Chinese Bulletin of Botany ›› 2012, Vol. 47 ›› Issue (3): 286-291.DOI: 10.3724/SP.J.1259.2012.00286

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Isolation of Zygotes and Proembryos in Doritis pulcherrima

Chenghou Wu1,2, Dongmei Li3, Huiqiao Tian1*   

  1. 1School of Life Sciences, Xiamen University, Xiamen 361005, China;

    2Guangzhou Institute of Landscape Gardening, Guangzhou 510405, China;

    3Shunde Polytechnic, Shunde 528300, China
  • Received:2011-10-28 Revised:2012-03-02 Online:2012-05-01 Published:2012-07-06
  • Contact: Huiqiao Tian

Abstract: The isolation of zygotes and proembryos would provide invaluable material for the study of plant fertilization and embryogenesis. We isolated zygotes and proembryos by manual microdissection from Doritis pulcherrima embryo sacs with only one layer of integument cells after pretreatment combining enzymatic maceration with shaking to the ovules. The enzymatic maceration was with 0.7%–1.3% cellulase, 0.6%–1.0% pectinase, and 10% mannitol, pH5.8, for one-half to 3 hours. Suspensor haustorium was discovered on the micropylar end from early proembryo to maturing spherical proembryo stages. The successful isolation of zygotes and proembryos in D. pulcherrima provides a novel approach to study embryogenesis and is a valuable experimental tool for further detailed research at the cellular and molecular levels.

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