储存条件对表达马铃薯甲虫腺苷高半胱氨酸水解酶基因dsRNA的大肠杆菌生物活性的影响
Storage conditions affect the toxicity of E. coli expressing S-adenosyl-L- homocysteine hydrolase (SAHase) gene dsRNA to potato beetles
李晓旭1, 2** 付开赟3 李国清3 王 刚2 吐尔逊1 何 江1 郭文超1***
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DOI：10.7679/j.issn.2095-1353.2015.145
作者单位：1. 新疆农业科学院植物保护研究所，乌鲁木齐 830091；2. 石河子大学农学院，石河子 832003；3. 南京农业大学植物保护学院 农业部病虫害监测与治理重点开放实验室，南京 210095
中文关键词：马铃薯甲虫，腺苷高半胱氨酸水解酶，dsRNA，RNA干扰，储存条件
英文关键词：Leptinotarsa decemlineata, S-adenosyl-L-homocysteine hydrolase, dsRNA, RNAi, storage conditions
中文摘要：
【目的】 将RNA干扰应用于害虫防治领域，通过饲喂表达dsRNA 的转基因植物或表达dsRNA的细菌，可沉默特定基因进而控制害虫，为农业害虫防治开辟一个新领域。而表达dsRNA的细菌能否有效储存是决定此项技术实际应用的关键。【方法】 用﹣20℃冻存的表达马铃薯甲虫腺苷高半胱氨酸水解酶（SAHase）dsRNA的大肠杆菌E.coli，饲喂马铃薯甲虫Leptinotarsa decemlineata（Say）2龄幼虫，检测不同时间储存、灭活与否的活性变化，明确其储存条件。【结果】 大肠杆菌﹣20℃储存24 h后生物活性优于新鲜菌，储存48 h后效果减弱，而载体大肠杆菌是否灭活对活性影响不大。【结论】 dsRNA大肠杆菌发酵液在﹣20℃条件下可以短暂储存。
英文摘要： [Objectives] By expressing double stranded RNA (dsRNA) in transgenic plants and in prokaryotic cells, RNA interference has potential applications in pest control. Determination of the influence of storage conditions on the toxic effects of dsRNA expressing E. coli is therefore of great importance for determining the effectiveness of such methods. [Methods] We stored living, or sterile, dsRNA expressing E. coli at ﹣20℃ for different periods, and compared their toxicity to Leptinotarsa decemlineata larvae. [Result] E. coli that had been stored for 24 h had higher biological activity than fresh E. coli, however, this difference sharply decreased after 48 h. No significant difference was found between living and sterile dsRNA expressing E. coli. [Conclusion] Twenty-four hours storage under ﹣20℃ has little influence on toxic effects of dsRNA expressing E. coli.