Abstract
Meiosis is a highly dynamic and precisely regulated process of cell division, leading to the production of haploid gametes from one diploid parental cell. In the crop plant barley (Hordeum vulgare), male meiosis occurs in anthers, in specialized cells called meiocytes. Barley meiotic tissue is scarce and not easily accessible, making meiosis study a challenging task. We describe here a new micro-proteomics workflow that allows sensitive and reproducible genome-wide label-free proteomic analysis of individual staged barley anthers. This micro-proteomic approach detects more than 4,000 proteins from such small amounts of material as two individual anthers, covering a dynamic range of protein relative abundance levels across five orders of magnitude. We applied our micro-proteomics workflow to investigate the proteome of the developing barley anther containing pollen mother cells in the early stages of meiosis and we successfully identified 57 known and putative meiosis-related proteins. Meiotic proteins identified in our study were found to be key players of many steps and processes in early prophase such as: chromosome condensation, synapsis, DNA double-strand breaks or crossover formation. Considering the small amount of starting material, this work demonstrates an important technological advance in plant proteomics and can be applied for proteomic examination of many size-limited plant specimens. Moreover, it is the first insight into the proteome of individual barley anther at early meiosis. The proteomic data have been deposited to the ProteomeXchange with the accession number PXD010887.
Highlights
Proteomics is a potent approach to identify and explore functionally important mechanisms and processes in diverse fields of biology
A consistent feature of many genome-wide proteomic studies is that they typically analyze populations of proteins extracted from tens of thousands to millions of cells derived from complex tissues comprised of multiple cell types, or from bulked small tissue samples isolated from multiple individuals
Sectioning, staining and cell counting revealed that approximately 10% of cells in the developing barley anther are meiocytes (Figure 1B), cells that undergo meiosis and subsequently develop into the haploid pollen grains that contain recombined parental chromosomes
Summary
Proteomics is a potent approach to identify and explore functionally important mechanisms and processes in diverse fields of biology. Proteomics studies have been relatively infrequently used in plant biology When they have been used successfully, global proteome studies have revealed a repertoire of up to 13,029 proteins in Arabidopsis by merging information from multiple tissue and organ samples (Baerenfaller et al, 2008) and, as an example, in targeted studies over 500 ubiquitylated proteins from different leaf tissue cells were identified after inhibition of the Ubiquitin-26S proteasome with the specific inhibitor Syringolin A followed by affinity enrichment of ubiquitinylated proteins (Svozil et al, 2014, 2015). While these represent significant successes, a major challenge that remains is how to best exploit the available and increasingly powerful proteomics technologies to understand cell and tissue specific processes or to identify and interrogate dysregulated cellular networks induced by variables that include developmental or environmental change, or genetic mutation (genotype)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
