Proteome Profiling of Flax (Linum usitatissimum) Seed: Characterization of Functional Metabolic Pathways Operating during Seed Development

Abstract

Flax (Linum usitatissimum L.) seeds are an important source of food and feed due to the presence of various health promoting compounds, making it a nutritionally and economically important plant. An in-depth analysis of the proteome of developing flax seed is expected to provide significant information with respect to the regulation and accumulation of such storage compounds. Therefore, a proteomic analysis of seven seed developmental stages (4, 8, 12, 16, 22, 30, and 48 days after anthesis) in a flax variety, NL-97 was carried out using a combination of 1D-SDS-PAGE and LC-MSE methods. A total 1716 proteins were identified and their functional annotation revealed that a majority of them were involved in primary metabolism, protein destination, storage and energy. Three carbon assimilatory pathways appeared to operate in flax seeds. Reverse transcription quantitative PCR of selected 19 genes was carried out to understand their roles during seed development. Besides storage proteins, methionine synthase, RuBisCO and S-adenosylmethionine synthetase were highly expressed transcripts, highlighting their importance in flax seed development. Further, the identified proteins were mapped onto developmental seed specific expressed sequence tag (EST) libraries of flax to obtain transcriptional evidence and 81% of them had detectable expression at the mRNA level. This study provides new insights into the complex seed developmental processes operating in flax.

PT-Flax (phenotyping and TILLinG of flax): development of a flax (Linum usitatissimumL.) mutant population and TILLinG platform for forward and reverse genetics

“…The flax genome has recently been sequenced [13], physical and genetic maps have been developed and SNP markers identified [10]. In addition, flax specific microarrays, proteomics and metabolomics [6,7,12,50,51] have also been used to increase our knowledge of flax biology. In this paper we report the development of a flax TILLinG platform for forward and reverse genetics in this economically important species.…”

Fusarium oxysporum mediates systems metabolic reprogramming of chickpea roots as revealed by a combination of proteomics and metabolomics

“…Gene‐specific primers were designed using Primer Express (v2.0) software (Applied Biosystems) and listed in Table S4. Real‐time PCR was carried out as earlier described (Barvkar et al ., ) using FastStart universal SYBR green master mix (Roche, Mannheim, Germany) with 7900HT Fast real‐time PCR system (Applied Biosystems). The Initiation factor 4α ( IF4 α) gene was used as internal standard or reference gene (Garg et al ., ).…”

Agriproteomics of Bread Wheat: Comparative Proteomics and Network Analyses of Grain Size Variation

“…2012;Kim et al, 2010;Skylas et al, 2005). Combined use of SDS-PAGE and LC-MS E techniques has been proposed to be more effective (Salekdeh and Komatsu, 2007) and previously utilized for profiling the complex flax seed proteome (Barvkar et al, 2012), insect gut proteome (Dawkar et al, 2011), and to understand the kiwi fruit ripening (Minas et al, 2012). To the best of our knowledge, this is the first report of the wheat grain proteomics using combination of SDS-PAGE and LC-MS E .…”