分离合子和原胚可以为植物受精和胚胎发生提供很好的研究材料, 因此具有重要意义。用酶解-振荡法分离出五唇兰(Doritis pulcherrima)受精后胚囊, 然后以显微解剖获得合子和原胚。酶解液组成为0.7%–1.3%纤维素酶、0.6%–1.0%果胶酶和10%甘露醇, pH值为5.8, 酶解时间0.5–3.0小时。在分离的早期原胚和接近成熟的球形胚中, 珠孔端均有较发达的胚柄吸器。实验获得了对五唇兰胚胎发育的新认识, 合子和原胚的成功分离也为进一步的细胞学和分子生物学研究打下基础。
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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