發(fā)布時(shí)間：2018-07-11 12:16

  本文選題：小蓬竹 + 土壤酶�。� 參考：《貴州大學(xué)》2015年碩士論文

【摘要】：小蓬竹作為貴州喀斯特山地適生性竹種,在IUCN紅色名錄中,被定為極危種,對(duì)喀斯特地區(qū)水源涵養(yǎng)、水土保持、養(yǎng)分平衡等生態(tài)功能的實(shí)現(xiàn)起著重要作用。本文以羅甸縣小蓬竹根際土壤為研究對(duì)象,采用稀釋平板法,挑選菌落形態(tài)差異的菌株,分離純化小蓬竹根際土壤細(xì)菌、真菌和放線菌,對(duì)其16S rDNA(細(xì)菌、放線菌)、ITS rDNA(真菌)序列進(jìn)行分析構(gòu)建系統(tǒng)發(fā)育樹,研究根際土壤可培養(yǎng)微生物多樣性。同時(shí),對(duì)小蓬竹根際土壤相關(guān)理化性質(zhì)進(jìn)行了研究,為小蓬竹的保護(hù)以及喀斯特山地土壤微生物多樣性研究,提供一定的理論依據(jù)和技術(shù)支撐。主要研究結(jié)果如下:(1)小蓬竹根際土壤酶研究發(fā)現(xiàn),過氧化氫酶活性8.74mL KMnO4?g-1?min-1;蔗糖酶的活性表示為2.64mg?g-1?d-1;脲酶的活性為0.04mg NH3-N?g-1?d-1;蛋白酶的活性表示為0.10mg NH2-N?g-1?d-1;堿性磷酸酶的活性高于中性磷酸酶的活性,分別為7.97mg P2O5?g-1?d-1、6.11mg P2O5?g-1?d-1。(2)小蓬竹根際土壤微生物數(shù)量為細(xì)菌數(shù)量放線菌數(shù)量真菌數(shù)量。每克干土中所含細(xì)菌、真菌、放線菌的平均數(shù)量分別為8.32×107 cfu·g-1、2.32×106 cfu·g-1、9.19×105 cfu·g-1。(3)采用稀釋平板法分離小蓬竹根際土壤微生物,共獲得菌落表型差異的細(xì)菌41株、放線菌20株、真菌33株。16S rDNA有效序列進(jìn)行BLAST顯示,41株細(xì)菌可劃分為26個(gè)分類單元、放線菌可劃分為15個(gè)分類單元均為鏈霉菌屬(Streptomyces);真菌的ITS rDNA有效序列分析,可劃分為24個(gè)分類單元。僅有1株真菌BLAST比對(duì)相似度僅為75%,可能為新種或?qū)?其余菌株相似性均≥97%。細(xì)菌和真菌的Shannon-Wiener多樣性指數(shù)H分別為1.0679、2.2180,豐富度指數(shù)S分別為7、12,均勻度指數(shù)J分別為0.5488、0.8926,及優(yōu)勢(shì)度指數(shù)D分別為0.5217、0.1295。(4)小蓬竹根際土壤細(xì)菌、放線菌基于16S rDNA序列建立系統(tǒng)發(fā)育樹表明,29株細(xì)菌屬于厚壁菌門(Firmicutes)(70.73%)(按單元種類數(shù)計(jì)算百分比),11株屬于變形菌門(Proteobacteria)(26.83%),1株屬于擬桿菌門(Bacteroidetes)2.44%,其中芽孢桿菌屬(Bacillus)占絕對(duì)優(yōu)勢(shì);分離獲得的20株放線菌均為鏈霉菌屬。ITS rDNA系統(tǒng)發(fā)育樹分析,小蓬竹根際土壤真菌類群子囊菌門類群(占真菌總菌株數(shù)的96.97%),青霉菌屬(Penicillium)、木霉屬(Trichoderma)、曲霉屬(Aspergillus)占絕對(duì)優(yōu)勢(shì),占總菌株數(shù)51.51%,只有1株真菌屬于擔(dān)子菌門的菌屬(Perenniporia),占真菌總菌株數(shù)的3.03%。(5)小蓬竹根際土壤中微生物的總數(shù)較多,細(xì)菌的數(shù)量上占優(yōu)勢(shì),真菌的多樣性指數(shù)較高,放線菌僅為1個(gè)屬但種類較多,總體上小蓬竹根際土壤中含有較為豐富的微生物多樣性。
[Abstract]:As a suitable species of bamboo in Guizhou karst mountain area, it is regarded as an extremely dangerous species in IUCN red list, which plays an important role in the realization of ecological functions such as water conservation, soil and water conservation, nutrient balance and so on. In this paper, the rhizosphere soil of Xiaopeng bamboo in Luodian County was studied. The bacteria, fungi and actinomycetes in rhizosphere soil were isolated and purified by dilution plate method, and 16s rDNA (bacteria) were isolated and purified. The phylogenetic tree of its rDNA (fungi) was constructed to study the diversity of culturable microorganisms in rhizosphere soil. At the same time, the physical and chemical properties of rhizosphere soil were studied in order to provide some theoretical basis and technical support for the protection of Xiaobang bamboo and the study of soil microbial diversity in karst mountains. The main results are as follows: (1) the soil enzymes in rhizosphere of Phyllostachys amurensis were found. The activity of catalase was 8.74 mL KMnO _ 4 / g ~ (-1) 路min ~ (-1); the activity of sucrase was 2.64 mg / g ~ (-1) 路d ~ (-1); the activity of urease was 0.04mg NH _ 3-Nng ~ (-1) / d ~ (-1); the activity of protease was expressed as 0.10mg NH _ 2-N _ (g-1) ~ (-1) d ~ (-1); the activity of alkaline phosphatase was higher than that of neutral phosphatase. The results showed that the number of microbes in rhizosphere soil was 7.97mg P _ 2O _ 5 路g ~ (-1) d ~ (-1) 6.11mg P _ 2O _ 5 路g ~ (-1) 路d ~ (-1) 路d ~ (-1). The average number of bacteria, fungi and actinomycetes per gram dry soil was 8.32 脳 10 ~ 7 cfu g ~ (-1) 2.32 脳 10 ~ (6) cfu g ~ (-1) ~ (-1) 9.19 脳 10 ~ 5 cfu ~ (-1) g ~ (-1) respectively. (3) A total of 41 strains of bacteria and 20 strains of actinomycetes were isolated from rhizosphere soil by dilution plate method. 33 strains of fungi. 16s rDNA effective sequence showed that 41 strains of bacteria could be divided into 26 taxonomic units, actinomycetes could be divided into 15 taxonomic units were Streptomyces, fungi its rDNA effective sequence analysis, can be divided into 24 taxonomic units. Only one fungus blast had only 75 similarity, which might be a new species or genus, and the other strains were more than 97% in similarity. The Shannon-Wiener diversity index (H) of bacteria and fungi was 1.0679 (2.2180), the richness index (S) was 7.12, the evenness index (J) was 0.5488 (0.8926), and the dominance index (D) was 0.5217 (0.1295). (4) the bacteria in rhizosphere soil. Phylogenetic tree of actinomycetes based on 16s rDNA sequence showed that 29 strains belong to Firmicutes (70.73%) (calculated by the number of unit species) 11 strains belong to Proteobacteria (26.83%) and 1 strain belongs to Bacteroidetes 2.444.Bacillus is the dominant strain. The 20 actinomycetes isolated were all streptomyces. Its rDNA phylogenetic tree analysis showed that ascomycetes of rhizosphere soil fungi group (96.97%), Penicillium, Trichoderma and (Aspergillus) were the dominant ones. Only one fungus belongs to Perenniporia, which accounts for 3.03 of the total strains. (5) the total number of microbes in rhizosphere soil is more, the number of bacteria is dominant, and the diversity index of fungi is higher. Actinomycetes belong to only one genus but there are many species. In general, there are abundant microbial diversity in rhizosphere soil of Phyllostachys vulgaris.
【學(xué)位授予單位】：貴州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S714.3

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