Chinese Bulletin of Botany ›› 2009, Vol. 44 ›› Issue (03): 345-350.DOI: 10.3969/j.issn.1674-3466.2009.03.012
• 技术方法 •
Mingxia Jian, Yanan Zhang, Yaying Wang, Huiqiao Tian*
Viable egg cells of Allium fistulosum were isolated by enzymatic digestion and mechanical dissection. The ovules were incu-bated in an enzymatic solution containing 0-0.8% pectolyse Y-23, 0-1.2% pectinase (Serva), 0-1.2% cellulase (Onozuka R-S), 0-1% hemicellulase (Sigma) and 6%-12% mannitol for 20-40 min to remove the outer integument. Then ovules were tr-ansferred into an isolating solution without enzymes for mechanical dissection. The ovules were cut from the middle part, and the micropylar part was gently extruded by use of a dissection needle. In general, the three cells of the egg apparatus spilled out from the cut end of the ovule. The enzymatic solution containing 0.2% pectolyse Y-23, 0.8% pectinase (Serva), 0.8% cellulase (Onozuka RS), 0.5%hemicellulase (Sigma) and 10% mannitol was best for isolating about 18 egg cells from 30 ovules in 2 h. The isolated egg cells could be used for in vitro fertilization to explore the fertilization mechanism and for egg development with molecular biological methods.
Mingxia Jian;Yanan Zhang;Yaying Wang;Huiqiao Tian*. Isolation of Egg Cells from Allium fistulosum[J]. Chinese Bulletin of Botany, 2009, 44(03): 345-350.
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