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.
|State||Published - Jan 2023|
Bibliographical noteFunding Information:
This research was funded by The Army Research Office which supported part of this project through Grant W911NF1910222 to ABD to purchase the Dionex HPLC. Hatch Grant 1019088 to ABD purchased all consumables.
© 2023 by the authors.
- raffinose family oligosaccharides
ASJC Scopus subject areas
- Molecular Biology