外源基因导入部分酶解的大麦愈伤组织和转基因植株的再生

INTRODUCTION OF FOREIGN GENE INTO BARLEY CALLI AFTER PARTIALLY ENZYMATIC DIGESTION AND REGENERATION OF TRANSGENIC PLANTS

采用PEG法、电击法及PEG加电击法,成功地将GUS与NPT-Ⅱ基因(pBI121)导入部分酶解的大麦愈伤组织,并获得了大麦转基因植株。从而表明,对于原生质体培养特别困难的大麦来说,以部分酶解愈伤组织作为外源基因转化的另一类受体系统,是一条切实可行的途径。

Embryogenic calli derived from immature inflorescences of barley 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. GUS,NPT-Ⅱ activity assay and Southern blot showed that the report genes were expressed and integrated in transformed calli. Transgenic plants were regenerated from calli possessing GUS activity. The results prove that the partially digested callus is a potential,feasible system as receptor for gene transfer, especially for plants which are difficult of protoplast culture and plant regeneration from protoplasts.