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楊樹(shù)根際高效解鉀細(xì)菌的分離篩選與鑒定

發(fā)布時(shí)間:2018-03-20 06:16

  本文選題:解鉀菌 切入點(diǎn):楊樹(shù) 出處:《南京林業(yè)大學(xué)》2016年碩士論文 論文類型:學(xué)位論文


【摘要】:我國(guó)的楊樹(shù)產(chǎn)業(yè)非常發(fā)達(dá),在全國(guó)分布廣泛、品種數(shù)量齊全,楊樹(shù)人工林面積超過(guò)其他國(guó)家人工林面積的總和,是我國(guó)速生用材的重要樹(shù)種。但是近50年來(lái),人工林的面積不斷的擴(kuò)大,輪伐期不斷縮短,造成林地地力衰退,生產(chǎn)力下降,這與林地土壤肥力下降密切相關(guān),其中鉀素供應(yīng)不足是突出而具普遍性的問(wèn)題。鉀是植物生長(zhǎng)發(fā)育的必需養(yǎng)分。植物缺鉀主要是由于土壤中能被其利用的有效鉀很少,若能將土壤礦物鉀轉(zhuǎn)化為有效鉀,則能有效改善目前缺鉀的現(xiàn)狀。解鉀菌(potassium bacteria)是指從土壤中分離出來(lái)的,能分解鉀長(zhǎng)石等硅鋁酸鹽礦物,把難溶性的鉀、磷、硅轉(zhuǎn)變?yōu)榭扇苄誀顟B(tài),促進(jìn)植物生長(zhǎng)發(fā)育,提高產(chǎn)量的一類微生物。由于不同植物根際的解鉀細(xì)菌的種類和數(shù)量存在差異,不同種類的解鉀菌或不同菌株之間的解鉀能力存在較大差異,所以從特定植物根際篩選高效解鉀細(xì)菌的工作顯得尤為重要。本研究從楊樹(shù)根際土壤分離并篩選到3株高效解鉀菌株JW-4、JW-5和JW-7,并進(jìn)行了鑒定,對(duì)楊樹(shù)的促生作用進(jìn)行了研究,最后對(duì)促生效果最好的菌株JW-7的發(fā)酵條件進(jìn)行了優(yōu)化,為其以后的開(kāi)發(fā)利用,解決我國(guó)土壤缺鉀現(xiàn)狀具有重要的現(xiàn)實(shí)意義,主要研究結(jié)果如下:利用硅酸鹽培養(yǎng)基,采用稀釋涂布平板法,初篩到13株解鉀菌。對(duì)這13株菌進(jìn)行搖瓶釋鉀試驗(yàn),測(cè)定其解鉀效率,從中篩選出3株高效解鉀菌JW-4、JW-5和JW-7,其解鉀效率分別達(dá)到26.30%、34.27%和38.70%。對(duì)3株高效解鉀細(xì)菌采用形態(tài)學(xué)觀察、生理生化特征及16SrDNA序列分析的方法進(jìn)行了鑒定。JW-4、JW-5、JW-7皆為桿菌;都是革蘭氏陰性菌、都沒(méi)有芽孢;菌落顏色呈乳白色;菌落表面凸起,濕潤(rùn);菌落圓形、光滑、不透明或半透明。結(jié)合生理生化特征和16SrDNA序列分析,JW-4初步鑒定屬于根瘤菌屬(Rhizobium sp.),JW-5屬于中華根瘤菌屬(Sinorhizobium sp.),JW-7屬于假單胞菌屬(Pseudomonas sp.)。以楊樹(shù)苗作為供試植物,接種Rhizobium sp.JW-4、Sinorhizobium sp.JW-5、Pseudomonas sp.JW-7于盆栽楊樹(shù)上,結(jié)果表明,JW-7對(duì)楊樹(shù)的促生效果最好。苗高和地徑分別比CK增長(zhǎng)了37.64%和19.44%,鮮重和干重分別比CK增長(zhǎng)了65.82%和110.71%。細(xì)菌、放線菌、硅酸鹽細(xì)菌的數(shù)量均有顯著升高,而真菌的數(shù)量都有不同程度的降低?傮w看,接種JW-4、JW-5、JW-7均能改善土壤微生物環(huán)境,促進(jìn)植物生長(zhǎng)。本試驗(yàn)采用單因素試驗(yàn)法,對(duì)解鉀培養(yǎng)基中的碳源、氮源和無(wú)機(jī)鹽的種類進(jìn)行了優(yōu)化。得出,適合JW-7菌株生長(zhǎng)的解鉀培養(yǎng)基的最佳碳源為麥芽糖,最適濃度為1%;最佳氮源為蛋白胨,最適濃度為0.2%;最佳無(wú)機(jī)鹽為磷酸氫二鉀,最適濃度0.05%。利用單因素試驗(yàn)與正交設(shè)計(jì)試驗(yàn)相結(jié)合的方法,對(duì)JW-7菌株的培養(yǎng)條件(裝液量、接種量、培養(yǎng)溫度、初始pH值、培養(yǎng)時(shí)間)進(jìn)行了優(yōu)化,結(jié)果顯示,JW-7菌株的最佳培養(yǎng)條件為:80mL的裝液量,7%的接種量,30℃培養(yǎng)溫度,7.5初始pH,72h培養(yǎng)時(shí)間。本研究結(jié)果表明,從楊樹(shù)根際土壤中篩選到的3株高效解鉀菌Rhizobium sp.JW-4、Sinorhizobium sp.JW-5、Pseudomonas sp.JW-7,對(duì)楊樹(shù)生長(zhǎng)均有促生作用,并能改善土壤中的微生物環(huán)境。
[Abstract]:Well developed poplar industry in China, in the country are widely distributed, the number of varieties complete, poplar plantation area of more than all other countries combined artificial forest area, is an important fast-growing timber tree species in China. But in the recent 50 years, expanding the plantation area, rotation is shortened and the cause of forest soil degradation with this, a decline in productivity, soil fertility decline is closely related to the supply of potassium deficiency is prominent and universal. Potassium is the essential nutrients for plant growth and development. The plant is mainly due to lack of potassium in soil can be the effective potassium rarely, if the soil mineral potassium into effective potassium then, can effectively improve the current situation of potassium deficiency. Potassiumbacteria (potassium bacteria) is separated from the soil, can decompose potassium feldspar aluminosilicate minerals, the solubility of potassium, phosphorus, silicon into soluble state, promote planting Growth and development, improve the yield. As a kind of microbial species and quantity of potassium bacteria in rhizosphere of different plants are different, there is a big difference in potassium solution ability between different kinds of bacteria or different strains, so the screening of high potassium bacterium from the specific plant rhizosphere is particularly important. This study from Yang the rhizosphere soil isolated and screened 3 strains of bacteria, strain JW-4, JW-5 and JW-7, and were identified on poplar growth promoting effect is studied, finally the fermentation conditions of the growth promoting effect of the best strains of JW-7 were optimized for the development and utilization of the future, has important practical significance to solve the lack of potassium China's soil status, the main results are as follows: medium using silicate, the dilution plate method, screening 13 strains of bacteria. Of the 13 strains of shake flask potassium release test, the determination of potassium in efficiency. Screening of 3 strains of bacteria, JW-4, JW-5 and JW-7, the potassium efficiency reached 26.30%, 34.27% and 38.70%. of 3 strains of high potassium bacteria with morphological observation, analysis of physiological and biochemical characteristics and 16SrDNA sequence were identified by.JW-4, JW-5, JW-7 are all bacteria; gram negative bacteria, no spore the milky white color; colony; wetting; colony colony surface bulge, round, smooth, opaque or translucent. Combined with the physiological and biochemical characteristics and 16SrDNA sequence analysis, identification of JW-4 belongs to the genus Rhizobium (Rhizobium sp.), JW-5 belongs to the genus Rhizobium (Sinorhizobium sp. JW-7), belonging to the genus Pseudomonas (Pseudomonas sp.). In poplar seedlings as tested plants inoculated with Rhizobium, sp.JW-4, Sinorhizobium sp.JW-5, Pseudomonas sp.JW-7 on potted poplar, the results show that the growth promoting effect of JW-7 on poplar. The best seedling height and diameter were increased than CK The 37.64% and 19.44%, fresh weight and dry weight were more than the CK growth of 65.82% and 110.71%. of bacteria, actinomycetes, quantity of silicate bacteria was significantly increased, while the number of fungi was decreased in different degree. Overall, JW-4 vaccination, JW-5, JW-7 could improve soil microbial environment, promote the growth of plants the experiment of single factor test, the carbon source medium potassium cultivation, nitrogen sources and inorganic salts were optimized. The suitable strain JW-7 growth solution of potassium culture medium was the best carbon source maltose, the optimal concentration is 1%; the best nitrogen source was peptone, the most optimum concentration was 0.2%; the best inorganic salt as two potassium hydrogen phosphate, the optimum concentration of 0.05%. method by single factor test and orthogonal design combined with the culture conditions of strain JW-7 (liquid volume, inoculation quantity, culture temperature, initial pH, incubation time) were optimized. The results showed that the JW-7 strain The optimal culture condition of liquid filled 80mL, inoculum 7%, 30 C 7.5 of the initial pH, culture temperature, culture time of 72h. The results of this study showed that from poplar rhizosphere soil screened 3 strains of bacteria, Rhizobium sp.JW-4, Sinorhizobium sp.JW-5, Pseudomonas sp.JW-7, has a promoting effect on the growth of poplar, and can improve the microorganism in the soil environment.

【學(xué)位授予單位】:南京林業(yè)大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2016
【分類號(hào)】:S792.11;S714.3

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