本文選題：辣椒疫病 + 枯草芽孢桿菌��； 參考：《福建農(nóng)林大學(xué)》2017年碩士論文

【摘要】：由辣椒疫霉菌(Phytophthora capsici)侵染引起的辣椒疫病,是辣椒生產(chǎn)上的重要病害之一,廣泛分布于世界各辣椒主產(chǎn)區(qū)。目前,在生產(chǎn)上控制辣椒疫病最易行且見效快的方法是施用化學(xué)藥劑,但長期施用藥劑使病原菌易產(chǎn)生一定抗藥性,且農(nóng)藥殘留問題突出,故尋找高效且不污染環(huán)境的方法對防治辣椒疫病具有重要意義,其中擁有廣泛前景的方法是利用生防菌進行生物防治。本研究利用實驗室早期分離獲得一株具有生防作用的枯草芽孢桿菌BS193為材料,以辣椒疫病為對象,研究該菌株抗菌譜、生防作用及其抗菌機制,為辣椒疫病的生物防治和該生防菌的深入開發(fā)應(yīng)用提供實踐依據(jù)。得出如下結(jié)論:1、枯草芽孢桿菌BS193的抗菌作用具有廣譜性。以平板對峙法對枯草芽孢桿菌BS193的抗菌譜進行測定,結(jié)果表明,枯草芽孢桿菌BS193對9個屬10種常見的植物病原菌均有抑制作用。2、枯草芽孢桿菌BS193對辣椒疫病具有良好的生防效果。在平板對峙試驗中,結(jié)果顯示出菌株BS193對4種辣椒疫霉菌小種抑制作用穩(wěn)定;在離體抑菌實驗中,表明經(jīng)BS193發(fā)酵液處理后抑制辣椒疫霉菌侵染葉片,其防效達到63.50%;在室內(nèi)盆栽防病試驗中,結(jié)果表明BS193菌體、無菌濾液、發(fā)酵液原液處理后24 h接種病原菌,9 d后各處理的防治效果分別為:30.75%、29.76%、52.56%;BS193發(fā)酵液、100倍稀釋液、1000倍稀釋液處理辣椒苗,接種病原菌9 d后的防效為:52.56%、34.56%、23.53%;BS193發(fā)酵液灌根后間隔0、24、48、72 h后接種病原菌,9 d后各處理的防治效果分別為:23.43%、41.25%、49.41%、51.44%。由此可知,枯草芽孢桿菌BS193對辣椒疫病的防病效果良好。同時,試驗中發(fā)現(xiàn)BS193發(fā)酵液對辣椒幼苗還具有很好的促生作用。用BS193發(fā)酵液灌根30 d后測量發(fā)現(xiàn),與對照組相比,辣椒苗株高、根長、鮮重和干重分別增加:50.04%、48.16%、73.27%和 48.83%。3、枯草芽孢桿菌BS193無菌濾液對辣椒疫霉菌菌絲生長、孢子囊形成、游動孢子萌發(fā)等具有顯著的抑制活性。試驗結(jié)果表明經(jīng)菌株BS193無菌濾液處理后菌絲生長速率顯著下降,在光學(xué)和電子顯微鏡下觀察到菌絲分枝增多、畸形膨大、扭曲變形、皺縮等;辣椒疫霉孢子囊和游動孢子萌發(fā)的數(shù)量均明顯減少,其抑制率都顯著上升。4、初步鑒定BS193所合成的抗菌物質(zhì):采用濃鹽酸沉淀法粗提抗菌物質(zhì),檢測抑菌活性并進行ULPC-QTOF/MS分析。試驗結(jié)果顯示抗菌粗提物對辣椒疫霉菌菌絲生長抑制效果顯著;經(jīng)ULPC-QTOF/MS分析出現(xiàn)多處明顯的質(zhì)譜響應(yīng)峰,核質(zhì)比m/z值比較,初步鑒定抗菌粗提物中含有Iturin(伊枯草菌素)、Feycin(豐原菌素)、Surfactin(表面活性素)這3類脂肽類抗菌物質(zhì)。
[Abstract]:Pepper blight caused by Phytophthora capsicii is one of the most important diseases in pepper production. At present, the most convenient and effective way to control pepper blight in production is to use chemical chemicals, but long-term application of insecticides makes pathogens easily develop resistance to certain drugs, and the pesticide residue problem is prominent. Therefore, it is of great significance to find efficient and non-polluting methods for the prevention and control of pepper blight, among which the biological control with biocontrol bacteria is one of the most promising methods. In this study, a biocontrol strain of Bacillus subtilis (BS193) was isolated in the early stage of laboratory. The antimicrobial spectrum, biocontrol effect and antibacterial mechanism of this strain were studied. It provides a practical basis for the biological control of pepper blight and the further development and application of this biocontrol bacterium. The results showed that the antimicrobial activity of Bacillus subtilis BS193 was broad spectrum. The antimicrobial spectrum of Bacillus subtilis BS193 was determined by plate confrontation method. Bacillus subtilis BS193 can inhibit 9 genera and 10 common plant pathogens. Bacillus subtilis BS193 has a good biocontrol effect on pepper blight. In the plate confrontation test, the results showed that the inhibition effect of the strain BS193 on the four strains of Phytophthora capsici was stable, and in vitro, the inhibition of Phytophthora capsici infection on leaves was shown by the treatment of BS193 fermentation broth. The control effect reached 63.50. The results showed that BS193 bacteria, aseptic filtrate, and sterility filtrate were used in pot experiment. The control effects of the fermentation broth after 24 h inoculation with pathogenic bacteria for 9 d were as follows: 1: 30.75 and 29.76% and 52.56% BS193 100 times diluent and 1000 times diluent to treat capsicum seedlings, respectively. 9 days after inoculation with pathogenic bacteria, the control effect was:% 52.56 and 34.560.The control effect of each treatment after inoculating the pathogen 9 days after inoculation with BS193 fermentation broth was 0.24 and 4872 hours after inoculation, and the control effects were respectively 51.441.49.41and 51.441.41.41.41.41.41.41.41.41.41.41.41.41.The control effect of each treatment was as follows: 59.41.41.41.44 after inoculating the pathogen for 9 days. It can be seen that Bacillus subtilis BS193 has a good effect on preventing pepper blight. At the same time, it was found that BS193 broth had a good effect on the growth of pepper seedlings. Compared with the control group, the plant height, root length, fresh weight and dry weight of capsicum seedlings were increased by 73.27% and 48.833.27% and 48.833.3.3respectively, respectively. The growth of Phytophthora capsici hyphae and the formation of sporangia were observed by BS193 aseptic filtrate of Bacillus subtilis. The germination of zoospore showed significant inhibitory activity. The results showed that the growth rate of mycelium decreased significantly after treatment with BS193 aseptic filtrate. The mycelium branching, deformity, distortion and shrinkage were observed under optical and electron microscopy. The number of sporangia and zoospore germination of Phytophthora capsici was significantly reduced, and the inhibition rate was increased significantly. The antimicrobial substances synthesized by BS193 were preliminarily identified. The antimicrobial substances were extracted by concentrated hydrochloric acid precipitation method, the antibacterial activity was detected and ULPC-QTOF/MS analysis was carried out. The results showed that the antimicrobial crude extract had significant inhibitory effect on the growth of Phytophthora capsici hyphae, and there were many obvious mass spectrum response peaks by ULPC-QTOF/MS analysis, and the ratio of nucleus to cytoplasm m / z was compared. Three kinds of lipopeptide antimicrobial substances Iturin (Iturin) Feycinin (surfacin) were identified in the crude antimicrobial extracts.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S436.418

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