本文選題：松嫩鹽堿地 + 纖維素降解菌�。� 參考：《東北師范大學(xué)》2017年碩士論文

【摘要】：本研究以松嫩草地不同鹽堿化程度的土壤為研究對(duì)象,對(duì)不同鹽堿化土壤中的纖維素降解菌的類群及其特性進(jìn)行了系統(tǒng)的研究與分析。研究中,分別從羊草草地(pH 8.68)、虎尾草草地(pH 9.47)和雜草草地(pH 10.03)篩選得到了纖維素降解菌,并對(duì)其進(jìn)行了形態(tài)學(xué)、生物化學(xué)特性以及16S rDNA核酸序列的分析與鑒定;確定了不同鹽堿化草地中纖維素降解菌類群;并在此前提下,對(duì)上述菌株的產(chǎn)酶特性及植物生長(zhǎng)特性進(jìn)行了系統(tǒng)研究。取得的研究成果如下:1.不同鹽堿化土壤中纖維素降解菌的鑒定通過對(duì)所篩選的纖維素降解菌的形態(tài)學(xué)、生物化學(xué)特性以及16S rDNA核酸序列分析,對(duì)分離菌株進(jìn)行了鑒定。對(duì)菌株的生長(zhǎng)特性分析表明,其都在0-4%NaCl及PH7-11條件下生長(zhǎng)良好。分離篩選共獲得35株菌經(jīng)16SrDNA核酸序列比對(duì),去除相同菌株,鑒定后得到27株纖維素降解菌。將所有菌株的16S rDNA核酸序列提交至GenBank共得到27株菌的登錄號(hào)。同時(shí),在核酸序列分析基礎(chǔ)上,構(gòu)建了所有菌株的系統(tǒng)發(fā)育樹。2.不同鹽堿化土壤中纖維素降解菌優(yōu)勢(shì)類群的分析在已分離鑒定的27株菌中,羊草草地土壤特有纖維素降解菌共10株,虎尾草草地土壤特有纖維素降解菌共9株,雜草草地土壤特有纖維素降解菌共4株。其余4株纖維素降解菌為三種不同草地土壤中交叉共有的重復(fù)菌株,其中,鏈霉菌KY007170.1和節(jié)桿菌KY007166.1為虎尾草草地以及羊草草地共有的菌株,鏈霉菌KY007169.1和節(jié)桿菌KY007185.1為三種草地中共有菌株。這說明在鹽堿土壤中,鏈霉菌和節(jié)桿菌分布范圍更廣,適應(yīng)性更強(qiáng)。采用CMC平板對(duì)三種不同草地中纖維素降解菌進(jìn)行分析,結(jié)果表明:在三種不同鹽堿化程度草地中,豐度最高的菌株均為鏈霉菌,豐度分別為羊草草地(71.4%)、虎尾草草地(71.4%)以及雜草草地(85.7%)。由此我們得出,鹽堿化草地中,鏈霉菌顯示出具有較強(qiáng)的鹽堿適應(yīng)能力,是優(yōu)勢(shì)的纖維素降解菌類群。3.分離菌株的纖維素降解特性(1)產(chǎn)纖維素酶特性采用剛果紅染色法對(duì)所分離菌株的纖維素酶活性進(jìn)行了初步分析,同時(shí)采用DNS法對(duì)分離菌株產(chǎn)生纖維素酶活性進(jìn)行了分析。結(jié)果表明,大多數(shù)菌株均產(chǎn)生胞外纖維素酶,其酶活性范圍在1.47~6.08×10~(-2)IU之間;鏈霉菌屬DS-22的纖維素酶活性最高,活性為6.08×10~(-2)IU。(2)產(chǎn)木聚糖酶特性采用DNS法對(duì)分離菌株產(chǎn)生木聚糖酶活性進(jìn)行了分析。結(jié)果表明,大多數(shù)菌都產(chǎn)生胞外木聚糖酶,其活性范圍在2.23~18.80×10~(-2)IU之間;微桿菌屬DS-28木聚糖酶活性最高,活性為18.80×10~(-2)IU。4.分離菌株的固氮特性采用無氮纖維素平板對(duì)分離菌株的固氮活性進(jìn)行了分析,結(jié)果表明,這些纖維素降解菌菌株在無氮源的纖維素平板上生長(zhǎng)良好,是可降解纖維素的固氮菌。通過乙炔還原法對(duì)分離菌株固氮酶活性進(jìn)行了檢測(cè)與分析,所有菌株均可產(chǎn)固氮酶,固氮酶活性范圍在36.29~112.08 nmoL·ml·h~(-1)之間。其中,鏈霉菌DS-18固氮酶活性最高,活性為112.08nmoL·ml·h~(-1)。5.分離菌株的促生特性對(duì)分離菌株的促生特性(產(chǎn)IAA、產(chǎn)鐵載體、溶磷以及抑菌能力)進(jìn)行了分析。結(jié)果表明:所有菌株均具有產(chǎn)IAA能力,范圍在2.4~80.69mg/L之間,其中微桿菌DS-34產(chǎn)IAA含量最大,為80.69mg/L;6株菌具備鐵載體產(chǎn)生能力,均屬于鏈霉菌屬;8株菌具備溶磷能力,包括7株為鏈霉菌屬以及1株為微桿菌屬;11株菌具抑菌活性,分別為7株為鏈霉菌屬,2株節(jié)桿菌屬,1株諾卡氏菌屬及1株黃桿菌屬。6.不同鹽堿化程度的土壤理化性質(zhì)及生物數(shù)量分析對(duì)不同鹽堿化程度土壤的理化特性及生物量進(jìn)行了分析。結(jié)果表明,隨著土壤的鹽堿程度增加,土壤中總氮以及有機(jī)碳含量不斷減少;三種不同鹽堿化程度草地土壤中可培養(yǎng)微生物數(shù)量分布情況均為:細(xì)菌放線菌纖維素降解菌真菌。7.分離菌株的植物促生實(shí)驗(yàn)從上述菌株中,挑選三株鏈霉菌DS-22、DS-2以及DS-30進(jìn)行盆栽實(shí)驗(yàn),研究它們對(duì)羊草的促生作用。盆栽實(shí)驗(yàn)結(jié)果表明,所有的單一菌株處理組的效果較對(duì)照組促生作用明顯,而混合菌株組的促生效果較單一菌株處理組促生作用更明顯。研究結(jié)果顯示了這些纖維素降解菌具有良好地促生作用。
[Abstract]:In this study, the groups and characteristics of cellulose degrading bacteria in different salinized soils were studied and analyzed in different saline alkaline soil. Cellulose degrading bacteria were screened from Leymus chinensis grassland (pH 8.68), pH 9.47 and pH 10.03. The morphology, biochemical characteristics and 16S rDNA nucleic acid sequence were analyzed and identified, and cellulose degrading bacteria groups in different saline alkali grassland were determined. Under this premise, the enzyme production and plant growth characteristics of the above strains were systematically studied. The results obtained are as follows: 1. fiber in different salinized soils The identification of the isolated strains of cellulose degrading bacteria was identified by the analysis of the morphology, biochemical characteristics and 16S rDNA nucleic acid sequence of the screened cellulose degrading bacteria. The analysis of the growth characteristics of the strain showed that all of the strains grew well under the conditions of 0-4%NaCl and PH7-11. The separation and screening of 35 strains were obtained by the 16SrDNA nucleic acid sequence ratio. After the identification of the same strain, 27 cellulose degrading bacteria were obtained. The 16S rDNA nucleic acid sequence of all the strains was submitted to GenBank to get the login number of 27 strains. On the basis of the nucleotide sequence analysis, the analysis of the dominant groups of cellulose degrading bacteria in the phylogenetic tree.2. of all strains was divided into the analysis of the dominant groups of cellulose degrading bacteria in different saline alkali soils. Among the 27 isolates, there were 10 endemic cellulose degrading bacteria in Leymus chinensis grassland, 9 strains of cellulose degrading bacteria endemic to the grassland soil, 4 strains of cellulose degrading bacteria in the weed grassland, and the remaining 4 strains of cellulose degrading bacteria in three different grassland soils, including Streptomyces KY007170.1 and ganglion The bacteria KY007166.1 is a common strain of the grass grassland and the Leymus chinensis grassland. The Streptomyces KY007169.1 and the Bacillus KY007185.1 are three kinds of grassland. This shows that in the saline alkali soil, the Streptomyces and Arthrobacter are more widely distributed and more adaptable. The results of the analysis of the cellulose degrading bacteria in three different grassland by using the CMC plate are analyzed. The results showed that among the three different salinity grassland, the highest abundance strains were Streptomyces, the abundance was 71.4% of Leymus chinensis grassland (71.4%), the grass grassland (71.4%) and the weedy grassland (85.7%). Thus, we concluded that the Streptomyces showed strong salt alkali adaptation ability and the dominant cellulose degrading bacteria.3. score in the saline alkali grassland. Cellulose degradation characteristics (1) produced cellulase from the strain (1), the cellulase activity of the isolated strain was preliminarily analyzed by the Congo red staining method. Meanwhile, the cellulase activity of the isolated strain was analyzed by the method of DNS. The results showed that most of the strains produced extracellular cellulase, and the activity range of the enzyme was in 1.47~6.. Between 08 * 10~ (-2) IU, the cellulase activity of Streptomyces DS-22 was the highest, the activity was 6.08 x 10~ (-2) IU. (2) xylanase, and the xylanase activity of the isolated strain was analyzed by DNS method. The results showed that most of the bacteria produced extracellular xylanase, and the activity range was between 2.23~18.80 * 10~ (-2) IU. The activity of the xylanase activity was the highest, the activity was 18.80 * 10~ (-2) IU.4. isolation strain. The nitrogen fixation activity of the isolated strain was analyzed by the nitrogen free cellulose plate. The results showed that the cellulose degrading bacteria grew well on the non nitrogen source cellulose plate and were the nitrogen fixing bacteria that degrade cellulose. The acetylene reduction method was used. The activity of nitrogen fixing enzyme of the isolated strain was detected and analyzed. All the strains could produce nitrogenase and the activity range of nitrogenase was between 36.29~112.08 nmoL ml. H~ (-1). Among them, the Streptomyces DS-18 nitrogenase activity was the highest, and the activity was the growth promoting characteristics of 112.08nmoL. Ml. H~ (-1).5. isolates. The results showed that all the strains had the ability to produce IAA, with the range of 2.4~80.69mg/L, of which the DS-34 produced the maximum IAA content, 80.69mg/L, and 6 strains possessed the ability to produce iron carriers, all of which belonged to the Streptomyces, and the 8 strains had the ability to dissolve phosphorus, including 7 strains of Streptomyces and 1 strains. Bacilli; 11 strains of bacteriostasis, 7 strains of Streptomyces, 2 strains of Arthrobacter, 1 strains of Nocardia and 1 strains of.6. in different salinization levels of soil physical and chemical properties and biomass analysis of different salinity soil physical and chemical properties and biomass were analyzed. The results showed that with the salinity of the soil, the degree of salt and alkali degree. The content of total nitrogen and organic carbon in the soil decreased continuously, and the number distribution of the cultivated microbes in the three different salinity grassland soils were all: the plant growth promoting experiment of bacterial actinomycetes cellulose degrading fungi.7. isolates from the above strains, three strains of Streptomyces, DS-2 and DS-30 were selected for pot experiments. The results of pot experiments showed that the effect of all single strain treatment group was more obvious than that of the control group, and the effect of the mixed strain group was more obvious than that of the single strain treatment group. The results showed that the cellulose degrading bacteria had a good promoting effect.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S812.2

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