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硼在维持植物细胞壁和细胞膜等方面有着重要的作用。以大豆品种吉林小粒7号为试验材料,研究硼浓度对大豆子叶节丛生芽的诱导的影响,并在不同硼元素浓度下采用农杆菌介导法将报告基因(GUS)导入大豆中,用组织化学法测定GUS基因的表达并计算瞬时表达率,最后采用PCR和斑点杂交做进一步的鉴定,以探讨硼介导的大豆转化效率及可行性。结果显示高硼和低硼培养基对大豆丛生芽诱导均有一定的抑制作用,尤其是高硼情况下丛生芽诱导率显著降低。30 mg·L-1硼酸环境下农杆菌介导的大豆转化效率较之正常硼酸环境下有显著的提高,GUS瞬时转化率高达35.92%。这一现象对建立新的大豆遗传转化体系的意义有待于进一步探究。
Boron plays an important role in maintaining plant cell wall and cell membrane. The effect of boron concentration on the induction of clump buds in soybean cotyledon was studied using soybean variety Jilin Xiaoyan No.7 as test material. The reporter gene (GUS) was introduced into soybean by Agrobacterium tumefaciens method under different boron concentrations. The expression of GUS gene was determined by chemical method and the transient expression rate was calculated. Finally, PCR and dot blot were used for further identification to explore the boron-mediated efficiency and feasibility of soybean transformation. The results showed that high boron and low boron medium inhibited the proliferation of soybean shoots to a certain degree, especially in the case of high boron, the induction rate of clustered shoots decreased significantly. Agrobacterium-mediated transformation efficiency of soybean under 30 mg · L-1 boric acid environment was significantly higher than that under normal boric acid environment, and the GUS transient conversion rate was as high as 35.92%. The significance of this phenomenon to establish a new soybean genetic transformation system needs to be further explored.