Detection of changes in the nuclear phase and evaluation of male germ units by flow cytometry during in vitro pollen tube growth in Alstroemeria aurea

概要

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This study aimed to analyze male gamete behavior from mature pollen to pollen tube growth in the bicellular pollen species Alstroemeria aurea. For mature pollen, pollen protoplasts were examined using flow cytometry. The protoplasts showed two peaks of DNA content at 1C and 1.90C. Flow cytometry of pollen tubes at different developmental stages that were cultured in vitro revealed changes in the nuclear phase at 9 h and 18 h after culture. Sperm cell formation occurred at 6–9 h after culture, indicating that the first change was due to the division of the generative cells into sperm cells. After sperm cell formation, the number of vegetative nucleus associations with sperm cell showed a tendency to increase. The association was suggested as male germ unit. When sperm cells, vegetative nuclei, and partial male germ units were separately collected from pollen tubes cultured for 18 h and analyzed using a flow cytometer, the sperm cells and vegetative nuclei contained 1C DNA, while the DNA content of partial male germ units was counted as 2C. Therefore, the second change in the nuclear phase, which resulted in an increase in 2C nuclei, was possibly related to the formation of male germ units.
Botanical Society of Japanの論文
2009-03-01