辣椒碱提取与剂型加工及对小菜蛾作用机制研究
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摘要
本论文系统研究了辣椒碱的提取与纯化、辣椒碱乳油的加工及辣椒碱乳油对小菜蛾的防效,并首次研究了辣椒碱对小菜蛾的作用机制。试验结果表明,辣椒碱的杀虫活性较高,对害虫具有触杀、胃毒等多种作用方式,而且对小菜蛾体内的一些靶标酶、代谢酯酶及酯酶同功酶都产生了不同程度的影响。本试验为辣椒碱的深入开发和研究奠定了理论基础,并对害虫的可持续治理具有重要的理论和实践意义。
采用丙酮温浸法、酸碱法和柱层析法相结合的方法对辣椒碱进行了提纯,最后制得了白色膏状的纯品辣椒碱。然后测定了纯品辣椒碱对小菜蛾的生物活性,结果表明,辣椒碱对小菜蛾表现出良好的拒食活性和一定的触杀活性。
采用实验室常规的乳油加工方法,对辣椒碱乳油的配方进行了筛选,最后所得的2%辣椒碱乳油的配方为:有效成分是2%的辣椒碱,助溶剂是20%的丙酮,乳化剂是10%的TW-80,润湿剂是2%的JFC润湿剂渗透剂,主要溶剂为二甲苯,添至100%。对此乳油配方的质量鉴定结果表明,各个指标测定结果均符合乳油标准。而且2%辣椒碱乳油对小菜蛾显示出较好的防治效果,在处理后的第9d,40mg·L~(-1)浓度下的校正虫口减退率及保叶效果分别为81.82%和66.67%。
采用常规的生物活性测定方法对辣椒碱的作用方式进行了研究。结果表明,辣椒碱具有一定的触杀和胃毒作用,较强的产卵忌避和拒食作用。触杀作用方式上,在1.00×10~6mg·L~(-1)浓度下,处理60h小菜蛾的校正死亡率达51.72%。胃毒作用方式上,在6.25×10~4mg·L~(-1)浓度下,处理60h小菜蛾的校正死亡率达44.44%。产卵忌避作用方式上,在6.25×10~4 mg·L~(-1)浓度下,处理24h辣椒碱对小菜蛾的非选择性产卵忌避率达96.55%,选择性产卵忌避率为84.30%。拒食作用方式上,在6.25×10~4mg·L~(-1)浓度下,处理48h辣椒碱对小菜蛾的非选择性拒食率达81.47%,选择性拒食率为69.69%。
采用常规的生化试验方法,研究了辣椒碱对小菜蛾体内酶系的影响。试验结果表明,辣椒碱对小菜蛾体内的乙酰胆碱酯酶、Na~+K~+-ATP酶、羧酸酯酶、谷胱甘肽-S-转移酶及酯酶同工酶都产生了不同程度的影响。经不同浓度的辣椒碱、不同时间处理后,短时间内不同浓度的辣椒碱都可以抑制乙酰胆碱酯酶的活性,而且高浓度的辣椒碱可以延长对乙酰胆碱酯酶抑制的时间;在受到辣椒碱长时间的刺激后,小菜蛾体内的Na~+K~+-ATP酶的活性会逐渐升高,而使小菜蛾处于兴奋状态;对羧酸酯酶的影响表现为先降后升,即处理后辣椒碱对小菜蛾体内的羧酸酯酶产生了一定的抑制作用,但随着处理时间的延长,羧酸酯酶的活性会逐渐增强。对谷胱甘肽-S-转移酶及酯酶同工酶的研究结果表明,辣椒碱能够抑制昆虫体内的一些酯酶,使这些酯酶的活性低于正常的水平,但不能使这些酶的活性得到完全的抑制。另外,辣椒碱对酯酶同工酶的数量没有影响,但对同工酶1的影响要高于对同工酶2的影响。
Extracting and purifying of capsaicin,formulation preparation and control effect of capsaicin emulsifiable concentrate,the action modes of capsaicin against Plutella xylostella L.,and effect of capsaicin against Plutella xylostella L.on enzymes activity were researched chiefly.The study provides the basis for the development and research thoroughly of capsaicin,and it has theoretical and practical meaning for the IPM.
Experiments were conducted to purifying capsaicin with acetone warm soaking method, acid base method,LCC method.The white cream pure capsaicin was obtained.Bioactivity studies showed that it had strong repellent activity and certain contact activity.
Experiments were conducted to screening formulation of capsaicin emulsifiable concentrate with conventional formulation preparation methods in laboratory.The screened formulation was,the percentage of active component was 2%capsaicin,the percentage of cosolvent was 20%acetone,the percentage of emulsifier was 10%TW-80,the percentage of wetting agent was 2%JFC,and the rest was xylene.The results of pot trails of 2%capsaicin emulsifiable concentrate showed that it could control Plutella xylostella L.effectively.2% capsaicin emulsifiable concentrate at 40 mg·L~(-1),the revised pest individual depress ratio was 81.82%after treatment 9 d,the rate of protecting leaves was 66.67%after treatment 9 d.
Bioactivity studies showed that capsaicin possessed strong oviposition deterrence activity and antifeedant activity,and it also had certain contact toxicity and stomach toxicity on Plutella xylostella L..In the test of contact toxicity,the corrected mortality was 51.72%after applying 60 hours at a capsaicin concentration of 1×10~6 mg·L~(-1).In the test of stomach toxicity, the corrected mortality was 44.44%after applying 60 hours at a capsaicin concentration of 6.25×10~4mg·L~(-1).In the test of oviposition deterrence activity,at a capsaicin concentration of 6.25×10~4mg·L~(-1),the nonchoice oviposition deterrence rate was 96.55%after applying 24 hours,the choice oviposition deterrence rate was 84.30%after applying 24 hours.In the test of antifeedant activity,At a capsaicin concentration of 6.25×10~4mg·L~(-1),the nonchoice antifeedant rate was 81.47%after applying 48 hours,the choice antifeedant rate was 69.69% after applying 48 hours.
Experiments were conducted to study the effects of capsaicin against Plutella xylostella L. on different enzymes activity with conventional methods in laboratory.The result showed that capsaicin could inhibit the activity of acetylcholinesterase after treated short time,and capsaicin could enhance the activity of Na~+K~+-ATPase after treated long time.Compared with the negative control,the activity of glutathione-S-transferase and carboxylesterase were decreased in different extent by capsaicin.And there were two different esterase isozymes in both treated group and control group.
采用丙酮温浸法、酸碱法和柱层析法相结合的方法对辣椒碱进行了提纯,最后制得了白色膏状的纯品辣椒碱。然后测定了纯品辣椒碱对小菜蛾的生物活性,结果表明,辣椒碱对小菜蛾表现出良好的拒食活性和一定的触杀活性。
采用实验室常规的乳油加工方法,对辣椒碱乳油的配方进行了筛选,最后所得的2%辣椒碱乳油的配方为:有效成分是2%的辣椒碱,助溶剂是20%的丙酮,乳化剂是10%的TW-80,润湿剂是2%的JFC润湿剂渗透剂,主要溶剂为二甲苯,添至100%。对此乳油配方的质量鉴定结果表明,各个指标测定结果均符合乳油标准。而且2%辣椒碱乳油对小菜蛾显示出较好的防治效果,在处理后的第9d,40mg·L~(-1)浓度下的校正虫口减退率及保叶效果分别为81.82%和66.67%。
采用常规的生物活性测定方法对辣椒碱的作用方式进行了研究。结果表明,辣椒碱具有一定的触杀和胃毒作用,较强的产卵忌避和拒食作用。触杀作用方式上,在1.00×10~6mg·L~(-1)浓度下,处理60h小菜蛾的校正死亡率达51.72%。胃毒作用方式上,在6.25×10~4mg·L~(-1)浓度下,处理60h小菜蛾的校正死亡率达44.44%。产卵忌避作用方式上,在6.25×10~4 mg·L~(-1)浓度下,处理24h辣椒碱对小菜蛾的非选择性产卵忌避率达96.55%,选择性产卵忌避率为84.30%。拒食作用方式上,在6.25×10~4mg·L~(-1)浓度下,处理48h辣椒碱对小菜蛾的非选择性拒食率达81.47%,选择性拒食率为69.69%。
采用常规的生化试验方法,研究了辣椒碱对小菜蛾体内酶系的影响。试验结果表明,辣椒碱对小菜蛾体内的乙酰胆碱酯酶、Na~+K~+-ATP酶、羧酸酯酶、谷胱甘肽-S-转移酶及酯酶同工酶都产生了不同程度的影响。经不同浓度的辣椒碱、不同时间处理后,短时间内不同浓度的辣椒碱都可以抑制乙酰胆碱酯酶的活性,而且高浓度的辣椒碱可以延长对乙酰胆碱酯酶抑制的时间;在受到辣椒碱长时间的刺激后,小菜蛾体内的Na~+K~+-ATP酶的活性会逐渐升高,而使小菜蛾处于兴奋状态;对羧酸酯酶的影响表现为先降后升,即处理后辣椒碱对小菜蛾体内的羧酸酯酶产生了一定的抑制作用,但随着处理时间的延长,羧酸酯酶的活性会逐渐增强。对谷胱甘肽-S-转移酶及酯酶同工酶的研究结果表明,辣椒碱能够抑制昆虫体内的一些酯酶,使这些酯酶的活性低于正常的水平,但不能使这些酶的活性得到完全的抑制。另外,辣椒碱对酯酶同工酶的数量没有影响,但对同工酶1的影响要高于对同工酶2的影响。
Extracting and purifying of capsaicin,formulation preparation and control effect of capsaicin emulsifiable concentrate,the action modes of capsaicin against Plutella xylostella L.,and effect of capsaicin against Plutella xylostella L.on enzymes activity were researched chiefly.The study provides the basis for the development and research thoroughly of capsaicin,and it has theoretical and practical meaning for the IPM.
Experiments were conducted to purifying capsaicin with acetone warm soaking method, acid base method,LCC method.The white cream pure capsaicin was obtained.Bioactivity studies showed that it had strong repellent activity and certain contact activity.
Experiments were conducted to screening formulation of capsaicin emulsifiable concentrate with conventional formulation preparation methods in laboratory.The screened formulation was,the percentage of active component was 2%capsaicin,the percentage of cosolvent was 20%acetone,the percentage of emulsifier was 10%TW-80,the percentage of wetting agent was 2%JFC,and the rest was xylene.The results of pot trails of 2%capsaicin emulsifiable concentrate showed that it could control Plutella xylostella L.effectively.2% capsaicin emulsifiable concentrate at 40 mg·L~(-1),the revised pest individual depress ratio was 81.82%after treatment 9 d,the rate of protecting leaves was 66.67%after treatment 9 d.
Bioactivity studies showed that capsaicin possessed strong oviposition deterrence activity and antifeedant activity,and it also had certain contact toxicity and stomach toxicity on Plutella xylostella L..In the test of contact toxicity,the corrected mortality was 51.72%after applying 60 hours at a capsaicin concentration of 1×10~6 mg·L~(-1).In the test of stomach toxicity, the corrected mortality was 44.44%after applying 60 hours at a capsaicin concentration of 6.25×10~4mg·L~(-1).In the test of oviposition deterrence activity,at a capsaicin concentration of 6.25×10~4mg·L~(-1),the nonchoice oviposition deterrence rate was 96.55%after applying 24 hours,the choice oviposition deterrence rate was 84.30%after applying 24 hours.In the test of antifeedant activity,At a capsaicin concentration of 6.25×10~4mg·L~(-1),the nonchoice antifeedant rate was 81.47%after applying 48 hours,the choice antifeedant rate was 69.69% after applying 48 hours.
Experiments were conducted to study the effects of capsaicin against Plutella xylostella L. on different enzymes activity with conventional methods in laboratory.The result showed that capsaicin could inhibit the activity of acetylcholinesterase after treated short time,and capsaicin could enhance the activity of Na~+K~+-ATPase after treated long time.Compared with the negative control,the activity of glutathione-S-transferase and carboxylesterase were decreased in different extent by capsaicin.And there were two different esterase isozymes in both treated group and control group.
引文
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