Cytological Techniques to Study Cytomixis in Plant Male Meiosis.

Abstract

In this chapter we describe cytological techniques to study cytomixis, a process of nuclear migration between plant cells, in squashed plant male meiocytes of Nicotiana tabacum and Secale cereale. To perform immunostaining or fluorescence in situ hybridization (FISH) on meiotic cells involved in cytomixis common protocols are modified. During preparation of specimens for subsequent cytological analysis, it is necessary not only to make DNA and proteins accessible to DNA probes and antibodies, but also to preserve cell cytoplasm. There are also some important modifications in the protocols applied for meiocytes of different plant species. Here we describe protocols for immunostaining and FISH in rigid tobacco male meiocytes with dense cytoplasm and thick callose wall, that tolerate hard squashing, and in soft rye male meiocytes, that are easily damaged upon squashing, both to study cytomixis.