【作者】 张朝辉；

【导师】 邱立友；

【作者基本信息】 河南农业大学 ， 微生物学， 2010， 硕士

【摘要】 本文应用分离自烟株根际的高效抗生菌B03、固氮菌N05、解磷菌P04、解钾菌K03,进行菌种鉴定,明晰其分类地位,制成PGPR肥料,用于烤烟漂浮育苗和大田生产,减少化肥、化学农药的使用量,提高烟叶产量和质量。取得四方面成果:1)PGPR菌肥菌种的鉴定应用16S rDNA序列分析构建系统发育树,结合生理指标、生化反应结果:抗生菌B03属于芽孢杆菌属(Bacillus),可能是蜡状芽孢杆菌(Bacillus cereus)、固氮菌N05属于产碱菌属(Alcaligenes),可能是粪产碱菌(Alcaligenes faecalis)、解磷菌P04属于肠杆菌属(Enterobacter),可能是阴沟肠杆菌(Enterobacter cloacae)、解钾菌K03亦属于肠杆菌属(Enterobacter),可能是阿氏肠杆菌(Enterobacter asburiae),均非病原菌,可用于菌肥生产。2)PGPR混合菌肥制备通过拮抗实验证明,本菌肥四个菌种,除抗生菌B03对解磷菌P04有较弱的拮抗作用外,其它菌株间没有拮抗反应,可制成混合菌肥。在菌肥制备后,检测活菌数,四种菌均不低于1亿个,符合菌肥质量的国家标准。3)PGPR混合菌肥应用于烤烟的菌糠基质漂浮育苗实验用菌糠替代草炭进行试验性育苗,结果证明:双孢菇菌糠能够替代草炭进行烤烟漂浮育苗,但是在育苗前的处理和育苗条件上还需要优化;平菇菌糠能否进行漂浮育苗需要进一步验证。PGPR菌肥的加入,能够提高出苗率,提幅达3%,能够提升烟苗的农艺性状,培育壮苗。其中菌肥对烟苗根生长的促进作用非常明显,加菌肥的双孢菇菌糠处理比对照增幅达101.7%。4)PGPR菌肥用于烤烟大田生产在烤烟移栽时施用PGPR混合菌肥,同时减少N、P和K肥使用量,研究施用菌肥和减施N、P和K肥对植烟土壤和烤烟生产的影响。试验表明:施用PGPR菌肥可显著提高现蕾期烤烟根际解磷菌的数量,降低放线菌的数量,对细菌、真菌、固氮菌、解钾菌影响不大,而减施N、P和K肥对各种菌数影响均不大。PGPR菌肥能提高烤烟根际微生物生物量碳含量,在烤烟生理活性强的现蕾期,常规施肥+菌肥处理微生物生物量碳含量提高22.2%,显著高于常规施肥对照(p<0.05);减施肥料能提高烤烟根际微生物生物量碳含量。PGPR的施入能提高矿质元素有效性,常规施肥+菌肥处理除B元素略低外N、P、K、Ca、Mg、Cu、Zn、Mn、Fe的有效性均明显高于常规施肥,提高幅度在8.19%～49.33%;N、P和K肥各减20%+菌肥处理与其对照相比,N、K、Cu、Zn、B、Fe 6种元素的有效性是提高的,提高幅度在4.46%～28.87%,略低于常规施肥+菌肥处理。PGPR菌肥能降低烤烟发病率,常规施肥+菌肥烤烟亩产量比常规施肥对照提高7.59%,产值显著提高14.8%;减施N、P和K肥会降低烟草的产量、质量和产值,而PGPR菌肥的加入能弥补减肥带来的不足,使N、P和K肥各减20%+菌肥的最终产值显著高于常规施肥,比减施N、P和K肥对照提高16.44%,略低于常规施肥+菌肥。综上所述,在烤烟漂浮育苗和栽培中应用菌肥,可有效地提高发芽率和成苗素质,降低化学肥料的施用量,改善烤烟根际微生物区系,提高土壤矿质元素的有效性,提高烟叶的产量、产值和上等烟率,是一项投入少、效益高的可持续生产发展措施。应大力推广使用PGPR菌肥,利用神农架自然原生态环境,以彰显烟叶地方特色,达到生态烟叶生产与自然环境的和谐发展。更多还原

【Abstract】 High-effective antibiotic bacterium B03, bacterium of nitrogen fixation N05, bacterium of dissolving phosphorite P04 and bacterium of dissolving potassium feldsper K03 were separated from the tobacco rhizosphere and mixed to PGPR fertilizer after strain identification to reduce the use of chemical fertilizer and poison, promote the output and quality of the tobacco.1) Strain identificationIt can be concluded that antibiotic bacterium B03, bacterium of nitrogen fixation N05, bacterium of dissolving phosphorite P04 and bacterium of dissolving potassium feldsper K03 belong to Bacillus and maybe Bacillus cereus, Alcaligenes and Alcaligenes faecalis, Enterobacter and Enterobacter cloacae, Enterobacter and Enterobacter asburiae, respectively with phylogenetic tree based on 16S rDNA sequences and the physiological and biochemical reaction. Four bacteria are all not pathogens, can be used for the production of PGPR fertilizer.2) The production of PGPR mixed fertilizerDuring the antagonistic experiment, no antagonism had been found except the antibiotic bacterium B03 exerts weak antagonistic reaction on bacterium of dissolving phosphorite P04. PGPR mixed fertilizer was made and the live bacteriums was no less than 100,000,000 which fits to the national standard.3) The application of PGPR mixed fertilizer in tobacco floating systemMushroom substrate were used to floating system and revealed that Agaricus Bisporus mushroom substrate could take the place of peat in tobacco seedling production after optimize the treatment before seedling and seedling condition, but required further confirm that whether the Pleurotus ostreatus mushroom substrate also can be used. The PGPR fertilizer improved rate of emergence by 3%, the agronomic characters of tobacco seedling as well as strong seedling. The fertilizer had been promoted the root growth greatly, for example, Agaricus Bisporus mushroom substrate with PGPR fertilizer raise to 101.7%.4) The application of PGPR mixed fertilizer in flue-cured tobacco fieldThe influence on the tobacco-planting soil and the production of flue-cured tobacco were studied employing PGPR fertilizer and reducing the use of the chemical fertilizer when flue-cured tobacco transplanted in plots. The results showed that application of PGPR fertilizer would increase the number of rhizo-bacterium of dissolving phosphorite significantly in budding period (p<0.05) and reduce amount of actinomycete during growth period and have no influence on total amount of bacteria, fungus, nitrogen-fixing bacteria and dissolving potassium bacteria. PGPR fertilizer also increased the tobacco rhizo-microbial biomass C contents, for example, 22.2% had been increased in budding period, a period of strong physiological activity. The use of PGPR fertilizer improved effectiveness of mineral element, such as N, P, K, Ca, Mg, Cu, Zn, Mn, Fe were all significantly higher than without GPGR fertilizer, range from 8.19% to 49.33%, while the effectiveness of B reduced. After PGPR fertilizer application, the disease incidence in flue-cured tobacco decreased and tobacco yield per 0.067 ha and production value increased up to 7.59% and 14.8%, respectively. It is well known that reducing N, P and K fertilizer would result for low yield, quality and production value of tobacco, but the production value of application of PGPR fertilizer while reducing N, P, K fertilizer increased up to 16.44%, it can be concluded that the use of PGPR fertilizer could make up for the decreased chemical fertilizer.更多还原