本文關(guān)鍵詞： 解磷微生物 鑒定 香蕉 促生效果　出處：《海南大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：解磷微生物或溶磷微生物(Phosphate-solubilizing microorganism)是指土壤里能夠?qū)⒘椎V粉等植物難以吸收利用的磷酸鹽轉(zhuǎn)化為可吸收利用形態(tài)的微生物�，F(xiàn)世界將近50%的農(nóng)業(yè)土壤存在缺磷問題,這里的土壤缺磷是“遺傳學(xué)缺磷”而非“土壤學(xué)缺磷”(康貽軍等,2006；王慶仁,2004)。土壤缺乏磷素使植物的生長受到嚴(yán)重影響。直接施加化學(xué)磷肥,一定程度緩解土壤缺磷現(xiàn)象,但現(xiàn)在植物對磷肥等化學(xué)肥料是吸收利用率很低,不足25%,造成資源浪費(fèi),并且還會造成土壤板結(jié)、酸堿化等問題。因此,微生物磷肥的研究是現(xiàn)農(nóng)業(yè)研究的熱點(diǎn)項(xiàng)目。本文以磷礦粉為底物,從香蕉根際土壤分離、篩選和鑒定一批高效解磷細(xì)菌,并研究了代表菌株解磷能力,優(yōu)化了培養(yǎng)條件和解磷菌劑對香蕉幼苗的促生效果。 1、篩選并鑒定了3株具有高效溶解磷礦粉能力的細(xì)菌。香蕉根際土壤經(jīng)過透明圈法和鉬銻抗比色法測定發(fā)酵液有效磷含量進(jìn)行初篩、復(fù)篩,得到了對磷礦粉溶解效果較好的解磷微生物B3-5-6,M-3-01和T1-4-01菌株。通過菌體形態(tài)觀察、生理生化實(shí)驗(yàn)和16S rDNA分子生物學(xué)鑒定,可以判定菌株B3-5-6為嗜氣芽孢桿菌(Bacillus aerophilus),M-3-01為蟲內(nèi)生沙雷氏菌(Serratia nematodiphila),T1-4-01為艾博麗腸桿菌(Enterobacter asburiae)。其中M-3-01菌株溶解磷礦粉效果最好,解磷能力為37.85mg/L,將其作為本實(shí)驗(yàn)解磷微生物代表菌株作更深入的研究。 2、優(yōu)化了M-3-01代表菌株的解磷發(fā)酵配方工藝。通過設(shè)計(jì)單因子試驗(yàn)和正交試驗(yàn)方法相結(jié)合。單因子實(shí)驗(yàn)篩選了最佳碳源為葡萄糖,氮源為草酸銨。再通過確定碳源、氮源、磷礦粉、無機(jī)鹽4組主要影響因子,進(jìn)行L9(34)正交試驗(yàn),篩選最優(yōu)培養(yǎng)基濃度配方,再通過使用單因子試驗(yàn),篩選最優(yōu)配方的起始pH、轉(zhuǎn)速、接種量等條件。最終確定了代表菌株M-3-01的高效解磷發(fā)酵配方。最終優(yōu)化結(jié)果為：葡萄糖15g/L,草酸銨1.5g/L,磷礦粉2.5g/L,無機(jī)鹽1.92g/L,2%接菌量,pH為6,150r/min,37℃。并對其配方進(jìn)行了驗(yàn)證試驗(yàn),優(yōu)化組有效磷含量達(dá)88.64mg/L,是CK有效磷含量的2.34倍。 3、將代表解磷菌株M-3-01制成菌劑,和化肥磷礦粉作對照,研究菌劑對香蕉幼苗的促生效果、盆栽實(shí)驗(yàn)研究發(fā)現(xiàn),菌株M-3-01菌劑+磷礦粉處理對香蕉幼苗促生效果最佳：該處理下香蕉植株的平均株高、莖圍、鮮重、干重分別是對照的1.26、1.16、1.38、1.47倍；且脂膜過氧化及質(zhì)膜透性滲透調(diào)節(jié)物質(zhì)含量及效果均顯著高于其它幾個處理。針對香蕉植株根際土壤的理化因子進(jìn)行分析,接解磷菌劑的處理組的有效磷含量顯著高于CK,有效K和全氮的含量明顯比CK的低,pH變化相對穩(wěn)定,接近于中性。
[Abstract]:Phosphate-solubilizing microorganism (Phosphate-solubilizing microorganism). It is a kind of microorganism that can transform phosphate which is difficult to be absorbed and utilized by plants such as phosphate rock powder into absorbable microorganism in soil. Nearly 50% of the world's agricultural soils are deficient in phosphorus. The soil phosphorus deficiency here is "genetic phosphorus deficiency" rather than "soil phosphorus deficiency" (Kang Yi-jun et al. 2006; Wang Qingren 2004.The plant growth was seriously affected by phosphorus deficiency in soil. Direct application of chemical phosphorus fertilizer to some extent alleviated the phenomenon of phosphorus deficiency in soil. But now plants to phosphorus fertilizer and other chemical fertilizer absorption efficiency is very low, less than 25 percent, resulting in waste of resources, and will also cause soil consolidation, acid and alkali problems. Microbial phosphate fertilizer is a hot research project in agriculture. In this paper, phosphate rock powder was used as substrate to isolate, screen and identify a number of high-efficient phosphate releasing bacteria from banana rhizosphere soil, and the phosphorus releasing ability of representative strains was studied. The effects of culture conditions and phosphorus-releasing bacteria on the growth of banana seedlings were optimized. 1. Three strains of bacteria with high ability to dissolve phosphate rock powder were screened and identified. The available phosphorus content of fermentation broth was determined by transparent circle method and molybdenum antimony colorimetric method in banana rhizosphere soil. Strains B3-5-6CM-3-01 and T1-4-01 were obtained, which had better dissolution effect on phosphate rock powder. The morphology of bacteria was observed. The strain B3-5-6 was identified as Bacillus aerophilus by physiological and biochemical experiments and 16s rDNA molecular biological identification. M-3-01 is Serratia nematodiphila. T1-4-01 was Enterobacter asburiaeae, in which M-3-01 was the most effective in dissolving phosphate rock powder. The phosphorus releasing ability was 37.85 mg / L, which was further studied as the representative strain of phosphorus releasing microorganism in this experiment. 2. The fermentation formula of the representative strain M-3-01 was optimized. The optimum carbon source was glucose by the combination of single factor test and orthogonal experiment. The nitrogen source was ammonium oxalate. By determining the main influencing factors of carbon source, nitrogen source, phosphate rock powder and inorganic salt, the orthogonal experiment was carried out to select the optimal medium concentration formula, and then the single factor test was used. The optimum conditions such as initial pH, rotation speed and inoculation amount were selected. The fermentation formula of the representative strain M-3-01 was determined. The final optimization result was: glucose 15g / L. Ammonium oxalate 1.5 g / L, phosphate rock powder 2.5 g / L, inorganic salt 1.92 g / L 2% inoculated at pH 6 ~ 150 r / min ~ 37 鈩，

本文編號：1478965