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The male gametophyte of Arabidopsis is a three-celled pollen grain that is thought to contain almost all the mRNAs needed for germination and rapid pollen tube growth. We generated a reference map of the Arabidopsis mature pollen proteome by using multiple protein extraction techniques followed by 2-DE and ESI-MS/MS. We identified 135 distinct proteins from a total of 179 protein spots. We found that half of the identified proteins are involved in metabolism (20%), energy generation (17%), or cell structure (12%); these percentages are similar to those determined for the pollen transcriptome and this similarity is consistent with the idea that in addition to the mRNAs, the mature pollen grain contains proteins necessary for germination and rapid pollen tube growth. We identified ten proteins of unknown function, three of which are flower- or pollen-specific, and we identified nine proteins whose RNAs were absent from the transcriptome, seven of which are involved in metabolism, energy generation, or cell wall structure. Our work complements and extends recent analyses of the pollen transcriptome.

The commercial value of the wheat crop is a function of the quality and amount of the storage protein and starch present in the grain, which in turn are influenced by environmental conditions during grain-fill. To understand how environment modifies the size and composition of wheat grains, we set out to identify the key metabolic and regulatory proteins in developing grain. We present results of initial studies aimed at establishing instrument conditions that will allow us to identify cytoplasmic proteins present in wheat endosperm. Proteins were isolated, separated by 2D gel electrophoresis and stained with Coomassie blue to visualize and quantify changes in protein expression. Mass spectrometry was used to identify protein spots in 2D gels by means of "peptide mass maps" of in-gel enzymatically digested protein spots. Because only about 30% of the proteins could be identified by "peptide mass mapping," we developed nano-flow LC-MS/MS techniques that allowed us to identify about 80% of the salt-soluble proteins in wheat endosperm.