We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Wheat egg in vitro fusion with wheat and green bristlegrass sperm.
- Authors
Li, Dong Xiao; Hu, Hai Yan; Ru, Zhen Gang; Tian, Hui Qiao
- Abstract
Summary: Isolated gametes can be used to investigate fertilization mechanisms, and probe distant hybridization between different species. Pollen grains of wheat and Setaria viridis are tricellular, containing sperm cells at anthesis. Sperm from these plants were isolated by breaking open pollen grains in a osmotic solution. Wheat ovules were digested in an enzyme solution for 20 min, and then transferred to an isolation solution without enzymes to separate egg cells from ovules. The fusion of wheat egg cells with wheat and S. viridis sperm was conducted using an electro-fusion apparatus. Under suitable osmotic pressure (10% mannitol), calcium concentration of 0.001% (CaCl2·2H2O), and a 30–35 V alternating electric field for 15 s, egg cells and sperm adhered to each other and became arranged in a line. Electroporation of the plasma membrane of egg cells and sperm using a 300–500 V direct-current electric field (45 µs amplitude pulse) caused them to fuse.
- Subjects
OVUM; SPERMATOZOA; WHEAT; POLLEN; OSMOTIC pressure; MANNITOL; OVALBUMINS
- Publication
Zygote, 2019, Vol 27, Issue 3, p126
- ISSN
0967-1994
- Publication type
Article
- DOI
10.1017/S0967199418000461