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LC-MS-Based Proteomics of Arabidopsis Phloem Sap

UNSPECIFIED (2009) LC-MS-Based Proteomics of Arabidopsis Phloem Sap. In: 20th International Conference for Arabidopsis Research, 30 June - 4 July 2009, Edinburgh, Scotland, United Kingdom.

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Abstract

We are interested in phloem as a dynamic compartment that transports not only photoassimilates but also many other metabolites and macromolecules, some of which have long-distance signalling functions. Here, we report preliminary characterisation of the phloem sap proteome of Arabidopsis thaliana, aiming to provide an essential data resource for future studies of macromolecule transport and signalling. Using samples derived by petiole exudation into EDTA solutions, tryptic digests were generated from total protein extracts or from excised portions of 1D-SDS PAGE separations. Nano-flow liquid chromatography-tandem MS (LC-MS-MS) was used to generate spectra from peptides, allowing conclusive identification of proteins. A total of 1353 peptides was matched from the MS-MS spectra when searched via MASCOT against the Arabidopsis databases. From these peptides, over 150 unique Arabidopsis proteins were identified. Some of these are homologs of proteins previously identified from phloem of Brassica, rice and cucurbits. However, a substantial proportion represent proteins not previously identified from phloem sap, including many with no functional annotations. Of those with functional annotations, the greatest over-representation was for proteins with roles in abiotic and biotic stress responses. We also developed a method for targeted analysis of the systemic protein hormone signal, FLOWERING LOCUS T (FT), using LC-MS with Multiple Reaction Monitoring (MRM). FT protein was detected in phloem or leaves of wild type and positive control FT-overexpressing plants, and was confirmed as absent from ft null mutant negative controls.

Item Type:Conference or Workshop Item (Poster)
Uncontrolled Keywords:Phloem sap, proteomics, Arabidopsis, FT
Subjects:Q Science > Q Science (General)
Divisions:Biosciences and Bioengineering (Formerly known)
ID Code:9243
Deposited By: Ms. Zaidah Rahmat
Deposited On:29 Aug 2009 00:30
Last Modified:02 Jun 2010 01:59

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