Antibacterial and Insecticidal Activities of Ethanol Plant Extracts
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摘要:
目的 研究5种植物乙醇提取物的抑菌杀虫活性，并对杀蝗活性最高的闹羊花提取物进行活性物质的分离，为进一步研究其杀蝗活性物质的结构和其作用机理提供参考。 方法 筛选铁棒锤Aconitum Pendulum、肉豆蔻Myristica fragrans Houtt、（紫丱子）紫铆Butea monosperma（Lan.）Kuntze、翼首草Pterocephalus hookeri和闹羊花Rhododendron mole 5种植物乙醇提取物，分别测定其对植物病原真菌的抑制活性，对蚜虫和东亚飞蝗的毒力；采用连续分级萃取法和柱层析法分离闹羊花的杀蝗活性物质。 结果 肉豆蔻提取物在1 000 mg·L-1质量浓度下对番茄绵腐病菌、苹果轮纹病菌和水稻纹枯病菌的抑制率分别为75.23%、77.78%和88.85%；48 h后闹羊花提取物在2 000 mg·L-1质量浓度下对蚜虫的致死率为55.38%，10 000 mg·L-1下的致死率为100%；闹羊花提取物在10 000 mg·L-1质量浓度下，24 h后对蝗虫的致死率为44.83%，96 h后的致死率为86.96%。从闹羊花提取物中分离得到的活性物质流分Ⅲ-2-2，在1 000 mg·L-1质量浓度下，24 h后对蝗虫的致死率为53.57%，120 h后的致死率为90.48%。 结论 肉豆蔻提取物的抑菌活性最高；闹羊花提取物对蚜虫和蝗虫的毒杀活性最高；闹羊花提取物中杀蝗活性物质集中在石油醚萃取物和二氯甲烷萃取物，从二氯甲烷萃取物中分离得到对蝗虫毒杀活性最高的流分Ⅲ-2-2。 Abstract:Objective Ethanol extracts of 5 species of plants were analyzed for possible antibacterial and/or insecticidal applications in disease control. Method Aconitum Pendulum, nutmeg (Myristica fragrans Houtt), Butea monosperma (Lan.) Kuntze, Pterocephalus hookeri, and Rhododendron molle were selected for the extraction. The inhibitory activities of the extracts against plant pathogenic fungi as well as the virulence to aphids and locusts were determined. Subsequently, the greatest locust-killing capacity demonstrated by the R. molle extract was subjected to a step-wise fractional extraction and column separations to isolate active substances. Result The nutmeg extract applied at 1 000 mg·L-1 level inhibited the growth of Pythium aphanidermatum, Physalospora berengeriana and Rhizoctonia solani at 75.23%, 77.78% and 88.85%, respectively. The R. molle extract showed the lethality on aphids at a rate of 55.38% with the dosage of 2 000 mg·L-1 and 100% with 10 000 mg·L-1in 48h; while that on locusts, 44.83% in 24h and 86.96% in 96h with 10 000 mg·L-1. At 1 000 mg·L-1 dosage, the active fraction, Ⅲ-2-2, isolated from the extract rendered the lethal rates on locusts of 53.57% in 24h and 90.48% in 120h. Conclusion The antibacterial activity of nutmeg extract was the highest among all extracts; the R. molle extract displayed the highest toxicity on aphids and locusts; most of the active substances in the extracts were petroleum ether and dichloromethane soluble; and, Ⅲ-2-2 obtained from the dichloromethane fraction had the highest potency toward locusts. -
表 1 不同植物乙醇提取物的相对抑菌率
Table 1. Antibacterial activity of extracts
(单位/%) 植物乙醇提取物
Ethanol extract小麦赤霉
Gibberella zeae苹果轮纹
Physalospora berengeriana棉花枯萎
Fusarium oxysporum玉米小斑
Helminthosporium maydis番茄绵腐
Pythium aphanidermatum马铃薯晚疫
Phytopthora infestans水稻纹枯
Rhizoctonia solani铁棒锤Aconitum Pendulum 2.59±0.17c 14.59±0.53c 10.02±0.60c 4.72±0.84c 24.77±1.54bc 3.46±0.73c 28.53±1.66bc 肉豆蔻 Myristica fragrans Houtt 52.78±2.03a 77.78±1.81a 65.49±2.67a 48.76±1.90a 75.23±2.71a 53.05±2.11a 88.85±2.83a 紫丱子 Butea monosperma(Lan.)Kuntze 10.00±1.90bc - 8.9±0.82c 4.72±0.23c 7.85±0.45d 7.08±0.32c 26.00±2.11c 翼首草 Pterocephalus hookeri 14.37±1.66b 34.5±2.47b 23.25±2.26b 15.67±1.03b 32.29±1.84b 19.38±2.23b 38.07±3.04b 闹羊花 Rhododendron molle 5.67±1.49c 15.78±1.56c 7.83±2.38c 20.00±1.60b 21.27±0.75c 12.49±1.88bc 11.78±1.91d 注：①“-”表示负数；②表内数据系3次重复的统计结果，供试药物质量浓度为1000 mg·L-1，同列数据后不同小写字母表示差异显著(P < 0.05)。表 2~7同。
Note:①“-”means negative；② The data was the statistical result of three repetitions, the concentration was 1 000 mg·L-1, different lower-case letters after the same column of data showed significant difference (P < 0.05).The same Table 2-7.
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表 2 乙醇提取物对蚜虫的致死率
Table 2. Lethality of ethanol extracts on aphids
(单位/%) 植物乙醇提取物
Ethanol extract质量浓度Concentration 2000 mg·L-1 5000 mg·L-1 10000 mg·L-1 铁棒锤 Aconitum Pendulum 29.74±3.54d 39.71±4.47cd 67.79±7.45c 肉豆蔻 Myristica fragrans Houtt 6.64±2.38e 42.79±6.81c 80.15±5.65b 紫丱子 Butea monosperma(Lan.)Kuntze 39.81±4.71c 38.46±4.37cd 39.14±5.54d 翼首草 Pterocephalus hookeri 10.33±3.23e 31.19±5.38d 46.67±5.02d 闹羊花 Rhododendron molle 55.38±7.26b 78.25±6.83b 100a 阳性对照Positive control 100a 100a 100a 注：阳性对照为质量浓度100 mg·L-1的氯虫菊酯。
Note: The positive control was permethrin with 100 mg·L-1.
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表 3 乙醇提取物对蝗虫的致死率
Table 3. 3 Lethality of ethanol extracts on locusts
(单位/%) 植物乙醇提取物
Ethanol extract校正死亡率Corrected mortality 24 h 48 h 72 h 96 h 铁棒锤 Aconitum Pendulum 24.14±2.53b 48.15±2.78b 60±3.54c 65.22±2.60c 肉豆蔻 Myristica fragrans Houtt 10.34±2.14c 29.63±2.43c 44±1.71d 47.83±3.14d 紫丱子 Butea monosperma(Lan.)Kuntze 17.24±1.37bc 33.33±2.32c 36±2.61d 39.13±2.38d 翼首草 Pterocephalus hookeri 13.79±1.64c 25.92±1.37c 40±2.87d 43.48±1.54d 闹羊花 Rhododendron molle 44.83±2.72a 74.07±3.49a 84±2.11b 86.96±2.66b 阳性对照Positive control 44.83±2.13a 81.48±3.02a 100a 100a 注：供试药物质量浓度为10 000 mg·L-1，阳性对照为40 mg·L-1的氟虫腈。
Note: The concentration was 10 000 mg·L-1, the positive control was fipronil with 40 mg·L-1.
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表 4 4种萃取物对蝗虫的致死率
Table 4. Lethality of extracts on locusts
(单位/%) 供试药物
Test drug校正死亡率Corrected mortality 24 h 48 h 72 h 96 h 石油醚萃取物Petroleum etherfraction 40.00±2.54c 55.17±3.32b 67.86±2.83b 70.37±2.71b 二氯甲烷萃取物Dichloromethanefraction 66.67±3.14a 86.21±2.32a 85.71±2.67a 88.98±2.65a 乙酸乙酯萃取物Ethyl acetate fraction 13.33±2.74d 13.79±2.75c 10.71±1.32d 14.81±2.04c 乙醇萃取物Ethanol fraction 3.33±1.60e 3.33±1.60d 7.14±2.56d 14.81±1.92c 提取物Extract 50.00±1.92b 51.72±2.23b 53.57±1.49c 70.37±3.78b 注：供试药物质量浓度为4 000 mg·L-1。
Note: The concentration was 4 000 mg·L-1.
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表 5 第一次柱层析各流分对蝗虫的致死率
Table 5. Lethality on locusts of substance separated from 1st column chromatography
(单位/%) 供试药物
Test drug校正死亡率Corrected mortality 24 h 48 h 72 h 96 h Ⅰ 3.33±1.03c 10.71±1.78c 17.86±2.32c 23.03±1.90c Ⅱ 13.33±1.73b 17.24±2.23c 23.08±2.14c 29.17±1.45c Ⅲ 40.00±2.11a 65.52±3.56a 88.46±2.81a 95.83±2.66a Ⅳ 3.33±0.54c 27.59±2.47b 34.62±1.91b 45.83±3.04b 注：供试药物质量浓度为2 000 mg·L-1。表 6同。
Note: The concentration was 2 000 mg·L-1.The same as Table 6.
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表 6 第二次柱层析各流分对蝗虫的致死活性
Table 6. Lethality on locusts of substance separated from 2nd column chromatography
(单位/%) 供试药物
Test drug校正死亡率Corrected mortality 24 h 48 h 72 h 96 h Ⅲ-1 0c 13.64±2.53b 23.08±2.65b 29.17±2.75c Ⅲ-2 76.67±2.53a 96.55±2.88a 100.00a 100.00a Ⅲ-3 9.33±1.26b 10.34±2.49b 30.77±2.32b 54.17±2.83b
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表 7 第三次柱层析杀蝗活性验证结果
Table 7. Lethality on locusts of substance separated from 3rd column chromatography
(单位/%) 供试药物
Test drug校正死亡率Corrected mortality 24 h 48 h 72 h 96 h 120 h Ⅲ-2-1 3.57±1.71b 33.33±2.38b 32±2.26b 30.43±2.84b 33.33±1.83b Ⅲ-2-2 53.57±2.71a 70.37±2.53a 72±2.81a 82.61±3.65a 90.48±2.49a 注：供试药物质量浓度为1 000 mg·L-1。
Note: The concentration was 1 000 mg·L-1.
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[1] 侯太平.植物农药研究进展[J].中国农业科技导报, 2006, 8(6):12-16. doi: 10.3969/j.issn.1008-0864.2006.06.003HOU T P. Research Progresses on Botanical Pesticides[J]. Journal of Agricultural Science and Technology, 2006, 8(6):12-16.(in Chinese) doi: 10.3969/j.issn.1008-0864.2006.06.003 [2] 何军, 马志卿, 张兴.植物源农药概述[J].西北农林科技大学学报(自然科学版), 2006, 34(9):79-85. doi: 10.3321/j.issn:1671-9387.2006.09.017HE J, MA Z Q, ZHANG X. Review of botanical pesticide[J]. Journal of Northwest A & F University(Natural Science Edition), 2006, 34(9):79-85.(in Chinese) doi: 10.3321/j.issn:1671-9387.2006.09.017 [3] 程东美, 胡美英.黄杜鹃的研究进展[J].天然产物研究与开发, 1999(5):109-113. http://d.old.wanfangdata.com.cn/Periodical/yjyzx201205001CHENG D M, HU M Y. Research advances in the reserch on Rhododendron molle G. Don[J]. Natural Product Research and Development, 1999(5):109-113.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/yjyzx201205001 [4] SHIN-FOON C, QIU Y T. Experiments on the application of botanical insecticides for the control of diamondback moth in South China[J]. Journal of Applied Entomology, 2010, 116(1-5):479-486. doi: 10.1111-j.1439-0418.1993.tb01224.x/ [5] 冯夏, 赵善欢.黄杜鹃提取物对几种害虫的生物活性及其作用机理的初步研究[J].华南农业大学学报, 1990, 11(2):68-76. http://www.cnki.com.cn/Article/CJFD1990-HNNB199002009.htmFENG X, ZHAO S H. Preliminary studies on the biological activity of extracts from Rhododendron molle against insect pests and their mode of action[J].Journal of South China Agricultural University, 1990, 11(2):68-76.(in Chinese) http://www.cnki.com.cn/Article/CJFD1990-HNNB199002009.htm [6] SHIN-FOON C, QIU Y T. Experiments on the application of botanical insecticides for the control of diamondback mothin South China[J].Journal of Applied Entomology, 1993, 116(1-5):479-486. doi: 10.1111/jen.1993.116.issue-1-5 [7] 肖波, 周俗, 张可君, 等.瑞香狼毒中灭菜青虫活性成分的提取与分离[J].四川大学学报(自然科学版), 2005, 42(3):605. doi: 10.3969/j.issn.0490-6756.2005.03.035XIAO B, ZHOU S, ZHANG K J, et al. The Separation and Isolation of the Extract from the Root of Stellera chamaejasme (L) Against Pieris rapae (L)[J]. Journal of Sichuan University(Natural Science Edition), 2005, 42(3):605.(in Chinese) doi: 10.3969/j.issn.0490-6756.2005.03.035 [8] 周丽, 高飞, 滕云, 等. 17种高原草地有毒植物对植物病原真菌生物活性的初步研究[J].四川大学学报(自然科学版), 2008, 45(1):214-218. doi: 10.3969/j.issn.0490-6756.2008.01.042ZHOU L, GAO F, TENG Y, et al. The bioactivity study of seventeen kinds of poisonous plants from grassland against four pathogenic fungi[J]. Journal of Sichuan University(Natural Science Edition), 2008, 45(1):214-218.(in Chinese) doi: 10.3969/j.issn.0490-6756.2008.01.042 [9] 高平, 刘世贵, 侯太平, 等.瑞香素对蚜虫生物活性的研究[J].植物保护学报, 2001, 28(3):265-268. doi: 10.3321/j.issn:0577-7518.2001.03.015GAO P, LIU S G, HOU T P, et al.BIOACTIVITY OF RUIXIANGSU TO APHIDS(APHIS GOSSYPⅡ GLOVER, MYZVS NICOTCANAE BLACKMAN, TOXOPTERA CITRICWA(KIRKALDY))[J]. Acta Phytophylacica Sinica, 2001, 28(3):265-268.(in Chinese) doi: 10.3321/j.issn:0577-7518.2001.03.015 [10] 曹威, 张可君, 龚晓霞, 等.瑞香狼毒素对蚜虫防治的初步试验[J].四川动物(自然科学版), 2005, 24(4):548. http://d.old.wanfangdata.com.cn/Periodical/scdw200504028CAO W, ZHANG K J, GONG X X, et al. Study on Control Effect of Daphneantoxin Against Aphis gossypii[J]. Sichuan Journal of Zoology(Natural Science Edition), 2005, 24(4):548.(in Chinese) http://d.old.wanfangdata.com.cn/Periodical/scdw200504028 [11] 赵成爱, 余梅燕, 吴现芳, 等.八宝景天乙醇提取物对菜青虫的生物活性[J].农药, 2012, 51(1):62-63, 70. doi: 10.3969/j.issn.1006-0413.2012.01.024ZHAO C A, YU M Y, WU X F, et al. Bioactivity of Sedum spectabile Boreau Extracts against Pieris rapae[J]. Agrochemicals, 2012, 51(1):62-63, 70.(in Chinese) doi: 10.3969/j.issn.1006-0413.2012.01.024 [12] 高嫚潞, 吴朗, 陈龙, 等.瑞香狼毒活性成分对东亚飞蝗的生物活性和病理学研究[J].四川大学学报(自然科学版), 2013(4):875-880. doi: 10.3969/j.issn.0490-6756.2013.04.038GAO M L, WU L, CHEN L, et al. Bioactivity and histopathology study of the active ingredients from Stellera chamaejasme L.against Locusta migratoria manilensis(Meyen)[J]. Journal of Sichuan University(Natural Science Edition), 2013(4):875-880.(in Chinese) doi: 10.3969/j.issn.0490-6756.2013.04.038 [13] 何秀玲.抗多种植物病原真菌的肉豆蔻木酚素的分离与抗菌活性研究[J].世界农药, 2008, 30(2):34-38. doi: 10.3969/j.issn.1009-6485.2008.02.007HE X L. Isolation and antibacterial activity of myristyl lignan against various phytopathogenic fungi[J]. World Pesticides, 2008, 30(2):34-38.(in Chinese) doi: 10.3969/j.issn.1009-6485.2008.02.007 [14] 程东美, 张志祥, 胡美英.黄杜鹃花粗提物及闹羊花素-Ⅲ对斜纹夜蛾幼虫杀虫活性的研究[J].长江蔬菜, 2007(5):47-49. doi: 10.3865/j.issn.1001-3547.2007.05.030CHENG D M, ZHANG Z X, HU M Y. Study on the Control Effect of the Crude Extract of Rhododendron molle and Rhodojaponin-Ⅲ to the Larvae of Spodoptera litura larvae[J]. Journal of Changjiang Vegetables, 2007(5):47-49.(in Chinese) doi: 10.3865/j.issn.1001-3547.2007.05.030 [15] 胡美英, 赵善欢.黄杜鹃花杀虫活性成分及其对害虫毒杀作用的研究[J].华南农业大学学报, 1992, 13(3):9-15. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=QK000001443042HU M Y, ZHAO S H. Studies on the insecticidal principles and biological activities of yellow azalea, Rhododendron molle G. don[J]. Journal of South China Agricultural University, 1992, 13(3):9-15.(in Chinese) http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=QK000001443042 [16] HU M Y, KOLCKE J A, CHIU S F. Response of five insects to botanical insecticides Rhodojaponin-Ⅲ[J]. J Econ Entomol, 1993, 86(3):706-711. doi: 10.1093/jee/86.3.706 -
