【摘要】：目前我國(guó)傳統(tǒng)農(nóng)業(yè)生產(chǎn)中長(zhǎng)期大量施用N、P、K元素肥料,雖然一定程度上緩解了我國(guó)糧食安全問(wèn)題,但也由此產(chǎn)生了一系列環(huán)境問(wèn)題,如土壤酸化、板結(jié)、鹽漬化等。適當(dāng)施用生物菌肥可以提高肥效、防治病害和修復(fù)土壤,是實(shí)現(xiàn)農(nóng)業(yè)可持續(xù)發(fā)展的必要途徑之一。生產(chǎn)低成本、高益生菌含量的微生物肥料具有廣闊的市場(chǎng)前景,也非常符合我國(guó)綠色農(nóng)業(yè)的發(fā)展策略。傳統(tǒng)益生菌液體發(fā)酵工藝因其工藝復(fù)雜、成本高,在一定程度上限制了其在農(nóng)業(yè)領(lǐng)域的應(yīng)用,因此開(kāi)發(fā)低成本、適用于農(nóng)業(yè)生產(chǎn)的益生菌劑及其衍生產(chǎn)品,是推動(dòng)微生物肥料領(lǐng)域技術(shù)進(jìn)步的關(guān)鍵環(huán)節(jié)。本文首先對(duì)4株益生芽孢桿菌進(jìn)行了形態(tài)學(xué)和分子生物學(xué)的鑒定及特性分析。隨后采用固體發(fā)酵手段研究開(kāi)發(fā)低成本、高密度益生芽孢桿菌生產(chǎn)工藝。通過(guò)單因素和正交工藝優(yōu)化,最終確定Bacillus subtilis L7固體發(fā)酵工藝條件為：糠醛渣、廢糖液和麥麩添加量分別為85%、10%和5%,初始水分60%,溫度30℃,初始pH 7.0,接種量5×106CFU/g,翻堆頻率為1次/d,L7發(fā)酵水平達(dá)到(活菌數(shù)和芽孢數(shù)分別為)2.48×1010CFU/g和2.40×1010CFU/g,芽孢率達(dá)到96.7%。隨后對(duì)L7固體發(fā)酵工藝進(jìn)行了100kg規(guī)模的中試放大,發(fā)酵后活菌數(shù)達(dá)到3.26×1010CFU/g,菌活水平比小試提高了31.1%,且發(fā)酵時(shí)間由6天縮短為3天。同時(shí)對(duì)其他三種益生芽孢桿菌進(jìn)行固體發(fā)酵小試實(shí)驗(yàn)。結(jié)果表明Bacillus licheniformis D6、Bacillus megaterium J3和Bacillus mucilaginosus G14發(fā)酵后總活菌數(shù)分別為2.48×1010CFU/g、1.02×1010CFU/g和8.08×109CFU/g,發(fā)酵水平高于相應(yīng)的液體工藝,且本方法成本低廉,具有明顯的競(jìng)爭(zhēng)優(yōu)勢(shì)。隨后我們將固體發(fā)酵菌劑與復(fù)合肥(NPK 16：16：8)復(fù)配制得益生菌含量為1.25×108 CFU/g的微生物肥料；以蘇州青油菜(Brassica campestris L. Sp)為實(shí)驗(yàn)樣本開(kāi)展盆栽實(shí)驗(yàn),研究微生物肥料對(duì)其作用效果。結(jié)果表明,所制備的復(fù)合微生物肥料B3可以有效的提高實(shí)驗(yàn)作物根系和莖葉干重,與對(duì)照相比分別提高46.1%和34.1%,以上結(jié)果說(shuō)明該復(fù)合微生物肥料能夠顯著促進(jìn)植株生長(zhǎng)。最后我們采用DGGE方法對(duì)盆栽作物根際土壤菌群進(jìn)行了分析。結(jié)果表明,添加30天后,微生物肥料對(duì)細(xì)菌和真菌種類和種群結(jié)構(gòu)都有明顯的影響。其中復(fù)合微生物肥料B3組細(xì)菌豐富度和多樣性指數(shù)與對(duì)照組相比分別降低206%和47.7%,真菌豐富度和多樣性指數(shù)分別降低233%和47.8%。添加60天后,微生物肥料對(duì)土壤中真菌結(jié)構(gòu)仍有顯著影響。其中復(fù)合微生物肥料B3與對(duì)照組相比,真菌豐富度和多樣性指數(shù)分別降低98.2%和26.7%。上述結(jié)果說(shuō)明經(jīng)固體發(fā)酵工藝所開(kāi)發(fā)的復(fù)合微生物肥料能夠明顯抑制土壤真菌,在防治作物各種土傳真菌病害方面具有廣闊的開(kāi)發(fā)前景。
[Abstract]:At present, the traditional agricultural production in our country has applied a large amount of fertilizer with NbPK element for a long time. Although it has alleviated the problem of food security in China to some extent, it has also resulted in a series of environmental problems, such as soil acidification, cementation, salinization and so on. Proper application of biofilm fertilizer can improve fertilizer efficiency, control diseases and repair soil, which is one of the necessary ways to realize the sustainable development of agriculture. The production of low cost and high probiotic microbial fertilizer has a broad market prospect and is also in line with the development strategy of green agriculture in China. The traditional liquid fermentation technology of probiotics, because of its complexity and high cost, limits its application in the field of agriculture to a certain extent, so it can develop low cost probiotics and its derivative products, which are suitable for agricultural production. It is the key link to promote the technical progress in the field of microbial fertilizer. In this paper, four strains of Bacillus probiotics were identified and characterized by morphological and molecular biology. Then the production process of low cost and high density probiotic bacillus was studied by solid-state fermentation. Through the optimization of single factor and orthogonal process, the technological conditions of Bacillus subtilis L7 solid fermentation were determined as follows: furfural residue, waste sugar solution and wheat bran were added to 85% and 5% respectively, the initial moisture content was 60%, the temperature was 30 鈩，

本文編號(hào)：2300042