micro-proteomics experiment for single meiotic-phase barley anthers

We describe here a new micro-proteomics workflow for the analysis of individual barley anthers. The workflow detects more than 4000 proteins from small amounts of material representing single or paired anther samples, covering a dynamic range of protein expression levels across five orders of magnitude. We consistently identified ~2800 proteins in a single 0.6mm anther and ~4000 proteins in a paired-anther sample. This demonstrates an important technological advance in plant proteomics, considering the limited amount of starting material. We employed our micro-proteomics workflow to investigate the proteome of the developing barley anther containing male gametes in the early stages of meiosis. We successfully identified several low abundant meiosis-related proteins, proving that our approach is highly sensitive and consequently powerful. To date, no work has been published previously on precisely staged individual plant anthers.