四倍体刺槐转甜菜碱醛脱氢酶基因的研究
SCIENTIA AGRICULTURA SINICA(2004)
Abstract
利用农杆菌(Agrobacterium tumefaciells conn)介导法,将山菠菜甜菜碱醛脱氢酶(BADH)基因转入四倍体刺槐(Robinia pseudoacacia L.).结果表明,以愈伤组织作为外植体、液体分化培养基(MS+6-BA 2.0 mg·L-1+IBA 0.2 mg·L-1)活化农杆菌、转化培养基中20 mg·L-1乙酰丁香酮、菌液浓度0D600值0.3~0.7、预培养1~2d、浸染4~8 min、共培养4~8 d、延迟15 d选择的条件下,400 mg·L-1头孢霉素可有效抑制农杆菌,卡那霉素为50 mg·L-1时愈伤组织上芽的白化率达97.3%.经PcR和PcR-Southern检测,外源基因已成功整合到植株基因组DNA中;转化植株丛生芽生长的Nacl相对抗性提高了2‰~3‰.
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