用根癌土壤杆菌和发根土壤杆菌转化马铃薯的初步研究  

POTATO GENETIC TRANSFORMATION USING AGROBACTERIUM TUMEFACIENS AND A. RHIZOGENES

作　　者：王春林 朱明凯[1] 程天庆[1] 

机构地区：[1]中国农科院蔬菜花卉研究所

出　　处：《马铃薯杂志》1990年第1期1-7,共7页Chinese Potato Journal

摘　　要：根癌土杆菌菌株T_(37),C_(58)和B_6S_3单独或与C_(58) CIpGV 3850::110(?)neo(带有一嵌合基因NOS—NPT使植物产生卡那霉素抗性)接种马铃薯块茎盘。每块茎盘肿瘤数因细菌菌株、马铃薯基因型和所取块茎盘在块茎上的位置而变化。T_(37)诱导的品种Desiree的肿瘤分化了芽。大部分肿瘤的分化芽合成胭脂碱,表明是经pTiT—DNA转化的。用发根土壤杆菌菌株A_4诱导的毛状根切段培养后再生成株。再生植株具有明显的表型变异,并有agropinc合成,证明是经pRiT—DNA转化的。Wild strains of Agrobacterium tumefaciens T_(37), C_(58) and B_6S_3 (with pTiT_(37) , pTiT_(58) and pTiB_6S_3 Plasmids resp.) independently or in combination with a “disarmed” A.tumefaciens strain C_(58)CI pGV3850:: 1103neo containing a chimaeric gene NOS—NPT confering plant lanamycin resistance were inoculated to and cocultured with tuber discs of tetraploid potato (Solanum tuberosum) cultivars DTO—33, Desiree, etc. Tumours with different habitus Were produced from surface of the tuber discs within l～2 weeks. Transformation effieiency in terms of the number of lumours per disc inoculated varied with bacterium strains, potato genotypes and position of discs dissected from tubers Mixed—inoculation of wild and disarmed strains deereased the number of tumours induced by T_(37) and C_(58) alone. High ratio of complete and partial kanamycin resistant tumours indicated a significant interaction between two A. tumefaciens strains when they transformed plant cells. Multiple shoots were regenerated from 4 of Desiree T_(37) induced tumours, of which 3 regenerated rootless and nopaline—positive shoots and 1 formed nopaline—negative shoots with roots.Tuber discs of Desiree inoeulated with A. rhizogenes A4 containing pRi A4 plasmids sprouted hairy roots after ineubated for 1～2 Weeks. Root segments developed green calli at one end of the segment when cultured on ZN medium (MS + ZT 1mg/l + NAA0.01 mg/l + Suerose10g/l) within 2 weeks. When callusing segment were cultured for further 2～3weets, then transfered to the shoot—indueing SH medium (MS + BA 2 mg/l + GA_3 5mg/l+Sucrose 20 g/1), multiple shoots appeared 2～3 weeks later. Randomly selected hairy root clones, their calli and regenerated shoots tested were all agropine—positive. This confirmed the transformation of hairy roots by pRi A4 T—DNA integration of the T—DNA into potato genome and stable transmition through mitosis.

关 键 词：根癌土壤杆菌 发根土壤杆菌 马铃薯 

分 类 号：S532.03[农业科学—作物学]