小花假泽兰杀菌作用研究
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摘要
天然产物的研究与开发是新农药创制的主要途径之一。小花假泽兰Mikania micrantha H.B.K号称为“植物杀手”,是一种非常危险的外来入侵植物,对其的防治及综合利用已是一件非常紧迫之事。前期研究发现,其提取物对多种植物病原菌均具较强的抑制活性,为了更有效地利用小花假泽兰植物资源,实现因势利导、趋利避害,为开发新型植物源杀菌剂提供理论依据,本论文在前期研究基础上主要从以下5个方面对小花假泽兰提取物的抑菌作用进行了较为系统的研究,主要结果如下：
1.采用活性跟踪法从小花假泽兰中分离得到17种抑菌活性化合物,鉴定出其中15种化合物的化学结构,分别为：薇甘菊内酯、去氧薇甘菊内酯、二氢薇甘菊内酯、桉烷-4(15),7(11)-二烯-12,8β-内酯、桉烷-4(15),7(11),8(9)-三烯-12,8β-内酯、Isabelin、scandenolide、Mikanokryptin、1β,6α-二羟基桉烷-4(15)-烯、Mikanin、泽兰黄酮、木犀草素、β-胡萝卜苷、豆甾醇、β-谷甾醇等化合物。主要为倍半萜内酯类化合物、黄酮类化合物和甾体及其苷类化合物。
2.系统测定了豆甾醇、去氧薇甘菊内酯、二氢薇甘菊内酯、Isabelin等化合物杀菌活性。豆甾醇对供试的12种病原菌菌丝生长均有一定的抑制活性,EC50在365.48-1438.59 mg/L之间,其中对番茄灰霉病菌的毒力最高,EC50为365.48mg/L；豆甾醇对不同病菌孢子萌发的抑制效果存在一定差异,对番茄叶霉病菌和番茄灰霉病菌孢子毒力最高,EC50分别为151.47和174.65mg/L；盆栽试验和组织法测试表明,豆甾醇对番茄灰霉病具有较好的防治效果,在供试浓度为1500mg／L时对番茄灰霉病的治疗效果为68.6%,对小麦白粉病也表现出较高的保护效果,并具一定的治疗作用,在剂量为1500mg/L时,对小麦白粉病的保护效果达到66.8%、治疗效果是43.1%。Isabenlin对番茄灰霉病菌、小麦纹枯病菌和辣椒疫霉病菌菌丝生长的EC50分别为221.25、432.21和714.69mg/L。去氧薇甘菊内酯对供试10种病原菌菌丝生长均具很高的抑制毒力,EC50在5.61～46.48mg/L之间,对小麦纹枯病菌菌丝生长的毒力最高,EC50是5.61mg/L；去氧薇甘菊内酯抑制不同病原菌孢子萌发效果差异明显,对番茄灰霉病菌孢子EC50为10.49 mg／L,抑制毒力最高。二氢薇甘菊内酯对辣椒疫霉病菌的毒EC50为12.84 mg/L,对5种供试病原菌孢子萌发的的EC50介于20-75mg/L之间。结合化合物的提取分离结果,分析认为,倍半萜内酯类化合物和甾醇类化合物是小花假泽兰主要的杀菌活性成分,而其中甾醇类化合物具有一定的直接开发利用前景。
3.系统测试并比较了水蒸汽蒸馏法(HD)、超临界CO2萃取法(SCE)及溶剂萃取法(PEE)等3种方法提取制备的小花假泽兰精油的化学成分。首先,采用正交设计试验提出小花假泽兰精油超临界C02萃取适宜工艺：先静态萃取20 min,然后动态萃取,动态萃取参数温度55℃、压力35 MPa、CO2体积40 mL／g；进而,结合前期研究结果,采用GC/MS法分析比较了HD法、SCE法及PEE法等3种方法制备的小花假泽兰精油的化学成分,并测试了3种提取物对小麦纹枯病菌和小麦全蚀病菌的毒力。结果表明：SCE法精油萃取率显著高于HD法及PEE法；小花假泽兰精油主要有烯烃类、萜类、醇类、脂肪酸、酯类、甾体类等化合物,不同方法提取物化学成分存在明显差异；小花假泽兰精油对不同病菌具明显的选择性抑制作用,对小麦赤霉病菌、小麦纹枯病菌和番茄灰霉病菌表现出很强的抑制作用,对棉花枯萎病菌的抑制作用较弱；3种提取方法所得精油的杀菌活性亦存在显著差异,SCE法精油对小麦赤霉病菌和小麦纹枯病菌的毒力亦显著高于HD法和PEE法提取物。
4.利用电镜技术开展了豆甾醇对小麦活体上小麦白粉病菌的影响初步研究,结果表明,豆甾醇可致小麦白粉病原菌菌丝扭曲、畸形,表面粗糙,生长点塌陷、老化加快；可致小麦白粉病菌分生孢子数量减少,孢子表面出现大面积凹坑；可致菌丝细胞内原生质电子致密度显著增加,各种细胞器解体,菌丝坏死；可导致病原菌菌丝细胞和吸器体液泡化严重,且出现空腔,液泡中有电子致密度高的沉积物；可致菌丝细胞内电子致密物不均匀,囊泡变大而数量减少；使小麦白粉病原菌吸器外间质膜不规则加厚。分析认为豆甾醇可能主要是对病原菌体器官的生物合成过程造成较大影响,致使其侵染能力下降,同时对病菌体内生物氧化过程亦具一定影响。对此,还有待于进一步深入探讨。
5.以小花假泽兰全株粗粉为原料,通过对表面活性剂、载体等助剂的筛选,研制出了70%小花假泽兰可湿性粉剂配方。大田药效试验结果表明,70%小花假泽兰可湿性粉剂在7500 mg/L剂量下,对番茄灰霉病的防治效果达70%以上。
本论文初步探明了小花假泽兰杀菌活性化学成分,发现了几个具有植物病原菌杀菌活性的化合物；探讨了豆甾醇、去氧薇甘菊内酯等化合物杀菌活性强度和杀菌谱,初步评价了其应用前景；初步研究了豆甾醇对小麦白粉菌形态和超微结构的影响；明确了小花假泽兰精油超临界C02萃取适宜工艺、不同提取方法效果差异及其主要化学成分；并对假泽兰可湿性粉剂制剂配方进行了初步探讨。研究结果为充分利用小花假泽兰这一植物资源,开发新型植物源农药提供了理论依据。
Research and development of natural products is one of primary and effective ways to develop novel pesticides. As dangerous invasive species from abroad, control and utilization of plants killer, Mikania micrantha H.B.K is urgently needed. Previous study has shown that the extract of Mikania micrantha H.B.K have a strong inhibition activity against many plant pathogens. In order to utilize Mikania micrantha H.B.K resource efficiently, make the best use of the situation while avoiding disadvantages, and provide theoretical basis for novel plant-derived fungicide developing, the chemical constituents and fungicidal activity of Mikania micrantha H.B.K were investigated on the basis of some previous study. The main result obtained from this study as follows:
1.17 compounds were isolated from Mikania micrantha H.B.K based on bioactivity-guided isolation, and the chemical structures of 15 compounds were identified. Those chemicals include Deoxymikanolide, Dihydromikanolide, Mikanolide, Isabelin, Scandenolide, Mikanokryptin,1β,6α-dihydroxy eudesmane-4(15)-alkene, eudesmane-4 (15), 7 (11)-12,8β-lactone, Mikanin, Stigmasterol,β-Sitosterol, eudesmane-4(15),7(11), 7(11)-triene-12,8β-lactone, luteolin,β-Daucosterol, Eupatolitin. The result shows that 15 identified compounds can be divided into 3 categories:sesquiterpene lactones; flavonoid compounds and steroidal saponins.
2. The fungicidal activities of Stigmasterol, Deoxymikanolide, Dihydromikanolide, and Isabelin were evaluated against plant pathothic fungi. The results showed that Stigmasterol can inhibit the mycelium growth of 12 pathogens, which EC50 value is within 365.48-1438.59 mg/L. Stigmasterol showed highest antifungal activities against Botrytis cinerea Pers with an EC50 value of 365.48 mg/L. The results also revealed that there are significant differences between the inhibition effects of Stigmasterol on different pathogens spore germination, Stigmasterol showed highest antifungal activities against Fulvia fulva and Botrytis cirerea Pers. et Tris with an EC50 value of 151.47 and 174.65mg/L, respectively. Furthermore, Stigmasterol displayed certain control effect (68.60% at 1500 mg/L) against B. cirerea in vivo. And Stigmasterol also displayed prominent systemic protective effect (66.80% at 1500 mg/L and 43.10% at 1000 mg/L) and control effect (50.60% at 1500 mg/L and 25.70% at 1000 mg/L) against wheat powdery mildew Blumeria graminis in vivo. Deoxymikanolide displayed a higher inhibition on mycelium growth of 10 pathogens, witch EC50 value was within 5.61-46.48 mg/L, Deoxymikanolide showed highest antifungal activities against Rhizoctonia cerealis Vander Hoeven with an EC50 value of 5.61 mg/L. The bioassay showed that there is a significant difference between the inhibition effect of Deoxymikanolide on different pathogens spore germination, Deoxymikanolide showed highest antifungal activities against Botrytis cirerea Pers. et Tris with an EC50 value of 10.49mg/L. The fungicidal activity of Dihydromikanolide was lower than Deoxymikanolide, Dihydromikanolide only showed a similar antifungal activities with Deoxymikanolide against Phytophthora capsici with an EC50 value of 12.84mg/L. There is a little difference between the inhibition effect of Dihydromikanolide on different pathogens spore germination, the EC50 value of Dihydromikanolide effects on spore germination of 5 pathogens was 20～75 mg/L. The EC50 value of Isabenlin against B. cirerea, R. cerealis and Phytophthora capsici was 221.25,432.21 and 714.69 mg/L, respectively. Based on the bioassay results combined with isolation and identification results of compounds, sesquiterpene lactones and steroidal saponins were considered to be primary fungicidical components in M. micrantha, and steroidal saponins has a certain prospect of developing and utilization.
3. In the basis of previous extraction technology of essential oils from M. micrantha with supercritical CO2, the chemical constituents of M. micrantha extracts acquired with 3 methods: hydrostillation(HD), SCE, and extracted with petrol ether(PEE) were analyzed by GC/MS, and the toxicity of those 3 extracts against Fusarium graminearum and Rhizoctonia cerealis was tested. The result showed that the optimized extraction condition was 35 MPa and 55℃with 40 mL/g supercritical carbon dioxide after 20-mintue static extraction, and the extraction rate of essential oil by SCE was higher than other 2 methods. The main components of essential oil from M. micrantha are polyolefins, terpenoids, alcohols, fatty acids, esters and steroidal compounds, and there are significant differences of chemical constituents between essential oil extracted with different methods. The essential oil of M. micrantha showed selective antifungal activities against different plant pathogens, the bioassay showed that essential oil has the highest antifungal activities against Fusarium graminearum and Rhizoctonia cerealis and Botrytis cinerea Pers and weaker inhibitory effect against Fusarium oxysporum f.vasinfectum (Atk.) Snyder et Hansen. There are significant differences of antifungal activities between essential oil acquired by different methods. The toxicity of M. micrantha essential oil extracted by SCE was higher than the essential oil extracted by HD and PEE against Fusarium graminearum and Rhizoctonia cerealis.
4. Effect of Stigmasterol against Blumeria graminis were observed by electron microscopy technology. The results showed that stigmasterol can lead to abnormal hyphal growth of B. graminis such as twisted and malformed mycelia, surface roughening, growing point collapse and faster aging. Stigmasterol can also reduce conidia quantity and lead to large area of surface pit in spore. Furthermore, stigmasterol treated fungi cause intracellular protoplasm electronic density to increase, organelles degenerated, mycelia putrescence, and cause mycelia and haustorial body heavy vacuolization and higher density substances sedimentary in mycelia cell. It could also lead to electronic density substances uneven in mycelia cell, enlarge vesicle and reduce quantity of vesicle. The outer plasma membrane haustorium of B. graminiwere was found to be irregulary thicken by stigmasterol. Those results showed that stigmasterol mainly effect biosynthetic process of plant pathogen, meanwhile, it can also effect biological oxidation of plant pathogen to a certain extent. Further research was needed to confirm those hypotheses.
5. The powder of M. micrantha has been formulated to 70% wettable powder through the surface active agent and carrier screening. The field trial showed that 70% M. micrantha WP can provide 70% control effect against B. cirerea at 7500 mg/L.
The dissertation preliminarily definited the chemical character of antifungal compounds from M.micrantha, and several novel compounds with fungicidal activity were found. The chemical constituent's character and antifungal activity of essential oil from M. micrantha have also been studied. The fungicidal spectrum and activities of Stigmasterol and Deoxymikanolide were investigated, and the application prospect as a novel control agent was evaluated. The formulation of wettable powder of M. micrantha was formulated and developed successfully. Those research results provide theoretical basis for utilization Mikania micrantha H.B.K resource efficiently and novel plant-derived fungicide developing.
1.采用活性跟踪法从小花假泽兰中分离得到17种抑菌活性化合物,鉴定出其中15种化合物的化学结构,分别为：薇甘菊内酯、去氧薇甘菊内酯、二氢薇甘菊内酯、桉烷-4(15),7(11)-二烯-12,8β-内酯、桉烷-4(15),7(11),8(9)-三烯-12,8β-内酯、Isabelin、scandenolide、Mikanokryptin、1β,6α-二羟基桉烷-4(15)-烯、Mikanin、泽兰黄酮、木犀草素、β-胡萝卜苷、豆甾醇、β-谷甾醇等化合物。主要为倍半萜内酯类化合物、黄酮类化合物和甾体及其苷类化合物。
2.系统测定了豆甾醇、去氧薇甘菊内酯、二氢薇甘菊内酯、Isabelin等化合物杀菌活性。豆甾醇对供试的12种病原菌菌丝生长均有一定的抑制活性,EC50在365.48-1438.59 mg/L之间,其中对番茄灰霉病菌的毒力最高,EC50为365.48mg/L；豆甾醇对不同病菌孢子萌发的抑制效果存在一定差异,对番茄叶霉病菌和番茄灰霉病菌孢子毒力最高,EC50分别为151.47和174.65mg/L；盆栽试验和组织法测试表明,豆甾醇对番茄灰霉病具有较好的防治效果,在供试浓度为1500mg／L时对番茄灰霉病的治疗效果为68.6%,对小麦白粉病也表现出较高的保护效果,并具一定的治疗作用,在剂量为1500mg/L时,对小麦白粉病的保护效果达到66.8%、治疗效果是43.1%。Isabenlin对番茄灰霉病菌、小麦纹枯病菌和辣椒疫霉病菌菌丝生长的EC50分别为221.25、432.21和714.69mg/L。去氧薇甘菊内酯对供试10种病原菌菌丝生长均具很高的抑制毒力,EC50在5.61～46.48mg/L之间,对小麦纹枯病菌菌丝生长的毒力最高,EC50是5.61mg/L；去氧薇甘菊内酯抑制不同病原菌孢子萌发效果差异明显,对番茄灰霉病菌孢子EC50为10.49 mg／L,抑制毒力最高。二氢薇甘菊内酯对辣椒疫霉病菌的毒EC50为12.84 mg/L,对5种供试病原菌孢子萌发的的EC50介于20-75mg/L之间。结合化合物的提取分离结果,分析认为,倍半萜内酯类化合物和甾醇类化合物是小花假泽兰主要的杀菌活性成分,而其中甾醇类化合物具有一定的直接开发利用前景。
3.系统测试并比较了水蒸汽蒸馏法(HD)、超临界CO2萃取法(SCE)及溶剂萃取法(PEE)等3种方法提取制备的小花假泽兰精油的化学成分。首先,采用正交设计试验提出小花假泽兰精油超临界C02萃取适宜工艺：先静态萃取20 min,然后动态萃取,动态萃取参数温度55℃、压力35 MPa、CO2体积40 mL／g；进而,结合前期研究结果,采用GC/MS法分析比较了HD法、SCE法及PEE法等3种方法制备的小花假泽兰精油的化学成分,并测试了3种提取物对小麦纹枯病菌和小麦全蚀病菌的毒力。结果表明：SCE法精油萃取率显著高于HD法及PEE法；小花假泽兰精油主要有烯烃类、萜类、醇类、脂肪酸、酯类、甾体类等化合物,不同方法提取物化学成分存在明显差异；小花假泽兰精油对不同病菌具明显的选择性抑制作用,对小麦赤霉病菌、小麦纹枯病菌和番茄灰霉病菌表现出很强的抑制作用,对棉花枯萎病菌的抑制作用较弱；3种提取方法所得精油的杀菌活性亦存在显著差异,SCE法精油对小麦赤霉病菌和小麦纹枯病菌的毒力亦显著高于HD法和PEE法提取物。
4.利用电镜技术开展了豆甾醇对小麦活体上小麦白粉病菌的影响初步研究,结果表明,豆甾醇可致小麦白粉病原菌菌丝扭曲、畸形,表面粗糙,生长点塌陷、老化加快；可致小麦白粉病菌分生孢子数量减少,孢子表面出现大面积凹坑；可致菌丝细胞内原生质电子致密度显著增加,各种细胞器解体,菌丝坏死；可导致病原菌菌丝细胞和吸器体液泡化严重,且出现空腔,液泡中有电子致密度高的沉积物；可致菌丝细胞内电子致密物不均匀,囊泡变大而数量减少；使小麦白粉病原菌吸器外间质膜不规则加厚。分析认为豆甾醇可能主要是对病原菌体器官的生物合成过程造成较大影响,致使其侵染能力下降,同时对病菌体内生物氧化过程亦具一定影响。对此,还有待于进一步深入探讨。
5.以小花假泽兰全株粗粉为原料,通过对表面活性剂、载体等助剂的筛选,研制出了70%小花假泽兰可湿性粉剂配方。大田药效试验结果表明,70%小花假泽兰可湿性粉剂在7500 mg/L剂量下,对番茄灰霉病的防治效果达70%以上。
本论文初步探明了小花假泽兰杀菌活性化学成分,发现了几个具有植物病原菌杀菌活性的化合物；探讨了豆甾醇、去氧薇甘菊内酯等化合物杀菌活性强度和杀菌谱,初步评价了其应用前景；初步研究了豆甾醇对小麦白粉菌形态和超微结构的影响；明确了小花假泽兰精油超临界C02萃取适宜工艺、不同提取方法效果差异及其主要化学成分；并对假泽兰可湿性粉剂制剂配方进行了初步探讨。研究结果为充分利用小花假泽兰这一植物资源,开发新型植物源农药提供了理论依据。
Research and development of natural products is one of primary and effective ways to develop novel pesticides. As dangerous invasive species from abroad, control and utilization of plants killer, Mikania micrantha H.B.K is urgently needed. Previous study has shown that the extract of Mikania micrantha H.B.K have a strong inhibition activity against many plant pathogens. In order to utilize Mikania micrantha H.B.K resource efficiently, make the best use of the situation while avoiding disadvantages, and provide theoretical basis for novel plant-derived fungicide developing, the chemical constituents and fungicidal activity of Mikania micrantha H.B.K were investigated on the basis of some previous study. The main result obtained from this study as follows:
1.17 compounds were isolated from Mikania micrantha H.B.K based on bioactivity-guided isolation, and the chemical structures of 15 compounds were identified. Those chemicals include Deoxymikanolide, Dihydromikanolide, Mikanolide, Isabelin, Scandenolide, Mikanokryptin,1β,6α-dihydroxy eudesmane-4(15)-alkene, eudesmane-4 (15), 7 (11)-12,8β-lactone, Mikanin, Stigmasterol,β-Sitosterol, eudesmane-4(15),7(11), 7(11)-triene-12,8β-lactone, luteolin,β-Daucosterol, Eupatolitin. The result shows that 15 identified compounds can be divided into 3 categories:sesquiterpene lactones; flavonoid compounds and steroidal saponins.
2. The fungicidal activities of Stigmasterol, Deoxymikanolide, Dihydromikanolide, and Isabelin were evaluated against plant pathothic fungi. The results showed that Stigmasterol can inhibit the mycelium growth of 12 pathogens, which EC50 value is within 365.48-1438.59 mg/L. Stigmasterol showed highest antifungal activities against Botrytis cinerea Pers with an EC50 value of 365.48 mg/L. The results also revealed that there are significant differences between the inhibition effects of Stigmasterol on different pathogens spore germination, Stigmasterol showed highest antifungal activities against Fulvia fulva and Botrytis cirerea Pers. et Tris with an EC50 value of 151.47 and 174.65mg/L, respectively. Furthermore, Stigmasterol displayed certain control effect (68.60% at 1500 mg/L) against B. cirerea in vivo. And Stigmasterol also displayed prominent systemic protective effect (66.80% at 1500 mg/L and 43.10% at 1000 mg/L) and control effect (50.60% at 1500 mg/L and 25.70% at 1000 mg/L) against wheat powdery mildew Blumeria graminis in vivo. Deoxymikanolide displayed a higher inhibition on mycelium growth of 10 pathogens, witch EC50 value was within 5.61-46.48 mg/L, Deoxymikanolide showed highest antifungal activities against Rhizoctonia cerealis Vander Hoeven with an EC50 value of 5.61 mg/L. The bioassay showed that there is a significant difference between the inhibition effect of Deoxymikanolide on different pathogens spore germination, Deoxymikanolide showed highest antifungal activities against Botrytis cirerea Pers. et Tris with an EC50 value of 10.49mg/L. The fungicidal activity of Dihydromikanolide was lower than Deoxymikanolide, Dihydromikanolide only showed a similar antifungal activities with Deoxymikanolide against Phytophthora capsici with an EC50 value of 12.84mg/L. There is a little difference between the inhibition effect of Dihydromikanolide on different pathogens spore germination, the EC50 value of Dihydromikanolide effects on spore germination of 5 pathogens was 20～75 mg/L. The EC50 value of Isabenlin against B. cirerea, R. cerealis and Phytophthora capsici was 221.25,432.21 and 714.69 mg/L, respectively. Based on the bioassay results combined with isolation and identification results of compounds, sesquiterpene lactones and steroidal saponins were considered to be primary fungicidical components in M. micrantha, and steroidal saponins has a certain prospect of developing and utilization.
3. In the basis of previous extraction technology of essential oils from M. micrantha with supercritical CO2, the chemical constituents of M. micrantha extracts acquired with 3 methods: hydrostillation(HD), SCE, and extracted with petrol ether(PEE) were analyzed by GC/MS, and the toxicity of those 3 extracts against Fusarium graminearum and Rhizoctonia cerealis was tested. The result showed that the optimized extraction condition was 35 MPa and 55℃with 40 mL/g supercritical carbon dioxide after 20-mintue static extraction, and the extraction rate of essential oil by SCE was higher than other 2 methods. The main components of essential oil from M. micrantha are polyolefins, terpenoids, alcohols, fatty acids, esters and steroidal compounds, and there are significant differences of chemical constituents between essential oil extracted with different methods. The essential oil of M. micrantha showed selective antifungal activities against different plant pathogens, the bioassay showed that essential oil has the highest antifungal activities against Fusarium graminearum and Rhizoctonia cerealis and Botrytis cinerea Pers and weaker inhibitory effect against Fusarium oxysporum f.vasinfectum (Atk.) Snyder et Hansen. There are significant differences of antifungal activities between essential oil acquired by different methods. The toxicity of M. micrantha essential oil extracted by SCE was higher than the essential oil extracted by HD and PEE against Fusarium graminearum and Rhizoctonia cerealis.
4. Effect of Stigmasterol against Blumeria graminis were observed by electron microscopy technology. The results showed that stigmasterol can lead to abnormal hyphal growth of B. graminis such as twisted and malformed mycelia, surface roughening, growing point collapse and faster aging. Stigmasterol can also reduce conidia quantity and lead to large area of surface pit in spore. Furthermore, stigmasterol treated fungi cause intracellular protoplasm electronic density to increase, organelles degenerated, mycelia putrescence, and cause mycelia and haustorial body heavy vacuolization and higher density substances sedimentary in mycelia cell. It could also lead to electronic density substances uneven in mycelia cell, enlarge vesicle and reduce quantity of vesicle. The outer plasma membrane haustorium of B. graminiwere was found to be irregulary thicken by stigmasterol. Those results showed that stigmasterol mainly effect biosynthetic process of plant pathogen, meanwhile, it can also effect biological oxidation of plant pathogen to a certain extent. Further research was needed to confirm those hypotheses.
5. The powder of M. micrantha has been formulated to 70% wettable powder through the surface active agent and carrier screening. The field trial showed that 70% M. micrantha WP can provide 70% control effect against B. cirerea at 7500 mg/L.
The dissertation preliminarily definited the chemical character of antifungal compounds from M.micrantha, and several novel compounds with fungicidal activity were found. The chemical constituent's character and antifungal activity of essential oil from M. micrantha have also been studied. The fungicidal spectrum and activities of Stigmasterol and Deoxymikanolide were investigated, and the application prospect as a novel control agent was evaluated. The formulation of wettable powder of M. micrantha was formulated and developed successfully. Those research results provide theoretical basis for utilization Mikania micrantha H.B.K resource efficiently and novel plant-derived fungicide developing.
引文
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