【摘要】：放線菌屬于革蘭氏陽性的一類原核微生物,它們分布于各種各樣的環(huán)境。由于產(chǎn)生有益的代謝物,它們在醫(yī)藥、食品、工農(nóng)業(yè)等多個領(lǐng)域上具有巨大的應(yīng)用價值和潛力,因而其研究得到了研究人員的廣泛關(guān)注,尤其是鏈霉菌。據(jù)報道,至2010年微生物產(chǎn)生的活性物質(zhì)已有超過30000種被發(fā)現(xiàn),其中由放線菌產(chǎn)生的具有生物活性的次級代謝產(chǎn)物達到13000多種,鏈霉菌產(chǎn)生的物質(zhì)約7600種。本研究從湘潭錳礦污染區(qū)(N27°53'-28°03',E112°45'-112°55')及周邊環(huán)境的土壤分離得到1200多株放線菌,通過結(jié)合菌落的培養(yǎng)特征和16S r RNA基因序列,最終得到51株菌落形態(tài)、特征不同的鏈霉菌。由此可知:湘潭錳污染土壤中存在豐富多樣的鏈霉菌。初步的研究結(jié)果表明:菌株LUSFXJ,MKLv,MK 44,MK 45和MK47可能為潛在的鏈霉菌新物種。本研究選取菌株LUSFXJ和MK 44進行了多相分類鑒定,研究結(jié)果如下:(1)菌株LUSFXJ,分離自錳礦污染區(qū)或周邊環(huán)境的土壤,16S r RNA基因序列以及孢子形態(tài)、結(jié)構(gòu)特征和培養(yǎng)特征均表明:菌株LUSFXJ屬于鏈霉菌。甲基萘醌包含MK-9(H_6),MK-9(H_8),MK-9(H_2)和MK-8(H_8)。磷脂類型包括雙磷脂酰甘油(DPG)和一個未鑒定的極性脂。脂肪酸主要成分為iso-C_(1 5:0),anteiso-C_(1 5:0),iso-C_(16:0),C_(16:0)和summed Feature 3。菌株LUSFXJ的細胞壁含有meso-DAP,細胞水解產(chǎn)物含有核糖,甘露糖和葡萄糖。菌株LUSFXJ與進化關(guān)系最近的參照菌株的雜交率遠遠低于70%。根據(jù)形態(tài)特征、生理生化特征以及DNA的分子雜交結(jié)果,確定菌株LUSFXJ是鏈霉菌的一個新種,并將其命名為Streptomyces(2)菌株MK 44,分離自錳礦污染區(qū)的土壤。根據(jù)菌落的形態(tài)、培養(yǎng)特征、生理生化特征的比較和16S r RNA基因序列分析,我們可以得出:菌株MK 44屬于鏈霉屬,同時明顯區(qū)別于同源性97%有效發(fā)表的種。其與選擇的菌株S.speciali JCM 16611~T和S.mayteni JCM 16957~T的雜交值分別為30.90±0.28%和29.86±3.51%遠遠低于Wayne et al(1987年)提出的70%作為新種的界限。因此可以判斷該菌為鏈霉菌的潛在新種,并將其命名為鶴嶺鏈霉菌Streptomyces heling sp.nov.(=GDMCC 4.137~T=KCTC 39920~T)。
[Abstract]:Actinomycetes belong to a class of Gram-positive prokaryotes, which are distributed in various environments. Due to the production of beneficial metabolites, they have great application value and potential in many fields, such as medicine, food, industry and agriculture, so their research has been widely concerned by researchers, especially Streptomyces. It is reported that more than 30000 active substances have been found by microbes in 2010, of which more than 13000 are bioactive secondary metabolites produced by actinomycetes and 7600 by Streptomyces. In this study, more than 1200 strains of actinomycetes were isolated from the contaminated area of Xiangtan Manganese Mine (N27 擄53 ~ 28 擄03 擄E 112 擄45 擄-112 擄55') and the surrounding environment. By combining the culture characteristics of the colony and 16s r RNA gene sequence, 51 strains of Streptomyces with different characteristics were obtained. It can be seen that there are abundant and diverse streptomyces in Xiangtan Manganese contaminated soil. The preliminary results showed that the strains LUSFXJ MKLv4, MK44, MK45 and MK47 might be potential new species of Streptomyces. The results were as follows: (1) strain LUSFXJ, 16s r RNA gene sequence and spore morphology were isolated from manganese ore contaminated area or surrounding environment. The structure and culture characteristics showed that the strain LUSFXJ belonged to Streptomyces. Methyl naphthoquinone contains MK-9 (H6) MK-9 (Hs _ 8), MK-9 (H _ s _ 2) and MK-8 (H _ s _ 8). Phospholipid types include diphosphatidylglycerol (DPG) and an unidentified polar lipid. The main components of fatty acids are iso-C _ (1 5:0) anteiso-C _ (1 5:0), iso-C _ (16:0) C _ (16:0) and summed Feature _ 3. The cell wall of strain LUSFXJ contains meso-DAP.The cell hydrolysate contains ribose, mannose and glucose. The hybridization rate of LUSFXJ with the nearest reference strain was far lower than 70%. According to the morphological, physiological and biochemical characteristics and the results of molecular hybridization of DNA, strain LUSFXJ was identified as a new species of Streptomyces and named Streptomyces (2) strain MK44, which was isolated from the soil of manganese ore contaminated area. According to the comparison of colony morphology, culture characteristics, physiological and biochemical characteristics and the sequence analysis of 16s r RNA gene, we can conclude that strain MK44 belongs to Streptomyces, and is obviously different from the effective species with 97% homology. The crossbreeding values with the selected strains S.speciali JCM 16611T and S.mayteni JCM 16957T were 30.90 鹵0.28% and 29.86 鹵3.51% respectively, which were much lower than that of Wayne et al (1987 (70%). Therefore, the strain can be identified as a potential new strain of Streptomyces and named Streptomyces Streptomyces heling sp.nov. (GDMCC 4.137~T=KCTC 39920T).
【學(xué)位授予單位】：湖南科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：X172;Q939.132

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