摘要
建立了油菜小孢子再生胚状体的实验体系，掌握了受体最佳发育时期，使再生频率高达２１．２个胚状体／花蕾。以油菜游离小孢子为受体，用微束激光穿刺法导入芜菁花叶病毒外壳蛋白基因（ＴｕＭＶ），成功地获得了转基因植株。研究结果表明，用预培养３天的小孢子进行转化得到的１５个胚状体，再生出一株绿苗，２株白苗。而未经预培养的小孢子未能得到转基因植株。用植物总ＤＮＡ以ＰＣＲ法扩增ＴｕＭＶ基因产物片断，然后进行电泳检测，证明该绿苗带有芜菁花叶病毒外壳蛋白基因。
The microspores embryogensis system of Brassica napus has been established.When acceptor materials were in appropriate development stage,the frequency of regeneration could reach to 21.1 embryos per flower buds.Using isolated microspores as receptor materials,TuMVCP gene has been introduced by laser microbeam puncture,and transgenic plants have been successfully obtained.The results showed that:15 embryos were acquired from microspores which were pre-cultured for 3 days before trnsformation,and one green and two white seedlings were regenerated from them.On the contrary,no seedlings were obtained from microspores which hadn't been precultured before transformation.Using total DNA extracted from green seedling as template,the PCR amplification analysis proved that the TuMVCP gene had been introduced into Brassica napus.
出处
《激光生物学报》
CAS
CSCD
1998年第2期103-107,共5页
Acta Laser Biology Sinica
