中药抑制番茄晚疫病原菌的配伍筛选及抑菌效果初步研究
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摘要
为得到对番茄晚疫病原菌有更好抑制作用的生物药剂,对15种中药进行筛选,并进行配伍抑菌研究,得出最佳三配伍抑菌组合为青蒿∶五味子∶乌梅=1∶4∶1,抑菌圈直径达到了56 mm,最佳配伍液的最小抑菌浓度(MIC)为7.82 mg/mL,最小杀菌浓度(MBC)为31.25 mg/mL,对生长曲线的抑制作用明显。将该中药配伍液稀释500～1 000倍时,其抑菌率明显高于典型化学抑菌剂。
In order to obtain a better biological agent for the suppression of late blight of tomato, the screening,compatibility and antibacterial experiments of 15 kinds of Chinese medicine herbs extracts were carried out. The results showed that the best combination of three Chinese herbal medicine was Artemisia annua ∶Fructus Schisandra chinensis∶dark plum fruit =1∶4∶1, and the diameter of bacteriostatic circle reached 56 mm. The minimum inhibitory concentration(MIC) of the best combined liquid was 7.82 mg/mL, the minimum bactericidal concentration(MBC) was31.25 mg/mL, the inhibitory effect on the growth curve was obvious. The bacteriostatic rate of the mixture was significantly higher than that of the typical chemical bacteriostatic agent when the combined Chinese herbs liquid was diluted to 500～1 000 times.
In order to obtain a better biological agent for the suppression of late blight of tomato, the screening,compatibility and antibacterial experiments of 15 kinds of Chinese medicine herbs extracts were carried out. The results showed that the best combination of three Chinese herbal medicine was Artemisia annua ∶Fructus Schisandra chinensis∶dark plum fruit =1∶4∶1, and the diameter of bacteriostatic circle reached 56 mm. The minimum inhibitory concentration(MIC) of the best combined liquid was 7.82 mg/mL, the minimum bactericidal concentration(MBC) was31.25 mg/mL, the inhibitory effect on the growth curve was obvious. The bacteriostatic rate of the mixture was significantly higher than that of the typical chemical bacteriostatic agent when the combined Chinese herbs liquid was diluted to 500～1 000 times.
引文
[1]袁高庆,黎起秦,王静,等.植物源杀菌剂研究进展Ⅰ:抑菌植物资源[J].广西农业科学,2010, 41(1):30-34.DOI:10.3969/j.issn.2095-1191.2010.01.010.
[2]袁高庆,黎起秦,王静,等.植物源杀菌剂研究进展Ⅱ:活性测定、活性物质及其应用[J].广西农业科学,2010,41(2):136-140.DOI:10.3969/j.issn.2095-1191.2010.02.013.
[3] SWAIN T. Secondary compounds as protective agents[J].Annual Review of Plant Physiology, 1997, 28:479-501. DOI:10.1146/annurev.pp.28.060177.002403.
[4]窦瑞木.植物源杀菌剂研究进展及存在的问题[J].河南农业,2009(5):49-51.DOI:10.3969/j.issn.1006-950X.2009.10.036.
[5]陈宇飞,韩桂珍,龚束芳.大棚番茄晚疫病药剂防治[J].北方园艺,2000(5):40-41.DOI:10.3969/j.issn.1001-0009.2000.05.038.
[6] STROUD J A, SHAW D S, HALE M D, et al. SSR assessment of Phytophthora infestans, populations on tomato and potato in British gardens demonstrates high diversity but no evidence for host specialization[J]. Plant Pathology, 2016, 65(2):334-341.DOI:10.1111/ppa.12407.
[7] DROBY S, CHALUTZ E, WILSON C L. Antagomsms as biological control agents of postharvest diseases of fruits and vegetables[J]. Postharvest News and Information, 1991, 2:169-173.
[8]王胤,王晓青,胡彬,等. 8种杀菌剂对番茄晚疫病菌的室内抑菌试验[J].农学学报,2016,6(2):48-51.DOI:10.3969/j.issn.1007-7774.2016.02.008.
[9]朱桂宁,黄福新,冯兰香,等.番茄晚疫病菌对甲霜灵霜脲氰和烯酰吗啉的敏感性检测[J].中国农业科学,2008,41(5):1355-1365. DOI:10.3864/j.issn.0578-1752.2008.05.013.
[10] BASHAN B, KADISH D, LEVY Y, et al. Infectivity to Potato,sporangial germination, and respiration of isolates of Phytophthora infestans from metalaxyl-sensitive and metalaxyl-resistant populations[J]. Phytopathology, 1989, 79(8):832-836. DOI:10.1094/Phyto-79-832.
[11] SUJKOWSKI L S, FRY B A, POWER R J, et al.Sensitivities of Mexican isolates of Phytophthora infestans to chlorothalonil, cymoxanil, and metalaxyl[J]. Plant Disease,1995, 79(11):1117-1120. DOI:10.1094/PD-79-1117.
[12] KATO M, MIZUBUTI E S, GOODWIN S B, et al. Sensitivity to protectant fungicides and pathogenic fitness of clonal lineages of Phytophthora infestans in the United States[J].Phytopathology, 1997, 87(9):973-978. DOI:10.1094/PHYTO.1997.87.9.973.
[13] SHATTOCK R C. Studies on the inheritance of resistance to metalaxyl in Phytophthora infestans[J]. Plant Pathology, 1988,37(1):4-11. DOI:10.1111/j.1365-3059.1988.tb02188.x.
[14] DEAHL K L, INGLIS D A, DEMUTH S P. Testing for resistance to metalaxyl in Phytophthora infestans, isolates from northwestern Washington[J]. American Potato Journal,1993, 70(11):779-795. DOI:10.1007/BF02849105.
[15] HERMANSEN A, HANNUKKALA A, NAERSTAD R H, et al. Variation in populations of Phytophthora infestans, in Finland and Norway:mating type, metalaxyl resistance and virulence phenotype[J]. Plant Pathology, 2000, 49(1):11-22.DOI:10. 1046/j.1365-3059.2000.00426.x.
[16] GOODWIN S B, SUJKOWSKI L S, FRY W E. Widespread distribution and probable origin of resistance to metalaxyl in clonal genotypes of Phytophthora infestans in the United States and Western Canada[J]. Phytopathology, 1996, 86(7):793-800. DOI:10.1094/Phyto-86-793.
[17] HANSON P, LU S F, WANG J F, et al. Conventional and molecular marker-assisted selection and pyramiding of genes for multiple disease resistance in tomato[J]. Scientia Horticulturae, 2016, 201:346-354.DOI:10.1016/j.scienta.2016.02.020.
[18]司红彬,梁松林,许桂芹,等. 4味中药及其与抗菌药的复方制剂的MIC测定[J].中国兽药杂志, 2006, 40(2):31-34.DOI:10.3969/j.issn.1002-1280.2006.02.010.
[19]李福娟,石学魁,张晓莉,等.苦参、黄连及其配伍的体外抑菌研究[J].牡丹江医学院学报, 2010,31(3):17-19.DOI:10.3969/j.issn.1001-7550.2010.03.008.
[20]郑海武,李正英.植物源抗番茄晚疫病菌的研究进展[J].食品工业科技,2016,37(23):387-390,395.DOI:10.13386/j.issn1002-0306.2016.23.064.
[21]韩雯雯,李正英,陈忠军.五味子和番泻叶中提取物的提取条件优化[J].食品工业科技, 2012, 33(14):295-298.
[22]宋琳琳,李正英,陈萍.五倍子抗蕃茄晚疫病原菌有效成分提取工艺研究[J].食品工业科技,2013,34(10):269-271.
[23]苏雅拉,李正英.青蒿中抗蕃茄晚疫病原菌的有效成分提取工艺的研究[J].内蒙古农业大学学报(自然科学版),2014(2):92-96. DOI:10.16853/j.cnki.1009-3575.2014.02.019.
[24]康帅,殷中琼,贾仁勇,等.乌梅等三味具有抑菌活性中药的最佳提取工艺研究[J].天然产物研究与开发,2014(26):2014-2021.DOI:10.16333/j.1001-6880.2014.12.023.
[25]李晓娟,李正英.黄芩抗蕃茄晚疫病菌抑菌成分的提取条件优选[J].中国酿造,2015,34(3):107-110. DOI:10.11882/j.issn.0254-5071.2015.03.025.
[26]孟赛楠.芦荟中抗蕃茄晚疫病原菌有效成分的提取分离及影响抗菌效果因素的研究[D].呼和浩特:内蒙古农业大学,2014.
[27]王孟,李昂,赵震宇,等.裸花紫珠叶抑菌性物质的ASE联合牛津杯和微孔刃天青法筛选[J].时珍国医国药,2017,28(1):43-45.
[28]祝若夫,胡宝龙,周德庆.微生物学实验教程[M].上海:复旦大学出版社,1993:192-196.
[29]张爱玉,李俊,黄秉智,等.韭菜汁对香蕉炭疽菌的抑制作用研究[J].保鲜与加工,2011,11(5):17-20. DOI:10.3969/j.issn.1009-6221.2011.05.006.
[2]袁高庆,黎起秦,王静,等.植物源杀菌剂研究进展Ⅱ:活性测定、活性物质及其应用[J].广西农业科学,2010,41(2):136-140.DOI:10.3969/j.issn.2095-1191.2010.02.013.
[3] SWAIN T. Secondary compounds as protective agents[J].Annual Review of Plant Physiology, 1997, 28:479-501. DOI:10.1146/annurev.pp.28.060177.002403.
[4]窦瑞木.植物源杀菌剂研究进展及存在的问题[J].河南农业,2009(5):49-51.DOI:10.3969/j.issn.1006-950X.2009.10.036.
[5]陈宇飞,韩桂珍,龚束芳.大棚番茄晚疫病药剂防治[J].北方园艺,2000(5):40-41.DOI:10.3969/j.issn.1001-0009.2000.05.038.
[6] STROUD J A, SHAW D S, HALE M D, et al. SSR assessment of Phytophthora infestans, populations on tomato and potato in British gardens demonstrates high diversity but no evidence for host specialization[J]. Plant Pathology, 2016, 65(2):334-341.DOI:10.1111/ppa.12407.
[7] DROBY S, CHALUTZ E, WILSON C L. Antagomsms as biological control agents of postharvest diseases of fruits and vegetables[J]. Postharvest News and Information, 1991, 2:169-173.
[8]王胤,王晓青,胡彬,等. 8种杀菌剂对番茄晚疫病菌的室内抑菌试验[J].农学学报,2016,6(2):48-51.DOI:10.3969/j.issn.1007-7774.2016.02.008.
[9]朱桂宁,黄福新,冯兰香,等.番茄晚疫病菌对甲霜灵霜脲氰和烯酰吗啉的敏感性检测[J].中国农业科学,2008,41(5):1355-1365. DOI:10.3864/j.issn.0578-1752.2008.05.013.
[10] BASHAN B, KADISH D, LEVY Y, et al. Infectivity to Potato,sporangial germination, and respiration of isolates of Phytophthora infestans from metalaxyl-sensitive and metalaxyl-resistant populations[J]. Phytopathology, 1989, 79(8):832-836. DOI:10.1094/Phyto-79-832.
[11] SUJKOWSKI L S, FRY B A, POWER R J, et al.Sensitivities of Mexican isolates of Phytophthora infestans to chlorothalonil, cymoxanil, and metalaxyl[J]. Plant Disease,1995, 79(11):1117-1120. DOI:10.1094/PD-79-1117.
[12] KATO M, MIZUBUTI E S, GOODWIN S B, et al. Sensitivity to protectant fungicides and pathogenic fitness of clonal lineages of Phytophthora infestans in the United States[J].Phytopathology, 1997, 87(9):973-978. DOI:10.1094/PHYTO.1997.87.9.973.
[13] SHATTOCK R C. Studies on the inheritance of resistance to metalaxyl in Phytophthora infestans[J]. Plant Pathology, 1988,37(1):4-11. DOI:10.1111/j.1365-3059.1988.tb02188.x.
[14] DEAHL K L, INGLIS D A, DEMUTH S P. Testing for resistance to metalaxyl in Phytophthora infestans, isolates from northwestern Washington[J]. American Potato Journal,1993, 70(11):779-795. DOI:10.1007/BF02849105.
[15] HERMANSEN A, HANNUKKALA A, NAERSTAD R H, et al. Variation in populations of Phytophthora infestans, in Finland and Norway:mating type, metalaxyl resistance and virulence phenotype[J]. Plant Pathology, 2000, 49(1):11-22.DOI:10. 1046/j.1365-3059.2000.00426.x.
[16] GOODWIN S B, SUJKOWSKI L S, FRY W E. Widespread distribution and probable origin of resistance to metalaxyl in clonal genotypes of Phytophthora infestans in the United States and Western Canada[J]. Phytopathology, 1996, 86(7):793-800. DOI:10.1094/Phyto-86-793.
[17] HANSON P, LU S F, WANG J F, et al. Conventional and molecular marker-assisted selection and pyramiding of genes for multiple disease resistance in tomato[J]. Scientia Horticulturae, 2016, 201:346-354.DOI:10.1016/j.scienta.2016.02.020.
[18]司红彬,梁松林,许桂芹,等. 4味中药及其与抗菌药的复方制剂的MIC测定[J].中国兽药杂志, 2006, 40(2):31-34.DOI:10.3969/j.issn.1002-1280.2006.02.010.
[19]李福娟,石学魁,张晓莉,等.苦参、黄连及其配伍的体外抑菌研究[J].牡丹江医学院学报, 2010,31(3):17-19.DOI:10.3969/j.issn.1001-7550.2010.03.008.
[20]郑海武,李正英.植物源抗番茄晚疫病菌的研究进展[J].食品工业科技,2016,37(23):387-390,395.DOI:10.13386/j.issn1002-0306.2016.23.064.
[21]韩雯雯,李正英,陈忠军.五味子和番泻叶中提取物的提取条件优化[J].食品工业科技, 2012, 33(14):295-298.
[22]宋琳琳,李正英,陈萍.五倍子抗蕃茄晚疫病原菌有效成分提取工艺研究[J].食品工业科技,2013,34(10):269-271.
[23]苏雅拉,李正英.青蒿中抗蕃茄晚疫病原菌的有效成分提取工艺的研究[J].内蒙古农业大学学报(自然科学版),2014(2):92-96. DOI:10.16853/j.cnki.1009-3575.2014.02.019.
[24]康帅,殷中琼,贾仁勇,等.乌梅等三味具有抑菌活性中药的最佳提取工艺研究[J].天然产物研究与开发,2014(26):2014-2021.DOI:10.16333/j.1001-6880.2014.12.023.
[25]李晓娟,李正英.黄芩抗蕃茄晚疫病菌抑菌成分的提取条件优选[J].中国酿造,2015,34(3):107-110. DOI:10.11882/j.issn.0254-5071.2015.03.025.
[26]孟赛楠.芦荟中抗蕃茄晚疫病原菌有效成分的提取分离及影响抗菌效果因素的研究[D].呼和浩特:内蒙古农业大学,2014.
[27]王孟,李昂,赵震宇,等.裸花紫珠叶抑菌性物质的ASE联合牛津杯和微孔刃天青法筛选[J].时珍国医国药,2017,28(1):43-45.
[28]祝若夫,胡宝龙,周德庆.微生物学实验教程[M].上海:复旦大学出版社,1993:192-196.
[29]张爱玉,李俊,黄秉智,等.韭菜汁对香蕉炭疽菌的抑制作用研究[J].保鲜与加工,2011,11(5):17-20. DOI:10.3969/j.issn.1009-6221.2011.05.006.