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Download fileCell-specific Analysis of the Tomato Pollen Proteome from Pollen Mother Cell to Mature Pollen Provides Evidence for Developmental Priming
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posted on 2013-11-01, 00:00 authored by Palak Chaturvedi, Till Ischebeck, Volker Egelhofer, Irene Lichtscheidl, Wolfram WeckwerthTomato is a globally important crop
grown and consumed worldwide.
Its reproductive activity is highly sensitive to environmental fluctuations,
for instance temperature and drought. Here, pollen development is
one of the most decisive processes. The present study aims for the
identification of cell-specific proteins during pollen developmental
stages of tomato. We have setup a protocol for stage-specific pollen
isolation including microsporocytes (pollen mother cells), tetrads,
microspores, polarized microspores, and mature pollen. Proteins were
extracted using phenol and prefractionated using SDS-PAGE followed
by protein digestion, peptide extraction, and desalting. Identification
and quantification of proteins were performed using nanoHPLC coupled
to LTQ-Orbitrap-MS. In total, 1821 proteins were identified. Most
of these proteins were classified based on their homology and designated
functions of orthologs. Cluster and principal components analysis
revealed stage-specific proteins and demonstrated that pollen development
of tomato is a highly controlled sequential process at the proteome
level. Intermediate stages such as tetrad and polarized microspore
are clearly distinguished by different functionality compared to other
stages. From the predicted functions, energy-related proteins are
increased during the later stages of development, which indicates
that pollen germination depends upon presynthesized proteins in mature
pollen. In contrast, heat stress-related proteins are highly abundant
in very early developmental stages, suggesting a dominant role in
stress protection. Taken together, the data provide a first cell-specific
protein reference set for tomato pollen development from pollen mother
cells to the mature pollen and give evidence for developmentally controlled
processes that might help to prepare the cells for specific developmental
programs and environmental stresses.