| 刘 露 1, 林志坚 1, 周 挺 2, 梁颁捷 3, 顾 钢 2*, 胡方平 1, 蔡学清 1*.植物青枯菌噬菌体保存方法的研究[J].植物保护,2021,47(4):155-160. |
| 植物青枯菌噬菌体保存方法的研究 |
| The preservation methods of Ralstonia pseudosolanacearum phage |
| 投稿时间：2020-04-01 修订日期：2020-05-14 |
| DOI：10.16688/j.zwbh.2020162 |
| 中文关键词: 青枯病菌 噬菌体 保存方法 |
| 英文关键词:Ralstonia pseudosolanacearum bacteriophage preservation method |
| 基金项目:国家重点研发计划(2018YFD0201300); 中国烟草总公司福建省公司科技计划项目(闽烟司科[2017]2号); 福建农林大学科技创新项目(KFA17541A) |
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| 中文摘要: |
| 噬菌体可用于防治植物青枯病, 但不易保存?本文对青枯病菌短尾噬菌体P1?P2?P3和P4株系的保存方法和条件进行了研究; 将各噬菌体原液?噬菌体原液∶青枯菌液=1∶1?噬菌体原液∶青枯菌液∶40%甘油=1∶1∶2?噬菌体原液∶青枯菌液∶80%甘油=1∶1∶2的处理分别置于-75?-25?4℃?室温下保存, 再分别于90?180?270?365 d后取样测定噬菌体的效价?结果表明, -75℃和-25℃下, 噬菌体原液∶青枯菌液∶40%或80%甘油=1∶1∶2保存的效果最好, 365 d后效价仅下降0~2个数量级; 噬菌体原液∶青枯菌液=1∶1在4℃和室温下保存效果也较好, 4℃保存365 d后效价仅下降1~3个数量级; 此处理在室温条件下保存也可达270 d?因此, 以资源长期保存为目的可采用噬菌体原液∶青枯菌液∶40%或80%甘油=1∶1∶2保存于-75℃或-25℃下; 短期保存可采用噬菌体原液∶青枯菌液=1∶1保存于室温下? |
| 英文摘要: |
| Bacterial phage could be used to control crop bacterial wilt caused by Ralstonia pseudosolanacearum, however, the phage is a living parasite and is not easy to be preserved. In this study, the preservation methods of Podoviridae phage strain P1, P2, P3 and P4 were screened from the following four treatments, which included bacteriophage suspension, bacteriophage suspension∶ R.pseudosolanacearum suspension at 1∶1, bacteriophage suspension∶ R.pseudosolanacearum suspension∶40% glycerol at 1∶1∶2 and bacteriophage suspension∶ R.pseudosolanacearum suspension∶80% glycerol at 1∶1∶2 were preserved at -75℃, -20℃, 4℃ and room temperature, respectively. The titer of the phage was detected by double-layer plate method after preservation for 90, 180, 270, 365 d. The results showed that the treatment of bacteriophage suspension∶ R.pseudosolanacearum suspension∶40% or 80% glycerol at 1∶1∶2 preserved at -75℃ and -25℃ was the best, and the phage titer reduced 0-2 magnitude after preservation for 365 d. At 4℃ and room temperature, the treatment of bacteriophage suspension∶ R.pseudosolanacearum suspension at 1∶1 had a good effect, the titer reduced only 1-3 magnitude after preservation for 365 d at 4℃, and the phage in this treatment could also keep alive for 270 d at room temperature. The results indicated that the treatment of bacteriophage suspension∶ R.pseudosolanacearum suspension∶40% or 80% glycerol at 1∶1∶2 preserved at -75℃ and -25℃ is a method for long time preservation. The treatment of bacteriophage suspension∶ R.pseudosolanacearum suspension at 1∶1 preserved at room temperature is a simple and reliable method. |
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