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.     

外源野生大豆DNA导入栽培大豆及RAPD分子验证

报道了利用大豆自花受粉后形成的花粉管通道,将外源高蛋白野生大豆总DNA直接导入栽培大豆的实验结果及对导入转化后代的分子验证。 结果显示:在D_1代所获7个单株中有1株发生明显变异,其结荚习性、株型、叶型、花色等与受体基本相同:百粒重、种皮色及茎杆强度介于双亲之间:单株荚数明显增多,连续3年测定蛋白质含量比受体提高12.5％,从D_1代起已稳定遗传3个世代。对该组合的受体、供体及转化后代进行RAPD分析,从150个引物中找到24个引物检测出DNA的多态性,证明外源DNA导入受体后引起后代基因组的显著变异,并推测大片段外源DNA同源重组可能是变异的主要原因。     

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.     

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。     

STUDY ON ACCUMULATION OF 11S AND 7S PROTEINS IN DEVELOPMENTAL SEED OF SOYBEAN BY MEANS OF IMMUNO-FLUORESCENCE

The accumulation courses of 11S and 7S proteins in the development seed of soybean were studied by means of immuno-fluorescence. Fifteen days after flowering, 7S proteins began to deposit and so did 11S proteins a little later. It was observed that 20—30 days after flowering, these two globulins or their precursor peptides had the tendency to transfer from tegument to the cotyledon. It was found that a few of the cotyledon cells in the mature seed did not accumulate 11S proteins, while in the high protein cultivar and high protein wild soybean all the cotyledon cells were filled with 11S proteins and other storage substances.     

利用免疫荧光法研究大豆种子发育过程中11S和7S蛋白的积累

本文利用免疫荧光法研究了种子发育过程中11S和7S两种球蛋白的积累过程。7S蛋白在子叶的沉积始于开花后15天,11S蛋白的沉积稍迟一些。开花后20—30天,这两种球蛋白增加非常迅速。观察到这两种蛋白或它们的类似物从珠被向子叶转运的趋势。还发现有的品种某些子叶细胞不积累11S蛋白,而在高蛋白品种和高蛋白野生大豆所有子叶细胞都充满11S蛋白和其它贮存物质。     

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。