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Isolation of male and female gametes, zygotes and proembryos of leek (Allium tuberosum Roxb)

Published online by Cambridge University Press:  03 April 2020

Yi Hua Lin ,
Mei Zhen Lin ,
Yu Qing Chen  and
Hui Qiao Tian Open the ORCID record for Hui Qiao Tian [Opens in a new window]
Yi Hua Lin
Affiliation:
Zhangzhou Health Vocational College, Zhangzhou363000, China
Mei Zhen Lin
Affiliation:
Zhangzhou Health Vocational College, Zhangzhou363000, China
Yu Qing Chen
Affiliation:
Zhangzhou Health Vocational College, Zhangzhou363000, China
Hui Qiao Tian*
Affiliation:
School of Life Sciences, Xiamen University, Xiamen361102, China
*
Address for correspondence: Hui Qiao Tian. School of Life Sciences. Xiamen University, Xiamen, Fujian361102, China. Tel: +11 86 592 2186486. E-mail: hqtian@xmu.edu.cn
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Summary

The isolation of male and female gametes is an effective method to study the fertilization mechanisms of higher plants. An osmotic shock method was used to rupture pollen grains of Allium tuberosum Roxb and release the pollen contents, including generative cells, which were mass collected. The pollinated styles were cut following 3 h of in vivo growth, and cultured in medium for 6–8 h, during which time pollen tubes grew out of the cut end of the style. After pollen tubes were transferred into a solution containing 6% mannitol, tubes burst and released pairs of sperm cells. Ovules of A. tuberosum were incubated in an enzyme solution for 30 min, and then dissected to remove the integuments. Following transfer to a dissecting solution free of enzymes, each nucellus was cut in the middle, and squeezed gently on the micropylar end, resulting in the liberation of the egg, zygote and proembryo from ovules at selected stages. These cells can be used to explore fertilization and embryonic development using molecular biological methods for each cell type and development stage.

Keywords

Allium tuberosum RoxbEggProembryoSpermZygote
Type
Research Article
Information
Zygote , Volume 28 , Issue 4 , August 2020 , pp. 278 - 285
Copyright
© Cambridge University Press 2020

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