Meiotic chromosome preparation techniques of pollen mother cells for laser micro-dissection in Populus spp.

Abstract

In order to isolate meiotic chromosomes of Populus species meiotic chromosome preparation techniques of pollen mother cells for laser micro-dissection were studied. Pollen mother cells at diakinesis of Populus canadensis Moench were used as samples. Two methods were used to prepare meiotic chromosomes: in the first, cell suspensions were dropped on polyethylene-naphthalate or polyester membrane slides which had just been incubated at −20°C; in the second method, cell suspensions were also dropped on polyethylene-naphthalate or polyester membrane slides, but spread with the aid of high temperatures. The cells did not completely spread by the first method and chromosomes at diakinesis could not be individually distinguished. In contrast, well-spread diakinesis chromosomes were obtained by the second method, where chromosomes, connected with their nucleolus, were successfully isolated with the laser micro-dissection system. As well, we discuss the prospect of applications of laser micro-dissection in cytogenetics and molecular genetics in Populus species.