拟南芥转化体筛选中Basta喷洒时期和浓度试验

Experiment on Basta Spraying Period and Concentration in Transgenic Arabidopsis Screening

为了更加方便有效地利用Basta来筛选带草铵磷(PPT)抗性Bar基因的转基因拟南芥,对不同时期喷洒不同浓度Basta的效果进行了研究。结果表明:拟南芥转化体刚长出2片真叶时对Basta比较敏感,此时开始喷施效果比较好;PPT含量为13.5%的情况下,稀释2 000～4 000倍喷施效果好,此时既可有效筛选出阳性苗,又不会抑制阳性苗的生长;第一次喷施后6天左右移栽阳性苗较好,此时可以明显区分出阳性苗,且移栽后缓苗快、长势好。

In order to efficiently screen transgenic Arabidopsis thaliana containing phosphinothricin-resistant Bar gene different concentrations of Basta were sprayed at different periods.The results showed that spraying Basta on Arabidopsis thaliana seedlings with two real leaves,which were sensitive to Basta,would have a good effect;when the PPT content of Basta was 13.5%,a dilution of 2 000~4 000 times was suitable to screen out the resistant plants from non-resistant ones without growth inhibition;it was suitable to transplant the resistant plants at 6th days after the first spraying,because obvious differences appeared between resistant and non-resistant plants and they grew fast and well after transplanted at this time.