拮抗微生物對列當(dāng)?shù)姆莱饔眉皺C理
發(fā)布時間：2018-07-16 23:36
【摘要】：根寄生雜草列當(dāng)對全球許多地區(qū)農(nóng)業(yè)生產(chǎn)造成嚴(yán)重危害。目前,有效防除列當(dāng)仍是一個世界性難題。微生物防治是防除列當(dāng)?shù)挠行緩街?但關(guān)于列當(dāng)生防微生物的研究目前多集中于列當(dāng)病原菌上。本研究以篩選自來源于健康土壤的上萬株微生物資源庫中對多種作物病害病原菌有拮抗作用的放線菌和真菌為試驗材料,通過培養(yǎng)皿內(nèi)種子萌發(fā)抑制試驗篩選向日葵列當(dāng)和瓜列當(dāng)?shù)纳谰?通過皿內(nèi)共培養(yǎng)試驗結(jié)合盆栽及田間試驗對篩選出的生防菌株防除列當(dāng)?shù)男ЧM行了驗證并對生防菌株防除列當(dāng)?shù)臋C理進行了研究;通過盆栽試驗研究了列當(dāng)生防菌株對寄主作物生長的影響。此外,本研究還通過皿內(nèi)種子萌發(fā)試驗從供試拮抗微生物中篩選具有誘導(dǎo)列當(dāng)種子萌發(fā)潛能的菌株。本研究所得主要結(jié)論如下:(1)向日葵列當(dāng)和瓜列當(dāng)拮抗微生物的篩選。本研究篩選出強烈抑制向日葵列當(dāng)種子萌發(fā)的放線菌和真菌各1株,分別為淡紫褐鏈霉菌(Streptomyces enissocaesilis,509)和灰黃青霉(Penicillium griseofulvum,CF3);同時還篩選出強烈抑制瓜列當(dāng)種子萌發(fā)的放線菌2株(密旋鏈霉菌S.pactum,12#;黃白鏈霉菌S.albidoflavus,T4)和真菌1株,CF3。在3.5 mg ml-1濃度下,509和CF3的無細(xì)胞發(fā)酵濾液對向日葵列當(dāng)及12#、T4和CF3的無細(xì)胞發(fā)酵濾液對瓜列當(dāng)種子萌發(fā)的抑制率均達(dá)到75%以上。在50.0 mg ml-1濃度下,509和CF3的菌體甲醇浸提液對向日葵列當(dāng)種子萌發(fā)的抑制率分別為14.8%和100.0%;而12#、T4和CF3的菌體甲醇浸提液對瓜列當(dāng)種子萌發(fā)的抑制率為50.7%~100.0%。在皿內(nèi)共培養(yǎng)試驗中,509和CF3的無細(xì)胞發(fā)酵濾液均能夠顯著抑制向日葵列當(dāng)種子的萌發(fā)。CF3的無細(xì)胞發(fā)酵濾液在皿內(nèi)共培養(yǎng)8天后對瓜列當(dāng)種子萌發(fā)的抑制率仍高達(dá)47.5%~80.3%。(2)放線菌防除列當(dāng)作用研究。盆栽試驗中,施加1.0 g kg-1的509菌劑使收獲期向日葵列當(dāng)?shù)某鐾翑?shù)量和寄生總數(shù)較對照分別降低了39.2%~47.5%和39.3%~62.4%。在楊凌盆栽試驗中,施加1.0 g kg-1的509菌劑使生長中期和收獲期向日葵列當(dāng)?shù)目偢芍嘏c對照相比分別降低了46.9%和36.7%。施加12#菌劑使盆栽試驗收獲期瓜列當(dāng)?shù)某鐾翑?shù)量、出土率和總干重較對照分別降低了85.7%、75.7%和55.4%。(3)真菌防除列當(dāng)作用研究。楊凌盆栽試驗中,施加1.0 g kg-1的CF3菌劑使收獲期向日葵列當(dāng)?shù)募纳倲?shù)和總干重較對照分別降低了25.2%和46.0%。施加CF3菌劑使盆栽試驗收獲期瓜列當(dāng)?shù)某鐾翑?shù)量和出土率較對照分別降低了76.2%和85.3%。進一步研究發(fā)現(xiàn)展青霉素是CF3抑制列當(dāng)種子萌發(fā)的主要活性物質(zhì)。展青霉素和CF3無細(xì)胞發(fā)酵濾液的乙酸乙酯萃取物在1.0 mg ml-1濃度下均能夠完全抑制向日葵列當(dāng)和瓜列當(dāng)種子的萌發(fā)。(4)拮抗微生物對列當(dāng)寄主的促生作用研究。在有列當(dāng)寄生的條件下,施加509和CF3菌劑顯著促進了盆栽向日葵的生長,增加了向日葵的生物量及籽粒干重。其中,拌土施加509和CF3菌劑使楊凌盆栽試驗收獲期向日葵的株高增加了39.1%~63.4%,地上干重增加了114.7%~179.0%。施加12#和CF3菌劑使盆栽試驗收獲期番茄產(chǎn)量與對照相比分別增加了51.6%和84.9%。在無列當(dāng)寄生條件下,施加1.0 g kg-1的509和CF3菌劑使盆栽試驗收獲期向日葵的產(chǎn)量較對照分別增加了79.2%和46.6%,而施加12#菌劑使收獲期番茄產(chǎn)量增加了57.3%。(5)拮抗微生物誘導(dǎo)列當(dāng)種子萌發(fā)研究。從供試拮抗微生物中分別篩選出能夠誘導(dǎo)瓜列當(dāng)種子萌發(fā)的放線菌(鏈霉菌Streptomyces sp.,Act13;球孢鏈霉菌球孢亞種S.globisporus subsp.globisporus,D141;加利利鏈霉菌S.galilaeus,G37)3株,能夠誘導(dǎo)向日葵列當(dāng)種子萌發(fā)的放線菌(球孢鏈霉菌S.globisporus,C28和鏈霉菌Streptomyces sp.,S8)和真菌(木霉Trichoderma spp.;M1,M2)各2株。試驗結(jié)果表明Act13、D141和G37的無細(xì)胞發(fā)酵濾液在0.35 mg ml-1濃度下可分別誘導(dǎo)14.7%、14.9%和15.4%的瓜列當(dāng)種子萌發(fā)。C28的無細(xì)胞發(fā)酵濾液在3.5×10-4 mg ml-1濃度下可誘導(dǎo)13.4%的向日葵列當(dāng)種子萌發(fā)。S8、M1和M2的菌體甲醇浸提液在50.0 mg ml-1濃度下分別誘導(dǎo)了23.4%、17.0%和15.5%的向日葵列當(dāng)種子萌發(fā)。(6)拮抗微生物防除列當(dāng)機理研究。試驗結(jié)果表明拮抗微生物主要通過以下3種機理來防除列當(dāng):(1)通過改變寄主根區(qū)土壤中微生物區(qū)系來抑制列當(dāng)?shù)募纳蜕L。盆栽試驗中,向日葵列當(dāng)?shù)目倲?shù)及出土數(shù)與向日葵根區(qū)土壤中放線菌和細(xì)菌的數(shù)量及比例呈顯著負(fù)相關(guān)關(guān)系。瓜列當(dāng)?shù)某鐾翑?shù)和出土率與放線菌與真菌的數(shù)量之比(A/F)、細(xì)菌與真菌的數(shù)量之比(B/F)呈負(fù)相關(guān)關(guān)系。施加1.0 g kg-1的509菌劑使盆栽試驗向日葵根區(qū)土壤中放線菌的數(shù)量較對照增加了76.7%~439.9%。施加CF3、509和12#也顯著增加了寄主根區(qū)土壤中A/F和B/F。(2)通過增強寄主植物的防御酶系統(tǒng)抗性來阻礙列當(dāng)?shù)募纳蜕L。固原盆栽試驗中向日葵列當(dāng)?shù)目倲?shù)、鮮重及干重均與向日葵根系中多酚氧化酶PPO活力呈顯著負(fù)相關(guān)。施加509菌劑使寄主向日葵根系中PPO活力與對照相比增加了43.1%。(3)通過抑制列當(dāng)種子的萌發(fā)、芽管的伸長,影響芽管的形態(tài)來直接防除列當(dāng)。在0.35 mg ml-1濃度下,509和12#的無細(xì)胞發(fā)酵濾液使向日葵列當(dāng)芽管長度較對照分別減少了86.2%和89.6%,而CF3和12#的無細(xì)胞發(fā)酵濾液也使瓜列當(dāng)芽管長度分別縮短了68.8%和55.6%。經(jīng)509無細(xì)胞發(fā)酵濾液和CF3菌體甲醇浸提液處理后,部分向日葵列當(dāng)?shù)难抗馨l(fā)生了褐變。
[Abstract]:Root parasitic weeds have caused serious harm to agricultural production in many parts of the world. At present, effective prevention and control is still a worldwide problem. Microbial control is one of the effective ways to prevent and control, but the research on Biocontrol microorganism is mainly concentrated on the pathogenic bacteria. This study is to screen from healthy soil. Strains of actinomycetes and fungi, which have antagonistic effects on the pathogens of various crop diseases, were used as experimental materials to screen the biocontrol strains of sunflower and cucumber by seed germination inhibition test in a Petri dish. The effect was verified and the mechanism of the anti-bacterial strain was studied. The effect of the Biocontrol Strain on the growth of the host crops was studied by a pot experiment. In addition, the study also screened the strains with the potential of the Seed Germination from the tested antagonist microorganism in the dish seed germination test. The following conclusions are as follows: (1) screening of the antagonistic microorganisms from the sunflower and the cucumber. This study screened out 1 strains of actinomycetes and fungi which strongly inhibited the germination of sunflower, Streptomyces enissocaesilis, 509, and Penicillium griseofulvum, CF3, respectively. When 2 strains of actinomycetes (Streptomyces S.pactum, 12#; Streptomyces yellow and white fungus S.albidoflavus, T4) and 1 strains of fungi were germinated, CF3. at the concentration of 3.5 mg ml-1, 509 and CF3 without cell-free filtrating filtrate of sunflower, 12#, T4 and CF3 on the germination of melon seeds reached more than 75%. In 50 mg The inhibitory rates of methanol extracts from 509 and CF3 on the germination of sunflower seeds were 14.8% and 100%, respectively, while the inhibition rate of methanol extracts from 12#, T4 and CF3 was 50.7%~100.0%. in the co culture test of 50.7%~100.0%. in the dish, and the 509 and CF3 filtrate free filtrates could significantly inhibit the sunflower seeded species. The inhibitory rate of the seedless fermentation filtrate of.CF3 on seed germination was still up to 47.5%~80.3%. (2) of actinomycetes in the germination of seed germination of the seedless fermentation filtrate of the seed of.CF3. In pot experiment, the number of unearthed and parasitism of the sunflower in the harvest period decreased by 39.2%~47.5% and 3, respectively, by applying the 509 agent of 1 g kg-1. In the Yangling pot experiment, the total dry weight of 1 g kg-1 was applied to the total dry weight of 1 g kg-1 and the total dry weight of sunflower in the mid growth and harvest period was reduced by 46.9% and 36.7%., respectively. The amount of unearthed melon in the harvest period of the pot experiment was reduced by 85.7%, 75.7% and 55.4%. (3), respectively. In the pot experiment in Yangling, the 1 g kg-1 inoculants were applied to reduce the parasitic total and total dry weight of sunflower in the harvest period by 25.2% and 46.0%. by applying CF3 bacteria, respectively. The amount of unearthed soil and the unearthed rate of the melon in the harvest period were reduced by 76.2% and 85.3%., respectively, by 76.2% and 85.3%. respectively. It was found that penicillin was the main active substance for CF3 inhibition of seed germination. The ethyl acetate extract of penicillin and CF3 without cell fermentation filtrate could completely inhibit the germination of sunflower and cucumber seeds at the concentration of 1 mg ml-1. (4) study on the growth promoting effect of antagonistic microorganisms on the host. The application of 509 and CF3 inoculants significantly promoted the growth of sunflower in potted plants and increased the biomass and dry weight of sunflower. Among them, 509 and CF3 inoculants in the mixed soil increased the plant height of the sunflower in the harvest period of Yangling, and the dry weight of the ground was increased by 114.7%~ 179.0%., and 12# and CF3 were applied to the harvest period of the pot experiment. Compared with the control, tomato yield increased by 51.6% and 84.9%., respectively, under the parasitic conditions. The 1 g kg-1 509 and CF3 inoculants increased the yield of sunflower in the harvest period by 79.2% and 46.6% respectively, while the 12# inoculant increased the yield of the harvested tomato with 57.3%. (5) antagonistic microorganisms to induce the seed germination. 3 strains of actinomycetes (Streptomyces Streptomyces sp., Act13, S.globisporus subsp.globisporus, D141, S.galilaeus, G37) of Streptomyces Galilee were screened from the tested antagonist microorganism to induce seed germination respectively, which could guide the actinomycetes (Streptomyces spores S.) as seed germination of sunflower (Streptomyces spores S.). 2 strains of globisporus, C28 and Streptomyces Streptomyces sp., S8) and fungi (Trichoderma Trichoderma spp.; M1, M2). The results showed that the cell-free filtrate of Act13, D141 and G37 could induce 14.7%, 14.9% and 15.4% of the seedless fermentation filtrate of seed germination respectively in the concentration of 3.5 * 10-4. The 13.4% sunflower seed germination was.S8, the methanol extract of M1 and M2 induced 23.4%, 17% and 15.5% of sunflower seed germination at the concentration of 50 mg ml-1. (6) the antagonistic microorganism defense mechanism was studied. The experimental results showed that the antagonistic microorganisms were mainly through the following 3 mechanisms to prevent and treat: (1) through change. Parasitism and growth of the soil in the soil of the main root zone were sent to inhibit the parasitism and growth of the column. In the pot experiment, the total number and unearthed number of sunflower were negatively correlated with the number and proportion of actinomycetes and bacteria in the soil of the sunflower root region. The number of unearthed soil and the ratio of unearthed soil to the number of actinomycetes and fungi (A/F), bacteria and true The number of bacteria was negatively correlated with the ratio of B/F. The number of 509 inoculants with 1 g kg-1 increased the number of actinomycetes in the soil of the sunflower root zone increased by 76.7%~439.9%. and CF3509 and 12# significantly increased A/F and B/F. (2) in the soil of the main root region. The total number, fresh weight and dry weight of sunflower in Guyuan pot experiment were all negatively correlated with the activity of polyphenol oxidase PPO in sunflower root system. The effect of 509 inoculant on PPO activity in the root of sunflower was increased by 43.1%. (3) by inhibiting the germination of the seed, the elongation of the bud tube and the shape of the bud tube. Under the concentration of 0.35 mg ml-1, 509 and 12# cell-free filtrates reduced the length of the sunflower bud tube by 86.2% and 89.6% respectively, while the CF3 and 12# cell-free filtrates shortened the length of the melon bud tube by 68.8% and 55.6%. through 509 fine cell fermentation filtrate and CF3 bacterial extract solution. After that, some of the sunflower bud tubes were browning.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S45
，
本文編號：2128060
[Abstract]:Root parasitic weeds have caused serious harm to agricultural production in many parts of the world. At present, effective prevention and control is still a worldwide problem. Microbial control is one of the effective ways to prevent and control, but the research on Biocontrol microorganism is mainly concentrated on the pathogenic bacteria. This study is to screen from healthy soil. Strains of actinomycetes and fungi, which have antagonistic effects on the pathogens of various crop diseases, were used as experimental materials to screen the biocontrol strains of sunflower and cucumber by seed germination inhibition test in a Petri dish. The effect was verified and the mechanism of the anti-bacterial strain was studied. The effect of the Biocontrol Strain on the growth of the host crops was studied by a pot experiment. In addition, the study also screened the strains with the potential of the Seed Germination from the tested antagonist microorganism in the dish seed germination test. The following conclusions are as follows: (1) screening of the antagonistic microorganisms from the sunflower and the cucumber. This study screened out 1 strains of actinomycetes and fungi which strongly inhibited the germination of sunflower, Streptomyces enissocaesilis, 509, and Penicillium griseofulvum, CF3, respectively. When 2 strains of actinomycetes (Streptomyces S.pactum, 12#; Streptomyces yellow and white fungus S.albidoflavus, T4) and 1 strains of fungi were germinated, CF3. at the concentration of 3.5 mg ml-1, 509 and CF3 without cell-free filtrating filtrate of sunflower, 12#, T4 and CF3 on the germination of melon seeds reached more than 75%. In 50 mg The inhibitory rates of methanol extracts from 509 and CF3 on the germination of sunflower seeds were 14.8% and 100%, respectively, while the inhibition rate of methanol extracts from 12#, T4 and CF3 was 50.7%~100.0%. in the co culture test of 50.7%~100.0%. in the dish, and the 509 and CF3 filtrate free filtrates could significantly inhibit the sunflower seeded species. The inhibitory rate of the seedless fermentation filtrate of.CF3 on seed germination was still up to 47.5%~80.3%. (2) of actinomycetes in the germination of seed germination of the seedless fermentation filtrate of the seed of.CF3. In pot experiment, the number of unearthed and parasitism of the sunflower in the harvest period decreased by 39.2%~47.5% and 3, respectively, by applying the 509 agent of 1 g kg-1. In the Yangling pot experiment, the total dry weight of 1 g kg-1 was applied to the total dry weight of 1 g kg-1 and the total dry weight of sunflower in the mid growth and harvest period was reduced by 46.9% and 36.7%., respectively. The amount of unearthed melon in the harvest period of the pot experiment was reduced by 85.7%, 75.7% and 55.4%. (3), respectively. In the pot experiment in Yangling, the 1 g kg-1 inoculants were applied to reduce the parasitic total and total dry weight of sunflower in the harvest period by 25.2% and 46.0%. by applying CF3 bacteria, respectively. The amount of unearthed soil and the unearthed rate of the melon in the harvest period were reduced by 76.2% and 85.3%., respectively, by 76.2% and 85.3%. respectively. It was found that penicillin was the main active substance for CF3 inhibition of seed germination. The ethyl acetate extract of penicillin and CF3 without cell fermentation filtrate could completely inhibit the germination of sunflower and cucumber seeds at the concentration of 1 mg ml-1. (4) study on the growth promoting effect of antagonistic microorganisms on the host. The application of 509 and CF3 inoculants significantly promoted the growth of sunflower in potted plants and increased the biomass and dry weight of sunflower. Among them, 509 and CF3 inoculants in the mixed soil increased the plant height of the sunflower in the harvest period of Yangling, and the dry weight of the ground was increased by 114.7%~ 179.0%., and 12# and CF3 were applied to the harvest period of the pot experiment. Compared with the control, tomato yield increased by 51.6% and 84.9%., respectively, under the parasitic conditions. The 1 g kg-1 509 and CF3 inoculants increased the yield of sunflower in the harvest period by 79.2% and 46.6% respectively, while the 12# inoculant increased the yield of the harvested tomato with 57.3%. (5) antagonistic microorganisms to induce the seed germination. 3 strains of actinomycetes (Streptomyces Streptomyces sp., Act13, S.globisporus subsp.globisporus, D141, S.galilaeus, G37) of Streptomyces Galilee were screened from the tested antagonist microorganism to induce seed germination respectively, which could guide the actinomycetes (Streptomyces spores S.) as seed germination of sunflower (Streptomyces spores S.). 2 strains of globisporus, C28 and Streptomyces Streptomyces sp., S8) and fungi (Trichoderma Trichoderma spp.; M1, M2). The results showed that the cell-free filtrate of Act13, D141 and G37 could induce 14.7%, 14.9% and 15.4% of the seedless fermentation filtrate of seed germination respectively in the concentration of 3.5 * 10-4. The 13.4% sunflower seed germination was.S8, the methanol extract of M1 and M2 induced 23.4%, 17% and 15.5% of sunflower seed germination at the concentration of 50 mg ml-1. (6) the antagonistic microorganism defense mechanism was studied. The experimental results showed that the antagonistic microorganisms were mainly through the following 3 mechanisms to prevent and treat: (1) through change. Parasitism and growth of the soil in the soil of the main root zone were sent to inhibit the parasitism and growth of the column. In the pot experiment, the total number and unearthed number of sunflower were negatively correlated with the number and proportion of actinomycetes and bacteria in the soil of the sunflower root region. The number of unearthed soil and the ratio of unearthed soil to the number of actinomycetes and fungi (A/F), bacteria and true The number of bacteria was negatively correlated with the ratio of B/F. The number of 509 inoculants with 1 g kg-1 increased the number of actinomycetes in the soil of the sunflower root zone increased by 76.7%~439.9%. and CF3509 and 12# significantly increased A/F and B/F. (2) in the soil of the main root region. The total number, fresh weight and dry weight of sunflower in Guyuan pot experiment were all negatively correlated with the activity of polyphenol oxidase PPO in sunflower root system. The effect of 509 inoculant on PPO activity in the root of sunflower was increased by 43.1%. (3) by inhibiting the germination of the seed, the elongation of the bud tube and the shape of the bud tube. Under the concentration of 0.35 mg ml-1, 509 and 12# cell-free filtrates reduced the length of the sunflower bud tube by 86.2% and 89.6% respectively, while the CF3 and 12# cell-free filtrates shortened the length of the melon bud tube by 68.8% and 55.6%. through 509 fine cell fermentation filtrate and CF3 bacterial extract solution. After that, some of the sunflower bud tubes were browning.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S45
，
本文編號：2128060
本文鏈接：http://sikaile.net/shoufeilunwen/nykjbs/2128060.html
最近更新
教材專著
