XIAO Wen-li,GUAN Da-wei,LI Jun,et al.Evaluation on the Competitiveness of Strains of Soybean Rhizobia Marking with gfp and rfp Genes[J].Soybean Science,2010,29(03):366-369.[doi:10.11861/j.issn.1000-9841.2010.03.0366]
采用gfp和rfp基因标记评价大豆根瘤菌竞争结瘤能力
- Title:
- Evaluation on the Competitiveness of Strains of Soybean Rhizobia Marking with gfp and rfp Genes
- 文章编号:
- 1000-9841（2010）03-0366-04
- Keywords:
- Fluorescent protein gene; Rhizobium; Competitive nodulation
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 采用二亲本接合方法，将绿色荧光蛋白基因（gfp）或红色荧光蛋白基因（rfp）分别导入与大豆品种中黄13相匹配的快生根瘤菌Sinorhizobium fredii SR25a、S.fredii USDA205、与慢生根瘤菌Bradyrhizobium japonicum 4222、B.japonicum 4230和B.japonicum 4534菌株中，得到6株标记成功的菌株，并检验标记菌株的外源基因在培养条件下和共生条件下的稳定性，以及对菌株结瘤性能的影响。进一步将带有不同荧光蛋白基因的供试菌株按等密度等体积配成9组，通过蛭石盆栽试验评价菌株的竞争结瘤能力。结果表明：外源基因能够遗传稳定，且不影响共生结瘤固氮行为，该标记方法可用于评价菌株竞争结瘤能力；慢生大豆根瘤菌的竞争能力明显高于快生大豆根瘤菌，其中慢生大豆根瘤菌B.japonicum 4534竞争能力最强，其占瘤率比慢生菌株B.japonicum 4230和B.japonicum 4222分别高出35％和90％。结果证明gfp和rfp双标记技术为根瘤菌竞争结瘤能力评价提供了直观、简便、准确的检测手段。
- Abstract:
- The green-fluorescent protein gene (gfp) or red-fluorescent protein gene (rfp) was transferred into Sinorhizobium fredii SR25a, S.fredii USDA205, Bradyrhizobium japonicum4222, B.japonicum 4230 and B.japonicum 4534 by double-parent hybridizing method respectively to obtain 6 marked strains. The genetic stability of the foreign genes were detected and their effects on the strains nodulation ability were evaluated. Nine pairs of strains from testing strains with different fluorescent protein gene were inoculated on the soybean cultivar Zhonghuang 13 for competitive nodulation experiment. The results showed that the foreign genes could transfer stably, and they had no impact on the symbiotic nitrogen fixation activity of strains. The competitive nodulation ability of B.japonicum was higher than S.fredii. Among the B.japonicum strains, nodule occupancy of B.japonicum 4534 was the highest and its nodule occupancy was 35％ and 90％ over B.japonicum 4230 and B.japonicum 4222, respectively. Marking with fluorescent protein gene provides one way to evaluate rapidly the competitive nodulation ability of rhizobial strains.
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备注/Memo
基金项目：现代农业产业技术体系建设专项资助项目(nycytx-004)；中央级公益性科研院所科研业务专项资助项目（2010-34）。