101种植物提取物对两种植物病原菌生物活性的研究
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摘要
植物是生物活性化合物的天然宝库,中国具有丰富的植物资源,从植物资源中寻找具有抑菌活性的物质,作为先导化合物开发植物源杀菌剂,是目前植物源农药研究的热点之一。华北地区具有丰富的植物资源,其在医药和临床医学上的应用研究比较广泛,而在农业植物病原菌的防治方面研究较少。本论文选取了华北地区46个科101种植物,以苹果腐烂病菌和棉花立枯病菌为供试菌,以丙酮、乙醇两种溶剂进行提取,对其室内抑菌生物活性进行了初步研究,并对效果较好的桑白皮进行了抑菌活性的初步研究。为新型植物源杀菌剂的研制提供了依据,对防治农作物真菌病害和农业的可持续发展具有重要的意义。研究结果如下:
101种植物丙酮提取物中,苦参、桑白皮、狼毒、藿香、贯众、白芍对苹果腐烂病菌的抑制率达到了80%以上,天明精、土木香、藿香、草石蚕、忽布筋骨草、苦参、白扁豆、虎杖、狼毒、淡花当药、贯众、地血香、蛇莓对棉花立枯病菌的抑制率达到了90%以上。101种植物乙醇提取物中,天明精、苦参、桑白皮对苹果腐烂病菌的抑制率达到了80%以上,土木香、苦参、桑白皮对棉花立枯病菌的抑制率达到了85%以上。
选取桑白皮作为进一步的研究对象,进行抑菌活性成分的追踪。桑白皮乙醇提取物依次用石油醚、乙酸乙酯、正丁醇进行萃取,生物活性测定结果表明,在10mg/ml浓度下,乙酸乙酯萃取物植物病原菌的抑制效果最高,对苹果腐烂病菌的抑制率为96.05%,对棉花立枯病菌的抑制率为97.29%,石油醚相对植物病原菌的抑制效果较好,对苹果腐烂病菌的抑制率为80.05%,对棉花立枯病菌的抑制率为81.12%。
对桑白皮乙酸乙酯萃取物采用柱层析分离进行活性追踪,结果表明,L11、L14、L17、L20对2种植物病原菌有较强的抑制作用,抑制率在70%以上,其它馏分的生物活性相对较弱。在10mg/ml的浓度下,馏分20对2种植物病原菌的抑制活性较高,对苹果腐烂病菌的抑制率达到了85.58%,对棉花立枯病菌的抑制率达到了90.12%,其次为馏分17,对苹果腐烂病菌和棉花立枯病菌的抑制率分别为78.38%和74.07%。采用TLC和柱层析分离对馏分20做进一步的分离并对各馏分进行了生物活性测定,结果表明,L20-2活性最强,对其进行了GC-MS测定,表明馏分中有效成分可能是酚类、酮类和苯甲酸酯类化合物的混合物。
In this dissertation, studies were focused on finding natural bio-active from the abundant resources of plants. It is one of the hot spots to isolate these compounds and to make use of them as new agrochemicals or leading structure now.
There are abundant resources of plants in North China, which can be used in clinical medicine, but rarely used in agriculture as fungicides. 46 families and 101 kinds of plants were selected. The acetone extracts and ethanol extracts were tested for the antifungal activities on Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn. Morus alba L. was primarily studied, which provides the basis for the new style of botanical fungicide. And it is a great significance for the prevention and treatment of fungal diseases of crops and the sustainable development of agriculture. The results are as follows:
Of the 101 kinds of acetone extracts, the inhibition rate of Sophora flavescens Ait. , Morus alba L., Stellera chamaejasme Linn., Agastache rugosa (Fisch. et Mey.) O. Ktze., Blechnum orientale Linn., Paeonia lactiflora Pall reached 80% or more for Valsa mali Miyabe et Yamada. The inhibition rate of Carpesium abrotanoides Linn., Inula helenium L., Agastache rugosa (Fisch. et Mey.) O. Ktze., Stachys sieboldii Miq., Ajuga lupulina Maxim., Sophora flavescens Ait., Semen dolichoris Album, Polygonum cuspidatum Sieb. et Zucc., Stellera chamaejasme Linn., Swertia diluta (Turcz.) Benth.et Hook. F., Blechnum orientale Linn., Kadsura heteroclita (Roxb.) Craib, Duchesnea indica (Andr.) Focke reached 90% or more for Rhizoctonia solani Kuhn. Of the 101 kinds of ethanol extracts, the inhibition rate of Carpesium abrotanoides Linn., Sophora flavescens Ait., Morus alba L. reached 80% or more for Valsa mali Miyabe et Yamada. The inhibition of Inula helenium L., Sophora flavescens Ait.and Morus alba L. reached 85% or more.
Morus alba L. was selected for a further study of antifungal activity. Methanol extracts were continuously extracted by petroleum ether, ethyl acetate and n-butyl alcohol. At the concentration of 10mg/ml, the ethyl acetate extract has the highest activities to Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn., and the growth inhibition ratios were 96.05% and 97.29%, respectively The petroleum ether extract has the good activities to Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn. with the inhibition 80.05% and 81.12%, respectively.
The ethyl acetate extract was isolated by the method of column chromatography. The results showed that L11, L14, L17, L20 have the highest activities to Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn and the inhibition is 70% or more. Other extracts have lower activities. At the concentration of 10 mg/ml, the inhibition of L20 is 85.58% for Valsa mali Miyabe et Yamada and the inhibition of L20 is 90.12%. The inhibition of L17 is 78.38% for Valsa mali Miyabe et Yamada and the inhibition of L17 is 74.07%. By means of TLC and column chromatography, L20 was isolated. The results showed that L20-2 had the highest activities and the active ingredient may be a mixture of phenols, ketones and benzoic acid esters by means of GC-MS.
101种植物丙酮提取物中,苦参、桑白皮、狼毒、藿香、贯众、白芍对苹果腐烂病菌的抑制率达到了80%以上,天明精、土木香、藿香、草石蚕、忽布筋骨草、苦参、白扁豆、虎杖、狼毒、淡花当药、贯众、地血香、蛇莓对棉花立枯病菌的抑制率达到了90%以上。101种植物乙醇提取物中,天明精、苦参、桑白皮对苹果腐烂病菌的抑制率达到了80%以上,土木香、苦参、桑白皮对棉花立枯病菌的抑制率达到了85%以上。
选取桑白皮作为进一步的研究对象,进行抑菌活性成分的追踪。桑白皮乙醇提取物依次用石油醚、乙酸乙酯、正丁醇进行萃取,生物活性测定结果表明,在10mg/ml浓度下,乙酸乙酯萃取物植物病原菌的抑制效果最高,对苹果腐烂病菌的抑制率为96.05%,对棉花立枯病菌的抑制率为97.29%,石油醚相对植物病原菌的抑制效果较好,对苹果腐烂病菌的抑制率为80.05%,对棉花立枯病菌的抑制率为81.12%。
对桑白皮乙酸乙酯萃取物采用柱层析分离进行活性追踪,结果表明,L11、L14、L17、L20对2种植物病原菌有较强的抑制作用,抑制率在70%以上,其它馏分的生物活性相对较弱。在10mg/ml的浓度下,馏分20对2种植物病原菌的抑制活性较高,对苹果腐烂病菌的抑制率达到了85.58%,对棉花立枯病菌的抑制率达到了90.12%,其次为馏分17,对苹果腐烂病菌和棉花立枯病菌的抑制率分别为78.38%和74.07%。采用TLC和柱层析分离对馏分20做进一步的分离并对各馏分进行了生物活性测定,结果表明,L20-2活性最强,对其进行了GC-MS测定,表明馏分中有效成分可能是酚类、酮类和苯甲酸酯类化合物的混合物。
In this dissertation, studies were focused on finding natural bio-active from the abundant resources of plants. It is one of the hot spots to isolate these compounds and to make use of them as new agrochemicals or leading structure now.
There are abundant resources of plants in North China, which can be used in clinical medicine, but rarely used in agriculture as fungicides. 46 families and 101 kinds of plants were selected. The acetone extracts and ethanol extracts were tested for the antifungal activities on Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn. Morus alba L. was primarily studied, which provides the basis for the new style of botanical fungicide. And it is a great significance for the prevention and treatment of fungal diseases of crops and the sustainable development of agriculture. The results are as follows:
Of the 101 kinds of acetone extracts, the inhibition rate of Sophora flavescens Ait. , Morus alba L., Stellera chamaejasme Linn., Agastache rugosa (Fisch. et Mey.) O. Ktze., Blechnum orientale Linn., Paeonia lactiflora Pall reached 80% or more for Valsa mali Miyabe et Yamada. The inhibition rate of Carpesium abrotanoides Linn., Inula helenium L., Agastache rugosa (Fisch. et Mey.) O. Ktze., Stachys sieboldii Miq., Ajuga lupulina Maxim., Sophora flavescens Ait., Semen dolichoris Album, Polygonum cuspidatum Sieb. et Zucc., Stellera chamaejasme Linn., Swertia diluta (Turcz.) Benth.et Hook. F., Blechnum orientale Linn., Kadsura heteroclita (Roxb.) Craib, Duchesnea indica (Andr.) Focke reached 90% or more for Rhizoctonia solani Kuhn. Of the 101 kinds of ethanol extracts, the inhibition rate of Carpesium abrotanoides Linn., Sophora flavescens Ait., Morus alba L. reached 80% or more for Valsa mali Miyabe et Yamada. The inhibition of Inula helenium L., Sophora flavescens Ait.and Morus alba L. reached 85% or more.
Morus alba L. was selected for a further study of antifungal activity. Methanol extracts were continuously extracted by petroleum ether, ethyl acetate and n-butyl alcohol. At the concentration of 10mg/ml, the ethyl acetate extract has the highest activities to Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn., and the growth inhibition ratios were 96.05% and 97.29%, respectively The petroleum ether extract has the good activities to Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn. with the inhibition 80.05% and 81.12%, respectively.
The ethyl acetate extract was isolated by the method of column chromatography. The results showed that L11, L14, L17, L20 have the highest activities to Valsa mali Miyabe et Yamada and Rhizoctonia solani Kuhn and the inhibition is 70% or more. Other extracts have lower activities. At the concentration of 10 mg/ml, the inhibition of L20 is 85.58% for Valsa mali Miyabe et Yamada and the inhibition of L20 is 90.12%. The inhibition of L17 is 78.38% for Valsa mali Miyabe et Yamada and the inhibition of L17 is 74.07%. By means of TLC and column chromatography, L20 was isolated. The results showed that L20-2 had the highest activities and the active ingredient may be a mixture of phenols, ketones and benzoic acid esters by means of GC-MS.
引文
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