海藻对不同促生菌生长的影响及应用
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摘要
海藻是生长在海洋中的低等隐花植物,由于其特殊的生长环境,营养成分与陆生植物存在很大的不同,海藻中含有一般陆生植物无法比拟的丰富的矿物元素及维生素等。目前海藻及其相关产品已被应用于多个领域。海藻可作为生物促生剂及肥料应用于有机农业中。本文以山东青岛岸边丰富的海带(Laminaria japonica)及浒苔(Entermorpha)为原材料,研究了海藻液态提取物对根际促生菌( PGPR, plant growth-promoting rhizobacteria)生长的影响,海藻有机肥生产,施用海藻有机肥对果园土壤理化性质、微生物数量、活性及种群结构的影响。
在LB液体培养基中添加1/100、1/200、1/500、1/1000体积比例的海藻提取液,接入自生固氮菌、解钾菌、有机磷细菌、无机磷细菌四种根际促生菌,在不同的时间点测定菌液OD值。结果表明,添加海藻提取液对有机磷细菌的生长无显著影响,可以促进自生固氮菌、无机磷细菌、解钾菌的生长,不同海藻提取液添加量对自生固氮菌和无机磷细菌的生长促进作用无显著差异,1/1000的海藻提取液添加量即可收到很好的促生效果。海藻提取液对解钾菌的促生作用随海藻提取液添加量的增大而增强,以1/100的添加量效果最好。
以山东青岛岸边的海带和浒苔为原料,通过高温发酵制作海藻有机肥,以山东烟台10年树龄的苹果园进行施肥试验,2008年秋季施肥,两种肥料的施肥量一致,分别以7.5、15、22.5kg/株三种不同的量在秋季施入果园中,正常管理,同时设不施有机肥的对照。为保持果树的产量,所有处理均施用等量化肥。次年的五月和十月采集土样。通过平板培养、测定土壤酶活性、构建微生物群落脂肪酸甲酯(FAME, fatty acid methyl ester)图谱的方法研究海藻有机肥对土壤微生物数量、活性及种群结构的影响。
结果表明,施用海带有机肥及浒苔有机肥可以显著提高果园土壤有机质、全氮、速效磷、速效钾含量,提高土壤pH值,改良酸性土壤。海带肥的效果优于浒苔肥。
在五月份的采样检测表明,海带有机肥及浒苔有机肥均可显著增加土壤中可培养细菌及放线菌的数量,细菌数量为对照土壤的2.81-9.30倍,放线菌数量为对照土壤的1.95-18.62倍。施用海带肥及浒苔肥对可培养真菌数量无显著影响。海带有机肥及浒苔有机肥均可显著提高土壤荧光素二乙酸酯(FDA,Fluorescein diacetate)酶及脲酶活性,FDA酶活性为对照土壤的1.25-1.88倍,脲酶活性为对照土壤的1.71-2.79倍。施用海带肥及浒苔肥对过氧化氢酶活性无显著影响。对微生物脂肪酸甲酯的分析表明,施用海带肥及浒苔肥均可显著提高微生物生物量,总的FAME的量为对照土壤的1.60-2.64倍。施用海带肥和浒苔肥可增加土壤微生物中单不饱和脂肪酸的含量比例,为对照土壤的1.14-1.64倍。对FAME数据的主成分分析表明,海带肥及22.5kg/株浒苔肥处理对土壤微生物种群结构的改变较大,而7.5kg/株及15kg/株浒苔肥对微生物种群结构的影响较小。在十月份的采样检测中,两种肥料对土壤微生物的影响已减弱。这些结果表明,施用海带有机肥和浒苔有机肥会对果园土壤质量产生有益的影响。
Seaweed is an inferior cryptogamous plant in the sea. The nutrients of seaweeds are greatly different from that of plants on land because of their particular environment. Seaweed contains rich minerals and vitamins that are much less in amount in ordinary plants on land. Seaweed and its products are now widely used in many fields. Seaweed has gained application in the field organic agriculture as biostimulants and fertilizer. In this article, the rich resources of Laminaria japonica and Entermorpha on the sealine of Tsingtao, Shandong Province are used as raw materials to study the effect of liquid seaweed extract on the growth of PGPR(plant growth-promoting rhizobacteria), the production process of seaweed organic fertilizer and the effect of the application of seaweed fertilizer on the chemical properties of orchard soil and the amount, activity and community structures of soil microorganisms.
1/100, 1/200, 1/500, 1/1000 proportional seaweed liquid extract was added to LB liquid medium (v/v). Four species of PGPR (plant growth-promoting rhizobacteria), Azotobacteria, organic phosphobacteria, inorganic phosphobacteria, and silicate-dissolving bacteria were inoculated to the medium and OD values were measured at different time points. The result indicates that adding seaweed liquid extract has no significant influence on the growth of organic phosphobacteria while promoting the growth of azotobacteria, inorganic phosphobacteria, silicate-dissolving bacteria. The different quantities of seaweed liquid extract added have no significantly different growth-promoting effect on azotobacteria and inorganic phosphobacteria. Adding 1/1000 proportional seaweed liquid extract would have significant growth-promoting effect. In the case of silicate-dissolving bacteria, the growth-promoting effect becomes stronger as the quantity of seaweed extract becomes greater, being best when adding 1/100 proportional seaweed extract.
Seaweed organic fertilizer was manufactured from Laminaria japonica and Entermorpha on the sealine of Tsingtao, Shandong Province, by high-temperature fertilization. An apple orchard planted with trees 10 years old in Yantai, Shandong Province was chosen for the experiment. The fertilizer was applied in the autumn of 2008. The two species of fertilizer were applied with the same amount, each applied with three different quantities, 7.5, 15, 22.5kg/plant to the orchard. The orchard was managed as usual. The control sites with no organic manure were established at the same time. To maintain the production of apple trees, chemical fertilizer were applied to all the experimental sites with the same amount. Soil samples were collected in May and October the next year. The effect of seaweed fertilizer on the amount, activity and community structure of soil microorganisms was studied using dilution plating, measuring soil enzyme activities and forming microbial community FAME (fatty acid methyl ester) profiles.
The result indicates that Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase the contents of organic matter, total nitrogen , available K and available P in orchard soil. Laminaria japonica organic fertilizer and Entermorpha organic fertilizer can increase the soil pH value and improve the quality of acidic soil. The Laminaria japonica organic fertilizer has better effect than Entermorpha organic fertilizer.
The analysis of soil samples collected in May indicates that Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase the amounts of culturable bacteria and actinomycetes in soil. The amount of bacteria is 2.81-9.30 times higher than that in control soil. The amount of actinomycetes is 1.95-18.62 times higher than that in control soil. The application of Laminaria japonica fertilizer and Entermorpha fertilizer has no significant influence on the amount of culturable fungi. Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase FDA (Fluorescein diacetate) enzyme activity and urease activity in soil. FDA enzyme activity is 1.25-1.88 times higher than that in control soil. Urease activity is 1.71-2.79 times higher than that in control soil. The application of Laminaria japonica fertilizer and Entermorpha fertilizer has no significant influence on catalase activity. The analysis of FAME indicates that both Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase microbial biomass. The total FAME amount is 1.60-2.64 times higher than that in control soil. The application of Laminaria japonica fertilizer and Entermorpha fertilizer increases the relative abundance of monounsaturated fatty acids in soil microorganisms, being 1.14-1.64 times higher than that in control soil. The principal components analysis on FAME data indicates that soil microbial community structures are much changed due to the application of Laminaria japonica organic fertilizer and 22.5kg/plant Entermorpha organic fertilizer. However, 7.5kg/plant and 22.5kg/plant Entermorpha organic fertilizer have little effect on microbial community structures. Both fertilizers have much less effect on soil microorganisms in the analysis of soil samples collected in October. These results indicate that the application of Laminaria japonica organic fertilizer and Entermorpha organic fertilizer has beneficial effect on orchard soil quality.
在LB液体培养基中添加1/100、1/200、1/500、1/1000体积比例的海藻提取液,接入自生固氮菌、解钾菌、有机磷细菌、无机磷细菌四种根际促生菌,在不同的时间点测定菌液OD值。结果表明,添加海藻提取液对有机磷细菌的生长无显著影响,可以促进自生固氮菌、无机磷细菌、解钾菌的生长,不同海藻提取液添加量对自生固氮菌和无机磷细菌的生长促进作用无显著差异,1/1000的海藻提取液添加量即可收到很好的促生效果。海藻提取液对解钾菌的促生作用随海藻提取液添加量的增大而增强,以1/100的添加量效果最好。
以山东青岛岸边的海带和浒苔为原料,通过高温发酵制作海藻有机肥,以山东烟台10年树龄的苹果园进行施肥试验,2008年秋季施肥,两种肥料的施肥量一致,分别以7.5、15、22.5kg/株三种不同的量在秋季施入果园中,正常管理,同时设不施有机肥的对照。为保持果树的产量,所有处理均施用等量化肥。次年的五月和十月采集土样。通过平板培养、测定土壤酶活性、构建微生物群落脂肪酸甲酯(FAME, fatty acid methyl ester)图谱的方法研究海藻有机肥对土壤微生物数量、活性及种群结构的影响。
结果表明,施用海带有机肥及浒苔有机肥可以显著提高果园土壤有机质、全氮、速效磷、速效钾含量,提高土壤pH值,改良酸性土壤。海带肥的效果优于浒苔肥。
在五月份的采样检测表明,海带有机肥及浒苔有机肥均可显著增加土壤中可培养细菌及放线菌的数量,细菌数量为对照土壤的2.81-9.30倍,放线菌数量为对照土壤的1.95-18.62倍。施用海带肥及浒苔肥对可培养真菌数量无显著影响。海带有机肥及浒苔有机肥均可显著提高土壤荧光素二乙酸酯(FDA,Fluorescein diacetate)酶及脲酶活性,FDA酶活性为对照土壤的1.25-1.88倍,脲酶活性为对照土壤的1.71-2.79倍。施用海带肥及浒苔肥对过氧化氢酶活性无显著影响。对微生物脂肪酸甲酯的分析表明,施用海带肥及浒苔肥均可显著提高微生物生物量,总的FAME的量为对照土壤的1.60-2.64倍。施用海带肥和浒苔肥可增加土壤微生物中单不饱和脂肪酸的含量比例,为对照土壤的1.14-1.64倍。对FAME数据的主成分分析表明,海带肥及22.5kg/株浒苔肥处理对土壤微生物种群结构的改变较大,而7.5kg/株及15kg/株浒苔肥对微生物种群结构的影响较小。在十月份的采样检测中,两种肥料对土壤微生物的影响已减弱。这些结果表明,施用海带有机肥和浒苔有机肥会对果园土壤质量产生有益的影响。
Seaweed is an inferior cryptogamous plant in the sea. The nutrients of seaweeds are greatly different from that of plants on land because of their particular environment. Seaweed contains rich minerals and vitamins that are much less in amount in ordinary plants on land. Seaweed and its products are now widely used in many fields. Seaweed has gained application in the field organic agriculture as biostimulants and fertilizer. In this article, the rich resources of Laminaria japonica and Entermorpha on the sealine of Tsingtao, Shandong Province are used as raw materials to study the effect of liquid seaweed extract on the growth of PGPR(plant growth-promoting rhizobacteria), the production process of seaweed organic fertilizer and the effect of the application of seaweed fertilizer on the chemical properties of orchard soil and the amount, activity and community structures of soil microorganisms.
1/100, 1/200, 1/500, 1/1000 proportional seaweed liquid extract was added to LB liquid medium (v/v). Four species of PGPR (plant growth-promoting rhizobacteria), Azotobacteria, organic phosphobacteria, inorganic phosphobacteria, and silicate-dissolving bacteria were inoculated to the medium and OD values were measured at different time points. The result indicates that adding seaweed liquid extract has no significant influence on the growth of organic phosphobacteria while promoting the growth of azotobacteria, inorganic phosphobacteria, silicate-dissolving bacteria. The different quantities of seaweed liquid extract added have no significantly different growth-promoting effect on azotobacteria and inorganic phosphobacteria. Adding 1/1000 proportional seaweed liquid extract would have significant growth-promoting effect. In the case of silicate-dissolving bacteria, the growth-promoting effect becomes stronger as the quantity of seaweed extract becomes greater, being best when adding 1/100 proportional seaweed extract.
Seaweed organic fertilizer was manufactured from Laminaria japonica and Entermorpha on the sealine of Tsingtao, Shandong Province, by high-temperature fertilization. An apple orchard planted with trees 10 years old in Yantai, Shandong Province was chosen for the experiment. The fertilizer was applied in the autumn of 2008. The two species of fertilizer were applied with the same amount, each applied with three different quantities, 7.5, 15, 22.5kg/plant to the orchard. The orchard was managed as usual. The control sites with no organic manure were established at the same time. To maintain the production of apple trees, chemical fertilizer were applied to all the experimental sites with the same amount. Soil samples were collected in May and October the next year. The effect of seaweed fertilizer on the amount, activity and community structure of soil microorganisms was studied using dilution plating, measuring soil enzyme activities and forming microbial community FAME (fatty acid methyl ester) profiles.
The result indicates that Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase the contents of organic matter, total nitrogen , available K and available P in orchard soil. Laminaria japonica organic fertilizer and Entermorpha organic fertilizer can increase the soil pH value and improve the quality of acidic soil. The Laminaria japonica organic fertilizer has better effect than Entermorpha organic fertilizer.
The analysis of soil samples collected in May indicates that Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase the amounts of culturable bacteria and actinomycetes in soil. The amount of bacteria is 2.81-9.30 times higher than that in control soil. The amount of actinomycetes is 1.95-18.62 times higher than that in control soil. The application of Laminaria japonica fertilizer and Entermorpha fertilizer has no significant influence on the amount of culturable fungi. Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase FDA (Fluorescein diacetate) enzyme activity and urease activity in soil. FDA enzyme activity is 1.25-1.88 times higher than that in control soil. Urease activity is 1.71-2.79 times higher than that in control soil. The application of Laminaria japonica fertilizer and Entermorpha fertilizer has no significant influence on catalase activity. The analysis of FAME indicates that both Laminaria japonica organic fertilizer and Entermorpha organic fertilizer significantly increase microbial biomass. The total FAME amount is 1.60-2.64 times higher than that in control soil. The application of Laminaria japonica fertilizer and Entermorpha fertilizer increases the relative abundance of monounsaturated fatty acids in soil microorganisms, being 1.14-1.64 times higher than that in control soil. The principal components analysis on FAME data indicates that soil microbial community structures are much changed due to the application of Laminaria japonica organic fertilizer and 22.5kg/plant Entermorpha organic fertilizer. However, 7.5kg/plant and 22.5kg/plant Entermorpha organic fertilizer have little effect on microbial community structures. Both fertilizers have much less effect on soil microorganisms in the analysis of soil samples collected in October. These results indicate that the application of Laminaria japonica organic fertilizer and Entermorpha organic fertilizer has beneficial effect on orchard soil quality.
引文
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