Metabolomics and Starch Quality

Laise Rosado-Souza et al. Curr Protoc Plant Biol. 2019 Dec.

Abstract

Cassava plays an important role as a staple food for more than 800 million people in the world due to its ability to maintain relatively high productivity even in nutrient-depleted soils. Even though cassava has been the focus of several breeding programs and has become a strong focus of research in the last few years, relatively little is currently known about its metabolism and metabolic composition in different tissues. In this article, the absolute content of sugars, organic acids, amino acids, phosphorylated intermediates, minerals, starch, carotenoids, chlorophylls, tocopherols, and total protein as well as starch quality is described based on multiple analytical techniques, with protocols specifically adjusted for material from different cassava tissues. Moreover, quantification of secondary metabolites relative to internal standards is presented using both non-targeted and targeted metabolomics approaches. The protocols have also been adjusted to apply to freeze-dried material in order to allow processing of field harvest samples that typically will require long-distance transport. © 2019 The Authors. Basic Protocol 1: Metabolic profiling by gas chromatography-mass spectrometry (GC-MS) Support Protocol 1: Preparation of freeze-dried cassava material Support Protocol 2: Preparation of standard compound mixtures for absolute quantification of metabolites by GC-MS Support Protocol 3: Preparation of retention-time standard mixture Basic Protocol 2: Determination of organic acids and phosphorylated intermediates by ion chromatography-mass spectrometry (IC-MS) Support Protocol 4: Preparation of standards and recovery experimental procedure Basic Protocol 3: Determination of soluble sugars, starch, and free amino acids Alternate Protocol: Determination of soluble sugars and starch Basic Protocol 4: Determination of anions Basic Protocol 5: Determination of elements Basic Protocol 6: Determination of total protein Basic Protocol 7: Determination of non-targeted and targeted secondary metabolites Basic Protocol 8: Determination of carotenoids, chlorophylls, and tocopherol Basic Protocol 9: Determination of starch quality.

Keywords: Manihot esculenta; chemotypes; metabolite classification; metabolite profiling; metabolomics.

PubMed Disclaimer

References

Literature Cited

    1. Alseekh, S., & Fernie, A. R. (2018). Metabolomics 20 years on: What have we learned and what hurdles remain? Plant Journal, 94, 933-942. doi: 10.1111/tpj.13950.
    1. Arrivault, S., Guenther, M., Ivakov, A., Feil, R., Vosloh, D., van Dongen, J. T., … Stitt, M. (2009). Use of reverse-phase liquid chromatography, linked to tandem mass spectrometry, to profile the Calvin cycle and other metabolic intermediates in Arabidopsis rosettes at different carbon dioxide concentrations. Plant Journal, 59, 826-839. doi: 10.1111/j.1365-313X.2009.03902.x.
    1. Bjarnholt, N., Neilson, E. H. J., Crocoll, C., Jorgensen, K., Motawia, M. S., Olsen, C. E., … Moller, B. L. (2018). Glutathione transferases catalyze recycling of auto-toxic cyanogenic glucosides in sorghum. Plant Journal, 94, 1109-1125. doi: 10.1111/tpj.13923.
    1. Bligh, E. G., & Dyer, W. J. (1959). A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology, 37, 911-917. doi: 10.1139/y59-099.
    1. Bradford, M. M. (1976). Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. doi: 10.1016/0003-2697(76)90527-3.

Publication types

LinkOut - more resources