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Liu Z.,Australian Department of Primary Industries and Fisheries | Liu Z.,Molecular Plant Breeding Co operative Research Center | Mouradov A.,Australian Department of Primary Industries and Fisheries | Mouradov A.,Molecular Plant Breeding Co operative Research Center | And 6 more authors.
Analytical Biochemistry | Year: 2011

Current methods for measuring fructan levels in plant tissues are time-consuming and costly. They often involve multiple or sequential extractions, enzymatic or acid hydrolysis of fructan polymers, and multiple HPLC runs to quantify fructan-derived hexoses. Here we describe a new method that requires a single extraction step, followed by selective precipitation of fructans by acetone, acid hydrolysis of the precipitate, and a short (10 min) HPLC run to complete the procedure. We used perennial ryegrass samples to show that the new method has similar sensitivity, but better reproducibility, than a more complex method that is widely used. We have used the new method to study developmentally related changes in fructan levels in glasshouse-grown perennial ryegrass plants. Copyright © 2011 Published by Elsevier Inc. All rights reserved.

Abeynayake S.W.,Australian Department of Primary Industries and Fisheries | Abeynayake S.W.,Molecular Plant Breeding Co operative Research Center | Abeynayake S.W.,La Trobe University | Panter S.,Australian Department of Primary Industries and Fisheries | And 7 more authors.
Plant Methods | Year: 2011

Background: Histochemical staining of plant tissues with 4-dimethylaminocinnamaldehyde (DMACA) or vanillin-HCl is widely used to characterize spatial patterns of proanthocyanidin accumulation in plant tissues. These methods are limited in their ability to allow high-resolution imaging of proanthocyanidin deposits.Results: Tissue embedding techniques were used in combination with DMACA staining to analyze the accumulation of proanthocyanidins in Lotus corniculatus (L.) and Trifolium repens (L.) tissues. Embedding of plant tissues in LR White or paraffin matrices, with or without DMACA staining, preserved the physical integrity of the plant tissues, allowing high-resolution imaging that facilitated cell-specific localization of proanthocyanidins. A brown coloration was seen in proanthocyanidin-producing cells when plant tissues were embedded without DMACA staining and this was likely to have been due to non-enzymatic oxidation of proanthocyanidins and the formation of colored semiquinones and quinones.Conclusions: This paper presents a simple, high-resolution method for analysis of proanthocyanidin accumulation in organs, tissues and cells of two plant species with different patterns of proanthocyanidin accumulation, namely Lotus corniculatus (birdsfoot trefoil) and Trifolium repens (white clover). This technique was used to characterize cell type-specific patterns of proanthocyanidin accumulation in white clover flowers at different stages of development. © 2011 Abeynayake et al; licensee BioMed Central Ltd.

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