Studies on the widening symbiotic specificity by the cell fusion of legume rhizobium

豆科植物根瘤菌细胞融合扩大共生特异性的研究

• 批准号: 62560271
• 负责人: IKEDA Hajime
• 金额: $ 1.22万
• 依托单位: Miyazaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560271/
• 关键词: rhizobium; alfalfa; common lespedeza; acid tolerance; spheroplast; 細胞融合

For the purpose of breeding more acid tolerant isolates of Rhizobium meliloti on alfalfa plant, cell fusion of R.meliloti with Rhizobium sp. (common lespedeza) was tried.The following results were obtained:1)When each or R.meliloti and R.sp.(common lespedeza) were cultured on YM agar media with various pHs ranging 4.0 to 8.0, rhizobium sp.proliferated beyond pH value of 4.0, while proliferation of R.meliloti was not observed until pH 5.0.2)As a result of comparing the method of spheroplast preparation for R.meliloti and R.sp., Rhizobium meliloti was spheroplasted by treating for 2 hrs with 200 mg/l lysozyme solution. On the contrary, R.sp.required 400 mg/l lysozyme for 2 hrs to obtain spheroplasts.3)After cell fusion of spheroplasts of R.meliloti into those of R.sp.with 40%(W/V) polyethylene glycol 6000, cell fused rhizobia were isolated by 200 units/ml penicillin. Rhizobia isolated by treating with penicillin once for 6 hr or 3 hr were stimulated on YM agar medium pH 4.0 and they formed root nodules along the root of alfalfa.

为了在紫花苜蓿上选育耐酸根瘤菌，对紫花苜蓿根瘤菌与胡枝子根瘤菌进行了细胞融合试验。得到了以下结果：1)小茴香和小茴香分别在不同的条件下。在pH值为4.0 ~ 8.0的YM琼脂培养基上培养，根瘤菌在pH值为4.0时开始增殖，而木耳在pH值为5.0.2时才开始增殖。用200 mg/l溶菌酶溶液处理2 h后，木耳根瘤菌成球。相反，rsp需要400 mg/l的溶菌酶作用2小时才能获得球质体。3)用40%（W/V）的聚乙二醇6000将木犀草球质体与木犀草球质体融合后，用200单位/ml青霉素分离融合后的根瘤菌。在pH为4.0的YM琼脂培养基上，分别用青霉素处理1次、6小时或3小时分离得到根瘤菌，根瘤菌沿苜蓿根部形成根瘤。