Alterations in Carbohydrate Quantities in Freeze-Dried, Relative to Fresh or Frozen Maize Leaf Disks

Lynnette M.A. Dirk, Tianyong Zhao, John May, Tao Li, Qinghui Han, Yumin Zhang, Mohammad R. Sahib, Allan Bruce Downie

Research output: Contribution to journalArticlepeer-review

Abstract

For various reasons, leaves are occasionally lyophilized prior to storage at −80 °C and preparing extracts. Soluble carbohydrate identity and quantity from maize leaf disks were ascertained in two separate years using anion exchange HPLC with pulsed electrochemical detection. Analyses were made from disks after freezing in liquid nitrogen with or without subsequent lyophilization (both years) or directly after removal from plants with or without lyophilization (only in the second year). By adding the lyophilizing step, galactose content consistently increased and, frequently, so did galactoglycerols. The source of the galactose increase with the added lyophilizing step was not due to metabolizing raffinose, as the raffinose synthase (rafs) null mutant leaves, which do not make that trisaccharide, also had a similar increase in galactose content with lyophilization. Apparently, the ester linkages attaching free fatty acids to galactoglycerolipids of the chloroplast are particularly sensitive to cleavage during lyophilization, resulting in increases in galactoglycerols. Regardless of the galactose source, a systematic error is introduced for carbohydrate (and, most likely, also chloroplast mono- or digalactosyldiacylglycerol) amounts when maize leaf samples are lyophilized prior to extraction. The recognition of lyophilization as a source of galactose increase provides a cautionary note for investigators of soluble carbohydrates.

Original languageEnglish
Article number148
JournalBiomolecules
Volume13
Issue number1
DOIs
StatePublished - Jan 2023

Bibliographical note

Publisher Copyright:
© 2023 by the authors.

Keywords

  • carbohydrate
  • galactose
  • glycerol–galactopyranoside
  • lyophilization
  • raffinose family oligosaccharides

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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