Establishment of an in vitro fertilization system in rice (Oryza sativa L.)
Establishment of an in vitro fertilization system in rice (Oryza sativa L.)
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DOI：
10.1007/s00425-007-0506-2
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发表时间：
2007-03
期刊：
影响因子：
4.3
通讯作者：
T. Uchiumi;I. Uemura;T. Okamoto
中科院分区：
文献类型：
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作者：
T. Uchiumi;I. Uemura;T. Okamoto
由于植物的雌配子体深埋在子房中，因此利用分离的雄配子和雌配子的体外受精（IVF）系统已被用于解剖被子植物中的受精诱导事件，例如卵激活、合子发育和早期胚胎发生。本研究建立了水稻体外受精体系，以充分利用水稻研究的丰富资源，研究水稻受精和早期胚胎发生的机制。采用改良的电融合方法进行配子融合，融合产物合子形成细胞壁和额外的核仁。融合后15-24 h，合子分裂为二细胞胚，融合后48 h，合子发育为球形胚，平均由15-16个细胞组成。IVF产生的受精卵与植物中产生的受精卵的发育过程的比较表明，IVF产生的受精卵以与植物中的受精卵高度相似的方式发育并生长成早期球形阶段的胚胎。虽然试管受精产生的球形胚没有发育成球形晚期或分化胚，而是发育成形状不规则的细胞团，但从细胞团再生出可育植株，从这些植株收获的种子正常萌发。本研究建立的水稻体外受精系统为研究被子植物早期胚胎发生的分子机制和培育新品种提供了有力的工具。
In vitro fertilization (IVF) systems using isolated male and female gametes have been utilized to dissect fertilization-induced events in angiosperms, such as egg activation, zygote development and early embryogenesis, as the female gametophytes of plants are deeply embedded within ovaries. In this study, a rice IVF system was established to take advantage of the abundant resources stemming from rice research for investigations into the mechanisms of fertilization and early embryogenesis. Fusion of gametes was performed using a modified electrofusion method, and the fusion product, a zygote, formed cell wall and an additional nucleolus. The zygote divided into a two-celled embryo 15–24 h after fusion, and developed into a globular-like embryo consisting of an average of 15–16 cells by 48 h after fusion. Comparison of the developmental processes of zygotes produced by IVF with those of zygotes generated in planta suggested that zygotes produced by IVF develop and grow into early globular stage embryos in a highly similar manner to those in planta. Although the IVF-produced globular embryos did not develop into late globular-stage or differentiated embryos, but into irregularly shaped cell masses, fertile plants were regenerated from the cell masses and the seeds harvested from these plants germinated normally. The rice IVF system reported here will be a powerful tool for studying the molecular mechanisms involved in the early embryogenesis of angiosperms and for making new cultivars.