噻嗪酮降解菌株的篩
本文選題:噻嗪酮 切入點(diǎn):降解 出處:《淮北師范大學(xué)》2017年碩士論文 論文類型:學(xué)位論文
【摘要】:噻嗪酮是一種新型的殺蟲劑,能夠通過抑制幾丁質(zhì)的合成干擾昆蟲的新陳代謝,主要用于對飛虱、粉虱、葉嬋及介殼蟲等同翅目害蟲有很好的防治作用。由于噻嗪酮在使用過程中的不規(guī)范儲藏,和在農(nóng)作物上的重復(fù)噴灑,對環(huán)境造成了危害。因此,如何治理環(huán)境中的噻嗪酮?dú)埩艟统蔀榱宋覀兗毙杞鉀Q的難題。可通過物理方法、化學(xué)方法、生物方法治理環(huán)境中的噻嗪酮污染,生物降解是治理的重要方法之一,高效降解菌能夠?qū)⑧玎和鳛樘嫉磳⑵浯x成小分子物質(zhì),在生物修復(fù)中起著重要的作用。遺憾的是,有關(guān)噻嗪酮降解菌的報道只有五株,有關(guān)其降解途徑的報道也少之又少。本研究從常年生產(chǎn)噻嗪酮農(nóng)藥的工廠排污口處采取水樣與土樣,于100 m L無機(jī)鹽培養(yǎng)基中,并添加噻嗪酮作為碳源,在30°C、160 rpm min-1搖床條件下經(jīng)富集馴化和分離純化,成功篩選到多個高效降解菌。對具有噻嗪酮降解能力的菌株做了降解能力的驗(yàn)證,選擇一株降解能力較好的菌株展開研究,將其命名為RX-3。通過形態(tài)學(xué)、生理生化實(shí)驗(yàn)及16S rRNA序列比對鑒定菌株RX-3為紅球菌屬(Rhodococcus sp.),其最適生長溫度為30°C,最適生長酸堿度值為7.0,且對金屬離子Ba2+、Zn2+和Cu2+有一定的耐受性。當(dāng)無機(jī)鹽中噻嗪酮濃度為60 mg L-1時,菌株RX-3在80 h內(nèi)能夠降解98.01%的噻嗪酮。使用二氯甲烷等體積萃取法提取到噻嗪酮的降解產(chǎn)物,并借助于氣相色譜質(zhì)譜聯(lián)用儀分析了9種中間代謝產(chǎn)物,初步剖析了噻嗪酮的降解途徑。在代謝途徑A中,噻嗪酮中的雜環(huán)發(fā)生水解生成中間代謝產(chǎn)物B;在代謝途徑B中,噻嗪酮的代謝起始于側(cè)鏈中叔丁基和異丙基鍵的斷裂,伴隨產(chǎn)物C,E和F的生成,隨后發(fā)生了水解反應(yīng)和氧化還原反應(yīng)。在代謝途徑C中,噻嗪酮的代謝起始于苯環(huán)與N之間化學(xué)鍵的斷裂,生成中間代謝產(chǎn)物D。本研究使用聚乙二醇4000對噻嗪酮降解菌和毒死蜱降解菌的原生質(zhì)體進(jìn)行融合,成功獲得其融合子。
[Abstract]:Thiazinone is a new type of insecticide that interferes with the metabolism of insects by inhibiting the synthesis of chitin. It is mainly used in the treatment of planthopper and whitefly. Homoptera pests such as Ye Chan and mesoptera insects have a good control effect. Because of the nonstandard storage of thiazinone in the process of use and repeated spraying on crops, it is harmful to the environment. How to control thiazinone residues in the environment has become a difficult problem that we need to solve urgently. The biodegradation is one of the most important methods to control thiazinone pollution in the environment by physical, chemical and biological methods. Highly efficient biodegradable bacteria can metabolize thiazines as carbon and nitrogen sources into small molecules and play an important role in bioremediation. Unfortunately, only five strains of thiazone degrading bacteria have been reported. In this study, water samples and soil samples were taken from the sewage outlets of factories that produce thiazinone pesticides all the year round, and thiazinone was added as carbon source in 100 mL inorganic salt medium. After enrichment, acclimation, purification and purification, a number of high efficient biodegradable bacteria were successfully screened under the condition of 30 擄CX 160 rpm min-1 shaking bed. The biodegradable ability of the strain with thiazinone degradation ability was verified, and a strain with better degradation ability was selected. It's named RX-3. it's morphologically, Physiological and biochemical experiments and 16s rRNA sequence alignment identified the strain RX-3 as Rhodococcus sp., the optimum growth temperature is 30 擄C, the optimum growth pH value is 7.0, and the strain is tolerant to metal ions Ba2 Zn 2 and Cu2. When the concentration is 60 mg L ~ (-1), The strain RX-3 could degrade 98.01% thiazinone within 80 h. The degradation products of thiazinone were extracted by dichloromethane isovolumetric extraction, and 9 intermediate metabolites were analyzed by gas chromatography-mass spectrometry. The degradation pathway of thiazinone was preliminarily analyzed. In metabolic pathway A, the heterocycles in thiazinone hydrolyzed to form intermediate metabolite B, and in metabolic pathway B, the metabolism of thiazinone originated from the breakage of tertiary Ding Ji and isopropyl bond in the side chain. With the formation of Con E and F, hydrolysis reaction and redox reaction took place. In metabolic pathway C, the metabolism of thiazinone begins with the breaking of the chemical bond between benzene ring and N. In this study, the protoplasts of thiazine-degrading bacteria and chlorpyrifos degrading bacteria were fused with polyethylene glycol 4000, and the fusion was successfully obtained.
【學(xué)位授予單位】:淮北師范大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:X172;X592
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