花椒精油提取及其生物活性测定研究

【作者】 庄世宏；

【导师】 李孟楼；

【作者基本信息】 西北农林科技大学 ， 森林保护学， 2002， 硕士

【摘要】 花椒精油是从芸香科花椒属植物花椒（Zanthoxylum bungeanum Maxim.）中提取的一种植物精油，国内外学者通过研究发现，花椒精油对粮食害虫麦蛾、玉米象、杂拟谷盗、赤拟谷盗及四纹豆象有较强的生物活性，且作用方式多元化，并对曲霉（Aspergillus SP.）和青霉（Penicillium SP.）等多种储粮致霉菌抑菌效果明显。本文在以上研究的基础上，对花椒精油的提取、花椒精油对某些农田害虫、林木害虫、植物病原菌和卫生病菌的生物活性及其部分生理生化反应进行了初步研究，得出如下结果。 1、花椒精油提取率的影响因素：花椒果实精油含量在幼果期较高，精油含量随着果实的生长而下降；新鲜花椒其精油提取率高于阴干的；采用循环水蒸汽蒸馏法可极大地提高精油提取率。 2、花椒精油的抑菌作用：对真菌的抑菌效果较细菌差，在真菌中，以霉菌受抑制程度最大，酵母菌最弱，对于细菌，对革兰氏阴性细菌的效果好于革兰氏阳性菌；在效价浓度以上，花椒精油随浓度增大而抑菌效果明显，在效价以下时，浓度变化对抑菌效果无影响；浓度变化对真菌的作用影响最强烈；49h对各菌抑菌作用最强。 3、花椒精油对试虫的生物活性： （1）对小菜蛾、粘虫和杨毒蛾表现出很强的熏蒸毒杀作用，其LC₅₀值分别是6.7491μl//L、8.3568μl//L、6.4508μl//L，熏蒸活性和时间呈正相关关系； （2）对粘虫、杨毒蛾有较好的触杀活性，其LD₅₀值分别是5.2555μl／g、1.1692μl／g，对杨毒蛾的触杀活性明显强于粘虫； （3）对杨毒蛾有极强的拒食活性，对粘虫拒食活性随时间延长而出现诱食现象；当花椒精油浓度小于0.05ml／ml时，杨毒蛾和粘虫表现出“波浪状拒食效应”，其拒食率并非随着时间而下降，而是忽而高、忽而低； （4）对粘虫和杨毒蛾均有很强的忌避活性，忌避率随时间延长而下降。 4、试虫的中毒症状：分别以19.35μl//L和15.48μl//L的浓度熏蒸处理粘虫和小菜蛾四龄幼虫，其中毒试虫活动缓慢、行动迟缓，有吐水及体表失水现象，无兴奋、痉挛症状，最终昏迷死亡，是呼吸毒剂中毒症状；以19.35μl//L的浓度熏蒸处理杨毒蛾四龄幼虫、以分别为1ml／ml和2ml／ml浓度的花椒精油1μl分别点滴处理粘虫和杨毒蛾四龄幼虫，试虫到处爬动，头部上扬，并不停扭动，持续一段时间之后，活动减弱，虫体瘫软、蜡缩，呈麻痹、昏迷状，并有抽搐、痉挛现象，最终虫体皱缩死亡，表现出神经性毒剂中毒症状。 5、花椒精油对试虫生理生化的影响： 门)经熏蒸处理，对粘虫五龄幼虫呼吸作用有显著影响，与对照相比，呼吸率提高而呼吸商明显下降，说明花椒精油导致试虫耗氧量增大，而试虫的氧化能力受到抑制； （2）经点滴处理，杨毒蛾四龄幼虫血淋巴的蛋白质含量比对照显著下降，而组织液中的蛋白含量则高于对照； （3）经点滴处理，对杨毒蛾四龄幼虫血淋巴及组织液中的酯酶同工酶的表达带来影响。 从整个研究结果看，花椒精油对害虫表现出拒食、熏蒸、触杀等多种作用方式，其作用机理多样，影响神经系统及呼吸系统，并具有较强的杀菌活性，是一种颇具开发潜力的杀虫杀菌剂。更多还原

【Abstract】 As a refined plant oil, domestic and abroad scientists found that Zanthoxylum oil which extracted from zanthoxylum bungednum, belong to zanthoxylum of retraced, had quite powerful and different biological active to grain pests (Such as bungeanum sitotroga cerealella, sitophilus zeamains,, Tribolium confusum, Tribolium castaneum and Callosobruchus maculatus ) and many mildew (such as Aspergillus sp. andpenicillium sp.). By studying on its extract, biological activity and some physiological reaction on certain farmland pests, forest pests, plant pathogenic and healthy bacteria’s, it showed that:1 immature fruit had higher content of zanthoxylum oil than others, with the growing of fruit, the centent of zanthoxylum oil began to decrease; Fresh fruit had higher collect efficiency than dry fruit; By using the method of water vapor cycle distillation, the abstract efficiency of refined oil can be improver extremely.2 Zanthoxylum oil had poorer bacteriostatic action effect on fungus than germs. Experiment showed that mildews was subjected to be more controlled by zanthoxylum oil than yeast. G germs was subjected to be more controlled by zanthoxylum oil than G+ germs; Above the effective concentration, the control effect by zanthoxylum oil on bacterium and fungus was increased with the improvement of the oil concentration; Below the effective concentration the controller effect was constant with the change of the oil concentration. Concentration variation of zanthoxylum oil had the most effective on fungus. However, both bacterium and fungus had been powerful restrained at 49h.3 Biological active experiment of zanthoxylum oil on pests showed that:(1) Zanthoxylum oil and quite powerful fumigation action effect on Plutella maculi pennis, Mythimna separata and Leucoma Candida, the LDso of the three kinds pest were 6.7491 u 1/L, 8.3568 u 1/L and 6.4508 u 1/L respectively. Suffocating active and time had a hotable direct relationship;(2) Zanthoxylum oil had fairly contact effect on Mythimna Separata and Leucoma Candidq, the LD50 of the two kinds pest were 5.2555 y 1/L and 1.1692 u 1/L; (3)Zanthoxylum oil had exceeding powerful resist food effect on Leucoma Candida,howere, with the extend of time, the seduce food effect began to appear for Mythimna Separata; When the concentration of zanthoxylum oil below 0.05ml/ml, both Leucoma Candida and Mythimna Separata showed a fluctuate resist food effect from.(4) Zanthoxylum oil had quite strong abstain from Mythimna Spearata and Leucoma Candida,\viih the extend of time, the avoid rate were began to decrease.4 The poisoning symptoms of experimental pests, which had been dialed with zanthoxylum oil (19.35 u 1/L and 15.48 u 1/L), were slow in action, spit out water, lose water on body surface, without excitement and convulsion, of which belong to the respiredpoisoning symptoms. However, the poisoning symptoms of Leucoma Candida, which had been dealed with zanthoxylum oil (19.35 p 1/L) were fast in action and run around, after a short while, the experimental pests were weaken inaction, totally paralyzed and twitched, at last, the pest’s body was wrinkled and died.5 Effect of zanthoxylum oil on experimental pest’s biology and physiology showed that:(1) Zanthoxylum oil had striking influence on the respiratory system of Mythimna Separata laboratory test showed that zanthoxylum oil can lead to the increase of oxygen consumption of experimental pests, which restrain their oxidants capable.(2) The protein content of hemolymph lymph in experimental pests, which had been dialed with zanthoxylum oil, was decreased obviously comparing to common pests but the protein content of organize liquid was higher than common pests.(3)The effects on expression of estterase isoenzyme which were in hemolymph and tissue fluid of Leucoma Candida which were four ages by contact application.whole studying showed that zanthoxylum oil had many effect ways on pests, which effected the pest’s harrows and respiratory systems, and also have some potential effect patterns on更多还原