Biology Reference
In-Depth Information
Chapter 30
Pollen Cultivation and Preparation for Proteomic Studies
Heidi Pertl-Obermeyer and Gerhard Obermeyer
Abstract
The quality of the collected experimental data very much depends on the quality of the biological starting
material. Especially the proteome analysis of a highly dynamic system like the germinating and tube-
growing pollen grain needs several precautions which allow an accurate and acceptable interpretation of
the obtained results. Optimized protocols for pollen collection, storage, and in vitro culture as well as
pollen organelle separations are described which help to obtain well-defi ned and reproducible experimental
conditions for the subsequent proteomic analysis.
Key words
Allergy, Germination,
Lilium longifl orum
, Organelle isolation, Marker enzyme, Pollen
grain, Pollen tube, Tip growth
1
Introduction
For several reasons the male gametophytes of plants (the pollen)
are interesting subjects for proteome research. Firstly, they are the
source of allergens affecting human health. Secondly, the germina-
tion of pollen grains and the subsequent pollen tube growth are
rapid and highly dynamic processes during plant reproduction
which, in turn, is the basis for food production for a growing
human population. Finally, the special growth process known as tip
growth, by which the pollen tubes elongate, is a complex network
of interacting proteins and signal transduction cascades. Table
1
presents some of the important pollen proteome studies. No pro-
teomic studies on pollen allergens are included because the majority
of these reports use commercially available pollen grains that are
hardly able to germinate anymore and mainly focus on the allergy-
causing properties of proteins but not on their physiological role.
To investigate the various biological questions related to pollen
allergy and pollen physiology, the source and the way how the
biological material is prepared are essential prerequisites for the
accuracy of the subsequent analysis. Here we describe several
methods to obtain and prepare defi ned protein fractions from
