A STUDY ON TUMOR FORMATION OF SOYBEAN AND GENE TRANSFER

From 15 strains of Agrobacterium tumefaciens 7 were selected as the best inductors oftumors on soybean and 94 tumor formation genotypes were screened from 1553 varieties andforms of Glycine soja, G. gracilis and G. max. The frequency of tumor formation was 6%.The bacteria-free calluses were obtained. The data of biochemical analysis verify that someof the above-mentioned calluses contained nopaline. This means that Ti-plasmid of Agrobac-terium tumefaciens can be used as a vector for the transfer of nopaline, a marker gene, intogenome of several species of soybean. In the soybean genome the transferred nopaline genewas integrated and expressed. The transformed gene can be stably integrated in the soybeangenome through vegetative propagation.     

大豆致瘤及基因转移研究

本文报道了致瘤农杆菌(Agrobacterium tumefaciens)的15个菌系对大豆的致瘤作用,筛选出致瘤效果较好的7个菌系.从野生大豆、半野生大豆和栽培大豆的1553个品种和品系中筛选出94个结瘤品种和品系,占筛选总数的6%,并获得了无菌愈伤组织.经生化鉴定证明,上述愈伤组织中有一部分含有胭脂碱(nopaline),证明Ti质粒可以做为载体,把胭脂碱基因转移到野生大豆、半野生大豆和栽培大豆的基因组中去整合和表达,并稳定地保存在大豆基因组中。     

ESTABLISHMENT OF T-DNA-CONTAINING SOYBEAN CELL LINES

This paper reports establishment of stable diploid cell lines containing nopaline synthase genes from callus containing T-DNA after 65 generations of successive suspension cultures. The strain C_(58) Agrobacterium tumefaciens was used for tumor inducing on variety "Siyuchuang". The T-DNA-containing tissue was separated in auxin-free MS medium by strictly screening three times from single cells. The presence of nopaline was identified by paper electrophoresis. The T-DNA-containing polyploid soybean cell lines were obtained at the same time. The nopaline synthase genes from T-DNA were stably integrated and expressed in soybean genome, which can be used in gene engineering with soybean.     

含T-DNA大豆细胞系的建立

本文报道了由致瘤农杆菌(Agrobacterium tumefaciens)C58菌株诱发的栽培大豆“四月黄”品种的瘤组织,经过无激素的MS培养基初步筛选和标记基因产物胭脂碱纸上电泳鉴定,筛选出含T-DNA的大豆愈伤组织。经过65代悬浮继代培养以及三次单细胞筛选,成功地获得了含T-DNA的稳定的大豆细胞系。同时还建立了一个含有T-DNA的多倍体细胞系。异源的T-DNA中的胭脂碱合成酶基因在大豆细胞基因组中稳定的整合和表达。为大豆的基因工程研究创造了良好的条件。