【作者】 何顺；

【导师】 曹永松；

【作者基本信息】 中国农业大学 ， 植物检疫与农业生态健康， 2014， 博士

【摘要】 番茄黄化曲叶病毒病(tomato yellow leaf curl disease, TYLCD)是目前严重威胁番茄生产的一种毁灭性病害。番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV)是单链环状DNA双生病毒,依靠烟粉虱传播,对寄主进行循环侵染。本论文开展TYLCV的化学防治研究,其研究内容包括以天然或抗病毒药物为先导的新抗病毒药物的制备及活性评价,防治媒介昆虫新制剂的制备及活性评价两个方面,该项研究可为TYLCV的防治提供翔实数据和科学依据。通过实验研究,获得了以下主要结果：1.以4-氯-3,5-二硝基三氟甲苯为母体结构,在缚酸剂存在下,合成了一系列氨基酸衍生物,并经核磁共振氢谱、元素分析确证其结构。温室盆栽试验表明,化合物1-7在2000mg/L时,对TYLCV的抑制率为36.26%,1-13在1000mg/L时,对TYLCV的抑制率为40.66%。田间大棚试验表明,化合物I-7和1-13均能较好抑制病害的发生,且增加番茄的产量,增产率可达35%左右。2.以利巴韦林为母体化合物,经过叉基保护、酯化和去保护反应对利巴韦林呋喃糖环进行了改造,制备了17个化合物,并经核磁共振氢谱、元素分析确证其结构。温室盆栽试验表明,经过结构改性的化合物,对番茄植株的药害较利巴韦林低。随着取代基碳链的增长,利巴韦林5’位的烷酰基取代产物对TYLCV的抑制作用增强,当取代基碳原子个数达到一定的数目(10)时,活性开始下降。5’芳酰基取代产物的抗TYLCV的活性比烷酰基取代的要高,氟代芳基取代产物的抗TYLCV活性比氯代芳基的要好。化合物Ⅱ-11、Ⅱ-16和Ⅱ-17在1000mg/L时,对TYLCV的抑制率分别为34.09%、28.41%和29.55%。田间大棚试验表明,化合物Ⅱ-11、Ⅱ-16和Ⅱ-17均能一定程度地抑制病害的发生,且增加番茄的产量,增产率可达30%左右。3.以小檗碱为母体化合物,经过去甲基、酯化反应对小檗碱进行改造,制备了17个化合物,并经核磁共振氢谱、元素分析确证。温室盆栽试验表明,随着取代基碳链的增长,小檗碱9位烷酰基取代产物对TYLCV的抑制作用减弱,当取代基碳原子个数达到一定的数目时,化合物对TYLCV基本没有抑制作用。9位芳酰基取代产物的抗TYLCV的活性比烷酰基取代的要强,氟代芳基取代产物的抗TYLCV活性比氯代芳基的要高。田间大棚试验表明,化合物Ⅲ-11、Ⅲ-16和Ⅲ-17均有一定的增产效果。4.以正硅酸乙酯(TEOS)为硅源,十六烷基三甲基溴化铵(CTAB)为表面活性剂,十六烷为稳定剂,通过微乳液法超声辅助制备了丁硫克百威二氧化硅纳米缓释剂。优化了制备条件,所制得的纳米缓释剂的负载率达到42%(w/w)。缓释动力学研究结果表明溶液体系的温度、pH值等影响制剂的缓释特性,在pH3.0-9.0范围内,体系的pH值越小,有效成分释放的速度越慢,缓释性能越好；在温度25-45℃范围内,随着体系温度的升高释放速度加快。温室番茄盆栽试验表明,施药后14d之前,乳油对烟粉虱的防效比缓释剂的好,14d开始,缓释剂的防效比乳油的防效好,药后第28天,carbosulfan-PHSNs的防效分别可以达到67.71、62.08、56.17%。5.以正硅酸乙酯为硅源,十六烷基三甲基氯化铵(CTAC)为表面活性剂,通过乳液聚合法制备了阿维菌素单层的二氧化硅微胶囊,再用3-氨丙基三乙氧基硅烷氨基化改性二氧化硅,在戊二醛作用下将所制备的壳聚糖纳米颗粒与氨基功能化的二氧化硅交联反应,制备二氧化硅-戊二醛-壳聚糖复合微胶囊。优化了制备条件,微胶囊的负载率可达到40%(w/w)。研究还表明,双层微胶囊能有效地防止阿维菌素光解,与单层微胶囊相比,双层微胶囊具有更好的控释特性。缓释动力学研究结果表明溶液体系的温度、pH值等影响制剂的缓释特性,在pH3.0-9.0范围内,体系的pH值越低,有效成分释放的速度越慢,缓释性能越好；在温度25-45℃范围内,随着体系温度的升高释放速度加快。温室番茄盆栽试验表明,施用9mg/L和4.5mg/L阿维菌素乳油7d后对烟粉虱的防效分别为82.12、76.31、68.09%,而相当浓度的阿维菌素微胶囊7d后的防效分别为64.91、52.61、40.87%。随着时间的延长,阿维菌素乳油的防效开始下降,而微胶囊的防效上升,直到药后第28天,阿维菌素微胶囊的防效分别可以达到81.02、68.30、63.29%。更多还原

【Abstract】 Tomato yellow leaf curl disease caused by tomato yellow leaf curl virus (TYLCV) is one of the most devastating viral diseases of cultivated tomato (Lycopersicon esculentum) in tropical and subtropical regions worldwide. TYLCV belongs to the genus Begomovirus (family Geminiviridae) and are transmitted in a circulative, persistent manner by the whitefly Bemisia tabaci (Gennadius) This dissertation provides detailed data and scientific basis for the chemical control of tomato yellow leaf curl virus (TYLCV) by inhibition of viral and control whitefly. Through experimental research, the thesis has obtained the following main results:1. A series of amino acid analogs was prepared using4-chloro-3,5-dinitrobenzotrifluoride as the parent structure, and the structures were identified by1H NMR and elemental analysis. Greenhouse trial showed that the highest inhibition rate was36.26%and40.66%for1-7at2000mg/L,I-13at1000mg/L respectively. Field trial showed that compound1-7and1-13could inhibit the occurrence of the disease effectively, and increase the yield of tomatoes up to35%.2. A series of ribavirin analogs was prepared using ribavirin as the parent compound, and the structures were identified by1H NMR and elemental analysis. Greenhouse trial showed that the phytotoxicity of ribavirin analogs was lower than that of ribavirin. The activities of anti-TYLCV increased with the linear carbon chain of the substituted groups. When the number of carbon atoms of the substituted group exceeded10, the activities of anti-TYLCV became lower. The activities of anti-TYLCV of analogs with an aryl group were better than that of with an alkyl group. The highest inhibition rate was34.09%,28.41%and29.55for Ⅱ-11,Ⅱ-16and Ⅱ-17at2000mg/L respectively. Field trial showed that compound Ⅱ-7,Ⅱ-16and Ⅱ-17could inhibit the occurrence of the disease effectively, and increase the yield of tomatoes up to30%.3. A series of berberine analogs was prepared using berberine as the parent compound, and the structures were identified by1H NMR and elemental analysis. Greenhouse trial showed that the activities of anti-TYLCV decreased with the linear carbon chain of the substituted groups. Just as ribavirin analogs, the activities of anti-TYLCV of analogs with an aryl group were better than that of with an alkyl group. The activities of anti-TYLCV of fluorinated aryl group were better than that of chlorinated. The highest inhibition rate was30.43%for Ⅲ-11at2000mg/L.4. Carbosulfan porous hollow silica nanoparticles (carbosulfan-PHSNs) was prepared using TEOS as the silica precursor, hexadecyl trimethyl ammonium bromide (CTAB) as a surfactant, hexadecane as a stabilizer, and carbosulfan as a model pesticide through microemulsion polymerization assisted by ultrasound. The results showed that the resulting carbosulfan-PHSNs had a remarkable loading ability for carbosulfan (about42%w/w). The release kinetics of carbosulfan-PHSNs showed that the factors such as temperature and pH affected the controlled release behaviors. Within the range of pH3.0-9.0, the lower the pH value was, the slower active ingredients released. Within the temperature range of25-45℃, the release rate speeded up with the temperature increasing. The efficacy of carbosulfan-PHSNs was evaluated by pot experiment using Bemisia tabaci as the target insect. The results showed that the efficacy of carbosulfan-PHSNs against Bemisia tabaci was better than carbosulfan emulsifiable concentrate (EC) after14days. The efficacy of different carbosulfan-PHSNs against Bemisia tabaci was67.71、62.08、56.17%respectively after28days.5. The single-shelled microcapsule was prepared via emulsion polymerization using tetraethyl orthosilicate (TEOS) as the silica precursor, hexadecyl trimethyl ammonium chloride (CTAC) as a surfactant and avermectin as the model pesticide. The silica shell was formed through the hydrolysis and polycondensation of TEOS at the oil droplet/water interface. Then the silica shell was modified with3-aminopropyl triethoxysilane. Chitosan nanospheres were prepared by adjusting pH value of the solution and then cross-linked with modified silica at the surface of silica shell in the presence of glutaraldehyde to form double-shelled avermectin microcapsules. The results showed that the resulting microcapsules had a remarkable loading ability for avermectin (about40%w/w) and can protect avermectin against photo-and thermal-degradation effectively. Compared to single-shelled microcapsules, the double-shelled ones had better controlled release properties under all conditions. The efficacy of resulting microcapsule was evaluated by pot experiment using Bemisia tabaci as the target insect. The results showed that the efficacy of the resulting microcapsule against Bemisia tabaci was64.91,52.61,40.87%after7days, while avermectin EC were82.12,76.31and68.09%respectively. The efficacy of microcapsules was better than that of avermectin EC after14days. The efficacy of different microcapsules against Bemisia tabaci was67.71,62.08and56.17%respectively after28days.更多还原