影响谷子愈伤组织基因枪转化的因素

Factors Influencing Foxtail Millet Embryogenic Calli Transformation by Particle Bombardments

通过 Ｇｕｓ 基因 在谷子 胚性愈伤 组织中 的瞬时表 达检测， 分析 了影 响谷 子愈 伤组 织基 因枪转化效 率的几个 因素。钨 粉的包 被质粒 Ｄ Ｎ Ａ 用量以 ３ μｇ／ ｍ ｇ 钨粉 ， Ｃａ Ｃｌ２ 浓度１５ ｍ ｏｌ／ Ｌ，亚 精胺浓度４０ ｍ ｍ ｏｌ／ Ｌ 为最 佳，过高 的 Ｄ Ｎ Ａ 、 Ｃａ Ｃｌ２ 和亚精 胺加入量 均显著降 低转 化率 ； Ｊ Ｑ７００ 基因 枪样品室高度 ７ ｃ ｍ ，轰 击速 度 ４００ ～４５０ ｍ ／ｓ 转 化， 较其 它 速度 和样 品 室高 度下 转 化 Ｇｕｓ 基因 表 达率高；每 次转化的 愈伤组织 用量对 转化效率 也有影 响， 以１ ～２ ｇ Ｇｕｓ 基 因表 达斑 最多； 玉米 ｕｂｉｑｕｉｔｉｎ启动子 较 Ｃａ Ｍ Ｖ３５ Ｓ 启动子 的瞬时表 达显著增 强。

Factors influencing foxtail millet (Setaria italica Beauv.)embryogenic calli transformation by particle bombardments were studied using the method of Gus transient expression with JQ 700 gene gun.Relatively high numbes of Gus transient expression unites were obtained when the quantity of plasmid DNA added to microprojectles was 4 μg/mg tungsten powder and the concentration of CaCl 2 and spermidine used to absorb DNA to microprojectles were 1.5 mol and 40 mmol respectively.The velocity of the microprojectles and the distance between the calli being bombarded and the stopping plate of JQ 700 gene gun optimized in this report were 400-450 m/s and 7 cm.The quantity of calli used per treatment was also a factor influencing gene delivery and 1 2 g of calli per treatment gave higher Gus transient expression.The promoter derived from maize ubiquitin produced significantly more Gus foci compared to the CaMV35 s Promoter.