Isolation of Egg Cells from Allium fistulosum

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  • School of Life Sciences, Xiamen University, Xiamen 361005, China

Received date: 2008-06-17

  Revised date: 2008-10-01

  Online published: 2010-11-03

Abstract

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.

Cite this article

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 . DOI: 10.11983/CBB28175

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