本文關(guān)鍵詞：貴州喀斯特土壤放線菌的分離鑒定及多樣性研究　出處：《東華理工大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 放線菌 分離 多樣性 菌株鑒定

【摘要】：貴州是東亞喀斯特片區(qū)中心,是世界三大喀斯特集中區(qū)之一。微生物是土壤中最活躍的部分,是生態(tài)系統(tǒng)中能量流動和物質(zhì)循環(huán)的主要推動者。而放線菌主要存在于土壤中,放線菌能夠產(chǎn)生大量的具有生物活性的次級代謝產(chǎn)物,因此對土壤放線菌的研究具有重要的價(jià)值。貴州喀斯特區(qū)域土壤放線菌資料目前還非常有限,亟需進(jìn)行深入細(xì)致的研究。本研究采用不同的分離方法對貴州萬峰林國家地質(zhì)公園、草海國家自然保護(hù)區(qū)、雙河洞國家地質(zhì)公園等8個(gè)地區(qū)的土壤放線菌進(jìn)行分離和鑒定,對其多樣性進(jìn)行研究,并對分離得到的菌株進(jìn)行抑菌活性的研究。具體實(shí)驗(yàn)結(jié)果如下:1.采用四種不同的方式預(yù)處理來處理106份土壤樣品,其中用微波干熱法的效果最佳。最終獲得了76株放線菌。2.采用9種培養(yǎng)基對放線菌進(jìn)行分離培養(yǎng),用LNMS分離出來的放線菌數(shù)量最多為每克土5.2×105株。燕麥瓊膠培養(yǎng)基分離出的放線菌種類數(shù)最多,為8種。3.在不同地區(qū)、不同植被類型的土壤分離中,耕地中分離出的放線菌數(shù)量最多可達(dá)4.65×105個(gè)/克土。4.在不同生境中,草海的邊坡、黃果樹林下和雙河洞的洞穴的土壤中分離出的放線菌所占比重都在65%以上,在小生境中溝邊放線菌種類比重最小為49%,而洞穴放線菌種類所占比重最大為76%。5.在高海拔2400 m時(shí)放線菌的種類數(shù)量減少,但稀有放線菌的數(shù)量有所增加,分別有羅氏菌屬、類諾卡氏菌屬等。6.采用濾紙片法對放線菌進(jìn)行抑菌活性測定。有10株放線菌對大腸埃希氏菌和金黃色葡萄桿菌的抑菌直徑達(dá)到了10 mm以上,其中3菌株對金黃色葡萄桿菌的抑菌直徑都在14 mm以上。有12株放線菌對真菌的拮抗性較強(qiáng),然后用生長速率法進(jìn)行活性復(fù)篩。至少對2種真菌有明顯的抑制作用。綜合抑細(xì)菌實(shí)驗(yàn)和抑真菌實(shí)驗(yàn)的結(jié)果可知,C1、E4這兩株放線菌對細(xì)菌和真菌均有很好的抑制作用,有繼續(xù)開發(fā)的價(jià)值。7.通過對菌株的形態(tài)學(xué)特征、培養(yǎng)特征和16S rDNA序列分析,可將拮抗性優(yōu)良的菌株1030、1051、1112、1095、1312歸屬于鏈霉菌屬。
[Abstract]:Guizhou is the center of karst region in East Asia and one of the three largest karst regions in the world. Microbes are the most active part of the soil. Actinomycetes mainly exist in soil, and actinomycetes can produce a large number of secondary metabolites with biological activity. Therefore, the study of soil actinomycetes is of great value. At present, the data of soil actinomycetes in Guizhou karst region is very limited. This study uses different separation methods to study the Wanfenglin National Geopark and the Caohai National Nature Reserve in Guizhou Province. Soil actinomycetes were isolated and identified from 8 areas such as Shuanghe Dong National Geopark and so on. The diversity of actinomycetes was studied. The antimicrobial activity of the isolated strains was studied. The results were as follows: 1. 106 soil samples were treated with four different pretreatment methods. Among them, microwave dry heating method was the best. Finally, 76 strains of actinomycetes were obtained. 9 kinds of media were used to isolate and culture actinomycetes. The maximum number of actinomycetes isolated by LNMS was 5.2 脳 10 ~ 5 strains per gram soil, and 8 species of actinomycetes were obtained from oat Agar medium, which were found in different regions. In the soil separation of different vegetation types, the maximum number of actinomycetes isolated from cultivated land was 4.65 脳 105 / g soil. The proportion of actinomycetes isolated from the soil of the caves of the Huangguoshu forest and Shuanghe Cave was above 65%, and the percentage of actinomycetes at the edge of the gullies was 49% in the microhabitat. The largest proportion of actinomycetes in caves was 76. 5. The number of actinomycetes decreased at 2400m above sea level, but the number of rare actinomycetes increased, respectively. The antimicrobial activity of actinomycetes was determined by filter paper method. The antibacterial diameter of 10 actinomycetes against Escherichia coli and Grape aureus was more than 10 mm. The inhibition diameter of 3 strains to grape aureus was more than 14 mm, and 12 strains of actinomycetes had strong antagonism to fungi. Then the activity was screened by growth rate method. At least two kinds of fungi were obviously inhibited. E4 two actinomycetes have a good inhibitory effect on bacteria and fungi, which has the value of further development. Through morphological characteristics, culture characteristics and 16s rDNA sequence analysis. The strain 1030, 1051, 1112, 1095 and 1312, which have good antagonism, can be assigned to Streptomyces.
【學(xué)位授予單位】：東華理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S154.3

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