褐飞虱遇险释放挥发性物质的提取及成分分析
Extraction of the volatile substance released by brown planthopper, Nilaparvata lugens (St?l) , in danger and its component analysis
李凯，陈列忠，陈建明，何月平，张珏锋
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作者单位：杭州师范大学 生命与环境科学学院;浙江省植物有害生物防控重点实验室省部共建国家重点实验室培育基地，浙江省农业科学院植物保护与微生物研究所
中文关键词：褐飞虱；挥发性物质；行为反应；成分分析
英文关键词：Nilaparvata lugens; volatile substance; behavioral response; component analysis
中文摘要：利用Y型嗅觉仪测定不同试验条件下褐飞虱对同类昆虫遭遇草间小黑蛛捕食时挥发性物质的行为反应，并利用固相微萃取、气相色谱与质谱技术分离鉴定挥发性物质。目的明确褐飞虱遇险释放挥发性物质的最佳提取条件，并分析该挥发性物质的成分。结果表明，用乙酸乙酯、甲醇和正己烷分别提取“褐飞虱2-3龄若虫+草间小黑蛛成蛛”共存体释放的挥发性物质，发现乙酸乙酯提取物能引起褐飞虱极显著的逃避行为。用不同褐飞虱虫量（100、300、600头/500 mL瓶）、不同提取时间（1、3、6小时）和不同溶剂温度（15、25、35℃）分别提取“褐飞虱2-3龄若虫+草间小黑蛛成蛛”共存体释放的挥发性物质，发现300-600头、3-6小时和25℃下的提取物能引起褐飞虱极显著的逃避行为，明确了褐飞虱遇险释放挥发性物质的最佳提取条件。利用固相微萃取法分别萃取“褐飞虱2-3龄若虫”释放的挥发性物质和“褐飞虱2-3龄若虫+草间小黑蛛成蛛”共存体释放的挥发性物质。气相色谱分析表明，保留时间16min、21.5min时，“褐飞虱”与“褐飞虱+草间小黑蛛”共存体释放的挥发物均出现新峰；保留时间10.94min时，仅“褐飞虱+草间小黑蛛”共存体释放的挥发物出现新峰。推测保留时间10.94min时出现的峰为褐飞虱被草间小黑蛛捕食时释放的示警挥发物。质谱分析进一步表明，这种挥发物具有与(E)-2-己烯醛相类似的化学结构，但其实际结构还需深入研究。
英文摘要：Behavior response of the brown planthopper (BPH), Nilaparvata lugens (Stål), to the volatile substance released by the same kinds of insects in danger at different experimental conditions was conducted using Y-tube olfactometer, the volatile substance was extracted with SPME method, and identified by GC-MS. The objectives were determination for optimum extraction conditions of the volatile substance released by brown planthopper (BPH), Nilaparvata lugens (Stål), in dangerous and analysis for the volatile component. The results showed that the volatile substance, released by the coexistence of “2nd to 3th instar nymphs of BPH + adults of Erigonidium graminicolum”, was extracted using ethyl acetate, methanol and n-hexane, respectively. We found the significant escape response of BPH caused by the extraction of ethyl acetate. The volatile substance released by the coexistence was further extracted at different BPH densities (100, 300, 600 insects in 500 mL bottle), different extraction times (1, 3, 6 hrs) and different extraction temperatures (15, 25, 35℃), respectively. We found the extraction at the conditions of 300-600 insects, 3-6 hours and 25℃ led to a significant escape behavior of BPH. The volatile substance, released by 2nd to 3th instar nymphs of BPH or the coexistence of “2nd to 3th instar nymphs of BPH + adults of E. graminicolum”, was extracted with method of solid phase micro-extraction (SPME). Gas chromatograph (GC) analysis showed a new peak appearance of the volatile released by BPH nymphs or the coexistence of “BPH nymphs + E. graminicolum adults” when retention time was 16 minutes and 21.5 minutes, respectively. Another new peak of the volatile released by the coexistence appeared only when retention time was 10.94 minutes. Analysis of mass spectrometry (MS) indicated chemical structure of the volatile was similar to the structure of (E)-2-hexenal, but its actual chemical structure still further determination.