发根农杆菌对骆驼刺的转化及转化组织的形态发生

将骆驼刺离体再生苗的茎切段经发根农杆菌A4菌株感染后,在含500mg/L头孢霉素的MS无激素培养基上培养,产生了转化的发根和愈伤组织.转化根在附加2mg/L2,4-D和0.5-1mg/L6BA的MS培养基上培养后,亦可诱导出愈伤组织.在含3mg/L6BA和0.5mg/LNAA的培养基上诱导出了苗的分化.冠瘿碱分析表明,在95%以上的发根和75%的转化愈伤组织及再生植株中都显示了T-DNA的整合和表达.染色体检查发现,约81%的发根细胞具有正常染色体数(2n=18),其余则存在染色体数目的变化,在继代培养一年之后,转化体仍维持旺盛的再生能力.

AGROBACTERIUM RHIZOGENES-MEDIATED TRANSFORMATION AND THE REGENERATION OF TRANSFORMANTS IN ALHAGI PSEUDALHAGI

The regenerated shoot segments of Alhagi pseudalhagi were sliced and infected with Agrobacterium rhizogmes strain A4. The hairy roots and transformed calli were obtained through selection on hormone free MS medium. The transformants were cultured on MS medium with 2mg/L 2, 4-dichlorophenoxy acetic acid (2,4-D) and 0.5 - 1mg/L 6-benzylaminopurine (6-BA) to induce calli. 3mg/L 6-BA and 0. 5mg/L naphthalene acetic acid (NAA) were applied for shoot differentiation. Shoots were planted on MS medium with 2mg/L indole-3-butyric acid (IBA) and produced roots. Opine analysis proved the integration and expression of T-DNA in over 95 % hairy roots, 75% transformed calli and transformed plantlets respectively. The 81 % hairy root cells had normal chromosome numbers ( 2n = 18). The alterations of chromosome number were observed. After one year of subculturing, the regeneration ability of transformants was maintained.