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Standardizing methods for measuring non-structural carbohydrates (NSCs) is crucial for understanding plant carbon balance. This study provides robust protocols to improve the accuracy and comparability of NSC measurements across laboratories.

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Area of Science:

  • Plant physiology
  • Plant metabolism
  • Biochemistry

Background:

  • Non-structural carbohydrates (NSCs) are vital for plant growth and respiration, but their measurement is inconsistent across studies.
  • Variability in NSC measurement methods leads to challenges in integrating plant carbon balance data.
  • Previous assessments showed significant discrepancies in NSC concentrations from common samples, with unclear sources of variation.

Purpose of the Study:

  • To assess the variability in Non-structural Carbohydrate (NSC) measurements across different quantification methods and laboratories.
  • To identify factors influencing NSC measurement accuracy, including sample handling, extraction solvents, and quantification techniques.
  • To develop and provide standardized protocols for reliable NSC extraction, digestion, and quantification in plant samples.

Main Methods:

  • Six laboratories measured nine plant types and two synthetic samples using a common protocol for sugar extraction and starch digestion.
  • Three distinct sugar quantification methods were employed: ion chromatography, enzymatic assay, and acid hydrolysis.
  • The study also evaluated the impact of sample handling, extraction solvent (water vs. ethanol), and centralized processing.

Main Results:

  • All three quantification methods yielded results within 11.5% of known values for synthetic samples.
  • Quantification method differences were the primary source of variation for natural plant samples due to varying NSC targets.
  • The acid method quantified a broader range of sugars, resulting in 2-5 times higher concentrations for some natural samples compared to other methods.

Conclusions:

  • Standardized protocols can achieve reasonable accuracy in NSC measurements despite using different quantification methods.
  • Ethanol extraction is preferable to water extraction for improved accuracy.
  • The provided detailed protocols aim to enhance the comparability and reliability of NSC measurements in plant science research.