三叶草根瘤菌SDS-PAGE分析及结瘤试验分子验证  

The analysis on Rhizobium of Trifolium spp.by SDS-PAGE and molecular verification of nodulation ability test

作　　者：刘晓云[1] 戴燕燕[1] 郭振国[1] 肖猛[1] 张斌[2] 

机构地区：[1]河北大学生命科学学院,河北保定071002 [2]西南林学院资源学院,云南昆明650224

出　　处：《草业科学》2010年第1期79-84,共6页Pratacultural Science

基　　金：国家自然科学基金项目(30570001)

摘　　要：全细胞蛋白SDS-PAGE是一种能快速、稳定、准确地反映细胞内蛋白分子类型的方法,可对根瘤菌进行初步分群,也可用于结瘤试验中结瘤菌株的验证。从云南、湖北、河北等6个地区采集的三叶草Trifo-lium spp.根瘤中分离得到120个根瘤菌菌株,采用SDS-PAGE分子标记法对其进行初步分群得到104个类型,在80%的相似性水平上分为6个类群,并依据菌株特征及采集地点等选取其中的42株进行结瘤试验。结果显示,接种根瘤菌的42株三叶草植株全部结瘤,结瘤率达100%,同时对部分代表菌株所结根瘤进行重新分离,得到二次分离菌株(以下简称结瘤菌株)。在相同的培养条件下对结瘤菌株与供试菌株进行全细胞蛋白SDS-PAGE分子标记试验,发现10株供试菌株与其对应结瘤菌株的蛋白分子图谱相似性极高,从而肯定了它们与三叶草的共生结瘤关系。The whole cell protein SDS-PAGE was a rapid, stable, accurate method to reflect protein molecules types in cells, it could be developed to perform primarily clustering of Rhizobium and verification of nodulation strains in nodulation ability test. 120 strains isolated from the root nodule of Trifolium spp. which were collected from six regions such as Yunnan, Hubei, Hebei, and so on. The clustering analysis of the whole cell protein SDS-PAGE molecular marker showed that there were 104 types which clustered six groups at a similarity level of 80%. 42 strains were selected by strains char-acteristic and collection site etc. to carry out the nodulation test. The results showed that 42 Trifoli-um spp. plants with rhizobia inoculation could nodulate, and the nodulation rate was 100%. At the same time, some re-isolated strains named nodulation strains gained by re-isolated root nodule of rep-resentative strains partly. Then, we carried out the whole cell protein molecular marker between orig-inal strains and re-isolates strains in same cultivation conditions. The clustering analysis demonstrated that 10 original strains had an identical protein maps with their re-isolated strains at higher similarity,it indicated that these strains could establish symbiotic nodulation connection with their original Tri-folium spp. hosts.

关 键 词：三叶草 根瘤菌 分子标记 全细胞蛋白 SDS-PAGE 

分 类 号：S541.9[农业科学—作物学]