【作者】 刘娜；

【导师】 韩梅； 何志刚；

【作者基本信息】 沈阳农业大学 ， 农业硕士（专业学位）， 2020， 硕士

【摘要】 微生物肥料能够增加有效养分含量,提高养分利用率,减少化肥用量,是一种无公害的绿色肥料,微生物肥料与传统的肥料的区别在于微生物肥料是有活性的,并且会产生自身的代谢产物。菌种资源的收集是发展微生物肥料的根本,单一的肥料微生物菌种仅仅提供单一养分,因此不同功能菌种的复配是实现微生物肥料不同菌种协同增效的有效途径之一。本研究采用稀释平板法从果园和菜园耕层土壤中初步分离获得解磷解钾固氮菌28株,其中解磷菌14株,解钾菌11株,固氮菌3株,通过透明圈半定量和液体培养全定量分析等方法对菌株进一步复筛,最终获得优良菌株10株;通过拮抗试验,确证10株菌无拮抗;将菌株发酵液等比例混合后与化肥配施应用于油菜的盆栽试验,发现菌肥+30%化肥的处理油菜多项指标最好。主要研究结果如下:(1)采用解磷固体培养基,通过稀释平板法从沈阳农业大学后山试验基地的菜园和果园耕层土壤中初筛获得解磷菌14株;采用难溶磷酸盐和有机磷液体培养进行菌株解磷能力的定量分析,筛选出优势解磷菌5株,分别为JDYB1、PSYB-1、HLS1、WJ5、YJ1;各菌在解无机磷和有机磷能力上有所侧重,JDYB1、PSYB-1、HLS1解无机磷能力较强,解无机磷的量分别为26.12μg/m L、26.27μg/m L和27.85μg/m L;WJ5和YJ1解有机磷能力较强,解有机磷量分别为23.91μg/m L和23.97μg/m L;对菌株进行产IAA能力检测,显色反应呈阳性的有2株,其中HLS1分泌IAA能力最强,产IAA的量为39.92μg/m L。(2)采用解钾固体培养基,通过稀释平板法从沈阳农业大学后山试验基地的果园和菜园耕层土壤中初筛获得解钾菌11株;采用钾长石液体培养进行菌株解钾能力的定量分析,筛选出优势菌4株,分别为JDYB1、BKR2、YTM和JK1,解钾量依次为12.95μg/m L、12.92μg/m L、12.20μg/m L和11.14μg/m L,其中JDYB1同时具有解磷解钾能力;对菌株进行产IAA能力的检测,显色反应呈阳性的有3株,其中JK1分泌IAA能力最强,产IAA的量为36.01μg/m L。(3)采用无氮固体培养基,通过稀释平板法从沈阳农业大学后山试验基地的菜园和果园耕层土壤中初筛获得固氮菌3株;通过定性和定量的方法进一步筛选产IAA菌株,显色反应呈阳性的有2株,分别为NCC1和ZRT-X,产IAA的量为37.81μg/m L和33.63μg/m L。(4)拮抗试验表明,解磷量高的解磷菌、解钾量高的解钾菌和产IAA量高的固氮菌间生长关系良好,无拮抗现象。(5)将解磷量高的解磷菌、解钾量高的解钾菌和产IAA量高的固氮菌分别发酵到对数期,按等体积混合发酵液,与不同量的化肥配合施用于油菜盆栽中,结果为菌肥配施30%化肥和菌肥配施70%化肥的处理的多项农艺指标优于或等于全量化肥处理,且菌肥配施30%化肥好于菌肥配施70%化肥的处理。综上所述,本研究共分离获得10株解磷解钾促生优良菌株,其中解磷菌5株(有2株产IAA),解钾菌4株(有3株产IAA),解磷解钾菌中有一株同时具有解磷解钾能力,产IAA的固氮菌2株;在油菜盆栽中菌肥配施30%化肥效果最好。更多还原

【Abstract】 Microbial fertilizer can increase the effective nutrient content,improve the nutrient utilization rate and reduce the amount of chemical fertilizer.It is a kind of pollution-free green fertilizer.The difference between microbial fertilizer and traditional fertilizer is that microbial fertilizer is active and will produce its own metabolic products.Collection of microbial strain resources is the key to the development of microbial fertilizer.A single microbial strain of fertilizer only provides a single nutrient.Therefore,the combination of different functional strains is one of the effective ways to realize the synergistic effect of different microbial strains of microbial fertilizer.In this study,28 strains of phosphorus and silicate nitrogen-fixing bacteria were preliminarily isolated from the soil of orchards and vegetable gardens by dilution plate method,including 14 strains of phosphorus and silicate bacteria,11 strains of silicate bacteria and 3 strains of nitrogen fixing bacteria.The strains were further screened through semi-quantitative analysis of transparent circle and full quantitative analysis of liquid culture,and finally 10 excellent strains were obtained.By antagonistic test,10 strains were confirmed to have no antagonism.The pot experiment of rapeseed was carried out by mixing bacterial strain fermentation liquid and chemical fertilizer.It was found that the treatment of rapeseed with bacterial fertilizer +30% chemical fertilizer had the best indexes.The main results are as follows:(1)14 strains of phosphorus solubilizing bacteria were obtained from the surface soil of vegetable garden and orchard in Houshan Test base of Shenyang Agricultural University by using solid phosphorus solubilizing medium and dilution plate method.By using insoluble phosphate and organophosphorus liquid culture to conduct quantitative analysis on the phosphorus hydrolysis capacity of the strain,5 dominant phosphorus hydrolysate strains were screened out,namely JDYB1,PSYB-1,HLS1,WJ5 and YJ1.JDYB1,PSYB-1 and HLS1 had strong ability to detoxiate inorganic phosphorus,and the quantities of inorganic phosphorus were 26.12μg/m L,26.27μg/m L and 27.85 μg/m L.WJ5 and YJ1 had strong organophosphorous ability,and their organophosphorous contents were 23.91 μg/m L and 23.97 μg/m L.The IAA production capacity of the strain was tested,and there were 2 strains with positive color reaction,among which HLS1 had the strongest IAA secretion capacity,and the IAA production capacity was 39.92 μg/m L.(2)The 11 strains of silicate bacteria were obtained from orchard and garden surface soil of Houshan Test base of Shenyang Agricultural University by dilution plate method.Four dominant strains,namely JDYB1,BKR2,YTM,and JK1,were selected by quantitative analysis of the potassium dissolving capacity of the strain.The potassium dissolving capacity of the strain was 12.95 μg/m L,12.92 μg/m L,12.20 μg/m L,and 11.14 μg/m L.The IAA production capacity of the strain was tested,and there were 3 strains with positive color reaction,among which JK1 had the strongest IAA secretion capacity and the IAA production capacity was 36.01μg/m L.(3)Three nitrogen-fixing bacteria strains were obtained from the surface soil of vegetable gardens and orchards in Houshan Test Base of Shenyang Agricultural University by dilution plate method using nitrogen-free solid medium;Qualitative and quantitative methods were used to further screen the IAA-producing strains.There were 2 strains with positive color reaction,namely NCC1 and ZRT-X,producing IAA at 37.81 μg/m L and 33.63 μg/m L.(4)Antagonistic test showed that there was a good growth relationship between the bacteria with high phosphorus hydrolysis amount,the bacteria with high potassium hydrolysis amount and the bacteria with high IAA production amount.(5)The phosphate-solubilizing measure between high phosphate-solubilizing bacteria,high releasing from bacteria and produce IAA content of nitrogen-fixing bacteria fermentation to the logarithmic phase respectively,according to the volume mix fermented liquid,such as cooperate with different amount of fertilizer applied in rape potted plant,the results of bacterial fertilizer with 30% and 70% chemistry fertilizer processing of many agronomic index is better than that of or the whole amount of chemical fertilizer treatment,and bacterial manure with 30% chemical fertilizer is better than bacterial manure with 70% chemical fertilizer processing.To sum up,10 strains were isolated in this study,among which 5 strains produced IAA(2 strains produced IAA),4 strains produced IAA(3 strains produced IAA),1 strain was capable of producing both phosphorus and potassium,and 2 strains were nitrogen-fixing bacteria producing IAA.The effect of 30% fertilizer combined with fungus fertilizer in rape pot is the best.更多还原