高羊茅组织培养再生体系及GUS基因瞬间表达研究  

Studies on Tissue Culture and Regeneration System of Festuca arundinacea and Transient Expression of GUS Gene

作　　者：高丽美[1] 徐子勤[1] 张永彦[1] 黄萱[1] 刘杨[1] 

机构地区：[1]西北大学生物技术省级重点实验室,西安710069

出　　处：《西北植物学报》2005年第1期40-45,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：陕西省自然科学基金重点项目 2001SM24 ;陕西高校重点实验室重点科研项目 陕教研2001-29-2 资助

摘　　要：以成熟种子为外植体,对高羊茅组织培养和植株再生体系进行了优化,分析了不同浓度2,4-D、6-BA和激动素对高羊茅愈伤组织诱导和愈伤组织分化成苗的影响,结果表明:9.0mg/L2,4-D对愈伤组织的诱导效果最佳,0.2mg/L激动素是愈伤组织分化成苗的最适浓度,二者的诱导率和分化率分别达到68.08%和45.83%.在愈伤组织继代培养基中附加1.0mg/L2,4-D、0.5mg/L6-BA和1.25mg/LCuSO4有利于胚性愈伤组织的形成,可以明显促进愈伤组织分化.同时,采用基因枪法将GUS基因导入高羊茅愈伤组织中,通过组织化学染色检测到了GUS瞬间表达活性;并对影响GUS基因瞬间表达的因素进行了分析,以期为提高基因枪法遗传转化效率提供参考.Tissue culture and plantlet regeneration system of tall fescue was optimized by using MS (Murashige and Skoog,1962) as basal medium and mature seeds as explants.Effects of different concentration of 2,4-D,6-BA (BAP)and kinetin on callus induction and plantlet regeneration in tall fescue were analyzed.It has shown that 9.0 mg/L 2,4-D was optimal for callus induction,and 0.2 mg/L kinetin for plantlet regeneration,and the frequencies of callus induction and plantlet regeneration were 68.08% and (45.83%) respectively.Embryonic calli could form easily on a modified medium containing 1.0 mg/L 2,4-D,0.5 mg/L 6-BA and 1.25 mg/L CuSO_4,which promoted plantlet regeneration obviously.Transient expression of GUS gene was observed through histochemical stainig of calli transformed with GUS gene by microprojectile bombardment.The factors influencing transient expression of GUS gene were discussed.The purpose of this work was aimed at improving the efficiency of plant transformation by microprojectile bombardment.

关 键 词：高羊茅 组织培养 基因枪 GUS基因 瞬间表达 

分 类 号：Q943.2[生物学—植物学]