本文選題：枯草芽孢桿菌 + 培養(yǎng)基��； 參考：《東北農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：枯草芽孢桿菌(Bacillus subtilis)是當(dāng)今研究最為熱門的生防菌,含有較為多樣的生理習(xí)性,容易被分離培養(yǎng),因能夠產(chǎn)生多樣抗性的內(nèi)生芽孢,所以被認(rèn)為是作為生防菌較好的一種選擇。沼液是沼氣在發(fā)酵后剩余的液體,不僅富含大量可快速被吸收的營(yíng)養(yǎng)成分,而且還含有其它供微生物生長(zhǎng)的物質(zhì),如氨基酸、多種微量元素、維生素和其他活性物質(zhì)。隨著我國(guó)農(nóng)村沼氣工程的規(guī)模不斷擴(kuò)大,沼液資源化安全處理與利用備受重視。利用的枯草芽孢桿菌D1是一株具有拮抗性的細(xì)菌,前期試驗(yàn)發(fā)現(xiàn)在牛糞沼液中可以生長(zhǎng),進(jìn)一步優(yōu)化該菌株利用沼液的能力,為沼液再利用提供了新的代謝途徑。該菌株對(duì)水稻的立枯病具有較高的拮抗作用,為了能將其更好的保存下來,通過選取優(yōu)良的載體和穩(wěn)定的保護(hù)劑,在高效利用沼液的同時(shí)將其保存,這也為后期在生物農(nóng)藥上利用提供技術(shù)參數(shù)。但想要將其應(yīng)用于菌劑的制備,菌體本身芽孢含量是影響實(shí)際應(yīng)用的關(guān)鍵因素,采用分批培養(yǎng)法優(yōu)化了枯草芽孢桿菌高效利用沼液的產(chǎn)孢培養(yǎng)基;利用比濁法和稀釋平板涂布法來檢測(cè)枯草芽孢桿菌的菌體數(shù)量及芽孢產(chǎn)率;利用火焰原子吸收光譜法分析了沼液中金屬離的含量,優(yōu)化產(chǎn)孢培養(yǎng)基成分后進(jìn)行菌劑載體和保護(hù)劑的初步研究,旨在高附值利用沼液,降低生產(chǎn)成本的同時(shí),枯草芽孢桿菌的芽孢產(chǎn)率得到提升,并得到穩(wěn)定的保存,為后續(xù)的實(shí)際應(yīng)用做出理論依據(jù)。得到的主要研究結(jié)論如下:(1)活化菌株并測(cè)定生長(zhǎng)曲線�？莶菅挎邨U菌D1是革蘭氏陽(yáng)性菌,對(duì)立枯病有顯著的拮抗作用,在19h時(shí)為生長(zhǎng)高峰期。(2)利用單因素試驗(yàn)確定枯草芽孢桿菌D1的最佳產(chǎn)孢培養(yǎng)基的最佳碳源是麩皮,最佳氮源是大豆粉;優(yōu)化后得到一組最優(yōu)解為培養(yǎng)基的成分為含量為2.45%的麩皮,含量為0.9%的大豆粉和含量為49.01%的沼液,芽孢產(chǎn)率為87.82%。(3)對(duì)沼液的性質(zhì)進(jìn)行測(cè)定。供試沼液中含N 1580 mg/Kg、P 379 mg/Kg、K 998 mg/Kg,該沼液的p H為7.52,總固體含量即TS%為5;沼液中含有能促進(jìn)枯草芽孢桿菌D1生長(zhǎng)的Ca~(2+)、Mn~(2+)、Mg~(2+)這三種離子,檢測(cè)了沼液中的這三種離子的含量,分別為:Ca~(2+)14.8 mg/L、Mg~(2+)0.151 mg/L、Mn~(2+)0.08 mg/L。(4)對(duì)優(yōu)化產(chǎn)孢培養(yǎng)基后獲得的高產(chǎn)菌株劑型的載體及保護(hù)劑進(jìn)行篩選試驗(yàn),確定枯草芽孢桿菌D1固體劑型的最適載體為硅藻土,同時(shí)篩選出最佳保護(hù)劑是液體石蠟油,最終有效活菌數(shù)量可達(dá)到89.9%。
[Abstract]:Bacillus subtilisis is one of the most popular biocontrol bacteria, which contains various physiological habits and is easy to be isolated and cultured. Because of its ability to produce endophytic spores with various resistance, Bacillus subtilisis is considered as a better choice for biocontrol. Biogas liquid is the residual liquid of biogas after fermentation. It not only contains a large amount of nutrients which can be quickly absorbed, but also contains other substances for microbial growth, such as amino acids, trace elements, vitamins and other active substances. With the increasing scale of rural biogas project in China, more and more attention has been paid to the safe treatment and utilization of biogas liquid. Bacillus subtilis D1 was an antagonistic bacterium, which was found to be able to grow in cow manure biogas liquid in previous experiments, and further optimized the ability of the strain to utilize biogas liquid, which provided a new metabolic pathway for the reuse of biogas liquid. The strain has a high antagonistic effect on rice bacterial blight. In order to preserve it better, it can be preserved by selecting excellent carrier and stable protective agent, using biogas liquid efficiently at the same time. This also provides technical parameters for the later use of biological pesticides. In order to be used in the preparation of bacterial agents, the spores content of Bacillus subtilis was the key factor affecting the practical application. The spore production medium of Bacillus subtilis was optimized by batch culture. The bacterial number and spores yield of Bacillus subtilis were determined by turbidimetry and dilution plate coating, and the content of metal dissociation in biogas liquid was analyzed by flame atomic absorption spectrometry. After optimizing the composition of spore producing medium, the preliminary study of fungicide carrier and protective agent was carried out. The purpose of this study was to use biogas liquid with high yield value and reduce the production cost, at the same time, the spores yield of Bacillus subtilis was improved and kept stably. To provide the theoretical basis for the subsequent practical application. The main results are as follows: 1) activated strain and measured growth curve. Bacillus subtilis D1 is a Gram-positive bacterium, which has a significant antagonistic effect on bacterial blight. It is the peak period of growth at 19 h.) the best carbon source for spores production medium of Bacillus subtilis D1 is wheat bran by single factor test. The optimum nitrogen source was soybean powder, and the optimum composition of the medium was wheat bran (2.45%), soybean powder (0.9%) and biogas liquid (49.01%), and the spore yield was 87.82%. The biogas liquid contained N 1580 mg / kg P 379 mg / K g / K 998 mg / kg, pH = 7.52, and total solid content was 5%. The biogas liquid contained three kinds of ions, Ca~(2 Mn-MN 2 mg 2, which could promote the growth of Bacillus subtilis D1), and the contents of the three ions in the biogas liquid were detected. The optimal carrier and protectant of high yield strain obtained by optimizing the sporulation medium were determined to be diatomite as the best carrier of Bacillus subtilis D1 solid dosage form, which was 0.151 mg 路L ~ (-1) mg / L ~ (-1) mg 路L ~ (-1) 路L ~ (-1)% 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1). At the same time, the best protective agent is liquid paraffin oil, and the effective live bacteria quantity can reach 89. 9%.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S476.1;TQ450.6

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉振華;邢雪琨;;微生物農(nóng)藥助劑研究進(jìn)展[J];基因組學(xué)與應(yīng)用生物學(xué);2016年08期

2 馮冠穎;;廢水中重金屬離子的火焰原子吸收光譜法測(cè)定研究[J];廣東化工;2016年05期

3 劉榮華;;微生物檢測(cè)中培養(yǎng)基的制備及注意事項(xiàng)[J];當(dāng)代臨床醫(yī)刊;2015年01期

4 劉振華;羅遠(yuǎn)嬋;張道敬;李元廣;;農(nóng)用微生物殺菌劑劑型研究進(jìn)展[J];農(nóng)藥學(xué)學(xué)報(bào);2014年05期

5 蘇紹輝;;微生物降解農(nóng)藥的研究現(xiàn)狀與發(fā)展趨勢(shì)[J];宿州教育學(xué)院學(xué)報(bào);2014年04期

6 花鏡;滕子言;陸小華;楊祝紅;王昌松;;沼液余熱回收對(duì)高溫發(fā)酵沼氣工程凈產(chǎn)氣率的影響[J];化工學(xué)報(bào);2014年05期

7 楊森;王金水;楊小佳;藺丹華;;枯草芽孢桿菌誘變育種研究進(jìn)展[J];農(nóng)產(chǎn)品加工(學(xué)刊);2014年05期

8 高亮;譚德星;;腐植酸生物菌肥對(duì)保護(hù)地次生鹽漬化土壤改良效果研究[J];腐植酸;2014年01期

9 田宜水;;2012年中國(guó)農(nóng)村能源發(fā)展現(xiàn)狀與未來趨勢(shì)[J];中國(guó)能源;2013年03期

10 杜娟;孫佰平;趙思峰;吳彩蘭;;枯草芽孢桿菌S37沼液發(fā)酵培養(yǎng)基優(yōu)化及其產(chǎn)物對(duì)棉苗立枯病的防治[J];西北農(nóng)業(yè)學(xué)報(bào);2013年07期

相關(guān)會(huì)議論文 前1條

1 劉錦霞;李晶;杜文靜;李娜;丁品;武建榮;張建軍;沈思遠(yuǎn);楊蘭廷;;枯草芽孢桿菌Bs-a高毒力、強(qiáng)抗逆生防菌株選育[A];第三屆全國(guó)微生物資源學(xué)術(shù)暨國(guó)家微生物資源平臺(tái)運(yùn)行服務(wù)研討會(huì)會(huì)議論文摘要集[C];2011年

相關(guān)博士學(xué)位論文 前3條

1 章博;枯草芽孢桿菌戊糖與半纖維素的高效利用及乙偶姻的生產(chǎn)[D];天津大學(xué);2014年

2 魏泉源;規(guī)�；託夤こ陶右�、沼渣減量化及資源化利用研究[D];北京化工大學(xué);2014年

3 李文濤;沼液對(duì)土壤改良作用研究[D];東北農(nóng)業(yè)大學(xué);2013年

相關(guān)碩士學(xué)位論文 前6條

1 殷麗麗;餐廚廢棄物產(chǎn)氫產(chǎn)甲烷梯度發(fā)酵試驗(yàn)研究[D];東北農(nóng)業(yè)大學(xué);2014年

2 范慧青;木材炭化防腐處理對(duì)微生物生存條件的影響研究[D];內(nèi)蒙古農(nóng)業(yè)大學(xué);2014年

3 孔令澤;枯草芽孢桿菌SYST2對(duì)田間作物生長(zhǎng)的影響[D];南京農(nóng)業(yè)大學(xué);2014年

4 齊建;粘球菌DK1622胞內(nèi)游離鈣離子濃度的測(cè)定及鈣結(jié)合蛋白基因的預(yù)測(cè)[D];山東大學(xué);2010年

5 徐守健;幾株海洋微生物次生代謝產(chǎn)物篩選農(nóng)用活性物質(zhì)的研究[D];山東農(nóng)業(yè)大學(xué);2006年

6 張麗霞;枯草芽孢桿菌B908發(fā)酵工藝優(yōu)化研究[D];內(nèi)蒙古農(nóng)業(yè)大學(xué);2006年

，

本文編號(hào)：1960402