A novel method for efficient in vitro germination and tube growth of Arabidopsis thaliana pollen.

In addition to its importance in studies of plant reproduction and fertility, pollen is as widely employed as a model system of cell growth and development. This work demands robust, reproducible methods to induce pollen germination and morphologically normal growth of pollen tubes in vitro. Despite numerous advantages of Arabidopsis thaliana as a model plant, such experiments on pollen germination and pollen tube growth have often proved challenging. Our new method employs a physical cellulosic membrane, overlying an agarose substrate. By modulating the substrate composition, we provide important insights into the mechanisms promoting pollen growth both in vitro and in vivo. This effective new technical approach to A. thaliana pollen germination and tube growth results in swift, consistent and unprecedented levels of germination to over 90%. It can also promote rapid growth of long, morphologically normal pollen tubes. This technical development demonstrates that exogenous spermidine and a cellulosic substrate are key factors in stimulating germination. It has potential to greatly assist the study of reproduction in A. thaliana and its closest relatives, not only for the study of germination levels and pollen tube growth dynamics by microscopy, but also for biochemical and molecular analysis of germinating pollen.