A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues
- PDF / 2,447,540 Bytes
- 16 Pages / 595.276 x 790.866 pts Page_size
- 50 Downloads / 174 Views
Plant Methods Open Access
METHODOLOGY
A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues Cheka Kehelpannala1* , Thusitha W. T. Rupasinghe1,2, Thomas Hennessy3, David Bradley3, Berit Ebert1 and Ute Roessner1
Abstract Background: The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds in solution at concentrations adequate for subsequent detection, quantitation and analysis through spectroscopic methods are required. To date, numerous methods are used to extract lipids from plant tissues. However, a comprehensive analysis of the efficiency and reproducibility of these methods to extract multiple lipid classes from diverse tissues of a plant has not been undertaken. Results: In this study, we report the comparison of four different lipid extraction procedures in order to determine the most effective lipid extraction protocol to extract lipids from different tissues of the model plant Arabidopsis thaliana. Conclusion: While particular methods were best suited to extract different lipid classes from diverse Arabidopsis tissues, overall a single-step extraction method with a 24 h extraction period, which uses a mixture of chloroform, isopropanol, methanol and water, was the most efficient, reproducible and the least labor-intensive to extract a broad range of lipids for untargeted lipidomic analysis of Arabidopsis tissues. This method extracted a broad range of lipids from leaves, stems, siliques, roots, seeds, seedlings and flowers of Arabidopsis. In addition, appropriate methods for targeted lipid analysis of specific lipids from particular Arabidopsis tissues were also identified. Keywords: Lipid extraction methods, Arabidopsis, Lipids, Mass spectrometry, QTOF, HPLC, LC–MS, Untargeted lipid analysis Background Lipids are a large group of highly diverse compounds present in all living organisms and cell types [1]. They play a myriad of crucial roles in biological systems as structural components of membranes [2], for energy storage [3], as signalling molecules in various biological pathways and modulators of cellular functions and diseases [4]. *Correspondence: [email protected] 1 School of BioSciences, The University of Melbourne, Melbourne, VIC 3010, Australia Full list of author information is available at the end of the article
Plant lipids are highly complex [5] and essential for plant growth and development [6]. However, little is known about their composition in different tissue types and related functions. To date, mass spectrometry is the most prevalent technique applied to detect and analyze lipids in biological samples due to its high sensitivity, mass accuracy and scan speed [7]. As a result of recent advancements in mass spectrometry and the development of accompanying lipid identification software, the detection a
Data Loading...
