崔晓东 王伊玲 李红霞 李小勇 苏鹏飞 潘瑞朋 刘琼光 胡蓉花

摘  要：了解不同噬菌体在不同环境中的存活情况，探索化学杀菌剂与噬菌体联合防治烟草青枯病的可行性。将P7-1、P1556-1及P1556-2等3种噬菌体与7种化学杀菌剂混合后，置于4 ℃和30 ℃培养箱，测定噬菌体效价随时间的变化规律。结果表明，3个噬菌体在7种杀菌剂中存活时间存在明显差异：3种噬菌体与氢氧化铜和王铜混合后1个月，其效价均为0;对其他5种杀菌剂的耐药由强到弱为P1556-2>P1556-1>P7-1，其中P1556-2在噻唑锌、中生菌素、春雷霉素、喹啉铜和辛菌胺醋酸盐中12个月测定的效价对数值分别为8.73、7.65、7.93、7.39和8.18。部分噬菌体在30 ℃可存活1～5个月，但在4 ℃下可存活6～12个月。噬菌体P1556-2和P1556-1在无致病力青枯菌Trp574中保存，其效价明显高于在无菌水中保存。可见，噬菌体在杀菌剂中的存活（效价）受时间、温度、药剂种类和噬菌体自身特性的影响，氢氧化铜与王铜制剂对噬菌体具有明显抑制作用，4 ℃（低温）下能够延长噬菌体的存活时间。无致病力青枯菌Trp574可用于噬菌体P1556-1和P1556-2的保存和大量培养。

关键词：烟草青枯病;杀菌剂;噬菌体;无致病力青枯菌;存活;效价

Abstract： The survival of bacteriophages of Ralstonia solanacearum in different environments and the feasibility of combined use of chemical agents and phages in disease control were studied. Three phages P7-1， P1556-1 and P1556-2 were mixed with seven kinds of bactericides and stored at 4 ℃ and 30 ℃ respectively， to determine the change of phage titer with time. The results showed that the survival time of the three phages were significantly different in the seven bactericides. The titers of the three phages were zero after mixing with copper hydroxide and copper oxychloride for one month. The resistance to bactericides of the three phages in the order from strong to weak is as follows， P1556-2 > P1556-1> P7-1， and the titer logarithms of P1556-2 were 8.73， 7.65， 7.93， 7.39 and 8.18 in thiazole zinc， zhongshengmycin， chunramycin， oxine-copper and octylamine acetate at 12 months， respectively. Some phages survived from 1-5 months at 30 ℃， but stayed alive for 6-12 months at 4 ℃. P1556-2 and P1556-1 had higher titers when they were stored in avirulent strain of R. solanacearum Trp574 than in sterile water. So， the titers were affected by time， temperature， bactericides and phages. Copper hydroxide and copper oxychloride showed obvious inhibition on phages. The survival time of phages can be prolonged at 4 ℃ （low temperature）. The avirulent strain of R. solanacearum Trp574 can be used for preservation and culture of P1556-1 and P1556-2.

Keywords： tobacco bacterial wilt; bactericide; bacteriophage; avirulent strain of Ralstonia solanacearum; activity; titer.

茄科勞尔氏菌（Ralstonia solanacearum）（俗称青枯菌），是引起植物青枯病的病原细菌，寄主范围广泛，能侵染54个科450多种植物[1-2]。烟草青枯病是我国烟草种植区普遍发生且危害严重的病害，其防治一直是生产上的难题。化学防治具有方便快速等优点，如20%噻唑锌悬浮剂、3%中生菌素可湿性粉剂等对青枯病有一定的防治效果[3-5]。但长期大量使用化学农药易导致环境污染和病菌抗药性等问题，由此，青枯病生物防治受到国内外重视[6-8]，其中包括青枯菌无致病力菌株的应用[9，13]，以及利用噬菌体防治作物青枯病[10-12]。……