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Assessment of the influence of different sample processing and cold storage duration on plant free proline content analyses

Tekli?, Tihana; �Poljarevi?, Marija; Stanisavljevi?, Aleksandar; ParadIkovi?, Nada; Andri?, Luka; Lisjak, Miroslav; Vinkovic, Tomislav; Hancock, John T.

Authors

Tihana Tekli?

Marija �Poljarevi?

Aleksandar Stanisavljevi?

Nada ParadIkovi?

Luka Andri?

Miroslav Lisjak

Tomislav Vinkovic



Abstract

Introduction - A method which is widely accepted for the analysis of free proline content in plant tissues is based on the use of 3% sulfosalicylic acid as an extractant, followed by spectrophotometric quantification of a proline-ninhydrin complex in toluene. However, sample preparation and storage may influence the proline actually measured. This may give misleading or difficult to compare data.Objective and Methodology - To evaluate free proline levels fresh and frozen strawberry (Fragaria × ananassa Duch.) leaves and soybean [Glycine max (L.) Merr.] hypocotyl tissues were used. These were ground with or without liquid nitrogen and proline extracted with sulfosalicylic acid. A particular focus was the influence of plant sample cold storage duration (1, 4 and 12 weeks at -20°C) on tissue proline levels measured.Results - The free proline content analyses, carried out in leaves of Fragaria × ananassa Duch. as well as in hypocotyls of Glycine max (L.) Merr., showed a significant influence of the sample preparation method and cold storage period. Long-term storage of up to 12 weeks at -20°C led to a significant increase in the measured proline in all samples analysed.Conclusion - The observed changes in proline content in plant tissue samples stored at -20°C indicate the likelihood of the over-estimation of the proline content if the proline analyses are delayed. Plant sample processing and cold storage duration seem to have an important influence on results of proline analyses. Therefore it is recommended that samples should be ground fresh and analysed immediately. Copyright © 2010 John Wiley & Sons, Ltd.The proline levels in fresh or frozen strawberry leaves and soybean hypocotyls, ground with or without liquid nitrogen, were quantified by spectrophotometry and the influence of sample storage duration (up to 12 weeks at -20°C) on measured proline was tested. It was found that storage of samples at -20°C had a significant influence on the amount of proline measured in the samples, and therefore it is recommended that samples should be ground fresh and analysed immediately. Copyright © 2010 John Wiley & Sons, Ltd.

Citation

Andrić, L., ParadIković, N., Stanisavljević, A., ŠPoljarević, M., Teklić, T., Lisjak, M., …Hancock, J. T. (2010). Assessment of the influence of different sample processing and cold storage duration on plant free proline content analyses. Phytochemical Analysis, 21(6), 561-565. https://doi.org/10.1002/pca.1233

Journal Article Type Article
Publication Date Nov 1, 2010
Journal Phytochemical Analysis
Print ISSN 0958-0344
Electronic ISSN 1099-1565
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 21
Issue 6
Pages 561-565
DOI https://doi.org/10.1002/pca.1233
Keywords cold storage, Fragaria ananassa Duch, glycine max (L.) Merr, hypocotyl, liquid nitrogen, leaf, proline analyses, sample processing
Public URL https://uwe-repository.worktribe.com/output/988690
Publisher URL http://dx.doi.org/10.1002/pca.1233