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Position: Home > Articles > Studies on the Genetic Transformation Factors of Photinia fraseri Using Microprojectile Bombardment Acta Horticulturae Sinica 2013,40 (11) 2280-2286

基因枪介导红叶石楠遗传转化因素分析

作  者：
刘翠兰;燕丽萍;毛秀红;孙超;夏阳;梁慧敏
单  位：
山东省林业科学研究院;江苏农林职业技术学院
关键词：
红叶石楠;rd29A;基因枪介导;转基因植株
摘  要：
以红叶石楠(Photinia fraseri)的胚性愈伤组织为受体材料,利用基因枪法导入外源抗寒基因rd29A,以期建立基因枪转化红叶石楠的技术体系。结果表明,基因枪转化红叶石楠的最佳组合条件为:胚性愈伤组织在诱导培养基MS+6-BA 2.0 mg·L-1+NAA 5.0 mg·L-1+2,4-D 0.5 mg·L-1+琼脂5.0g·L-1+蔗糖20 g·L-1里,用0.3 mol·L-1甘露醇处理15~16 h;基因枪轰击时添加12.5 mol·L-1氯化钙50μL+0.1 mol·L-1亚精胺50μL,每枪含金粉300μg+质粒DNA3.0μg,轰击距离9 cm,轰击压力1 100 psi,轰击次数1次。经多次筛选获得了6株转基因植株,转化植株经PCR检测和Southern分析,证实了rd29A已经整合到红叶石楠基因组中。
译  名：
Studies on the Genetic Transformation Factors of Photinia fraseri Using Microprojectile Bombardment
作  者：
LIU Cui-lan;YAN Li-ping;MAO Xiu-hong;SUN Chao;XIAYang;LIANG Hui-min;Shandong Provincial Academy of Forestry;Shandong Provincial Key Laboretory of Forest Genetic Improvement Tree;Jiangsu AgricμLture and Forestry Profession Technology College;
关键词：
Photinia fraseri;;rd29A;;microprojectile bombardment;;transgenic plants
摘  要：
The study has transformed the cold resistant gene rd29A into the acceptor,the embryo callus derived from Photinia fraseri leaf,in order to establish the transformation system by using particle bombardment method. The results indicate that the optimal transformation scheme for Photinia fraseri leaf callus using particle bombardment method was treating embryogenic callus tissue within 0.3 mol · L-1mannitol for 15–16 h,in MS medium supplemented with 2.0 mg · L-16-benzy aminopurine,5.0 mg · L-1napthaleneacetic acid,0.5 mg · L-12,4-dichlorophenoxyacetic acid,5.0 g · L-1agar powder and 20 g · L-1glucose. When particle bombardmenting,in addition of 12.5 mol · L-1CaCl250 μL and 0.1 mol · L-1spermidine 50 μL,and bombarding the samples once a time,at 9 cm bombardment distance,at 1 100 psi heli μm pressure and with the dosage of gold powder 300 μg and plasmid DNA 3.0 μg per bombardment.Six transgenic Photinia fraseri plants were obtained in the study and it was confirmed that the rd29A gene was integrated into Photinia fraseri genomes according to the PCR and Southern analysis of plant DNA.

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