小麦成熟胚再生体系及基因枪转化的初步研究  

A preliminary study of regeneration system of wheat mature embryo and its particle biolistic transformation

作　　者：林德书[1] 王艳丽[2] 庄振宏[1] 鲁国东[3] 叶兴国[2] 王宗华[1] 

机构地区：[1]福建农林大学生命科学学院,福建福州350002 [2]中国农业科学院作物科学研究所,北京10081 [3]福建农林大学教育部生物农药与化学生物学重点实验室,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2005年第2期141-144,共4页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30270870);福建省自然科学基金资助项目(B0410010)

摘　　要：以小麦品种晋麦2148的成熟胚为外植体,消毒后用解剖刀刨碎,接种于MS+VB5+11.2g·L-1葡萄糖+2mg·L-12,4D+8g·L-1琼脂糖的培养基中诱导愈伤组织,将诱导3周的成熟胚愈伤组织接种到胚状体成熟培养基(MS+VB5+11.2g·L-1葡萄糖+0.2mg·L-1IAA+8g·L-1琼脂糖)中培养4-8周,然后转接到再生培养基(1/2MS+VB5+30g·L-1蔗糖+8g·L-1琼脂糖)中进行再生成苗.接种1200块成熟胚,得到1123块愈伤组织,最终形成258株再生苗,再生率为21.5%.在建立了再生体系的基础上,用基因枪介导法将GUS基因导入成熟胚愈伤组织,Xgluc染色表明,GUS基因已经在小麦成熟胚愈伤组织中表达.将100块愈伤组织用Xgluc染色,29块愈伤组织出现肉眼可见的蓝色小点.Mature e mbryos of a common wheat variety Jinmai 2148 was used as explants t o induce regeneration and transformation, respectively. The embryos were isol ated from surface-sterilized mature caryopses, cut into pieces, and fil ter ed through a sterile nylon mesh, then inoculated on the MS medium with 2 mg·L ^-1 2,4 -dichlorophenoxyacetic acid to initiate calli. After 3 weeks, the embryogenic calli were transferred into the MS medium with 0.2 mg·L^-1 IAA an d cultured for 4-8 w eeks for plant regeneration. Following the above protocol, 1123 calli and 258 plantlets were obtained from 1200 embryos in total,and the regeneration rate wa s 2 1.5% Furthermore,plasmid DNA of pAHC25 containing GUS gene as the d onor of alie n gene was transformed into the mature embryo calli by bombardment. The result from X-gluc staining showed that GUS gene gave transient expression success full y in the transformed calli. Blue spots were observed in 29 calli out of 100 cul tures.

关 键 词：再生体系 成熟胚 基因枪转化 愈伤组织 再生培养基 GUS基因 琼脂糖 小麦品种 基因导入 葡萄糖 外植体 接种 体成熟 再生苗 再生率 介导法 2g 诱导 成苗 肉眼 

分 类 号：S682.11[农业科学—观赏园艺] S511.21[农业科学—园艺学]