大豆不定胚悬浮培养基因型筛选及基因枪遗传转化的研究

Screening of Soybean Genotypes Suitable for Suspension Culture with Adventitious Embryos and Genetic Transformation by Particle Bombardment

对32个不同基因型大豆的幼胚进行不定胚诱导,30 d时统计分析初生胚诱导率、次生胚诱导率和诱导效率,经综合比较得出适合体细胞诱导不定胚的4种基因型为L155、垦丰16、绥农25和吉育91。然后以不定胚诱导率较高的L155为材料建立大豆悬浮培养不定胚繁殖体系,并用基因枪轰击的方法将GUS表达盒片段转入不定胚细胞团。X-gluc染色后显微观察到基因蓝色产物,说明GUS基因通过微弹轰击法导入大豆不定胚细胞并表达。

In this work,we used the immature cotyledons of 32 soybean genotypes to induce adventitious embryos and com- pared the percentage of primary embryo induction, percentage of secondary embryo induction and induction efficiency after 30 days inducing culture. The results showed that L155, Kenfeng 16, Suinong 25 and Jiyu 91 had higher frequency of adventitious embryos induction. Furthermore, L155 was selected to further establish the soybean suspension culture system. The secondary embryogenesis systems were used for GUS gene transformation by particle gun bombardment method. The GUS staining results showed that the GUS gene was imported into soybean adventitious embryos cells and successfully expressed.