长期施肥对温室和大田土壤微生物的影响
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摘要
华北是我国重要的作物产区,大田及日光温室蔬菜种植是该地区重要的生产方式。施肥是农业生产中的主要管理措施之一,合理施肥能提高肥料利用率、改善土壤条件、并保障作物的稳产和增产。土壤微生物作为衡量土壤质量、肥力以及健康程度的重要指标,可作为参照用以制定农业措施,保证农作物生产的可持续进行。本试验以位于河北省邯郸市曲周县中国农业大学实验站的长期定位试验为依托,以小麦-玉米轮作和日光温室蔬菜种植体系为研究对象,通过2012年连续5次采集土壤样品并进行分析,对不同施肥和种植模式下的土壤理化性质、土壤微生物数量、土壤微生物群落多样性和群落结构变化进行深入研究,旨在探讨不同施肥处理和种植模式对土壤微生物的影响,为保护土壤生物多样性,建立合理的日光温室和大田施肥制度及管理模式,维持土壤质量提供理论依据。主要结论如下：
(1)不同种植模式对温室土壤养分的影响
日光温室长期定位实验共设置3个处理：有机种植模式(ORG),无公害种植模式(LOw)和常规种植模式(CON)。在0-20cm和20-40cm土层,土壤有机质、全氮、速效磷、速效钾含量和pH值在三种种植模式下差异显著。与CON相比,ORG和LOW均提高了土壤OM、TN、AP、AK的含量；而ORG比LOW在提高效果更加明显。在0-20cm土层,OM在ORG和LOW处理下比CON分别增长了25.1g·kg-1和9.4g·kg-1; TN分别增长1.4g·kg-1和0.5g· kg-1; AP分别增长194.7mg· kg-1和125.6mg·kg-1; AK分别增长289.9mg· kg-1和91.3mg·kg-1。在20-40cm土层,OM在ORG和LOW处理下比CON分别增长6.6g·kg-1和2.2g·kg-1；TN分别增长0.5g·kg-1和0.2g·kg-1；AP分别增长184.7mg·kg-1和76.7mg·kg-1;AK分别增长222.7mg·kg-1,85.3mg·kg-1。
(2)不同施肥处理对大田土壤养分的影响
大田长期定位实验共设置4个不同种类肥料投入处理：EM堆肥(EM)；传统堆肥(OF)；化肥(CF)；不施肥对照(CK)。土壤样品分析结果显示：不同施肥处理对0-20cm和20-40cm土壤有机质(OM)、全氮(TN)、速效磷(AP)、速效钾(AK)的含量和土壤pH值的影响差异显著。4种施肥处理均可以提高土壤OM、TN、AP、 AK的含量而降低pH值,与CF相比,EM和OF处理下土壤各养分指标提高的效果更加明显。在0-20cm土层,OM在EM、OF和CF处理下比CK的分别增加了8.4g·kg-1、5.3g·kg-1、1.6g·kg-1; TN分别增加了为0.7g· kg-1、0.5g·kg(?)1和0.2g·kg-1；AP分别增加了63.6mg· kg-1、59.2mg· kg-1和26.6mg· kg-1; AK分别增加了199.7mg· kg-1,142.1mg· kg(?)1和91.3mg· kg-1,在20-40cm土层,OM在EM, OF和CF处理下较CK分别增加了2.1g·kg-1、1.0g·kg-1和0.5g·kg-1；TN分别增加了0.2g· kg-1、0.2g· kg-1和O.1g· kg-1; AP分别增加了28.3mg· kg-1,11.7mg· kg-1和4.8mg· kg-1;与CK相比,在EM和OF处理下AK别分增加了39.8mg· kg-1和26.1mg· kg-1,而在CF处理下降低了5.7mg·kg-1。
(3)不同种植模式对温室土壤微生物学特征的影响
对日光温室种植体系中的土壤微生物研究的结果显示,土壤细菌和真菌在不同种植模式下差异显著。ORG处理的细菌16s rRNA和真菌ITS基因拷贝数数量均显著高于CON处理。不同种植模式下的土壤细菌16s rRNA基因拷贝数为6.63×109-1.13×1012拷贝数每克干土,高于真菌的ITS基因拷贝数(3.30×107-1.14×1010拷贝数每克干土)。用T-RFLP方法对土壤细菌群落结构和多样性进行了研究。结果表明,ORG可以增加土壤微生物多样性,各处理土壤细菌主要分布在厚壁菌门、放线菌门、拟杆菌门和变形菌门；ORG、LOW和CON处理下的优势真菌类群分别为132和368bp,138、154、334bp和347bp,134、150、374和466bp。表明不同种植改变了温室蔬菜土壤细菌和真菌的群落结构。
(4)不同施肥处理对大田土壤微生物学特征的影响
对土壤微生物的研究结果显示,土壤细菌和真菌在不同处理间有显著差异。其中EM堆肥处理的细菌16s rRNA和真菌ITS基因拷贝数数量显著高于其它处理,而在CF处理下两个指标显著降低。不同处理下,细菌16s rRNA基因拷贝数在1.83×108-2.33×1011拷贝数每克干土,高于真菌的ITS基因拷贝数(1.88×106-2.28×109)。用T-RFLP方法对土壤细菌群落结构进行了研究。结果表明,EM堆肥处理可以显著增加土壤微生物的多样性。经过对酶切片段鉴别分类,土壤细菌主要分布在厚壁菌门、放线菌门、拟杆菌门、变形菌门、蓝藻门和柔膜菌门。处理间相对丰度差异显著的真菌酶切片段包括了106、108、134、136、152和173bp。EM和OF处理可以增加片段134、173、152和1136bp的相对丰度,而降低106和108bp的相对丰度。上述结果表明长期施肥处理改变了大田土壤细菌和真菌的群落结构。
本研究表明,土壤微生物对长期施肥处理的不同响应特性在检测土壤质量变化时起到重要的指示作用。土壤中大量微生物的存在也显示了其在生化过程中的潜在价值,为进一步研究其在养分循环中的作用和地位提供了坚实基础。
The crop production in farmland and vegetable production in greenhouse are important modes of production in northern China, which is the important grain-producing region. Fertilization is one of the main agricultural measures, and reasonable fertilization can improve fertilizer use efficiency and soil conditions to guarantee grain yield stable or increasing. As an important indicator for valuating soil quality, fertility and health degree, the acquaintance of soil microbe can be used to help develop agriculture measures to ensure the continuous production of crops. In this study, on the basis of the long-term experiment located in China Agricultural University Experiment Station of Quzhou County, Handan City, Hebei Province, five consecutive times soil samples from wheat-maize rotation and greenhouse vegetable systems were taken for analysis in2012. A deep studies on the soil physic-chemical properties, the microbial number, community diversity and community structure evolution patterns under different fertilization and planting mode were conducted to investigate the effects of different fertilizer application treatments and planting patterns on soil microbes, expecting to maintain the biodiversity in soil, establish a reasonable greenhouse and field fertilization and management regime, and provide a theoretical basis for maintaining soil quality. The main conclusions are as follows:
(1) Effects of different vegetable production systems on greenhouse soil nutrients
The greenhouse long-term experiment includes3treatments:organic greenhouse (ORG), low input greenhouse (LOW) and conventional greenhouse (CON). Under different vegetable production systems, the soil organic matter, total nitrogen, available phosphorus, potassium content and pH in0-20cm and20-40cm soil layer had the significant different. Compared with CON, ORG and LOW could improve the soil OM, TN, AP and AK content. While compared with LOW, ORG had more obvious effects. In the0-20cm soil layer, compared with CON, OM under ORG and LOW increased by25.1g· kg-1and9.4g· kg-1, respectively; TN by1.4g· kg-1and0.5g· kg-1, respectively; AP by194.7mg· kg-1and125.6mg· kg-1, respectively; AK by289.9mg· kg-1and91.3mg· kg-1, respectively. In20-40cm soil layer, compared with CON, OM under ORG and LOW increased by6.6g· kg-1and2.2g· kg-1, respectively; TN by0.5g· kg-1and0.2g· kg-1, respectively; AP by184.7mg· kg-1and76.7mg· kg-1, respectively; AK by222.7mg· kg-1and85.3mg· kg-1, respectively.
(2) Effects of different fertilizer treatments on farmland soil nutrient
Long-term fertilization experiment includes four treatments:EM compost (EM); tranditional compost (OF); chemical fertilizer (CF) and unfertilizer (CK). In March, May, June, August and October of2012, soil samples were collected for5times. The results showed that different fertilization treatments had significant effects on soil organic matter (OM), total nitrogen (TN), available phosphorus (AP), available potassium (AK) and pH in0-20cm and20-40cm soil layers. Compared with CK, EM, OF and CF could improve soil OM, TN, AP, AK content and reduce pH. Compared with CF, EM and OF could make more increase of the soil nutrient. In0-20cm soil layer, compared with CK, OM under EM, OF and CF increased by8.4g· kg-1,5.3g· kg-1and1.6g· kg-1, respectively; TN by0.7 g· kg-1,0.5g· kg-1and0.2g· kg-1, respectively; AP by63.6mg· kg-1,59.2mg· kg1and26.6mg· kg"1, respectively; AP by199.7mg· kg-1,142.1mg· kg-1and91.3mg· kg-1, respectively. In20-40cm soil layer, compared with CK, OM under EM, OF and CF increased by2.1g· kg-1,1.0g· kg-1and0.5g· kg-1, respectively; TN by0.2g· kg-1,0.2g· kg-1and O.lg· kg-1, respectively; AP by28.3mg· kg-1,11.7mg· kg-1and4.8mg· kg-1, respectively; compared with CK, AK under EM and OF increased by39.8mg· kg-1and26.1mg· kg-1, respectively, while reduced by5.7mg· kg-1under CF.
(3) Effects of different planting patterns on greenhouse soil microbial characteristics
The results from microbiological studies in greenhouse showed that the soil bacteria and fungi were significantly different under different treatments. The number of bacterial16s rRNA and ITS fungal gene copy numbers under ORG were significantly higher than CON. The bacterial16s rRNA gene copy numbers under different treatments ranged from6.63x109to1.13x1012copies per gram dry soil, higher than fungi ITS gene copies number (3.30x107-1.14x1010copies per gram dry soil). The community structure and diversity of soil bacteria were studied by the T-RFLP approach. The results show that ORG could increase microbial diversity. Firmicutes, Thallobacteria, Bacteroidetes and Proteobacteria are mainly bacteria. Under ORG, LOW and CON, the privileged fungal taxa were132and368bp,138,154,334bp and347bp,134,150,374and466bp, respectively. These showed that different vegetable production systems have different soil microorganism community structure.
(4) Effect of different fertilization on farmland soil microbial characteristics
The results on soil microbiology showed that the soil bacteria and fungi had significant differences under different treatments. The bacteria16s rRNA and fungi ITS gene copy numbers under EM were significantly highest among treatments, and while lowest under CK. Under4treatments, bacterial16s rRNA gene copy numbers ranged from1.83x108to2.33x1011copies per gram dry soil, higher than the fungal ITS gene copy numbers (1.88x106-2.28x109). T-RFLP method was adopted to study the soil bacterial community structure. The results showed that EM compost could significantly increased soil microbial diversity. The results from identification and classification of restriction fragment showed that:Firmicutes, Thallobacteria, Bacteroidetes, Proteobacteria, Cyanobacteria and Tenericutes are mainly bacteria. The relative abundance of fungal fragments with significant differences under different treatments included106,108,134,136,152and173bp. EM and OF could increase the relative abundance of134,173,152and136bp fragment, and reduce the relative abundance of106and108bp. These showed that long-term fertilization treatment could change the soil bacteria and fungi community structures.
It is concluded that the different microbial characteristics responding to long-term fertilization on for the detection of changes in soil quality can function as the important indicator. And the existence of large number of microorganisms in soil predicts its potential value in the biochemical processes. This makes solid foundation for the future studies on their roles and statuses in the nutrient cycling process.
(1)不同种植模式对温室土壤养分的影响
日光温室长期定位实验共设置3个处理：有机种植模式(ORG),无公害种植模式(LOw)和常规种植模式(CON)。在0-20cm和20-40cm土层,土壤有机质、全氮、速效磷、速效钾含量和pH值在三种种植模式下差异显著。与CON相比,ORG和LOW均提高了土壤OM、TN、AP、AK的含量；而ORG比LOW在提高效果更加明显。在0-20cm土层,OM在ORG和LOW处理下比CON分别增长了25.1g·kg-1和9.4g·kg-1; TN分别增长1.4g·kg-1和0.5g· kg-1; AP分别增长194.7mg· kg-1和125.6mg·kg-1; AK分别增长289.9mg· kg-1和91.3mg·kg-1。在20-40cm土层,OM在ORG和LOW处理下比CON分别增长6.6g·kg-1和2.2g·kg-1；TN分别增长0.5g·kg-1和0.2g·kg-1；AP分别增长184.7mg·kg-1和76.7mg·kg-1;AK分别增长222.7mg·kg-1,85.3mg·kg-1。
(2)不同施肥处理对大田土壤养分的影响
大田长期定位实验共设置4个不同种类肥料投入处理：EM堆肥(EM)；传统堆肥(OF)；化肥(CF)；不施肥对照(CK)。土壤样品分析结果显示：不同施肥处理对0-20cm和20-40cm土壤有机质(OM)、全氮(TN)、速效磷(AP)、速效钾(AK)的含量和土壤pH值的影响差异显著。4种施肥处理均可以提高土壤OM、TN、AP、 AK的含量而降低pH值,与CF相比,EM和OF处理下土壤各养分指标提高的效果更加明显。在0-20cm土层,OM在EM、OF和CF处理下比CK的分别增加了8.4g·kg-1、5.3g·kg-1、1.6g·kg-1; TN分别增加了为0.7g· kg-1、0.5g·kg(?)1和0.2g·kg-1；AP分别增加了63.6mg· kg-1、59.2mg· kg-1和26.6mg· kg-1; AK分别增加了199.7mg· kg-1,142.1mg· kg(?)1和91.3mg· kg-1,在20-40cm土层,OM在EM, OF和CF处理下较CK分别增加了2.1g·kg-1、1.0g·kg-1和0.5g·kg-1；TN分别增加了0.2g· kg-1、0.2g· kg-1和O.1g· kg-1; AP分别增加了28.3mg· kg-1,11.7mg· kg-1和4.8mg· kg-1;与CK相比,在EM和OF处理下AK别分增加了39.8mg· kg-1和26.1mg· kg-1,而在CF处理下降低了5.7mg·kg-1。
(3)不同种植模式对温室土壤微生物学特征的影响
对日光温室种植体系中的土壤微生物研究的结果显示,土壤细菌和真菌在不同种植模式下差异显著。ORG处理的细菌16s rRNA和真菌ITS基因拷贝数数量均显著高于CON处理。不同种植模式下的土壤细菌16s rRNA基因拷贝数为6.63×109-1.13×1012拷贝数每克干土,高于真菌的ITS基因拷贝数(3.30×107-1.14×1010拷贝数每克干土)。用T-RFLP方法对土壤细菌群落结构和多样性进行了研究。结果表明,ORG可以增加土壤微生物多样性,各处理土壤细菌主要分布在厚壁菌门、放线菌门、拟杆菌门和变形菌门；ORG、LOW和CON处理下的优势真菌类群分别为132和368bp,138、154、334bp和347bp,134、150、374和466bp。表明不同种植改变了温室蔬菜土壤细菌和真菌的群落结构。
(4)不同施肥处理对大田土壤微生物学特征的影响
对土壤微生物的研究结果显示,土壤细菌和真菌在不同处理间有显著差异。其中EM堆肥处理的细菌16s rRNA和真菌ITS基因拷贝数数量显著高于其它处理,而在CF处理下两个指标显著降低。不同处理下,细菌16s rRNA基因拷贝数在1.83×108-2.33×1011拷贝数每克干土,高于真菌的ITS基因拷贝数(1.88×106-2.28×109)。用T-RFLP方法对土壤细菌群落结构进行了研究。结果表明,EM堆肥处理可以显著增加土壤微生物的多样性。经过对酶切片段鉴别分类,土壤细菌主要分布在厚壁菌门、放线菌门、拟杆菌门、变形菌门、蓝藻门和柔膜菌门。处理间相对丰度差异显著的真菌酶切片段包括了106、108、134、136、152和173bp。EM和OF处理可以增加片段134、173、152和1136bp的相对丰度,而降低106和108bp的相对丰度。上述结果表明长期施肥处理改变了大田土壤细菌和真菌的群落结构。
本研究表明,土壤微生物对长期施肥处理的不同响应特性在检测土壤质量变化时起到重要的指示作用。土壤中大量微生物的存在也显示了其在生化过程中的潜在价值,为进一步研究其在养分循环中的作用和地位提供了坚实基础。
The crop production in farmland and vegetable production in greenhouse are important modes of production in northern China, which is the important grain-producing region. Fertilization is one of the main agricultural measures, and reasonable fertilization can improve fertilizer use efficiency and soil conditions to guarantee grain yield stable or increasing. As an important indicator for valuating soil quality, fertility and health degree, the acquaintance of soil microbe can be used to help develop agriculture measures to ensure the continuous production of crops. In this study, on the basis of the long-term experiment located in China Agricultural University Experiment Station of Quzhou County, Handan City, Hebei Province, five consecutive times soil samples from wheat-maize rotation and greenhouse vegetable systems were taken for analysis in2012. A deep studies on the soil physic-chemical properties, the microbial number, community diversity and community structure evolution patterns under different fertilization and planting mode were conducted to investigate the effects of different fertilizer application treatments and planting patterns on soil microbes, expecting to maintain the biodiversity in soil, establish a reasonable greenhouse and field fertilization and management regime, and provide a theoretical basis for maintaining soil quality. The main conclusions are as follows:
(1) Effects of different vegetable production systems on greenhouse soil nutrients
The greenhouse long-term experiment includes3treatments:organic greenhouse (ORG), low input greenhouse (LOW) and conventional greenhouse (CON). Under different vegetable production systems, the soil organic matter, total nitrogen, available phosphorus, potassium content and pH in0-20cm and20-40cm soil layer had the significant different. Compared with CON, ORG and LOW could improve the soil OM, TN, AP and AK content. While compared with LOW, ORG had more obvious effects. In the0-20cm soil layer, compared with CON, OM under ORG and LOW increased by25.1g· kg-1and9.4g· kg-1, respectively; TN by1.4g· kg-1and0.5g· kg-1, respectively; AP by194.7mg· kg-1and125.6mg· kg-1, respectively; AK by289.9mg· kg-1and91.3mg· kg-1, respectively. In20-40cm soil layer, compared with CON, OM under ORG and LOW increased by6.6g· kg-1and2.2g· kg-1, respectively; TN by0.5g· kg-1and0.2g· kg-1, respectively; AP by184.7mg· kg-1and76.7mg· kg-1, respectively; AK by222.7mg· kg-1and85.3mg· kg-1, respectively.
(2) Effects of different fertilizer treatments on farmland soil nutrient
Long-term fertilization experiment includes four treatments:EM compost (EM); tranditional compost (OF); chemical fertilizer (CF) and unfertilizer (CK). In March, May, June, August and October of2012, soil samples were collected for5times. The results showed that different fertilization treatments had significant effects on soil organic matter (OM), total nitrogen (TN), available phosphorus (AP), available potassium (AK) and pH in0-20cm and20-40cm soil layers. Compared with CK, EM, OF and CF could improve soil OM, TN, AP, AK content and reduce pH. Compared with CF, EM and OF could make more increase of the soil nutrient. In0-20cm soil layer, compared with CK, OM under EM, OF and CF increased by8.4g· kg-1,5.3g· kg-1and1.6g· kg-1, respectively; TN by0.7 g· kg-1,0.5g· kg-1and0.2g· kg-1, respectively; AP by63.6mg· kg-1,59.2mg· kg1and26.6mg· kg"1, respectively; AP by199.7mg· kg-1,142.1mg· kg-1and91.3mg· kg-1, respectively. In20-40cm soil layer, compared with CK, OM under EM, OF and CF increased by2.1g· kg-1,1.0g· kg-1and0.5g· kg-1, respectively; TN by0.2g· kg-1,0.2g· kg-1and O.lg· kg-1, respectively; AP by28.3mg· kg-1,11.7mg· kg-1and4.8mg· kg-1, respectively; compared with CK, AK under EM and OF increased by39.8mg· kg-1and26.1mg· kg-1, respectively, while reduced by5.7mg· kg-1under CF.
(3) Effects of different planting patterns on greenhouse soil microbial characteristics
The results from microbiological studies in greenhouse showed that the soil bacteria and fungi were significantly different under different treatments. The number of bacterial16s rRNA and ITS fungal gene copy numbers under ORG were significantly higher than CON. The bacterial16s rRNA gene copy numbers under different treatments ranged from6.63x109to1.13x1012copies per gram dry soil, higher than fungi ITS gene copies number (3.30x107-1.14x1010copies per gram dry soil). The community structure and diversity of soil bacteria were studied by the T-RFLP approach. The results show that ORG could increase microbial diversity. Firmicutes, Thallobacteria, Bacteroidetes and Proteobacteria are mainly bacteria. Under ORG, LOW and CON, the privileged fungal taxa were132and368bp,138,154,334bp and347bp,134,150,374and466bp, respectively. These showed that different vegetable production systems have different soil microorganism community structure.
(4) Effect of different fertilization on farmland soil microbial characteristics
The results on soil microbiology showed that the soil bacteria and fungi had significant differences under different treatments. The bacteria16s rRNA and fungi ITS gene copy numbers under EM were significantly highest among treatments, and while lowest under CK. Under4treatments, bacterial16s rRNA gene copy numbers ranged from1.83x108to2.33x1011copies per gram dry soil, higher than the fungal ITS gene copy numbers (1.88x106-2.28x109). T-RFLP method was adopted to study the soil bacterial community structure. The results showed that EM compost could significantly increased soil microbial diversity. The results from identification and classification of restriction fragment showed that:Firmicutes, Thallobacteria, Bacteroidetes, Proteobacteria, Cyanobacteria and Tenericutes are mainly bacteria. The relative abundance of fungal fragments with significant differences under different treatments included106,108,134,136,152and173bp. EM and OF could increase the relative abundance of134,173,152and136bp fragment, and reduce the relative abundance of106and108bp. These showed that long-term fertilization treatment could change the soil bacteria and fungi community structures.
It is concluded that the different microbial characteristics responding to long-term fertilization on for the detection of changes in soil quality can function as the important indicator. And the existence of large number of microorganisms in soil predicts its potential value in the biochemical processes. This makes solid foundation for the future studies on their roles and statuses in the nutrient cycling process.
引文
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