不同形態(tài)氮鹽對(duì)銅綠假單胞菌SD-1種群穩(wěn)定性的影響及機(jī)制研究
發(fā)布時(shí)間：2018-11-01 19:13
【摘要】：銅綠假單胞菌SD-1是一株高效雜環(huán)化合物降解菌株,可通過生物強(qiáng)化的方式在污染物治理中發(fā)揮重要作用。已有研究提示,強(qiáng)化菌株的穩(wěn)定性對(duì)于生物強(qiáng)化系統(tǒng)穩(wěn)定運(yùn)行具有重要意義。雖然有大量報(bào)道關(guān)注了強(qiáng)化菌株在生態(tài)系統(tǒng)中的種間競(jìng)爭(zhēng),但目前尚未有研究涉及強(qiáng)化菌株的種內(nèi)生態(tài)穩(wěn)定性。鑒于此,本論文以銅綠假單胞菌SD-1作為試驗(yàn)菌株,通過連續(xù)傳代的方式,探究了廢水處理系統(tǒng)中不同形態(tài)氮鹽對(duì)菌株種群穩(wěn)定性的影響;對(duì)比野生株和群體感應(yīng)欺騙株在不同形態(tài)氮鹽中的生長差異,從代謝角度解析了不同形態(tài)氮鹽對(duì)菌株種群穩(wěn)定影響的群體感應(yīng)機(jī)制;此外,通過對(duì)比不同形態(tài)氮鹽條件下分離的進(jìn)化合作者群體感應(yīng)效應(yīng)差異,進(jìn)一步從進(jìn)化角度探究了不同形態(tài)氮鹽對(duì)菌株種群穩(wěn)定影響的群體感應(yīng)機(jī)制。該研究對(duì)于豐富群體感應(yīng)理論、拓展群體感應(yīng)理論在生物強(qiáng)化系統(tǒng)中的應(yīng)用、優(yōu)化生物強(qiáng)化系統(tǒng)的運(yùn)行策略具有重要意義。所獲得的主要結(jié)果如下:(1)長期進(jìn)化過程中,部分組別菌群密度下降了 1-2個(gè)數(shù)量級(jí),出現(xiàn)種群崩潰現(xiàn)象。菌株在外加銨鹽,硝酸鹽、亞硝酸鹽和不外加氮鹽的對(duì)照組別中崩潰的機(jī)率分別為0%,33.3%,66.6%和11.1%。穩(wěn)定組別中,欺騙子比例范圍為0%-53%,崩潰組別中欺騙子比例為51%-81%。崩潰組別中,蛋白酶含量顯著下降。不同形態(tài)的外加氮鹽對(duì)菌株SD-1種群穩(wěn)定性產(chǎn)生顯著影響,外加亞硝酸鹽和硝酸鹽組別中種群更容易崩潰。(2)對(duì)不同培養(yǎng)條件下菌株SD-1氮源利用情況進(jìn)行對(duì)比,顯示氨氮、硝氮的變化趨勢(shì)相同,但亞硝氮的變化趨勢(shì)存在差異。在硝酸鹽和亞硝酸鹽培養(yǎng)基中,菌株SD-1公共物質(zhì)分泌要較其他組別中高2-3倍。在外加硝酸鹽的葡萄糖培養(yǎng)基中,野生株和欺騙子利用氨氮的能力相同;但在外加硝酸鹽的葡萄糖培養(yǎng)基中,欺騙子前12 h比合作者生長更快;而外加亞硝酸鹽的葡萄糖培養(yǎng)基中,欺騙子的生長則始終高于合作者。硝酸鹽和亞硝酸鹽使合作者菌株肩負(fù)了更多的公共物質(zhì)代謝負(fù)擔(dān),同時(shí)使得欺騙子菌株獲得了較之合作者更高的生存優(yōu)勢(shì),即群體感應(yīng)介導(dǎo)的代謝差異是外加亞硝酸鹽和硝酸鹽條件下種群更容易崩潰的根本原因。(3)從長期進(jìn)化試驗(yàn)中篩選了 24株進(jìn)化合作者,其群體感應(yīng)均發(fā)生了顯著變化。雖然進(jìn)化菌株與初始合作者菌株的生長無顯著差異,但是所有進(jìn)化菌株的30C12-HSL,C4VHSL和蛋白酶產(chǎn)量不同程度地低于初始合作者菌株,變得更加"謹(jǐn)慎";而所有進(jìn)化菌株的綠膿素合成不同程度地高于初始合作者菌株,穩(wěn)定維持機(jī)制被加強(qiáng)。合作行為"謹(jǐn)慎"進(jìn)化以及維穩(wěn)能力提升是菌群在長期進(jìn)化中維持穩(wěn)定的重要原因。
[Abstract]:Pseudomonas aeruginosa SD-1 (Pseudomonas aeruginosa) is a high efficient heterocyclic compound degrading strain which can play an important role in pollutant treatment through biointensification. It has been suggested that the stability of the enhanced strain is of great significance for the stable operation of the biological strengthening system. Although there have been a lot of reports on the inter-species competition of the enhanced strains in the ecosystem, there are no studies on the intraspecific ecological stability of the enhanced strains. In this paper, using Pseudomonas aeruginosa SD-1 as the experimental strain, the effects of different nitrogen salts in wastewater treatment system on the population stability of the strains were studied by means of continuous passage. By comparing the growth differences of wild plants and colony sensing deceptive strains in different forms of nitrogen salts, the effects of different forms of nitrogen salts on the population stability of strains were analyzed from the point of view of metabolism. In addition, by comparing the population induction effects of evolutionary collaborators with different nitrogen salts, the population sensing mechanism of different forms of nitrogen salts on the population stability of strains was further explored from the perspective of evolution. This study is of great significance to enrich the theory of group induction, expand the application of group induction theory in biological reinforcement system, and optimize the operation strategy of biological reinforcement system. The main results are as follows: (1) during the long-term evolution, the density of some groups of bacteria decreased by 1-2 orders of magnitude, resulting in the phenomenon of population collapse. The probability of the strain collapsing in the control group with added ammonium salt, nitrate, nitrite and no nitrogen salt was 66.6% and 11.1%, respectively. In the stable group, the deceptive sub-ratio ranges from 0 to 53, and in the collapse group the deceptive sub-ratio is 51-81. In the collapse group, the protease content decreased significantly. The population stability of strain SD-1 was significantly affected by different forms of nitrogen salts, and the population in the groups of nitrite and nitrate was more likely to collapse. (2) the nitrogen source utilization of strain SD-1 was compared under different culture conditions. The results showed that the change trend of ammonia nitrogen and nitrite nitrogen was the same, but the change trend of nitrite nitrogen was different. In nitrate and nitrite medium, the secretion of common substance of the strain SD-1 was 2-3 times higher than that of other groups. In glucose medium supplemented with nitrate, wild plant and deceiver had the same ability to utilize ammonia nitrogen, but in glucose medium with nitrate added, deceiver grew faster than collaborator 12 hours before the addition of nitrate. In glucose medium supplemented with nitrite, the growth of deceivers was always higher than that of collaborators. Nitrate and nitrite make the co-strains bear more burden of common substance metabolism, and make the deceptive strains gain higher survival advantages than the collaborators. That is, the metabolic difference mediated by population induction is the root cause of population collapsing more easily under the conditions of nitrite and nitrate. (3) 24 evolutionary collaborators were selected from the long-term evolutionary experiment, and their population induction changed significantly. Although there was no significant difference between the growth of the evolutionary strain and the initial co-strain, the production of 30C12-HSLC4VHSL and protease of all the evolutionary strains was lower than that of the initial co-strain, and it became more "cautious". The pyocyanin synthesis of all the evolutionary strains was higher than that of the initial co-strains to some extent, and the stability and maintenance mechanism was strengthened. The evolution of cooperative behavior and the improvement of stability are the important reasons for the stability of the microflora in the long-term evolution.
【學(xué)位授予單位】：浙江工商大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：X172
[Abstract]:Pseudomonas aeruginosa SD-1 (Pseudomonas aeruginosa) is a high efficient heterocyclic compound degrading strain which can play an important role in pollutant treatment through biointensification. It has been suggested that the stability of the enhanced strain is of great significance for the stable operation of the biological strengthening system. Although there have been a lot of reports on the inter-species competition of the enhanced strains in the ecosystem, there are no studies on the intraspecific ecological stability of the enhanced strains. In this paper, using Pseudomonas aeruginosa SD-1 as the experimental strain, the effects of different nitrogen salts in wastewater treatment system on the population stability of the strains were studied by means of continuous passage. By comparing the growth differences of wild plants and colony sensing deceptive strains in different forms of nitrogen salts, the effects of different forms of nitrogen salts on the population stability of strains were analyzed from the point of view of metabolism. In addition, by comparing the population induction effects of evolutionary collaborators with different nitrogen salts, the population sensing mechanism of different forms of nitrogen salts on the population stability of strains was further explored from the perspective of evolution. This study is of great significance to enrich the theory of group induction, expand the application of group induction theory in biological reinforcement system, and optimize the operation strategy of biological reinforcement system. The main results are as follows: (1) during the long-term evolution, the density of some groups of bacteria decreased by 1-2 orders of magnitude, resulting in the phenomenon of population collapse. The probability of the strain collapsing in the control group with added ammonium salt, nitrate, nitrite and no nitrogen salt was 66.6% and 11.1%, respectively. In the stable group, the deceptive sub-ratio ranges from 0 to 53, and in the collapse group the deceptive sub-ratio is 51-81. In the collapse group, the protease content decreased significantly. The population stability of strain SD-1 was significantly affected by different forms of nitrogen salts, and the population in the groups of nitrite and nitrate was more likely to collapse. (2) the nitrogen source utilization of strain SD-1 was compared under different culture conditions. The results showed that the change trend of ammonia nitrogen and nitrite nitrogen was the same, but the change trend of nitrite nitrogen was different. In nitrate and nitrite medium, the secretion of common substance of the strain SD-1 was 2-3 times higher than that of other groups. In glucose medium supplemented with nitrate, wild plant and deceiver had the same ability to utilize ammonia nitrogen, but in glucose medium with nitrate added, deceiver grew faster than collaborator 12 hours before the addition of nitrate. In glucose medium supplemented with nitrite, the growth of deceivers was always higher than that of collaborators. Nitrate and nitrite make the co-strains bear more burden of common substance metabolism, and make the deceptive strains gain higher survival advantages than the collaborators. That is, the metabolic difference mediated by population induction is the root cause of population collapsing more easily under the conditions of nitrite and nitrate. (3) 24 evolutionary collaborators were selected from the long-term evolutionary experiment, and their population induction changed significantly. Although there was no significant difference between the growth of the evolutionary strain and the initial co-strain, the production of 30C12-HSLC4VHSL and protease of all the evolutionary strains was lower than that of the initial co-strain, and it became more "cautious". The pyocyanin synthesis of all the evolutionary strains was higher than that of the initial co-strains to some extent, and the stability and maintenance mechanism was strengthened. The evolution of cooperative behavior and the improvement of stability are the important reasons for the stability of the microflora in the long-term evolution.
【學(xué)位授予單位】：浙江工商大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：X172
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