Somatic Embryogenesis and Plant Regeneration from Immature Persimmon (Diospyros kaki Thunb.) Embryos
Somatic Embryogenesis and Plant Regeneration from Immature Persimmon (Diospyros kaki Thunb.) Embryos
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DOI：
10.21273/hortsci.43.1.211
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发表时间：
2008-02
期刊：
影响因子：
1.9
通讯作者：
A. Sugiura;Yoshiko Matsuda-Habu;M. Gao;T. Esumi;R. Tao
中科院分区：
文献类型：
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作者：
A. Sugiura;Yoshiko Matsuda-Habu;M. Gao;T. Esumi;R. Tao
在柿子中，培养细胞的植株再生通常通过不定芽形成。如果体细胞胚胎发生成为可能，将大大提高大规模繁殖和基因工程的效率。我们尝试从未成熟胚诱导体细胞胚胎发生，并从诱导胚诱导植株再生。将未成熟的‘扶玉’和‘次子’种子的下胚轴和子叶在含结冷胶固化的Murashige和Skoog培养基中进行暗培养，培养基中添加不同浓度的2,4-二氯苯氧乙酸（2,4- d）和6-苄基腺氨酸（BA）。愈伤组织形成始于培养约2周，2,4- d和BA在3 μM或10 μM组合时愈伤组织形成率最高。最初形成的愈伤组织逐渐变成褐色或黑色，随后出现白色胚性愈伤组织。培养约8周后，这些EC形成球状胚，并持续到培养20周。“扶玉”的球形胚形成率高于“二郎”，尤其是下胚轴。将具有球形胚的EC转移到不含植物生长调节剂的新鲜培养基中，约70%的EC在6周内发育为鱼雷型胚期。形成的鱼雷型胚在含或不含玉米素的琼脂培养基中发芽生根，数周后不进入休眠状态。发芽生根后，移栽到相同的培养基中，再驯化4周。由于胚萌发和生根同时进行，植株易于在盆中生长，移栽时无震荡。本文首次报道了柿子体细胞胚再生植株。
In persimmon, plant regeneration from cultured cells usually takes place through adventitious bud formation. If somatic embryogenesis were possible, the efficiency of mass propagation and genetic engineering would be greatly improved. We attempted to induce somatic embryogenesis from immature embryos and plant regeneration from the induced embryos. Hypocotyls and cotyledons from immature 'Fuyu' and 'Jiro' seeds were cultured in the dark in Murashige and Skoog medium solidified with gellan gum and supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) at various concentrations. Callus formation started at ≈2 weeks of culture, and the callus formation rate was highest at 3 or 10 μM combinations of 2,4-D and BA. The initially formed calli gradually became brown or black from which white embryogenic calli (EC) appeared secondarily. After ≈8 weeks of culture, globular embryos were formed from these EC, and the formation proceeded until 20 weeks of culture. Formation of globular embryos was higher with 'Fuyu' than 'Jiro', especially with hypocotyls. When EC with globular embryos were transferred to fresh medium with no plant growth regulators, ≈70% developed to the torpedo-type embryo stage in 6 weeks. The torpedo-type embryos thus formed were germinated and rooted in agar medium with or without zeatin in several weeks without entering dormancy. After germination and rooting, the plantlets were transferred to the same medium and acclimatized for another 4 weeks. As the embryos germinated and rooted simultaneously, the plantlets were easy to grow in pots without transplanting shock. This is the first report on plant regeneration through somatic embryogenesis of persimmon.