丁吡吗啉对辣椒疫霉菌的作用方式及作用机理探讨
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摘要
丁吡吗啉是中国农业大学、中国农业科学院植物保护研究所和江苏耕耘化学有限公司联合开发的丙烯酰胺类杀菌剂,本文研究了丁啉吗啉对辣椒疫霉的菌丝生长、孢子囊形成、孢子囊萌发、释放游动孢子的影响及室内盆栽和室外大田实验,并探讨了丁吡吗啉抑制辣椒疫霉的作用机制。研究结果如下:
(1)丁吡吗啉对辣椒疫霉菌丝生长、孢子囊的形成有较强的抑制作用,其有效中浓度(EC50)分别为2.92μg/mL、3.07μg/mL,对辣椒疫霉菌丝生长量的EC50为6.92μg/mL,但对孢子囊萌发的抑制作用较弱,EC50为20.19μg/mL。
(2)扫描电镜和透射电镜观察的结果表明,丁吡吗啉能够使辣椒疫霉的菌丝形态发生变化,菌丝的分枝增加,分枝间距减小,菌丝的表面有小的突起。利用透射电镜对丁吡吗啉引起的辣椒疫霉菌丝超微形态变化进行了研究。结果表明丁吡吗啉处理辣椒疫霉菌丝造成最明显的超微形态变化是细胞壁变厚,细胞内含物减少,细胞核变形,液泡及细胞内部结构的破坏,并且这种破坏作用与培养基中丁吡吗啉的浓度成正比。
(3)ATP是生物能的存在形式,不同浓度的ATP(1μg/mL和5μg/mL)都可促进辣椒疫霉菌丝的生长。丁吡吗啉在不同浓度条件下由于ATP的存在,使其菌丝生长的抑制率减小,并且抑制率的趋势明显降低。
(4)用电导率法测辣椒疫霉菌细胞膜的通透性,结果发现丁吡吗啉能够使辣椒疫霉细胞膜通透性发生改变,随着药剂浓度的加大,时间的延长,电导率值逐渐升高,细胞膜的通透性增加。改变的原因可能是丁吡吗啉破坏了辣椒疫霉细胞膜的超微结构,从而造成了细胞内成分的外泄。
(5)丁吡吗啉对辣椒疫霉菌体可溶性蛋白合成的影响,随着药剂浓度的增加,抑制作用逐渐增强。丁吡吗啉抑制菌丝的生长与其抑制蛋白质合成有关,具体抑制蛋白质合成的机理还有待于进一步研究。
(6)用氧电极法,从呼吸代谢的角度来研究丁吡吗啉的作用机制,结果发现丁吡吗啉对辣椒疫霉菌菌丝和孢子呼吸速率的抑制作用效果不明显。加入丙二酸、碘乙酸、磷酸钠三种典型代谢抑制剂,发现丁吡吗啉与磷酸钠的叠加率最小,初步说明丁吡吗啉的作用途径是磷酸戊糖途径。
(7)室内盆栽活体试验和大田试验结果表明,丁吡吗啉对辣椒疫病有较好的防治效果。
Pyrimorph is a novel acryl amide fungicide developed by China Agicultural University , Plant Protection Institute of CAAS and Jiangsu Cultivation Chemical Co. LTD. The effects include mycelial growth, sporangium formation, and sporangium germination.were studied in this paper. The effects of pyrimorph on Phytophthora capsici Leonian indoors pot experiment and outdoors field experiment were also investigated , and the research of Pyrimorph on mechanism of action of Phtophthore capsici were discussed. Research results are as follows:
The results showed that pyrimorph possessed strong inhibition on mycelial growth, sporangium production with the EC50 values of 2.92、3.07μg/mL, respectively, however,the inhibiting activity on sporangium germination was feeblish,the EC50 value was 20.19.μg/mL.
By light microscopy, scanning electron microscopy (SEM) and TEM, the investigations showed that pyrimorph could cause a series of morphological and structural alterations of Phytophthora capsici Leonian hyphae , These changes included excessive branching, the interval between branches shortened, protuberance on the surface of hyphae. The investigations made by transmission electron microscope showed that the most obvious ultrastructure changes of Phytophthora capsici Leonian treated with pyrimorph were cell wall becomes thick, the plasmlemma shrinking,malformed nuclei and damage of vacuole and cells inter structure , and the damage is proporitional the concentration of pyrimorph in substrate.
ATP is the existence of bio-energy formation, different concentrations of ATP can promote the growth of mycelium; it can be antagonistic against pyrimorph, and proves that pyrimorph affected the production of ATP.
Pyrimorph can chage membrane permeability of Phytophthora capsici, the reason which changed membrane permeabilithy was deduced that pyrimorph damaged the membrane permeability of Phytophthora capsici ultra-structure, caused the loss of cell components. The reason might also be considered as this specific ion-transprort carrier, the particular reason would be further studied.
The effect of pyrimorph on dissoluble protein of phytophthora capsici indicated that with the increasing of concentration of doses, the inhibition promoted increasingly .But the specific inhibiting protein synthesis mechanism remains to further study.
Mycelial respiration was affected. Pyrimorph exhibited certain inhibition on metabolic approaches of Embden-Meyerhof-Parnas (EMP), tricarboxylic acid cycle (TCA), and hexosemonophosphate (HMP) by measuing the oxygen consumption of pyrimorph combining with three representative inhibitors to the metabolic approaches.The results indicated that pyrimorph could inhibit the approach of HMP significantly.
The results of indoors pot experiment and outdoors experiment indicated that the control effects of pyrimorph on Phytophthora capsici Leonian is good.
(1)丁吡吗啉对辣椒疫霉菌丝生长、孢子囊的形成有较强的抑制作用,其有效中浓度(EC50)分别为2.92μg/mL、3.07μg/mL,对辣椒疫霉菌丝生长量的EC50为6.92μg/mL,但对孢子囊萌发的抑制作用较弱,EC50为20.19μg/mL。
(2)扫描电镜和透射电镜观察的结果表明,丁吡吗啉能够使辣椒疫霉的菌丝形态发生变化,菌丝的分枝增加,分枝间距减小,菌丝的表面有小的突起。利用透射电镜对丁吡吗啉引起的辣椒疫霉菌丝超微形态变化进行了研究。结果表明丁吡吗啉处理辣椒疫霉菌丝造成最明显的超微形态变化是细胞壁变厚,细胞内含物减少,细胞核变形,液泡及细胞内部结构的破坏,并且这种破坏作用与培养基中丁吡吗啉的浓度成正比。
(3)ATP是生物能的存在形式,不同浓度的ATP(1μg/mL和5μg/mL)都可促进辣椒疫霉菌丝的生长。丁吡吗啉在不同浓度条件下由于ATP的存在,使其菌丝生长的抑制率减小,并且抑制率的趋势明显降低。
(4)用电导率法测辣椒疫霉菌细胞膜的通透性,结果发现丁吡吗啉能够使辣椒疫霉细胞膜通透性发生改变,随着药剂浓度的加大,时间的延长,电导率值逐渐升高,细胞膜的通透性增加。改变的原因可能是丁吡吗啉破坏了辣椒疫霉细胞膜的超微结构,从而造成了细胞内成分的外泄。
(5)丁吡吗啉对辣椒疫霉菌体可溶性蛋白合成的影响,随着药剂浓度的增加,抑制作用逐渐增强。丁吡吗啉抑制菌丝的生长与其抑制蛋白质合成有关,具体抑制蛋白质合成的机理还有待于进一步研究。
(6)用氧电极法,从呼吸代谢的角度来研究丁吡吗啉的作用机制,结果发现丁吡吗啉对辣椒疫霉菌菌丝和孢子呼吸速率的抑制作用效果不明显。加入丙二酸、碘乙酸、磷酸钠三种典型代谢抑制剂,发现丁吡吗啉与磷酸钠的叠加率最小,初步说明丁吡吗啉的作用途径是磷酸戊糖途径。
(7)室内盆栽活体试验和大田试验结果表明,丁吡吗啉对辣椒疫病有较好的防治效果。
Pyrimorph is a novel acryl amide fungicide developed by China Agicultural University , Plant Protection Institute of CAAS and Jiangsu Cultivation Chemical Co. LTD. The effects include mycelial growth, sporangium formation, and sporangium germination.were studied in this paper. The effects of pyrimorph on Phytophthora capsici Leonian indoors pot experiment and outdoors field experiment were also investigated , and the research of Pyrimorph on mechanism of action of Phtophthore capsici were discussed. Research results are as follows:
The results showed that pyrimorph possessed strong inhibition on mycelial growth, sporangium production with the EC50 values of 2.92、3.07μg/mL, respectively, however,the inhibiting activity on sporangium germination was feeblish,the EC50 value was 20.19.μg/mL.
By light microscopy, scanning electron microscopy (SEM) and TEM, the investigations showed that pyrimorph could cause a series of morphological and structural alterations of Phytophthora capsici Leonian hyphae , These changes included excessive branching, the interval between branches shortened, protuberance on the surface of hyphae. The investigations made by transmission electron microscope showed that the most obvious ultrastructure changes of Phytophthora capsici Leonian treated with pyrimorph were cell wall becomes thick, the plasmlemma shrinking,malformed nuclei and damage of vacuole and cells inter structure , and the damage is proporitional the concentration of pyrimorph in substrate.
ATP is the existence of bio-energy formation, different concentrations of ATP can promote the growth of mycelium; it can be antagonistic against pyrimorph, and proves that pyrimorph affected the production of ATP.
Pyrimorph can chage membrane permeability of Phytophthora capsici, the reason which changed membrane permeabilithy was deduced that pyrimorph damaged the membrane permeability of Phytophthora capsici ultra-structure, caused the loss of cell components. The reason might also be considered as this specific ion-transprort carrier, the particular reason would be further studied.
The effect of pyrimorph on dissoluble protein of phytophthora capsici indicated that with the increasing of concentration of doses, the inhibition promoted increasingly .But the specific inhibiting protein synthesis mechanism remains to further study.
Mycelial respiration was affected. Pyrimorph exhibited certain inhibition on metabolic approaches of Embden-Meyerhof-Parnas (EMP), tricarboxylic acid cycle (TCA), and hexosemonophosphate (HMP) by measuing the oxygen consumption of pyrimorph combining with three representative inhibitors to the metabolic approaches.The results indicated that pyrimorph could inhibit the approach of HMP significantly.
The results of indoors pot experiment and outdoors experiment indicated that the control effects of pyrimorph on Phytophthora capsici Leonian is good.
引文
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