本文選題：內(nèi)生菌 + 促生長防病機(jī)能��； 參考：《云南農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：目前,農(nóng)業(yè)生產(chǎn)上提高作物單產(chǎn)及防控病蟲害的主要方式為增施化肥及使用化學(xué)農(nóng)藥。然而它們?cè)诒Ｕ霞Z食增產(chǎn)的同時(shí),也引發(fā)了環(huán)境污染,生態(tài)平衡破壞及食品安全等問題。微生物菌肥已被證實(shí)具有提高作物產(chǎn)量且能減輕病害的危害程度,因此,開展有益微生物篩選并開發(fā)成微生物菌肥的研究工作具有重要的意義。本研究圍繞篩選促生長防病的生防菌株、生防菌微生物菌肥的開發(fā)及其生物效益等方面開展研究,獲得了如下結(jié)果:1.通過本實(shí)驗(yàn)室先前分離并保存的幾株玉米內(nèi)生細(xì)菌的IAA合成、固氮解磷活性及盆栽玉米葉片葉綠素含量的測(cè)定,篩選出具有良好應(yīng)用潛力的內(nèi)生細(xì)菌。結(jié)果發(fā)現(xiàn)解淀粉芽孢桿菌Y19菌株合成IAA的能力和固氮解磷活性均優(yōu)于Y13和Y10菌株,盆栽玉米的葉綠素含量也更高。因此,將菌株Y19作為后續(xù)研究的供試菌株。2.通過自然轉(zhuǎn)化方式,將含有綠色熒光蛋白基因gfpmut3a的質(zhì)粒p HAPII導(dǎo)入Y19,獲得具有良好熒光表型的標(biāo)記菌株Y19-GFPmut3a。室內(nèi)平板對(duì)峙、玉米盆栽促生定殖試驗(yàn)的結(jié)果表明,質(zhì)粒的引入對(duì)Y19的拮抗病原及促生效果并無明顯的影響。在溫室玉米盆栽試驗(yàn)中,激光共聚焦顯微鏡觀察結(jié)果顯示,接菌后的第9天,可觀察到其在玉米根、莖及葉部組織的定殖;接種后的第100 d,仍能在玉米、茄子、白菜的根際土壤、根、莖及葉組織中檢測(cè)到標(biāo)記菌株。這些結(jié)果表明Y19在不同植物體內(nèi)具有良好的定殖及運(yùn)輸性能。3.使用含有終濃度為0.03%黃原膠的Y19-GFPmut3a(106CFU/m L)發(fā)酵液噴施煙草葉面,和(或)直接使用Y19-GFPmut3a(106CFU/m L)發(fā)酵液進(jìn)行灌根處理,結(jié)果表明灌根和葉面噴施的雙重處理組中煙草的株高、鮮重和干重明顯增加。接種后的第30天,仍能在煙草中檢測(cè)到標(biāo)記菌株,這表明Y19在煙草體內(nèi)具有良好的定殖能力,且Y19的定殖并不影響煙草葉內(nèi)及葉表的細(xì)菌種類和數(shù)量。4.菌株Y19分別與實(shí)驗(yàn)室保存的促生防病效果顯著的解淀粉芽孢桿菌B9601-Y2、具有溶磷活性的洋蔥伯克氏菌1-2、YN201305分別兩兩混合培養(yǎng)的結(jié)果表明彼此之間無拮抗作用。在此基礎(chǔ)上以單獨(dú)培養(yǎng)的發(fā)酵液為對(duì)照,將上述的兩兩混合培養(yǎng)后的發(fā)酵液分別與基質(zhì)復(fù)配(體積質(zhì)量比為1:2)后,種植玉米、蘿卜、白菜和油菜。試驗(yàn)結(jié)果表明,單一的Y19發(fā)酵液與基質(zhì)復(fù)配促生長效果最好。5.將Y19發(fā)酵液(106CFU/m L)與不同含量的氮肥、磷肥或鉀肥的基質(zhì)均勻混合后,用于漂浮育苗水稻和盆栽玉米的促生試驗(yàn)。結(jié)果發(fā)現(xiàn)Y19發(fā)酵液與含有3~4%的磷肥的基質(zhì)復(fù)配的促生長效果最好。盆栽30 d后,與對(duì)照相比,玉米鮮重可增加448.05%-487.60%。這表明Y19可與含有化學(xué)肥料的基質(zhì)復(fù)配且制成的復(fù)合基質(zhì)能顯著地促進(jìn)玉米幼苗生長發(fā)育。6.玉米根際土壤微生物群落功能樣性及土壤微生物量的變化檢測(cè)結(jié)果發(fā)現(xiàn)施入Y19微生物菌肥后,玉米根系細(xì)菌、放線菌的數(shù)量增加,而真菌的數(shù)量卻下降,特別是鐮刀菌的數(shù)量。此外,Y19微生物菌肥也提高了根際微生物群落的碳源利用率(AWCD),主要提高了對(duì)碳水化合物類、氨基酸和羧酸類化合物的利用。這些數(shù)據(jù)說明Y19微生物菌肥可通過改善根際土壤微環(huán)境,從而發(fā)揮促生和防病的雙重功能。7.Y19微生物菌肥菌株貨架期檢測(cè)結(jié)果發(fā)現(xiàn)Y19在純基質(zhì)及含3%磷肥的基質(zhì)中存活率最高。在溫度為28℃、濕度為50%的環(huán)境條件下保存1年,其活菌量維持在107CFU/g以上。
[Abstract]:At present, the main way to increase crop yield and prevent and control diseases and pests in agricultural production is to increase chemical fertilizer and use chemical pesticides. However, while ensuring the increase of grain yield, they also cause environmental pollution, ecological balance damage and food safety. Microbial fertilizer has been proved to increase crop yield and reduce the damage of disease. Therefore, it is of great significance to carry out beneficial microorganism screening and development of microbial microbial fertilizer. This study focuses on the screening of biocontrol strains for growth prevention and control of growth prevention, the development of microbial fertilizer for biocontrol bacteria and its biological benefits. The following results are obtained: 1. through several previous isolation and preservation in our laboratory The IAA synthesis of endophytic bacteria, the activity of nitrogen fixing and the determination of the content of chlorophyll in the leaves of the potted maize were measured, and the endophytic bacteria with good application potential were screened out. The results showed that the ability to synthesize IAA and the phosphorus fixing activity of the Bacillus amylobacter Y19 strain were superior to those of the Y13 and Y10 strains, so the content of chlorophyll in the potted maize was also higher. The strain Y19 was used as a follow-up study strain.2. to transfer the plasmid P HAPII containing the green fluorescent protein gene gfpmut3a into Y19 through natural transformation, and to obtain the labelled strain Y19-GFPmut3a. with good fluorescent phenotype, Y19-GFPmut3a. indoor flat confrontation, and the results of maize plant growth promoting colonization test showed that the plasmid was introduced to the antagonism of Y19. In a greenhouse maize pot experiment, the results of laser confocal microscopy showed that the colonization of the roots, stems and leaves could be observed in the ninth days after the inoculation, and the 100th d after inoculation could still be detected in the rhizosphere soil, root, stem and leaf tissues of maize, eggplant and cabbage. These results show that Y19 has good colonization and transport properties in different plants,.3. using Y19-GFPmut3a (106CFU/m L) fermented solution containing 0.03% yellowish original glue to spray tobacco leaf surface and (or) direct use of Y19-GFPmut3a (106CFU/m L) fermentation broth for root irrigation. The results show the double treatment group of root irrigation and foliar spraying. The plant height, fresh weight and dry weight of the tobacco increased obviously. Thirtieth days after inoculation, the labelled strains could still be detected in tobacco, which showed that Y19 had good colonization in tobacco, and the colonization of Y19 did not affect the species of bacteria in the leaf and leaf surface of tobacco and the number of.4. strain Y19, respectively. Bacillus amyloid B9601-Y2, the results of mixed culture of burkderia onions 1-2 and YN201305 with phosphorus soluble activity, showed no antagonism between 22 mixed cultures. On this basis, the fermentation broth of the 22 mixed culture was mixed with the substrate (volume mass ratio 1:2), respectively. Planting corn, radish, cabbage and rapeseed. The results showed that the best combination of Y19 fermentation liquid and matrix was the best.5., and the Y19 fermenting solution (106CFU/m L) was mixed with different content of nitrogen fertilizer, phosphate fertilizer or potash fertilizer. The results were used to promote the growth of floating seedling rice and potted jade rice. The results showed that the Y19 fermentation liquid and 3~4% contained 3~4%. The substrate compound of phosphate fertilizer has the best growth promoting effect. After 30 d potted plants, the fresh weight of corn can be increased by 448.05%-487.60%. compared with the control, which indicates that Y19 can be combined with the matrix containing chemical fertilizer and can significantly promote the microbial community function and soil microbial biomass in the maize seedling growth and development of maize rhizosphere soil. The results showed that the number of bacteria and actinomycetes increased, but the number of fungi decreased, especially the number of Fusarium. In addition, Y19 microbial fertilizer also improved the carbon source utilization (AWCD) of the rhizosphere microbial community (AWCD), which mainly improved carbohydrates, amino acids and carboxylic acids. Using these data, these data show that Y19 microbial fertilizer can improve the microenvironment of Rhizosphere Soil by improving the shelf life detection results of the dual function.7.Y19 microbial fertilizer strain for promoting and preventing disease. It is found that Y19 has the highest survival rate in the pure matrix and the matrix containing 3% phosphate fertilizer. At the temperature of 28, and the humidity of 50%, 1 is preserved. In the year, the amount of living bacteria was more than 107CFU/g.
【學(xué)位授予單位】：云南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S144

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