INTRODUCTION, EXPRESSION AND REGENE-RATION OF THE TiT-DNA GENES IN CULTURED CELLS FROM Hyoscyamus muticus +Nicotiana tabacum SOMATIC HYBRIDS  

作　　者：许耀 贾敬芬 郑国锠 

机构地区：[1]Biotechnology Research Centre, Sun Yat-sen University, Guangzhou 510275, PRC [2]Department of Biology, Lanzhou University, Lanzhou 730001. PRC

出　　处：《Science China Chemistry》1989年第11期1342-1349,共8页中国科学（化学英文版）

基　　金：Project supported by the Science Fund of the State Educational Committee. This work was finished in Lanzhou University.

摘　　要：After the cultured cells from Hyoscyamus muticus + Nicotiana tabacum somatic hybridswere cocultivated with different virulent strains of Agrobacterium tumefaciens harboringoctopine- type Ti plasmid or nopaline- type Ti plasmid using the transformation procedurein vitro developed in the present investigation, the TiT- DNA genes were introducedinto the host cells. The onc genes and ocs or nos genes located on TiT- DNA were expres-sed in transformed colonies derived from the cocultivated cells. Although the platingefficiencies of recipient cells were reduced by the agrobacterial treatment, the frequenciesof phytohormone autotrophy ranged from 33.9 to 76 .8% in the cells infected with viru-lent strains in hormone- free conditions, and the frequencies of opine synthase activityamounted to 9.7- 47 .5%. Teratomatous shoots were regenerated from the transformed col-onies. During the course of culture the shoots were no longer to lengthen when theygrew up to 1 -3 cm in length, and they could not be rooted. Following maintenance ?ofthe excised shoots on medium lacking hormones, further tumorous tissues were formedfrom their base sections. Some traits coded for by TiT- DNA were still retained in the leaftissues of regenerated shoots and the secondary tumorous tissues.After the cultured cells from Hyoscyamus muticus + Nicotiana tabacum somatic hybridswere cocultivated with different virulent strains of Agrobacterium tumefaciens harboringoctopine- type Ti plasmid or nopaline- type Ti plasmid using the transformation procedurein vitro developed in the present investigation, the TiT- DNA genes were introducedinto the host cells. The onc genes and ocs or nos genes located on TiT- DNA were expres-sed in transformed colonies derived from the cocultivated cells. Although the platingefficiencies of recipient cells were reduced by the agrobacterial treatment, the frequenciesof phytohormone autotrophy ranged from 33.9 to 76 .8% in the cells infected with viru-lent strains in hormone- free conditions, and the frequencies of opine synthase activityamounted to 9.7- 47 .5%. Teratomatous shoots were regenerated from the transformed col-onies. During the course of culture the shoots were no longer to lengthen when theygrew up to 1 -3 cm in length, and they could not be rooted.

关 键 词：cell transformation T-DNA transfer opine SYNTHASE HORMONE autonomy AGROBACTERIUM TUMEFACIENS 

分 类 号：O6[理学—化学]