J Integr Plant Biol. ›› 2010, Vol. 52 ›› Issue (12): 1043-1058.DOI: 10.1111/j.1744-7909.2010.00996.x

• Cell and Developmental Biology • Previous Articles     Next Articles

Isobaric Tags for Relative and Absolute Quantification- based Comparative Proteomics Reveals the Features of Plasma Membrane-Associated Proteomes of Pollen Grains and Pollen Tubes from Lilium davidii

Bing Han1,4†, Sixue Chen2†, Shaojun Dai3†, Ning Yang1,4 and Tai Wang1*   

  1. 1Research Center for Molecular & Developmental Biology, Key Laboratory of Photosynthesis & Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, and National Center for Plant Gene Research, Beijing 100093, China
    2Department of Biology, Genetics Institute and Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, Florida 32610, USA
    3Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field, Ministry of Education, Northeast Forestry University, Harbin 150040, China
    4Graduate University of the Chinese Academy of Sciences, Beijing 10049, China
  • Received:2010-06-30 Accepted:2010-09-06 Published:2010-09-22
  • About author:*Corresponding author Tel: +86 10 6283 6210; Fax: +86 10 6259 4170; E-mail: twang@ibcas.ac.cn
    These authors contributed equally to this work.

Abstract:

Mature pollen grains (PGs) from most plant species are metabolically quiescent. However, once pollinated onto stigma, they quickly hydrate and germinate. A PG can give rise to a vegetative cell-derived polarized pollen tube (PT), which represents a specialized polar cell. The polarized PT grows by the tip and requires interaction of different signaling molecules localized in the apical plasma membrane and active membrane trafficking. The mechanisms underlying the interaction and membrane trafficking are not well understood. In this work, we purified PG and PT plasma-membrane vesicles from Lilium davidii Duch. using the aqueous two-phase partition technique, then enriched plasma membrane proteins by using Brij58 and KCl to remove loosely bound contaminants. We identified 223 integral and membrane-associated proteins in the plasma membrane of PGs and PTs by using isobaric tags for relative and absolute quantification (iTRAQ) and 2-D high-performance liquid chromatography-tandem mass spectrometry. More than 68% of the proteins have putative transmembrane domains and/or lipid-modified motifs. Proteins involved in signal transduction, membrane trafficking and transport are predominant in the plasma-membrane proteome. We revealed most components of the clathrin-dependent endocytosis pathway. Statistical analysis revealed 14 proteins differentially expressed in the two development stages: in PTs, six upregulated and eight downregulated are mainly involved in signaling, transport and membrane trafficking. These results provide novel insights into polarized PT growth.

Han B, Chen S, Dai S, Yang N, Wang T (2010) Isobaric tags for relative and absolute quantification-based comparative proteomics reveals the features of plasma membrane-associated proteomes of pollen grains and pollen tubes from Lilium davidii. J. Integr. Plant Biol. 52(12), 1043–1058.

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