本文關(guān)鍵詞：堆肥中新菌的分離鑒定及其鉻還原特性研究　出處：《上海海洋大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 堆肥 新種 分離 鑒定 鉻還原

【摘要】：微生物是地球上分布最為廣泛的生命形式,其物種多樣性非常豐富。據(jù)估計,目前人類發(fā)現(xiàn)的微生物物種僅占微生物物種總數(shù)的10%左右,大量的微生物物種還沒有被發(fā)現(xiàn)。微生物廣泛存在于人類的生活環(huán)境中,通過發(fā)現(xiàn)微生物新物種及其功能,為人類探究大自然奧秘和利用微生物造福人類提供更多的機會。鉻作為工業(yè)生產(chǎn)中廣泛應(yīng)用的一種重金屬,其六價離子對細(xì)胞具有很高的毒性。傳統(tǒng)治理鉻污染的方式成本高、效果差,而微生物法治理鉻污染作為一種新興的方法,克服了上述缺點,提供了新的解決辦法。目前人類已經(jīng)發(fā)現(xiàn)了一些具有鉻還原功能的微生物,但是大量尚未被發(fā)現(xiàn)的微生物物種資源吸引著人類去進行更多更深入的研究。本研究從兩個不同采樣來源的堆肥土壤中分離出兩個疑似新菌,運用多相分類學(xué)方法,對菌株進行了系統(tǒng)鑒定,并對得到的新菌GSS04T進行了鉻還原特性的研究。主要結(jié)論如下:(1)從大寶山礦區(qū)堆肥中分離得到了菌株GSS04T。GSS04T能形成直徑為1-2 mm的淺黃色、圓形、表面光滑、微微向上隆起的不透明菌落。GSS04T為兼性厭氧的革蘭氏陽性菌,在無氧條件下生長相對緩慢。菌體細(xì)胞寬度為0.8μm,長度為2.6-3.0μm,有鞭毛,能產(chǎn)生橢圓的內(nèi)生孢子。在LB液體培養(yǎng)基中,GSS04T的生長溫度范圍為15-50℃,最適生長溫度為30-37℃;生長p H范圍為5.5-10.0,最適生長p H為7.0;可以在0-3%Na Cl濃度范圍內(nèi)生長,在1.0%的鹽濃度下生長最好。GSS04T的呼吸醌主要為甲基萘醌7(MK-7),主要脂肪酸為C15:0(42.9%)和anteiso-C15:0(24.1%)。GSS04T主要極性脂包括雙磷脂酰甘油(DPG)、磷脂酰甲基乙醇胺(PME)、磷脂酰甘油(PG)、磷脂酰乙醇胺(PE)以及三種未識別出的磷脂和兩種未知脂質(zhì)。GSS04T的DNA G+C含量為41.1 mol%。GSS04T在進化樹中的位置支持其屬于芽孢桿菌屬的結(jié)論。在形態(tài)特征、生理生化特征和分子生物學(xué)特征數(shù)據(jù)的基礎(chǔ)上,GSS04T被認(rèn)定為芽孢桿菌屬的一個新種,命名為Bacillus dabaoshanensis。典型菌株GSS04T保藏于中國典型培養(yǎng)物保藏中心(保藏號為CCTCC AB 2013260T)和韓國典型培養(yǎng)物保藏中心(保藏號為KCTC 33191T)。(2)從廣東省東莞市堆肥中分離得到了菌株GSS05T。GSS05T能形成直徑為0.5-1 mm的淺黃色、圓形、表面光滑、無隆起的不透明菌落。GSS05T為革蘭氏陽性菌,菌體細(xì)胞寬度為0.3-0.8μm,長度為1.1-2.6μm,具有運動性,有周生鞭毛,能產(chǎn)生橢圓的內(nèi)生孢子。在LB液體培養(yǎng)基中,GSS05T生長溫度為15-50℃,最適生長溫度為30-37℃,該菌能夠在5.5-10.0的p H條件下生長,p H=7.5為其最佳生長Ph,可以在0-3%Na Cl濃度范圍內(nèi)生長,在1.0%的鹽濃度下生長最好。生理鑒定結(jié)果表明,GSS05T氧化酶實驗呈現(xiàn)陽性,過氧化氫酶實驗為陰性。該菌可以將酪蛋白進行水解,可以產(chǎn)生D-巖藻糖和糖原。GSS05T的細(xì)胞壁肽聚糖類型為A4β,呼吸醌主要類型為甲基萘醌7(MK-7),這些結(jié)果與其模式菌的主要呼吸醌相同,并且符合鳥氨酸芽孢桿菌屬的特征,從另一個方面證明GSS05T屬于鳥氨酸芽孢桿菌屬。GSS05T的主要脂肪酸為C15:0(38.1%)和anteiso-C15:0(31.1%)。其在進化樹中的位置支持將GSS05T歸為鳥氨酸芽孢桿菌屬的結(jié)論。通過GSS05T與其模式菌的生理生化特征進行對比,可以將GSS05T與其他同屬菌株分別開來。在形態(tài)特征、生理生化特征和分子生物學(xué)特征數(shù)據(jù)的基礎(chǔ)上,GSS05T被認(rèn)定為鳥氨酸芽孢桿菌屬的一個新種,命名為Ornithinibacillus composti。典型菌株GSS05T保藏在位于武漢的中國典型培養(yǎng)物保藏中心(保藏號為CCTCC AB 2013261T)和位于韓國的韓國典型培養(yǎng)物保藏中心(保藏號為KCTC 33192T)。(3)GSS04T在有氧和厭氧條件下,均具有還原六價鉻離子的能力,而GSS05T沒有這些特性。GSS04T的六價鉻離子最小抑制濃度為600 mg l-1。GSS04T最佳鉻還原條件為37℃,p H=7,初始六價鉻離子濃度為100 mg l-1。在最佳鉻還原條件下,GSS04T可以在120h內(nèi)將100 mg l-1六價鉻離子完全還原。在鉻還原過程中,添加乙酸鈉、蔗糖、丙酮酸、乳酸、草酸鈉、葡萄糖作為電子供體,有利于GSS04T鉻還原速率的提高,含氧陰離子SO42-、NO3-、PO43-的添加則對GSS04T鉻還原過程沒有促進或者抑制作用。GSS04T和GSS05T的發(fā)現(xiàn)增加了對微生物物種多樣性的了解,豐富了對堆肥中微生物資源的認(rèn)識。GSS04T鉻還原特性的發(fā)現(xiàn)和研究,在理論方面為進一步探究微生物鉻還原原理提供研究基礎(chǔ),同時在環(huán)境治理應(yīng)用方面,為鉻污染治理提供更多的鉻還原微生物選擇。
[Abstract]:Microorganisms are the most widely distributed on earth life forms, the species diversity is very rich. It is estimated that the microbial species found in human microbial species accounted for only about 10% of the total, a large number of microbial species has not been found. Microorganisms widely exists in the human living environment, through the discovery of new microbial species and its function. For human being to explore the mystery of nature and the use of microorganisms to provide more opportunities for the benefit of mankind. A kind of heavy metal chromium is widely used in industrial production, the six valence ions have high toxicity to the cells. The cost of traditional way of high chromium pollution control, poor effect, and microbial treatment of chromium pollution as a new method. To overcome these shortcomings, provides a new solution. At present people have found some with chromium to restore the function of microorganisms, but has not yet been found a large number of micro Biological species resources attracted humans to make further research. This study from two different sampling sources in compost soil isolated two new suspected bacteria, using polyphasic taxonomy methods, the strains are systematically identified, and the new strain GSS04T obtained were studied. The main conclusions of the chromium reduction characteristics are as follows: (1) the strain GSS04T.GSS04T can form 1-2 diameter mm light yellow, round, smooth surface separated from the compost Baoshan mine, gram positive bacteria slightly upward uplift of the opaque colony.GSS04T is a facultative anaerobic, relatively slow growth under anaerobic conditions. The cell width is 0.8 m. The length of 2.6-3.0 m, a flagellum, can produce oval endospores. In LB liquid medium, growth temperature range of GSS04T to 15-50 DEG C, the optimum growth temperature is 30-37 DEG C; the growth of P H is in the range of 5.5-10.0, the optimum growth p H is 7; 0-3%Na can grow in the concentration range of Cl, the best.GSS04T growth at 1% salt concentration under the main respiratory quinone menadione (MK-7), 7 major fatty acids were C15:0 (42.9%) and anteiso-C15:0 (24.1%).GSS04T polar lipids including cardiolipin phosphatidylcholine (DPG). Methyl diethanolamine (PME), phosphatidyl glycerol (PG), phosphatidylethanolamine (PE) and DNA G+C content of three kinds of unrecognized phospholipid and two unknown lipid.GSS04T was 41.1 mol%.GSS04T in the evolutionary tree in position to support its belonging to the genus Bacillus. In conclusion the morphological characteristics, physiological and biochemical basis the characteristics and characteristics of molecular biology data, GSS04T was identified as a new species of the genus bacillus, named Bacillus dabaoshanensis. GSS04T typical strains preserved in China type culture collection (CCTCC AB accession number 2013260T) and South Korea type culture collection (Center for the preservation of KCTC 33191T). (2) the strain GSS05T.GSS05T can form 0.5-1 diameter mm light yellow, round, smooth surface isolated from Guangdong city of Dongguan Province in the compost, no uplift of the opaque colony.GSS05T gram positive bacteria, the cell width is 0.3-0.8 ~ m, length of 1.1-2.6 m. With the movement, a week flagella, can produce oval endospores. In LB liquid medium, GSS05T growth temperature is 15-50 DEG C, the optimum growth temperature of 30-37 DEG C, the bacteria can grow in 5.5-10.0 P H P H=7.5 conditions, the best growth Ph, can grow in 0-3%Na the concentration range of Cl, the best growth in the 1% salt. The results showed that the physiological identification, GSS05T oxidase test positive, catalase test was negative. The bacteria can be casein hydrolysis, cell wall peptidoglycan can produce D- fucose sugar and raw.GSS05T Type A4 beta, the main types of respiratory quinone menadione (MK-7 7), these results mainly respiratory and pattern bacteria and accord with the characteristics of ornithine quinone, bacillus, from another aspect that GSS05T belongs to the main fatty acids ornithine Bacillus.GSS05T for C15:0 (38.1%) and anteiso-C15:0 (31.1% in the evolutionary tree). The position will support GSS05T belongs to the genus Bacillus. Conclusion ornithine by comparing the physiological and biochemical characteristics of GSS05T and its pattern of bacteria, can be GSS05T and other strains belong to separate. In the morphological characteristics, physiological and biochemical characteristics and molecular biological features of the data, GSS05T was identified as a new species of the genus bacillus ornithine, named as Ornithinibacillus composti. typical strains preserved in GSS05T culture collection center in Wuhan (China typical preservation number is CCTCC AB 2013261T) In South Korea and South Korea type culture collection (KCTC accession number 33192T). (3) GSS04T under aerobic and anaerobic conditions, have the ability to restore six chromium ions, but not GSS05T six chromium ion minimum inhibitory concentration of these properties of.GSS04T 600 mg l-1.GSS04T the best conditions for the reduction of chromium 37 C, P H=7, six valence chromium ion concentration of 100 mg l-1. reduction in the initial optimal chromium conditions, GSS04T in 120h 100 mg L-1 six chromium ion. In the complete reduction of chromium reduction process, adding sodium acetate, sucrose, pyruvic acid, lactic acid, sodium oxalate, glucose as electron donor to GSS04T the reduction rate of chromium increased, oxygen anion SO42-, NO3-, PO43- of GSS04T added chromium reduction process did not promote or inhibit.GSS04T and GSS05T are found to increase the diversity of microbial species in the solution enriched in the compost. The recognition of material resources, the discovery and research of Cr reduction characteristics in.GSS04T, provide theoretical basis for further exploring the principle of microbial chromium reduction in theory, and provide more chromium reduction microbial selection for environmental remediation.

【學(xué)位授予單位】：上海海洋大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S141.4;X172

【參考文獻】

相關(guān)碩士學(xué)位論文 前1條

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本文編號：1399064