影响辣椒T-DNA标签突变体库构建的转化因素探讨

Study on Factors of Transformation in Construction of Capsicum T-DNA Insertional Mutagenesis

以辣椒自交系H8124无菌苗子叶和下胚轴为外植体,采用根癌农杆菌侵染转化法,将T-DNA标签导入辣椒,以探讨影响转化效率的主要因素,包括外源植物生长调节剂浓度、农杆菌类型、外植体材料、预培养时间、侵染时间等。结果表明,采用苗龄10-15 d的子叶为外植体,预培养2 d后用农杆菌GV3101侵染5 min,外源植物生长调节剂为5.0mg/L 6-BA,辣椒的T-DNA转化可以获得较高的转化效率。

Using calli from cotyledon and hypocotyl of pepper inbred line H8124 as explants, T-DNA tag was inserted into pepper through agrobacterium-mediated transformation. Six effect factors such as hormone concentration, agrobacterium type, type of explants, development stage, transformed times and pre-cuhured were detemined in this experiment. The results showed that when growth regulator 2 days pre-cuhured cotyledons which seedling age was 10 - 15 days old were mixed with celt suspension of agrobacterium in 5 minutes, the high transformation frequency was obtained.