Ti质粒介导的B、t、k-δ内毒素蛋白基因转化大豆的初步研究

STUDIES ON TRANSFERRING B.t. k-DELTA ENDOTOXIN GENE INTO SOYBEAN WITH Ti-PLASMID PRIMARIL

以Ｔｉ质粒为介导，将ｐＫＴ５４Ｂ７Ｃ５质粒上的Ｂ、ｔ、ｋ－δ内毒素蛋白基因导入东北大豆“黑农３７”、“黑农３９”等品种。采用多种外植体和感染方法，从胚轴和子叶节诱导出丛生芽与再生植株。经卡那霉素筛选和冠瘿碱检测，初步证明外源基因导入大豆基因组中。共获得８１株再生植株，其中成活３０株，仅３株结７个荚，得到７粒种子。ＰＣＲ检测和ＤＮＡ分子杂交，鉴定这７株再生植株呈阳性反应，证明外源基因已导入并整合到大豆的基因组中。７粒种子均已萌发。

The pKT 5 4B 7C 5 plasmid with B.t.k.-delta endoxin gene were trnsferred into Heinong 37 and Heinong 39 et. al soybean cultivars by inducting with various explant methods. Adventitious buds and regeneration plant were obtained from hypocotyl, cotylendon node. They were detected by kanamycine selection and alkaoids and primarily proved that foreign gene were transferred into soybean genome. Only 30 of 81 regenerated plant survived. Only 3 plants with 7 pods were obtained from the survived regenerated plants. It proved that 7 plants of the regenerated plants were transferred with forgein gene by PCR technique and dot hybrization. The 7 seeds sprouted, plant growth is normal.