花粉管通道法介导il-4基因转化大白菜(Brassica pekinensis Rupr.)的研究初探

Primary Study on Transgenic Chinese Cabbage with Interleukin-4 Gene via Pollen-tube Pathway

通过花粉管通道法介导il-4基因转化大白菜,利用PPT抗性筛选、PCR扩增和PCR-Southern杂交技术检测转化后代。本文针对品种、人工授粉时间和转化操作时机可能对il-4转化率产生的影响进行了研究。试验结果表明,il-4基因对品种有偏爱,哈白二号的转化率高;不同的人工授粉时期对转化率有影响,开花期授粉效果好;不同的转化时机对转化率也有影响,人工授粉后24h左右进行转化操作效果好。哈白二号在花期授粉后24h左右进行转化操作(AH),il-4的转化率达1.15%。对il-4基因在大白菜T2代中的传递规律进行观察,发现PPT的抗性分离情况,除哈白二号-2单株的后代外,其余均不符合抗性植株数:非抗性植株数=3:1的孟德尔分离规律;而il-4的分离情况,也不符合阳性植株数:阴性植株数=3:1的孟德尔分离规律。

Interleukin-4 gene was transformed into Chinese cabbage via pollen-tube pathway. Resistant selection by PPT, PCR amplification and PCR-Southern hybridization were used to detect whether transgene was in transformed progenies. This paper studied the effect of different variety, pollination period and transforming time on il- 4 transformation rate. The experiment results indicated that il-4 had bias towards variety. The transformation rate of Habai 2 was the highest. Different pollination period had effect on transformation rate. Florescence pollination was better. Different transforming time also had effect on transformation rate. It was better to transform at the time of 24h after pollination. The transformation rate of Habai 2 at the time of 24h after florescence pollination was 1.15%. This paper analysed the segregation in T2 generation of transgenic Chinese cabbage. The segregation of resistant selection by PPT didn＇t accord with Mendelian model except Habai 2-2. The segregation of il-4 was not in accordance with Mendelian model.