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旱稻仿野87-707品系具有多年生趋势,是我院研究多年生旱稻的一个材料。我们用它进行原生质体培养,想为研究多年生旱稻基因改良打下基础。现用该品系的成熟胚诱导的愈伤组织,在含有氨基酸丰富的AA培养基中进行细胞悬浮培养。取该细胞悬浮培养物游离的原生质体,用琼脂糖包埋于改良的KM8P培养基中,发生了持续分裂。经一系列变换不同分化培养基的培养再生成完整植株并移栽成活。对游离和培养方法做了如下改进:1)悬浮细胞培养基中加入ABA有利于原生质体的存活和分裂;2)掌握好原生质体游离前悬浮细胞的状态是游离成功的关键;3)琼脂糖包理培养可大大提高植板率;4)用较高渗透压的培养基培养原生质体再生的细胞团及愈伤组织,有利于提高植株再生能力;5)变换使用不同的分化培养基可明显提高植株的再生频率。这是迄今国内外第一个由旱稻原生质体再生植株的报道。
Upland 87-007 Upland Rice has a perennial trend and is one of the materials used by our institute to study perennial dry rice. We use it for protoplast culture and want to lay the foundation for studying genetic improvement of perennial upland rice. The callus induced by the mature embryos of this line is cultured in suspension in AA-rich AA medium. The free protoplasts of this cell suspension culture were stained with Sepharose in a modified KM8P medium and continued to divide. After a series of transformation of different culture medium differentiation and regeneration of the entire plant transplanting survival. The following improvements have been made in terms of free and culture methods: 1) Addition of ABA to suspension cell culture medium is favorable for the survival and division of protoplasts; 2) the key to successful dissociation of protoplast free suspension cells; 3) agarose Inclusion culture can greatly improve the plant plating rate; 4) Cultivation protoplasts regenerated cell mass and callus with higher osmotic pressure medium is conducive to improve plant regeneration ability; 5) Transforming using different differentiation medium can be obvious Improve plant regeneration frequency. This is by far the first report from Upland protoplast regenerating plants at home and abroad.