SYAUP-770对灰葡萄孢作用机理初步研究及其盐的合成与活性筛选

发布时间：2018-01-01 02:30

本文关键词：SYAUP-770对灰葡萄孢作用机理初步研究及其盐的合成与活性筛选　出处：《沈阳农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：嘧啶胺广泛应用于农药领域,是一类重要的杂环类化合物,该类化合物具有除草、杀虫、杀菌等生物活性。前期,本实验室合成了一系列嘧啶胺类化合物,在此基础上筛选出活性高的化合物SYAUP-770,进一步研究其作用机制,为开发防治灰霉病的新型杀菌剂、丰富杀菌剂品种提供理论基础和依据,为后续结构优化,筛选,研发奠定理论基础。本论文对SYAUP-770进行了生物抑制活性测定,防治番茄灰霉病的内吸输导性测定及对灰葡萄孢形态及超微结构影响的测定。此外,为优化SYAUP-770的杀菌活性,本实验进行了 SYAUP-770盐的合成及筛选。结论如下:本文研究了 SYAUP-770对四种抗嘧霉胺的灰霉菌的菌丝生长的抑制作用、菌丝干重的影响、孢子的萌发、菌核产生等不同生育阶段的抑制作用。结果表明:SYAUP-770对灰葡萄孢的菌丝生长和孢子形成以及孢子萌发抑制作用十分明显。从EC50及EC80值可以看出,SYAUP-770对四个抗性菌株的杀菌活性均好于对照药剂嘧菌环胺和嘧霉胺。此外在PDA培养基中,SYAUP-770对FS-09,HLD-15和CY-09菌株的EC50值均小于1μg/mL。此结果表明,在PDA和FGA培养基上,SYAUP-770对三种抗性菌株都表现出了很高的抑菌活性。同时试验也发现药剂量加大后不仅菌核数减少,菌核块也变小,颜色变淡,由黑色变浅灰色,当SYAUP-770的处理药剂浓度在5 μg/mL时,PDA含药平板上已无菌核产生。而对照药剂嘧菌环胺在40 μg/mL时,抑制率为100%,但是随着药剂浓度的降低,对菌核的抑制率也逐渐降低。当浓度为5 μg/mL时,已经不抑制菌核的产生,相反会促进菌核的产生。也就是说高浓度时抑制菌核的产生,低浓度时促进菌核的产生。采用活体盆栽喷雾法,测定了 SYAUP-770对灰葡萄孢菌寄主的保护与治疗作用,以及SYAUP-770对黄瓜植株的内吸作用、对番茄灰霉病的持效性测定。结果表明:SYAUP-770对灰霉病兼具保护和治疗作用,且在番茄植株上的治疗作用明显,黄瓜植株上的保护效果明显且优于治疗效果,其防治效果高达85.8%;内吸作用结果表明,SYAUP-770可能具有横向内吸性能;从持效性测定结果看,开始SYAUP-770对灰霉病的效果没有嘧菌环胺的明显,但是随着药剂处理时间的延长,药剂SYAUP-770处理的番茄植株发病率逐渐减低,并且长势优于对照药剂嘧菌环胺处理的番茄植株。本文所述的新型嘧啶胺类化合物SYAUP-770对灰葡萄孢菌丝形态和超微结构的影响与现有杀菌剂嘧菌环胺略有不同。SYAUP-770可以抑制灰葡萄孢孢子萌发,萌发数变少;菌丝的分枝增多,菌丝体萎缩、塌陷、变形;荧光显微试验可以证实:菌体及孢子体的生物膜受损伤。SYAUP-770处理菌丝体3d后,超微结构下菌体细胞壁外部物质发生变化,细胞壁结构被破坏,细胞囊泡化严重,但部分菌体线粒体清晰可见,且数目增多,线粒体的脊变大。综上可以初步推测,一方面细胞膜受到损伤,另一方面细胞壁及细胞壁外层物质受到损害,导致细胞防护能力降低,加速细胞死亡;细胞内囊泡化增多,这些都加速了病菌的死亡。本试验合成了 38个SYAUP-770的盐,对其进行了离体与活体活性筛选。该类化合物对三种灰霉病菌的离体活性普遍较高,尤其是对CY-09和HLD-15两个菌种在50μg/mL的抑制率基本在90%以上,其中有四个化合物对XM菌株的活性在80%以上。此外,38个化合物对水稻纹枯病和黄瓜褐斑病的杀菌活性也均在90%左右。在黄瓜活体盆栽试验中,所有化合物对灰霉病菌均表现出了防治效果,高于对照药剂嘧霉胺与嘧菌环胺,其中SYAUP-770的间三氟甲基苯甲酸盐18和2-甲氧基苯甲酸盐19的防效高于SYAUP-770。
[Abstract]:Pyrimidinamine is widely used in the field of pesticide, is a kind of important heterocyclic compounds, the compounds with herbicidal and insecticidal, sterilization and other biological activities. A series of pre pyrimidinamine compounds were synthesized in this laboratory, based on the screening of high activity compound SYAUP-770, further study of the mechanism for the development of new fungicide the prevention and control of Botrytis cinerea, rich varieties of fungicides to provide theoretical foundation and basis for the subsequent screening, structure optimization, development, lay the theoretical foundation of this thesis. The biological inhibition activity for SYAUP-770 determination to control Botrytis cinerea bioaccumulation determination and effect on the morphology and ultrastructure of Botrytis cinerea. In addition, for optimization of SYAUP-770 bactericidal activity, we studied the synthesis and screening of SYAUP-770 salt. The conclusions are as follows: This paper studies the SYAUP-770 mycelium of four pyrimethanil against Botrytis cinerea The growth inhibition effect, the dry weight of mycelium, spore germination, inhibition of production of sclerotia in different stages. The results showed that SYAUP-770 mycelium growth and spore of Botrytis cinerea formation and spore germination inhibition effect is very obvious. It can be seen from the EC50 and EC80 value, the bactericidal activity of four SYAUP-770 resistant strains were better than the control Cyprodinil and pyrimethanil agents. In addition in the PDA medium, SYAUP-770 of FS-09, HLD-15 and CY-09 strains of EC50 values were less than 1 g/mL. the results showed that in PDA and FGA medium, SYAUP-770 of three resistant strains showed high antibacterial activity at the same time. The test also found that the dose increased not only reduce the number of sclerotia of Sclerotium, is also smaller, lighter in color from black, light gray, when the concentration of SYAUP-770 in the treatment of 5 g/mL, containing PDA plate has been produced. While the control of nuclear sterile Medicament Cyprodinil at 40 g/mL, the inhibition rate was 100%, but with lower concentrations, the inhibition rate of sclerotia decreased gradually. When the concentration is 5 g/mL, has no inhibition of sclerotial formation, on the contrary will promote the sclerotial formation. That is to say were inhibited at high concentration, promote sclerotium the low concentration. The in vivo spray method, protective and therapeutic effects of SYAUP-770 on Botrytis cinerea host were determined, and SYAUP-770 on Cucumber in the suction effect and the effect of holding on the determination of Botrytis cinerea. The results show that SYAUP-770 has protective and therapeutic effects on Botrytis cinerea, and the therapeutic effect in the tomato plants obvious protective effect on cucumber plants and significantly better than the treatment effect, the control effect was 85.8%; systemic effect. The results showed that SYAUP-770 may have a lateral suction performance; results from persistence of determination Start, the effect of SYAUP-770 on Botrytis cinerea was not Cyprodinil but with longer treatment time, pharmacy, pharmacy SYAUP-770 processing tomato incidence rate decreased gradually, and the tomato plants grew better than the control treatment. The effect of reagent cy of the new type SYAUP-770 pyrimidinamine compounds on mycelial morphology and ultrastructure with the existing fungicide Cyprodinil slightly different.SYAUP-770 can inhibit the germination of Botrytis cinerea spore germination, fewer; hyphal branching increases, mycelia atrophy, collapse and deformation; fluorescence microscopic test can confirm:.SYAUP-770 damage processing mycelium 3D biofilm bacteria and sporophyte after external material cell wall ultrastructure changes in cell wall structure was destroyed, cell vesicles seriously, but some cell mitochondria are clearly visible, and the increase of the number of mitochondria The ridge becomes larger. As can be inferred, on the one hand, the cell membrane damage, on the other hand, cell wall and cell wall outer material damage, reduce cell protection, accelerate cell death; vesicles of increased, all these accelerate the death of bacteria. 38 SYAUP-770 salt was synthesized in this experiment and has carried on the in vitro and in vivo activity. The compounds of three kinds of Botrytis cinerea in vitro activity is generally high, especially for CY-09 and HLD-15 two strains in the inhibition of 50 g/mL rate in 90% above, there are four active compounds on XM strains in more than 80%. In addition, the bactericidal activity of 38 compounds against Rice Sheath Blight and cucumber spot disease were also around 90%. In vivo of Cucumber in the pot experiment, all the compounds of Botrytis cinerea showed the control effect is higher than that of the control drug pyrimethanil and azoxystrobin cyclic amine, the The control effect of three fluoromethyl benzoate 18 and 2- methoxy benzoate 19 in the middle SYAUP-770 is higher than that of SYAUP-770.

【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S481

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