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以 5株慢生型花生根瘤菌和天府 3号花生为材料 ,用 AFLP技术研究了慢生型花生根瘤菌 Spr2 - 9、Spr3- 3、Spr3- 5、Spr4- 5和 Spr7- 1的遗传特性和竞争结瘤能力。结果显示 ,供试条件下 ,传代次数对菌株的遗传性状无明显影响 ,2 8℃培养条件下 ,花生根瘤菌连续传 96代 ,其 AFLP指纹未发生明显变化 ;37℃培养 ,仅 Spr3- 3和 Spr3- 5能够存活并正常生长 ,其 AFLP指纹也未发生明显改变 ,然而其它菌株不能生长。将供试慢生型花生根瘤菌分别接种天府 3号花生 ,光照培养 30 d后 ,随机各取 4个根瘤 ,从根瘤中提取类菌体 DNA进行 AFLP分析 ,各根瘤类菌体 DNA的 AFLP指纹图谱与该菌株纯培养物 AFLP指纹相同。将 5个菌株混合接种天府 3号花生 ,不同菌株的占瘤率存在差异 ,Spr3- 3和 Spr3- 5的竞争结瘤能力最强 ,两菌株的占瘤率之和为 85.4% ;Spr4- 5的占瘤率为 1 2 .2 % ;Spr7- 1为 2 .4% ;而 Spr2 - 9的竞争结瘤能力最差。本试验结果说明 ,AFLP技术用于根瘤菌生态和竞争结瘤能力研究 ,具有下列优点 :简易、快速、准确 ;直接取豆科植物的根瘤提取 DNA,进行原位研究 ;在不改变菌株遗传特性 ,即不使用突变株的前提下 ,可以直接测定已知菌株的竞争结瘤能力
The genetic characteristics of the slow - growing peanut rhizobia Spr2 - 9, Spr3 - 3, Spr3 - 5, Spr4 - 5 and Spr7 - 1 were studied by using AFLP technique with five slow - growing peanut rhizobia and Tianfu 3 peanut. And competitive nodulation ability. The results showed that under the conditions of the test, the number of passages did not affect the genetic traits of the strains. Under the culture medium of 2 8 ℃, the peanut rhizobia were continuously passaged for 96 generations, and the AFLP fingerprints did not change significantly. Only cultured at 37 ℃, only Spr3-3 And Spr3-5 were able to survive and grow normally, their AFLP fingerprints did not change significantly, however, other strains could not grow. The slow-growing peanut rhizobia were inoculated with Tianfu No.3 peanut. After 30 days of light culture, four root nodules were randomly selected. The genomic DNA was extracted from nodules for AFLP analysis. AFLP fingerprinting The map is identical to the AFLP fingerprint of this strain pure culture. Five strains were inoculated with Tianfu No.3 peanut. The differences of tumor-bearing rates among different strains were different. Spr3-3 and Spr3-5 had the strongest competition nodulation ability, and the two strains accounted for 85.4%. Spr4-5 Accounted for 12.2%; Spr7- 1 was 2.4%; and Spr2-9 had the lowest competitive nodulation capacity. The results of this experiment show that AFLP technology is applied to the study of rhizobia ecology and competitive nodulation ability, which has the following advantages: simple, rapid and accurate; direct extraction of DNA from nodules of leguminous plants for in situ studies; without changing the genetic characteristics of strains , That is, without the use of mutant strains under the premise of the direct determination of competitive strains of nodulation ability