CN1204813C - 一种可在植物叶面形成特定拓扑结构的无毒抑菌剂 - Google Patents
一种可在植物叶面形成特定拓扑结构的无毒抑菌剂 Download PDFInfo
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Abstract
本发明是一种可在植物叶面形成特定拓扑结构的无毒抑菌剂，其特点是将无水乙醇、正硅酸乙酯及氨水混合后加热至15～50℃，再补加部分正硅酸乙酯并用水稀释，除去乙醇，最后加入0.005%吐温-20水溶液稀释即得到SiO2浓度为89～178的20～200nm的水溶性纳米结构氧化硅抑菌剂。本发明抑菌剂可控制真菌病害，而且不会对自然环境造成污染，对非靶标生物不会造成毒害，特别是对于蔬菜、果树和小麦具有重要的应用价值，是一种具有开发前景的绿色环保型抑菌剂，是纳米结构材料在农业上的直接应用。
Description
本发明属于农药技术领域，更确切地说，是一种水溶性的纳米结构氧化硅抑菌剂。
许多真菌病害如麦类白粉病(Blumeria graminis)，水稻稻瘟病(Magnaporthe grisea)，谷类作物炭疽病(Colletotrichum graminicola)和其他的温室蔬菜真菌病害如单丝壳菌(Sphaerotheca fuliginea)引起的黄瓜白粉病，终级腐霉菌(Pythium ultimum)引起的十字花科根腐病，瓜果腐霉菌(Pythium aphanidermatum)引起的番茄绵腐病，灰葡萄菌孢(Botrytis cinera)引起的黄瓜灰霉病和菜豆灰霉病，泻根亚隔孢壳菌(Didvmella bryoniae)引起的黄瓜蔓枯病，白粉菌(Erysiphecichoracearum)引起的苦荬菜白粉病和葡萄钩丝壳菌(Uncinula necato)引起的葡萄白粉病，近年来已成为影响我国主要农作物生产的病害，如小麦白粉病(Blumeria graminis f.sp.tritici)，1990和1991年在我国大流行时，两年发生面积均超过0.12亿公顷，年损失小麦32亿公斤。
对病害的防治主要采用抗病品种和杀菌剂两种措施。近年来，抗病基因的品种已丧失抗病性，目前生产上推广的品种大多为感病品种，因此在病害的防治上，有机药剂(如三唑酮)防治成了一种很重要的手段，但在药剂防治中出现了大量的抗性菌株，而且其频率在一些地区有明显的上升；另外有机药剂对非靶标生物毒性大，一些不易降解的农药或有代谢物在环境或动植物体内累积。
当一个真菌病原体在植物叶表着陆后，它所面临的最大障碍是如何进入叶组织。细菌感染植株是通过气孔，叶片表面的创口或其它自然通道，而真菌则是撕裂叶角质层直接进入植物体内。对于部分真菌种类，在侵染点上可明显地看到酶的活性，表明叶角质层在感病之前被溶解。许多专一性的真菌侵染寄主植物时，具有精确和独特的拓扑表面识别。例如，条锈(Uromyces appendiculatus)附着孢在气孔周围的精确定位，表明真菌可以识别叶表微小的拓扑变化，从而决定是否侵染寄主(H.C.Hoch等，Science235，1659，1987)。麦类白粉菌和水稻稻瘟病等是高度专一的寄主/病原菌相互作用的活体营养型(Biotrophic)真菌。病原菌侵染植物时真菌孢子首先与寄主表面接触，20秒内发生超分子识别反应(T.L.W.Carver等，Mycol.Res.103，547，1999)。孢子快速和持久地粘附在疏水的寄主叶表面上，表明未萌发的孢子与叶片初始接触后能够立即识别来自寄主叶片的信号，同时作出快速反应。同时，在孢子萌发之前释放大量胞外物质(ECM)紧紧地粘在疏水的寄主叶表面上(J.E.Hamer等，Science 239，288，1988)。孢子粘附是孢子/寄主界面多元协同粘附过程的第一步，孢子管的后期发育，初生芽管/寄主表面接触和附着孢分化之间的时间长短与界面疏水性(hydrophobicity)有关(K.A.Nielsen等，Physiological and MolecularPhant Pathology 56，63，2000)，Bechinger等测定了谷物作物炭疽病(Colletotrichum graminicola)侵染钉施加于叶片力的大小相当于汽车轮胎压力的30～40倍。因此附着孢必须紧紧地粘在叶表面，因为如果粘附力小于侵入力，附着孢将从叶片上抬起而脱落(C.Bechinger等Science 285，1896，1999)。
本发明的目的是提供一种抑制真菌病害的无毒抑菌剂。
本发明抑制真菌病害的原理：利用无机纳米氧化硅粒子修饰植物叶表面，改变植物叶表面原有的拓扑结构和疏水特性，形成特殊的双亲性表面，影响真菌胞外基质的释放，阻断真菌孢子和寄主表面的高度专一性的超分子识别过程的第一步反应，改变早期的侵染过程，从而达到抑制靶向真菌病害的目的。
本发明的主要技术方案：合成20-200nm可在水中分散的无毒氧化硅纳米粒子，进一步用化学方法如表面活性剂修饰，从而得到不同尺寸、不同表面形态的无毒氧化硅纳米粒子，将其均匀地分散在0.005％吐温-20的水溶液中。
本发明的抑菌剂的制备方法包括以下步骤：
(1)将无水乙醇、正硅酸乙酯(TEOS)、H2O和25～28％(重)的氨水，以50～100∶1.0～3.0∶1～3∶1～2体积比的量加入容器中，充分混合；
(2)将步骤(1)的混合物加热15-50℃，缓慢搅拌2-4小时后，补加步骤(1)加入的同等量的TEOS，继续反应2-4小时后，用5～10倍的水稀释；
(3)然后减压蒸馏除去乙醇，并用高纯水渗析至pH＝7为止；
(4)将步骤(3)所得的溶液用溶液重量的25-50倍的0.005％吐温-20的水溶液稀释，混匀后即得SiO2浓度为89～178ppm的20～200nm的水溶性氧化硅纳米粒子。
本发明所述的原料均是市售产品。
本发明产品的SiO2粒径的测定方法：将溶液滴到铜网上，干燥后在透射电子显微镜下观测。
本发明的抑菌剂可应用于各种温室和大田作物，使用方法：早晨或傍晚无风时喷洒，应保证12小时内不浇水，施用量80-100ml/m2。
本发明的优点和效果：本发明抑菌剂的作用机制不同于其他有机化学制剂，它通过水基性氧化硅纳米粒子对叶片表面进行修饰，从而改变了叶表面的疏水性和介观拓扑结构，进而影响真菌孢子胞外物质的释放以及病菌孢子的早期粘附作用以及孢子管和附着孢的形成。本发明对非靶标生物不会造成毒害，不会给自然环境带来污染，特别是在蔬菜、果树和小麦上具有重要的应用价值。综上所述，本发明由于氧化硅纳米粒子因其安全、廉价、无污染，是一种具有开发前景的绿色环保型抑菌剂，是纳米结构材料在农业上的直接应用。本发明是国家重点基础研究发展规划项目(G1999011707)的部分内容。
下面通过具体实例进一步说明本发明的特点：
实例1
将无水乙醇50ml、正硅酸乙酯(TEOS)1.0ml、H2O 1ml、NH3·H2O(28wt％)2ml，充分混合；将混合物用水浴18℃缓慢搅拌3小时后，补加1mlTEOS，继续反应3小时后，所得样品加入100ml的高纯水，然后减压蒸馏除去乙醇，并用高纯水渗析至pH＝7为止；将所得溶液用0.005％吐温-20的水溶液稀释25倍，混匀后即得浓度为178ppm的水溶性100nm氧化硅纳米粒子的抑菌剂。
实例2
将无水乙醇50ml、正硅酸乙酯3.0ml、H2O 1ml、NH3·H2O(25wt％)2ml，充分混合；将混合物用水浴加热40℃，缓慢搅拌3h后，补加3.0ml TEOS，继续反应3小时后，所得样品加入100ml的高纯水，然后减压蒸馏除去乙醇，并用高纯水渗析至pH＝7为止；将所得溶液用0.005％吐温-20的水溶液稀释50倍，混匀后即得SiO2浓度为89ppm的水溶性的60nm氧化硅纳米粒子的抑菌剂。
实例3
用实例1的产品防治小麦白粉病。
将配制好的药品以80ml/m2的用量喷施，以0.005％吐温-20的水溶液为对照。借助接触角测定仪测定Si源修饰的叶表面的接触角17°6′，而小麦叶正面的接触角是147°37′，测定温度：22℃，水滴大小：0.87ul。在室内对小麦白粉病的抑制作用进行了测定，结果表明抑制效果为75％，而用0.005％吐温-20水溶液不具抑制白粉病的作用。同时研究实例1的产品影响孢子在修饰的亲水模拟叶表面的早期粘附及孢子管和附着孢的形成。将小麦白粉菌孢子悬浮在10mM磷酸缓冲液中，调节PH6.5，取含有孢子的磷酸缓冲液100μl滴到2×2cm的Teflon-PFA膜和干净的显微镜盖玻片，在室温下培养。利用phase-contrast光学显微镜(×200)观察孢子萌发。形成孢子管的孢子是对照的5％。
实例4
用实例2的产品防治黄瓜白粉病。
将配制好的药品以100ml/m2的用量喷施，以0.005％吐温-20的水溶液为对照。借助接触角测定仪测定Si源修饰的叶表面的接触角为10°12′，而黄瓜叶正面的接触角是130°30′，测定温度：22℃，水滴大小：0.87ul。在室内对黄瓜白粉病的抑制作用进行测定，结果表明抑制效果为85％。利用气流室(flow chamber)研究孢子在实例2的产品修饰的叶表面的粘附强度。将大约含有5×105个孢子的1ml磷酸缓冲液重新悬浮在灭菌的蒸馏水中，将溶液滴到叶片或Teflon-PFA膜上后置于气室中，设置不同气流速度(flow rate)后发现80％孢子不能粘附在叶表面。借助低温电子扫描显微镜(SEM)技术观察孢子在与实例2的产品修饰的叶表面作用时胞外物质(ECM)释放被阻断。所使用的气流室研究方法见“Hamer等，Science，239，288，1988”。
Claims (1)
1.一种可在植物叶面形成特定拓扑结构的无毒抑菌剂，其特征在于是由下述方法制得
(1)将无水乙醇、正硅酸乙酯、H2O和25～28％的氨水，以50～100∶1.0～3.0∶1～3∶1～2体积比的量加入到容器里，充分混合，所述的氨水浓度为重量百分浓度，
(2)将步骤(1)的混合物加热至15-50℃，缓慢搅拌2～4小时后，补加步骤(1)加入的同等量的正硅酸乙酯，继续反应2～4小时后，用5～10倍的水稀释，
(3)然后减压蒸馏除去乙醇，并用高纯水渗析至pH＝7为止，
(4)将步骤(3)所得的溶液用溶液重量的25-50倍的0.005％吐温-20的水溶液稀释，混匀后即得到SiO2浓度为89-178ppm的60-100nm的在水中分散的水溶性氧化硅纳米粒子的无毒抑菌剂。
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| Application Number | Priority Date | Filing Date | Title |
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| CN 01109424 CN1204813C (zh) | 2001-03-09 | 2001-03-09 | 一种可在植物叶面形成特定拓扑结构的无毒抑菌剂 |
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| CN 01109424 CN1204813C (zh) | 2001-03-09 | 2001-03-09 | 一种可在植物叶面形成特定拓扑结构的无毒抑菌剂 |
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| CN1319338A CN1319338A (zh) | 2001-10-31 |
| CN1204813C true CN1204813C (zh) | 2005-06-08 |
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| DE102007060320A1 (de) * | 2007-12-12 | 2009-06-18 | Stiftung Nano Innovations, Olten | Schutzschicht für Pflanzen und Bäume, deren Herstellung und Verwendung |
| US11283754B2 (en) | 2018-09-19 | 2022-03-22 | Cisco Technology, Inc. | Unique identities of endpoints across layer 3 networks |
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