外源基因导入部分酶解的水稻小细胞团及其转基因植株的再生

INTRODUCTION OF FOREIGN GENE INTO SMALL CELL GROUPS DIGESTED PARTIAL-ENZYMICALLY IN RICE AND REGENERATION OF TRANSGENIC PLANTS

采用 PEG 法、电击法以及 PEG-电击法,成功地将 GUS 及 NPT-11基因(pBI 121)转入部分酶解的水稻小细胞团并获得了水稻转基因植株。从而表明绕过原生质体,以部分酶解小细胞团作为水稻外源基因转化的另一类受体系统是切实可行的。由于细胞团易于操作,成活率高,对于某些原生质体培养特别困难的植物类型或基因型不失为一条行之有效的途径。

Suspension cultures of small cell groups (SCG; ca. 50—100 cells per group) were estab- lished from calli of Japonica rice Fang 7 and H124. The SCG were partially digested and transformed by plasmid pBI121 harboring the NPT-Ⅱ (neomycin phosphotransferase) and GUS (betaglucuronidase) genes. Plasmid DNA was introduced into cells' by PEG, electroporation and PEG plus electroporation. NTP-Ⅱ and GUS activity assay showed that the report genes were expressed in transformed cells. Transgenic plants were regeneiated possessing GUS activity due to the integration of intact foreign DNA into their genome as evidanced by hybridization. The results prove that the partially digested SCG is a potential, feasible system as receptor for gene transfer, especially for plants which are difficult for protoplast culture and plant regeneration from protoplasts.