Bar基因的玉米花粉管通道法转化

Introduction of Herbicide Resistant Gene(bar) into Maize(Zea mays L.) by Pollen Tube Pathway

通过花粉管通道法利用抗除草剂基因(bar)转化3个玉米自交系,D0代获得35株GUS阳性植株,其中17株PCR阳性植株,除草剂筛选获得抗性植株13株。研究发现DNA溶液浓度影响转化率,当浓度为100μg/ml时,转化率最高,为2.61%。DNA导入时间距人工授粉时间的延长,结实率升高,但转化率显著下降,人工授粉后1d导入DNA产生的转化株最多,为19株,平均转化率为1.35%。

Bar gene was introduced into 3 maize inbred lines by pollen tube pathway. Twenty four D0 maize plants from three inbred lines showed GUS positive reaction , among which seventeen plants showed positive PCR reaction. And thirteen plants resisted to herbicide. DNA concentration and transformation duration had obviously influenced transformation rate, and with the increase of DNA concentration the number of seeds on corn ear was reduced; with the duration away from manual pollination prolonged, the seed-set rate was raised, while transformation rate was decreased sharply. ,The best transforming condition was DNA concentration 100μg/ml and to transform at one day after pollination.