摘要
以苹果砧木八楞海棠（Ｍａｌｕｓｍｉｃｒｏｍａｌｕｓ）为实验材料，用微束激光穿刺法将菜豆几丁酶基因导入苹果叶片，成功地获得了转基因植株。实验结果表明，以苗龄２６天的小苗上部刚展开的叶片再生频率可达１００％。经高渗缓冲液预处理后用激光微束穿刺导入外源ＤＮＡ可得到较高的转化频率。在１５０个再生叶片中经卡那霉素筛选得到２５株绿苗，经ＰＣＲ扩增检测，３株呈阳性结果，表明外源几丁酶基因已整合到苹果的基因组ＤＮＡ中。
The Bean Chitinase gene was introduced into apple rootstock(Malus micromalus)via laser microbeam puncture and transgenic plants were acquired in this research. The results showed that the regeneration freqency could attain 100% when the fully extended leaves on the upper part of 26 days old seelings were used as explants. High transformation frequency could be obtained by pretreating receptors in high osmltic buffer before laser microbean irradiation. 25 green seedlings were got from 150 regenerated leaf discs after selected by Km. 3 of them were verified to be transgenic plants through PCR analysis.. It was proved that Bean Chitinase gene have been integrated into the genome of apple rootstock.
出处
《激光生物学报》
CAS
CSCD
1998年第3期163-167,共5页
Acta Laser Biology Sinica
基金
国家自然科学基金
关键词
苹果砧木
几丁酶基因
微束激光
遗传转化
Apple rootstock(Malus micromalus) Chitinase gene Laser microbeam Genetic transformation.
