本文關(guān)鍵詞： 對(duì)羥基苯甲酸 抗氧化酶 土壤酶 原兒茶酸 Ensifer adhaerens Streptomyces griseoruber Streptomyces canus　出處：《山東農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：黃瓜是我國(guó)主要栽培蔬菜之一,溫室栽培的連續(xù)種植方式容易引起連作障礙。對(duì)羥基苯甲酸(p-hydroxybenzonic acid,PHBA)是引起連作障礙的主要自毒物質(zhì)之一。本實(shí)驗(yàn)從連作黃瓜根際土中篩選PHBA降解菌,研究降解菌的生理特性,降解菌對(duì)黃瓜抗氧化酶、土壤酶活性的影響,評(píng)估其對(duì)作物連作的緩解作用。具體研究結(jié)果如下,(1)篩選出對(duì)PHBA降解能力高的3株菌株。結(jié)合生理生化和分子鑒定結(jié)果,放線菌GLY-P1和GLY-P2分別被鑒定為Streptomyces griseoruber和Streptomyces canus,細(xì)菌GLY-P3被鑒定為Ensifer adhaerens。(2)Streptomyces griseoruber GLY-P1和Streptomyces canus GLY-P2在32?C、p H10、PHBA初始濃度為0.5g/L條件下降解率最高。在偏堿土壤中,Streptomyces griseoruber GLY-P1和Streptomyces canus GLY-P2降解率較高。Ensifer adhaerens GLY-P3最適降解條件為溫度30?C,p H7,PHBA初始濃度為0.5g/L,氮源為硫酸銨,接種量8%(8×109cfu)。在最優(yōu)降解條件下,12h內(nèi)Ensifer adhaerens GLY-P3降解PHBA99.2%。(3)推測(cè)Streptomyces griseoruber GLY-P1、Streptomyces canus GLY-P2和Ensifer adhaerens GLY-P3代謝PHBA途徑可能為原兒茶酸途徑。在32?C、p H7、PHBA初始濃度為0.5g/L條件下,Streptomyces griseoruber GLY-P1和Streptomyces canus GLY-P2發(fā)酵液中原兒茶酸的最大含量分別為3.2mg/L和3.8mg/L。(4)將菌株Streptomyces griseoruber GLY-P1和Streptomyces canus GLY-P2置于PHBA環(huán)境中,體內(nèi)SOD、CAT、DHAR和APX活性發(fā)生明顯改變,菌株Ensifer adhaerens GLY-P3處于PHBA環(huán)境中,體內(nèi)SOD、CAT、DHAR和MDHAR活性發(fā)生明顯改變,說(shuō)明在響應(yīng)PHBA脅迫中,SOD、CAT、DHAR和APX在菌株Streptomyces griseoruber GLY-P1和Streptomyces canus GLY-P2起主要作用,SOD、CAT、DHAR和MDHAR在菌株Ensifer adhaerens GLY-P3起主要作用。(5)當(dāng)在每克土壤中施加220微克PHBA時(shí),黃瓜葉片中H2O2、O2.-、MDA含量和SOD、POD、APX、DHAR活性都明顯升高,當(dāng)土壤中施加GLY-P3時(shí),黃瓜中H2O2、O2.-、MDA含量和SOD、CAT、POD活性都明顯降低,表明PHBA對(duì)黃瓜造成脅迫作用,GLY-P3緩解了PHBA對(duì)黃瓜的脅迫作用。土壤中PHBA的施加,使黃瓜根際土壤磷酸酶、脲酶和蔗糖酶活性都普遍的升高了而過(guò)氧化氫酶活性降低。GLY-P3的施加使過(guò)氧化氫酶活性明顯的升高了,土壤磷酸酶、脲酶和蔗糖酶活性都普遍的降低了,緩解了PHBA對(duì)土壤酶活的影響。結(jié)果表明Ensifer adhaerens GLY-P3能通過(guò)調(diào)節(jié)黃瓜葉片抗氧化酶、土壤的酶活性,緩解黃瓜的連作障礙。
[Abstract]:Cucumber is one of the main cultivated vegetables in China. Continuous planting in greenhouse is easy to cause continuous cropping obstacle. P-hydroxybenzonic acidonic acid (p-hydroxybenzoate) is one of the main autotoxic substances causing continuous cropping. In this experiment, PHBA degrading bacteria were screened from the rhizosphere soil of continuous cropping cucumber, and the physiological characteristics of degrading bacteria were studied. The effects of degrading bacteria on antioxidant enzymes and soil enzyme activities of cucumber were evaluated. The results were as follows: 1) three strains with high degradation ability of PHBA were screened, and the results of physiological, biochemical and molecular identification were combined with the results of physiological, biochemical and molecular identification. Actinomycetes GLY-P1 and GLY-P2 were identified as Streptomyces griseoruber and Streptomyces Canus, respectively. Bacterial GLY-P3 was identified as Ensifer adhaerens.(2)Streptomyces griseoruber GLY-P1 and Streptomyces canus GLY-P2. The degradation rate was the highest at the initial concentration of 0.5 g / L. the optimum conditions for degradation of Streptomyces griseoruber GLY-P1 and Streptomyces canus GLY-P2 were 30? The initial concentration of Ensifer adhaerens GLY-P3 was 0.5 g / L, the nitrogen source was ammonium sulfate, and the inoculation amount was 8 脳 10 9 cfu.In the optimal degradation conditions, the degradation of PHBA99.2. 3 by Ensifer adhaerens GLY-P3 within 12 hours was deduced that Streptomyces griseoruber GLY-P 1 Streptomyces canus GLY-P2 and Ensifer adhaerens GLY-P3 metabolism PHBA pathway might be protocatecholic acid pathway. The maximum contents of protocatechuic acid in the fermentation broth of Streptomyces griseoruber GLY-P1 and Streptomyces canus GLY-P2 were 3.2 mg / L and 3.8 mg / L respectively at the initial concentration of 0.5 g / L). The strains Streptomyces griseoruber GLY-P1 and Streptomyces canus GLY-P2 were placed in PHBA environment. The activities of SODCATDHAR and APX were significantly changed. The Ensifer adhaerens GLY-P3 was in the PHBA environment, and the activities of MDHAR and DHAR were obviously changed. The results showed that in response to PHBA stress, APX and APX played a major role in Streptomyces griseoruber GLY-P1 and Streptomyces canus GLY-P2. Streptomyces griseoruber CATHAR and MDHAR played a major role in Ensifer adhaerens GLY-P3.) when 220 渭 g PHBA was applied to the soil per gram, the contents of H _ 2O _ 2O _ 2.-malondialdehyde and the activity of SODPOD _ (2) -malondialdehyde (PHBA) in cucumber leaves were significantly increased. When GLY-P3 was applied to the soil, the content of H _ 2O _ 2O _ 2.-malondialdehyde (MDA) and the activity of POD in cucumber were significantly decreased, which indicated that PHBA caused stress on cucumber and GLY-P3 alleviated the stress of PHBA on cucumber. The application of PHBA in soil resulted in phosphatase in cucumber rhizosphere. The activities of urease and sucrase increased generally, but the activity of catalase decreased. GLY-P3 significantly increased the activity of catalase, and the activities of phosphatase, urease and sucrase in soil were all decreased. The effect of PHBA on soil enzyme activity was alleviated. The results showed that Ensifer adhaerens GLY-P3 could alleviate the obstacle of continuous cropping by regulating the antioxidant enzyme activity and soil enzyme activity in cucumber leaves.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S642.2;S182

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