Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2004 Aug;135(4):2241-60.
doi: 10.1104/pp.104.041947. Epub 2004 Aug 6.

Proteome reference maps of vegetative tissues in pea. An investigation of nitrogen mobilization from leaves during seed filling

Séverine Schiltz  1 , Karine Gallardo, Myriam Huart, Luc Negroni, Nicolas Sommerer, Judith Burstin
Affiliations

Proteome reference maps of vegetative tissues in pea. An investigation of nitrogen mobilization from leaves during seed filling

Séverine Schiltz et al. Plant Physiol. 2004 Aug.

Abstract

A proteomic approach was used to analyze protein changes during nitrogen mobilization (N mobilization) from leaves to filling seeds in pea (Pisum sativum). First, proteome reference maps were established for mature leaves and stems. They displayed around 190 Coomassie Blue-stained spots with pIs from 4 to 7. A total of 130 spots were identified by mass spectrometry as corresponding to 80 different proteins implicated in a variety of cellular functions. Although the leaf proteome map contained more abundant spots, corresponding to proteins involved in energy/carbon metabolism, than the stem map, their comparison revealed a highly similar protein profile. Second, the leaf proteome map was used to analyze quantitative variations in leaf proteins during N mobilization. Forty percent of the spots showed significant changes in their relative abundance in the total protein extract. The results confirmed the importance of Rubisco as a source of mobilizable nitrogen, and suggested that in pea leaves the rate of degradation of Rubisco may vary throughout N mobilization. Correlated with the loss of Rubisco was an increase in relative abundance of chloroplastic protease regulatory subunits. Concomitantly, the relative abundance of some proteins related to the photosynthetic apparatus (Rubisco activase, Rubisco-binding proteins) and of several chaperones increased. A role for these proteins in the maintenance of a Rubisco activation state and in the PSII repair during the intense proteolytic activity within the chloroplasts was proposed. Finally, two 14-3-3-like proteins, with a potential regulatory role, displayed differential expression patterns during the massive remobilization of nitrogen.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
N mobilization in vegetative nodes during pea seed development. Nitrogen content (%) in vegetative nodes 7 and 8 (nodes below the first flowering node) is represented on the left axis, dry weight (g) and water content (×100%) in the whole seeds are represented on the right axis. For the vegetative tissues, DAP corresponded to the DAP on the second flowering node. The seed-filling phase started at about 10 DAP and ended at about 24 DAP. Leaf yellowing started at 24 DAP. A, Phases of seed development. B, Stages associated with leaf senescence/break down of the vegetative nodes.
Figure 2.
Figure 2.
Two-dimensional protein patterns of leaves obtained from a constant fresh-tissue load. An equal volume of 80 μL corresponding to 240 mg of fresh weight of leaves harvested at 4, 12, and 24 DAP of the second flowering node was loaded on linear 24-cm IPG strips (pH 4–7). Gels were Coomassie Blue stained. The figure shows representative experiments carried out at three times. A, Enlarged image of massive degradation of rbcL between 4 and 24 DAP. B, Enlarged image represents some less abundant protein spots (labeled by arrows) with constant levels between 4 and 24 DAP.
Figure 3.
Figure 3.
Two-dimensional reference maps obtained from mature pea leaves (A) and stems (B) at 4 DAP of the second flowering node. Two-dimensional gels were performed from 200 μg of proteins using 24-cm IPG strips (linear pH 4–7) and Coomassie Blue stained. Two examples of protein profile similarities are shown in enlarged images in (C), (a), and (b) from both leaf and stem proteome maps. The figure shows representative experiments carried out five times. The labeled proteins are listed in Table I.
Figure 4.
Figure 4.
Assignment of the identified proteins to functional categories by using the classification described by Bevan et al. (1998). A total of 130 spots representing 80 different proteins were classified. The same spot identified by both MS methods (MALDI-TOF and LC-MS/MS) in both leaf and stem extracts was counted once.
Figure 5.
Figure 5.
Relative abundance of the different classes of identified proteins determined according to Bevan et al. (1998), in leaves (A) and stems (B). Abundance of protein categories was expressed as the percentage of the total volume of all detected spots in 2-DE gels.
Figure 6.
Figure 6.
Characterization of leaf proteins whose relative abundance varies significantly between 4 and 19 DAP. A, Coomassie Blue 2-DE gel of total proteins (pI 4–7) from leaves at 4 DAP. B, C, and D are enlarged. B, Spots 174 and 178 (anther-specific proteins) whose relative abundance decreased after 12 and 4 DAP, respectively. C, Spots 76 (chlorophyll a/b-binding protein type III precursor) and 78 (1,4-benzoquinone reductase-like), whose relative abundance transiently increased at 12 DAP. D, Increased relative abundance of spots 150 (Rubisco activase) and 154 (actin) from 4 to 19 DAP.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Adam Z, Clarke AK (2002) Cutting edge of chloroplast proteolysis. Trends Plant Sci 7: 451–456 - PubMed
    1. Bantignies B, Seguin I, Muzac I, Dedaldechamp F, Gulick P, Ibrahim R (2000) Direct evidence for ribonucleolytic activity of a PR-10-like protein from white lupin roots. Plant Mol Biol 42: 871–881 - PubMed
    1. Bardel J, Louwagie M, Jaquinod M, Jourdain A, Luche S, Rabilloud T, Macherel D, Garin J, Bourguignon J (2002) A survey of the plant mitochondrial proteome in relation to development. Proteomics 2: 880–898 - PubMed
    1. Bevan M, Bancroft I, Bent E, Love K, Goodman H, Dean C, Bergkamp R, Dirkse W, Van Staveren M, Stiekema W, et al (1998) Analysis of 1.9 Mb contiguous sequence from chromosome 4 of Arabidopsis thaliana. Nature 39: 809–821 - PubMed
    1. Bhalerao R, Keskitalo J, Sterky F, Erlandsson R, Björkbacka H, Birve SJ, Karlsson J, Gardeström P, Gustafsson P, Lundeberg J, et al (2003) Gene expression in autumn leaves. Plant Physiol 131: 430–442 - PMC - PubMed

Publication types

LinkOut - more resources