八角尺蠖粪便及八角叶片提取物的生物活性研究
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摘要
八角尺蠖(Dilophodes elegans sinica Prout)是危害八角树的主要食叶害虫,大发生时造成不可估量的经济损失,严重阻碍了八角产业的发展。为实现对该害虫的有效管理与利用,进一步提高八角产业的附加值,作者对八角尺蠖幼虫粪便及其寄主植物提取物的生物活性进行了初步研究。结果如下：
1、采用固相微萃取技术和气相色谱-质谱联用(GC-MS)技术检测出八角尺蠖幼虫虫粪挥发物有6种主要成分,分别为反式茴香醚(83.24%)、α-合金欢烯(2.95%)、2,6-二甲基-6-(4-甲基-3-戊烯基)-庚二烯(4.20%)、十一酸甲酯(0.71%)、十九酸乙酯(1.37%)、11-十八酸甲酯(2.40%),占总峰面积的94.87%。
2、八角尺蠖雌、雄成虫对幼虫粪便挥发物具有强烈的EAG反应。其中,雌、雄成虫对反式茴香醚的EAG反应较强,对十九酸乙酯的EAG反应最弱；反式茴香醚表现出性别差异,其余成分未表现出性别差异。剂量反应曲线表明,在测定的浓度范围内,雌、雄成虫对十一酸甲酯、反式茴香醚、11-十八酸甲酯等3种化合物的EAG反应与浓度的变化呈正相关；对a-合金欢烯的EAG反应呈“U”型；对十九酸乙酯的触角电位反应值在0.01(v/v)浓度下达到最大。雌、雄成虫对不同成分混合物的EAG反应表明,对于雌虫,各成分对反式茴香醚没有增效作用；对于雄虫,a-合金欢烯和十一酸甲酯对反式茴香醚存在增效作用。
3、幼虫虫粪不同溶剂提取物对不同菌种的抑制作用不同,其中丙酮提取物对细菌(如金黄色葡萄球菌、大肠杆菌)的抑菌作用较强,蒸馏水提取物对真菌(如枯青霉、根霉菌)的抑菌作用较强。
4、幼虫虫粪不同溶剂的萃取物对三种自由基均有一定的清除能力。其中,幼虫虫粪萃取物对DPPH自由基的清除能力较小,明显小于同浓度下八角叶片萃取物的清除能力；氯仿相和乙酸乙酯相对·OH自由基的清除能力较强,石油醚相、正丁醇相和水相对·OH自由基的清除能力均小于八角叶片萃取物的清除能力；幼虫虫粪萃取物对O2-自由基的清除能力均小于相同条件下八角叶片萃取物的清除能力。
5、八角叶的乙醇、丙酮、乙酸乙酯和石油醚4种溶剂提取物对榕管蓟马成虫均有较强的驱避作用和熏蒸作用,其中丙酮提取物的驱避作用和熏蒸作用最强,石油醚最弱。在丙酮提取物的各种萃取物中,石油醚萃取物的熏蒸作用最强。石油醚萃取层主要包括烯烃类、醇类、酸类、酮类等21种化学成分,其中,含量最多的成分为大茴香醛、反式茴香脑。
6、八角叶片乙醇和丙酮两种溶剂提取物对榕母管蓟马成虫体内3种保护酶(CAT、POD和SOD)活性均表现为处理初期有促进作用,而在处理后期有抑制作用。两种提取物处理榕母管蓟马2h以后,成虫体内酯酶活性显著升高。丙酮提取物处理6h后,试虫酯酶活性迅速下降到低于对照的水平；而乙醇提取物处理的试虫酯酶活性在6h后继续上升,在较长时间内维持较高的水平。两种提取物处理6h以前对GSTs的活力起促进作用,而处理6h以后则对GSTs的活力起抑制作用。
Dilophodes elegans sinica Prout was the main defoliator in Illicium verum. The development of Illicium verum industry was seriously hindered, and the major economic was immeasurably lost when this insect attack severely occurring. In order to realize the effective management and make use of the pests, and improve value-added of Illicium verum industry, the author preliminary studied on the biological activity of Dilophodes elegans sinica Prout frass and its host plant extracts. The results were summarized as follows:
1. The main volatile matters of Dilophodes elegans sinica Prout frass were detected by means of Solid Phase microextraction and Gas Chromatography-Mass Spectrometry (GC-MS), they were anisole p-propenyl(83.24%),α-farnesene, (2.95%), bicycle[3.1.1]hept-2-ene,2,6-dimethyl-6-(4-methyl-3-pentenyl)(4.20%), undecanoic acid, methyl ester (0.71%), nonadecanoic acid, ethyl ester(1.37%),11-octadecenoic acid methyl ester(2.40%) respectively. The six main volatile matters occupied 94.87% of the total in the peak area.
2. Dilophodes elegans sinica Prout adult showed the strong EAG responses to the volatile matters of Dilophodes elegans sinica Prout frass. Among the five compounds tested, EAG responses of Dilophodes elegans sinica Prout adult to Anisole, p-propenyl were stronger than that of other compounds tested, but the weakest to nonadecanoic acid, ethyl ester, all of the compounds no representation sex difference except anisole, p-propenyl. Dosage response curves of EAG of Dilophodes elegans sinica Prout adult showed that the male and the female adult showed positive correlation EAG response to concentration's varying of undecanoic acid, methyl ester and anisole, p-propenyl and 11-octadecenoic acid, methyl ester, showed "U" model EAG response to the a-farnesene, showed the maximum value of EAG response to nonadecanoic acid, ethyl ester in 0.01 (v/v). The EAG responses of female and male adults to different compositions mixture showed that there were no synergy effects between anisole, p-propenyl and other compositions to female. But the males showed synergy effects between anisole, p-propenyl and a-farnesene, undecanoic acid, methyl ester.
3. The bacteriostasis effect of different extracts of larvae frass to different strains existed some difference. Acetone extracts had a stronger effect to bacterium which including S. aureus and E. coli, and distilled water extracts had a stronger effect to fungus which including P. curinum and R. stolonifer.
4. All of the different extracts from the larvae frass in this paper had scavenging activity to the free radicals including DPPH free radical,-OH free radical,and O2- free radical. Concretely speaking, the extracts from the larvae frass had a weak scavenging activity to DPPH free radical, which was significantly weaker than the extract from Illicium verum leaves under the same concentration. The extracts of chloroform and ethyl acetate had a much stronger scavenging activity to the·OH free radical, while the scavenging activity of extracts of Petroleum ether, butanol and distilled water to·OH free radical was weaker than that of extracts from Illicium verum leaves. The scavenging activity from larvae frass to the O2- free radical was weaker than that of extracts from I. verum leaves under the same conditions.
5. The ethanol, acetone, ethyl acetate and petroleum ether, etc. four extracts all had a strong repellent and fumigate effect, and the repellent and fumigate effect of the acetone extract was strongest while that of petroleum ether extract was weakest among them. In various extracts of acetone extract, the petroleum ether extracts had the best fumigate action, which included 21 kinds of chemical composition of olefine, alcohols, acids, ketones and so on, and the most content were p-anisaldehyde and anethole among all kinds of chemical composition.
6. The ethanol and acetone extracts of I, verum leaves showed a stimulative effect to three protective enzymes (CAI、POD and SOD) of adults of G. ficorum in the early treatment and a inhibition effect in the late treatment. The carboxylesterase activity in adults of G. ficorum significantly increased when the two solvent extracts treated on G. ficorum after two hours. The CAT activity of the treated G. ficorum rapidly decreased down to lower than untreated group when the acetone extract treated after six hours. On the contrary, the CAT activity kept to rise during a long time to maintain a high level when the ethanol extract treated after six hours. The two solvent extracts above showed a stimulative effect to GSTs six hours before treatment and showed a inhibition effect six hours after treatment.
1、采用固相微萃取技术和气相色谱-质谱联用(GC-MS)技术检测出八角尺蠖幼虫虫粪挥发物有6种主要成分,分别为反式茴香醚(83.24%)、α-合金欢烯(2.95%)、2,6-二甲基-6-(4-甲基-3-戊烯基)-庚二烯(4.20%)、十一酸甲酯(0.71%)、十九酸乙酯(1.37%)、11-十八酸甲酯(2.40%),占总峰面积的94.87%。
2、八角尺蠖雌、雄成虫对幼虫粪便挥发物具有强烈的EAG反应。其中,雌、雄成虫对反式茴香醚的EAG反应较强,对十九酸乙酯的EAG反应最弱；反式茴香醚表现出性别差异,其余成分未表现出性别差异。剂量反应曲线表明,在测定的浓度范围内,雌、雄成虫对十一酸甲酯、反式茴香醚、11-十八酸甲酯等3种化合物的EAG反应与浓度的变化呈正相关；对a-合金欢烯的EAG反应呈“U”型；对十九酸乙酯的触角电位反应值在0.01(v/v)浓度下达到最大。雌、雄成虫对不同成分混合物的EAG反应表明,对于雌虫,各成分对反式茴香醚没有增效作用；对于雄虫,a-合金欢烯和十一酸甲酯对反式茴香醚存在增效作用。
3、幼虫虫粪不同溶剂提取物对不同菌种的抑制作用不同,其中丙酮提取物对细菌(如金黄色葡萄球菌、大肠杆菌)的抑菌作用较强,蒸馏水提取物对真菌(如枯青霉、根霉菌)的抑菌作用较强。
4、幼虫虫粪不同溶剂的萃取物对三种自由基均有一定的清除能力。其中,幼虫虫粪萃取物对DPPH自由基的清除能力较小,明显小于同浓度下八角叶片萃取物的清除能力；氯仿相和乙酸乙酯相对·OH自由基的清除能力较强,石油醚相、正丁醇相和水相对·OH自由基的清除能力均小于八角叶片萃取物的清除能力；幼虫虫粪萃取物对O2-自由基的清除能力均小于相同条件下八角叶片萃取物的清除能力。
5、八角叶的乙醇、丙酮、乙酸乙酯和石油醚4种溶剂提取物对榕管蓟马成虫均有较强的驱避作用和熏蒸作用,其中丙酮提取物的驱避作用和熏蒸作用最强,石油醚最弱。在丙酮提取物的各种萃取物中,石油醚萃取物的熏蒸作用最强。石油醚萃取层主要包括烯烃类、醇类、酸类、酮类等21种化学成分,其中,含量最多的成分为大茴香醛、反式茴香脑。
6、八角叶片乙醇和丙酮两种溶剂提取物对榕母管蓟马成虫体内3种保护酶(CAT、POD和SOD)活性均表现为处理初期有促进作用,而在处理后期有抑制作用。两种提取物处理榕母管蓟马2h以后,成虫体内酯酶活性显著升高。丙酮提取物处理6h后,试虫酯酶活性迅速下降到低于对照的水平；而乙醇提取物处理的试虫酯酶活性在6h后继续上升,在较长时间内维持较高的水平。两种提取物处理6h以前对GSTs的活力起促进作用,而处理6h以后则对GSTs的活力起抑制作用。
Dilophodes elegans sinica Prout was the main defoliator in Illicium verum. The development of Illicium verum industry was seriously hindered, and the major economic was immeasurably lost when this insect attack severely occurring. In order to realize the effective management and make use of the pests, and improve value-added of Illicium verum industry, the author preliminary studied on the biological activity of Dilophodes elegans sinica Prout frass and its host plant extracts. The results were summarized as follows:
1. The main volatile matters of Dilophodes elegans sinica Prout frass were detected by means of Solid Phase microextraction and Gas Chromatography-Mass Spectrometry (GC-MS), they were anisole p-propenyl(83.24%),α-farnesene, (2.95%), bicycle[3.1.1]hept-2-ene,2,6-dimethyl-6-(4-methyl-3-pentenyl)(4.20%), undecanoic acid, methyl ester (0.71%), nonadecanoic acid, ethyl ester(1.37%),11-octadecenoic acid methyl ester(2.40%) respectively. The six main volatile matters occupied 94.87% of the total in the peak area.
2. Dilophodes elegans sinica Prout adult showed the strong EAG responses to the volatile matters of Dilophodes elegans sinica Prout frass. Among the five compounds tested, EAG responses of Dilophodes elegans sinica Prout adult to Anisole, p-propenyl were stronger than that of other compounds tested, but the weakest to nonadecanoic acid, ethyl ester, all of the compounds no representation sex difference except anisole, p-propenyl. Dosage response curves of EAG of Dilophodes elegans sinica Prout adult showed that the male and the female adult showed positive correlation EAG response to concentration's varying of undecanoic acid, methyl ester and anisole, p-propenyl and 11-octadecenoic acid, methyl ester, showed "U" model EAG response to the a-farnesene, showed the maximum value of EAG response to nonadecanoic acid, ethyl ester in 0.01 (v/v). The EAG responses of female and male adults to different compositions mixture showed that there were no synergy effects between anisole, p-propenyl and other compositions to female. But the males showed synergy effects between anisole, p-propenyl and a-farnesene, undecanoic acid, methyl ester.
3. The bacteriostasis effect of different extracts of larvae frass to different strains existed some difference. Acetone extracts had a stronger effect to bacterium which including S. aureus and E. coli, and distilled water extracts had a stronger effect to fungus which including P. curinum and R. stolonifer.
4. All of the different extracts from the larvae frass in this paper had scavenging activity to the free radicals including DPPH free radical,-OH free radical,and O2- free radical. Concretely speaking, the extracts from the larvae frass had a weak scavenging activity to DPPH free radical, which was significantly weaker than the extract from Illicium verum leaves under the same concentration. The extracts of chloroform and ethyl acetate had a much stronger scavenging activity to the·OH free radical, while the scavenging activity of extracts of Petroleum ether, butanol and distilled water to·OH free radical was weaker than that of extracts from Illicium verum leaves. The scavenging activity from larvae frass to the O2- free radical was weaker than that of extracts from I. verum leaves under the same conditions.
5. The ethanol, acetone, ethyl acetate and petroleum ether, etc. four extracts all had a strong repellent and fumigate effect, and the repellent and fumigate effect of the acetone extract was strongest while that of petroleum ether extract was weakest among them. In various extracts of acetone extract, the petroleum ether extracts had the best fumigate action, which included 21 kinds of chemical composition of olefine, alcohols, acids, ketones and so on, and the most content were p-anisaldehyde and anethole among all kinds of chemical composition.
6. The ethanol and acetone extracts of I, verum leaves showed a stimulative effect to three protective enzymes (CAI、POD and SOD) of adults of G. ficorum in the early treatment and a inhibition effect in the late treatment. The carboxylesterase activity in adults of G. ficorum significantly increased when the two solvent extracts treated on G. ficorum after two hours. The CAT activity of the treated G. ficorum rapidly decreased down to lower than untreated group when the acetone extract treated after six hours. On the contrary, the CAT activity kept to rise during a long time to maintain a high level when the ethanol extract treated after six hours. The two solvent extracts above showed a stimulative effect to GSTs six hours before treatment and showed a inhibition effect six hours after treatment.
引文
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