茶园土壤微生物量、硝化和反硝化作用研究
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摘要
本文选择我国浙江绿茶产区为主的代表性茶园,包括不同理化性状、茶园管理水平和管理方式等典型土壤为研究材料,以不同生产力水平和植茶年龄茶园,及附近森林和蔬菜土壤为重点,采用田间调查与实验室培养相结合,综合运用传统和现代微生物分析技术,对茶园上壤微生物量、硝化作用强度、硝化微生物的种类及N2O释放特点等进行了系统全面的研究。获得以下主要研究结果：
1.浙江75个茶园土壤微生物量碳(C)在38.1-607.2mg/kg之间,平均为208.1mg/kg；微生物量C占总有机C的比例在0.21%-4.55%之间,平均为1.25%。微生物量氮(茚三酮反应态N)在1.76-76.14mg/kg之间,平均为20.67mg/kg；微生物量N占全N的比例在0.26%-9.09%之间,平均为1.80%。微生物量磷(P)在未检出到60.07mg/kg之间,平均为14.93mg/kg；微生物量P占全P的比例在0-10.20%之间,平均为3.06%。与多数耕地或森林土壤相比,茶园土壤微量物量C和N,以及微生物量C占总有机C的比例和微生物量N占全N的比例均偏低,但微生物量P与其它土壤相当,微生物量P占全P的比例高于其它土壤。
2.茶园土壤微生物量受生产力水平、植茶年龄、pH、施肥量和茶树种植模式的显著影响。随着茶园生产力水平和植茶年龄的提高,土壤pH明显降低,有机C、全N、全P和有效态营养元素含量显著提高。微生物C、N和PLFAs含量表现为森林>低产>高产>中产茶园：土壤微生物C和ATP含量表现为50年>9年>森林>90年生茶园,微生物量N及微生物量N占全N量的比例表现为森林≥9年>50年>90年生茶园。茶园土壤pH低是导致微生物量少和生长代谢弱的最重要原因之一,除微生物量P外,微生物量C和N、微生物商、土壤基础呼吸速率和代谢商均与pH呈极显著正相关。适量施P有利于提高土壤微生物量P,但过量施N降低微生物量C和N含量；随着茶园施氮量的提高,土壤微生物商C、N和P均呈极显著降低的趋势。对磷酯脂肪酸(PLFA)的分析表明茶园土壤微生物群落结构随着茶园生产力水平的提高和树龄的增加呈连续有规律的变化；PLFA标记革兰氏阳性细菌的数量显著高于革兰氏阴性细菌和真菌的数量；茶园土壤PLFA标记真菌和细菌的比例显著高于森林土壤,且PLFA标记真菌与细菌比例与代谢商呈显著负相关(R2=0.93,p<0.05)。茶园由常规种植向有机种植转换有利于提高土壤pH和有机C含量,有机茶种植年限越长,pH和有机C含量越高。茶园有机种植方式能明显提高土壤微生物量C、N和P的含量及微生物商C、N和P的比例,改善茶园土壤质量。
3.尽管土壤pH很低,茶园土壤存在较强的硝化作用。浙江为主共130个茶园土壤的NO3--N含量在0～286.8mg/kg之间,平均为41.7mg/kg。其中低于20mg/kg的样品占41%,超过100mg/kg的样品占12%。茶园上壤净硝化速率在-6.08-6.54mg/kg-d之间,平均1.62mg/kg-d,约有10%的土壤处于N固定化状态。极大多数NO3--N含量特别高的土壤施N量高、pH低于4.0,净硝化速率呈负数,土壤中积累的NO3-是大量施氮的结果。15个茶园土壤总硝化速率在0.75～11.56mg/kg.d之间,平均为4.66mg/kg.d。土壤微生物对NO3--N的同化速率在0.29～7.67mg/kg.d之间,平均为3.01mg/kg.d,占总硝化速率的33.7%～76.7%,平均为61.5%；茶园土壤与pH呈中性的蔬菜土壤差异不大,但显著高于森林土壤。
4.土壤硝化速率的高低主要取决于土地使用类型(土壤植被)及其管理水平,与茶树种植模式和植茶树龄也有较大的关系,与土壤pH的关系则相对较小。蔬菜、茶园和森林三种土壤比较,蔬菜土壤的硝化速率最高,茶园土壤次之,森林土壤最低；不同生产力水平茶园比较：高产>中产>低产茶园。施肥对土壤硝化速率有十分重要的影响。随着施N量的提高,土壤硝化作用随之增强,氮肥是刺激和促进硝化微生物数量和活性的重要因素。加入到蔬菜土壤的NH4+被迅速硝化,使土壤硝化速率随培养时间的变化表现为一级动力学方程,而加入到茶园和森林土壤中的NH4+被逐渐缓慢硝化,在35天的培养期内硝化的NH4+占60%～80%,硝化速率随培养时间的变化呈直线方程。适当降低茶园施N量,平衡供应土壤养分对于提高N素利用率,减少土壤NO3--N积累以及由此产生的环境污染有重要意义。不同种植模式土壤的净硝化速率表现为有机茶园>常规茶园>荒芜茶园,不同植茶年龄土壤的总硝化速率表现为50年>90年>9年生茶园>森林。土壤中添加不同剂量的CaCO3后,总硝化速率没有随pH的提高而增加。
5.导致酸性茶园土壤具有较强硝化活性的主要微生物是硝化古菌,而非硝化细菌。传统培养法没有检测到高生产力水平茶园土壤氨氧化细菌。DNA检测表明茶园土壤中硝化古菌(AO A) amoA基因拷贝数在5.5×104至2.4×107/g之间,平均为1.36×107/g,茶园土壤明显高于森林土壤；土壤硝化细菌(AOB) amoA基因拷贝数在未检出至4.40×106/g之间,平均为1.20×106/g。土壤硝化势与硝化古菌amoA基因拷贝数呈极显著正相关(p<0.001),但与硝化细菌amoA基因拷贝数的相关性不明显;土壤pH与硝化古菌amoA基因拷贝数的相关性不明显,但与硝化细菌amoA基因拷贝数却呈极显著正相关(p<0.001)；AOA与AOB amoA基因拷贝数之比随土壤pH的降低呈指数曲线显著提高,表明AOA是酸性茶园土壤硝化作用的主要微生物。茶园土壤AOA和AOB群落组成随上壤pH和施N量发生变化,不同石灰施用量茶园土壤间AOA群落组成、高N与低N施肥土壤间AOB群落组成有显著区别。不同基因型的AOA和AOB种类对土壤pH和施N量具有不同的要求,说明不同种类的AOA和AOB对土壤生态环境具有不同的要求。
6.茶园土壤具有较高的N20释放速率,施氮量高是最重要的原因之一。浙江142个茶园土壤N20释放速率在0～4960μg/m2.h之间,平均为467μg/m2.h。不同土壤类型间表现为：高产>中产>低产茶园和菜园>森林土壤。土壤N20释放速率的年周期变化与气温(地温)变化一致。随着施N量的提高,N20释放速率显著增加。茶园土壤N20-N占施N量的比例在1.43%-3.44%之间,平均为2.28%。N20释放速率随土壤含水量的提高明显增加,土壤含水量与施N量存在一定的正交互作用,上壤有机质含量高的土壤释放的N20也显著高于有机质含量低的土壤,但pH对土壤N20释放速率的影响不大。土壤中添加杀菌剂会明显降低N20释放速率,N20释放速率与硝酸还原酶活性呈正相关,表明茶园土壤释放的N20主要是生物反硝化产生的。
Tea is a globally important crop and is unusual because it both requires and acidifies the soil in which it grows. In spite of the low pH, high NO3-accumulation in and N2O emission from tea soils have been found, resulting in low N use efficiency and a great potential for diffuse pollution. In this study, represent tea soils with different basic properties, from tea gardens with varied management levels and planting models were collected from Zhejiang province, eastern China. A special attention was focused on tea soils with different production levels and tea stand ages, and their adjacent forest or/and vegetable soils in which tea soil originated. A series of field investigation and laboratory experiments were carried out to study soil microbial properties, biomass C, N and P, microbial quotient, net and gross nitrification, microbial nitrifiers, denitrification and N2O emission rate and their impact factors. The main results are as follows:
1. The microbial biomass C from75tea soils ranged from38.1to607.2mg/kg with an average of208.1mg/kg, microbial quotient C (ratio of microbial biomass C to total organic C) ranged from0.21%to4.55%with a mean of1.25%; the microbial biomass ninhydrin-N ranged from1.76to76.14mg/kg with an average of20.67mg/kg, microbial quotient ninhydrin-N ranged from0.26%to9.09%with a mean of1.80%; the microbial biomass P ranged from undetected to60.07mg/kg with a mean of14.93mg/kg, microbial quotient P ranged from0to10.20%with a mean of3.06%. Compared to other arable, grass and forest soils in literature, the microbial biomass, microbial quotient C and ninhydrin-N in tea soils were lower. However, the microbial biomass P was higher.
2. The microbial biomass content in tea garden soils was strongly influenced by tea cultivation intensity (production level), tea planting duration, soil pH, fertilizer application and tea planting model. Soil pH was highest in the forest soil and decreased with increasing productivity and age of tea stand. Soil total organic C, N, P and available nutrients significantly increased. Microbial biomass C, ninhydrin-N, PLFAs declined in the order forest> low> high> middle production, and biomass C and ATP declined in the order tea stand age50> age9> forest> age90. Microbial biomass ninhydrin-N and microbial quotient N declined in the order forest≥age9> age50> age90. Soil pH had a strong influence on the microbial biomass, demonstrated by positive linear correlations with: microbial biomass C, microbial biomass ninhydrin-N, PLFAs, the microbial quotient C, ninhydrin-N and P, the basal respiration rate and specific respiration rate. Appropriate fertilization could increase microbial biomass P. However, over application of N fertilizers reduced microbial biomass C, microbial biomass ninhydrin-N. With the increase of N application, the ratio of biomass C to total organic C, biomass ninhydrin-N to total N and biomass P to total P all significantly decreased. A principal component (PC) analysis of PLFA data showed consistent shift in the community composition with productivity level and stand age. The Gran positive bacterial PLFA biomarkers were significantly higher than Gran negative bacterial and fungal PLFA biomarkers. The ratio of fungal:bacterial PLFA biomarkers was negatively and linearly correlated with specific respiration rate in the soils (R2=0.93, p<0.05). Converting tea fields from conventional to organic increased soil pH and total organic C, the longer under organic cultivation, the higher pH and total organic C. Organic tea fields had higher microbial biomass C, ninhydrin-N, P and their microbial quotients than conventional fields.
3. In spite of low pH, tea garden soils had strong nitrification. The NO3--N concentrations in130tea field soils ranged from0to286.8mg/kg with an average of41.7mg/kg. About41%and12%of soil samples below20and above100mg/kg, respectively. Soil net nitrification rate ranged from-6.08to6.54mg/kg-d with an average of1.62mg/kg-d. About10%soils were in N immobilization. The NO3--N and net nitrification rate increased with increase of N application rate. The majority of soils with very higher NO3-accumulation had higher N application rate, pH below4.0, and even negative net nitrification rate. The gross nitrification rate from15soils ranged from0.75to11.56mg/kg.d with a mean of4.66mg/kg.d. Soil microbial NO3--N consumption rate ranged from0.29to7.67mg/kg.d with a mean of3.01mg/kg.d, accounting for33.7%-76.7%(mean61.5%) of gross nitrification rate. Nitrificaton rate in the tea soils had no significantly different with the neutral pH vegetable soil, but significant higher than the forest soil.
4. Type of land use and its management, tea planting model and duration were more effective to impact soil nitrification rate than soil pH. The net and gross nitrification rate was highest in vegetable soil, followed by tea soils, and the lowest in the forest soil. Net nitrification rate decreased in the order forest> age50> age90> age9tea stand, and organic> conventional> abandoned soils. Gross nitrification rate declined in the order tea age50> age90> age9> forest. The nitrification rate increased with increasing tea production and N application rates. The NH4+added to the vegetable soil was immediately and completely nitrified, resulted in nitrification pattern in first-order kinetics, but that in the tea and forest soils was nitrified more slowly with nearly linear nitrification pattern. About60%-80%of the added NH4+in the tea and forest soils was nitrified during35days of incubation, indicating nitrifiers do exist in these soils. High nitrogen application rate is the main cause of NO3-accumulation in tea soils and should be reduced to increase N use efficiency and minimize NO3-pollution of water resources. Liming did not increase soil gross nitrification rate.
5. The key microorganism for strong nitrification in tea soils were ammonia-oxidizing archaea (AOA), instead of ammonia-oxidizing bacteria (AOB). No AOB were found by traditional incubation method in the soil of high tea production field. Soil DNA sequence analysis after quantitative polymerase chain reaction (Q-PCR) and terminal-restriction fragment length polymorphism (T-RFLP) showed the abundance of AOA and AOB, and nitrification potentials were significantly higher in the tea soils than adjacent forest soils. The AOA amoA gene copy numbers ranged from5.5×104to2.4×107/g with an average of1.36X107/g, and the AOB amoA gene copy numbers ranged from below detection limits to4.40×106/g with a mean of1.20×106/g. A significant relationship between AOA abundance and nitrification potential (p<0.001) was found, but not between the AOB abundance and nitrification potential. Soil pH had a significantly negative relationship with AOB abundance, but not with AOA abundance. There was an exponential increase in the ratio of AOA to AOB amoA gene copies with decreasing soil pH values in the tea gardens. The AOB copy numbers were below detection limits in some highly acidic tea soils, but these soils still had high nitrification rates. These results suggest that nitrification is mainly driven by AOA and not AOB in highly acidic tea soils. We also found that different genotypes of AOA and AOB adapt to a particular soil pH and N content which suggest that functional microbial populations also follow niche based community assembly.
6. High N2O emission rate was found in tea garden soils, high N application rate was the main cause. The N2O emission rate in142tea soils ranged from0to4960μg/m2.h with an average of467μg/m2.h. It decreased in the order high> middle> low tea production and vegetable> forest soils. Its annual change was in accordance with the soil temperature. N2O emission rate was significantly increased with the increase of N application rate and of soil moisture contents, which had a synergistic effect on N2O emission rate. The annual fertilizer-induced emission factor ranged from1.43%to3.44%with a mean of2.28%. High soil organic C increased N2O emission, but no effect of soil pH on N2O emission was found. There were a significant reduction of N2O emission by bactericides and significant relationship between N2O emission rate and nitrate reductase, indicating that N2O emitted by microbial denitrification.
1.浙江75个茶园土壤微生物量碳(C)在38.1-607.2mg/kg之间,平均为208.1mg/kg；微生物量C占总有机C的比例在0.21%-4.55%之间,平均为1.25%。微生物量氮(茚三酮反应态N)在1.76-76.14mg/kg之间,平均为20.67mg/kg；微生物量N占全N的比例在0.26%-9.09%之间,平均为1.80%。微生物量磷(P)在未检出到60.07mg/kg之间,平均为14.93mg/kg；微生物量P占全P的比例在0-10.20%之间,平均为3.06%。与多数耕地或森林土壤相比,茶园土壤微量物量C和N,以及微生物量C占总有机C的比例和微生物量N占全N的比例均偏低,但微生物量P与其它土壤相当,微生物量P占全P的比例高于其它土壤。
2.茶园土壤微生物量受生产力水平、植茶年龄、pH、施肥量和茶树种植模式的显著影响。随着茶园生产力水平和植茶年龄的提高,土壤pH明显降低,有机C、全N、全P和有效态营养元素含量显著提高。微生物C、N和PLFAs含量表现为森林>低产>高产>中产茶园：土壤微生物C和ATP含量表现为50年>9年>森林>90年生茶园,微生物量N及微生物量N占全N量的比例表现为森林≥9年>50年>90年生茶园。茶园土壤pH低是导致微生物量少和生长代谢弱的最重要原因之一,除微生物量P外,微生物量C和N、微生物商、土壤基础呼吸速率和代谢商均与pH呈极显著正相关。适量施P有利于提高土壤微生物量P,但过量施N降低微生物量C和N含量；随着茶园施氮量的提高,土壤微生物商C、N和P均呈极显著降低的趋势。对磷酯脂肪酸(PLFA)的分析表明茶园土壤微生物群落结构随着茶园生产力水平的提高和树龄的增加呈连续有规律的变化；PLFA标记革兰氏阳性细菌的数量显著高于革兰氏阴性细菌和真菌的数量；茶园土壤PLFA标记真菌和细菌的比例显著高于森林土壤,且PLFA标记真菌与细菌比例与代谢商呈显著负相关(R2=0.93,p<0.05)。茶园由常规种植向有机种植转换有利于提高土壤pH和有机C含量,有机茶种植年限越长,pH和有机C含量越高。茶园有机种植方式能明显提高土壤微生物量C、N和P的含量及微生物商C、N和P的比例,改善茶园土壤质量。
3.尽管土壤pH很低,茶园土壤存在较强的硝化作用。浙江为主共130个茶园土壤的NO3--N含量在0～286.8mg/kg之间,平均为41.7mg/kg。其中低于20mg/kg的样品占41%,超过100mg/kg的样品占12%。茶园上壤净硝化速率在-6.08-6.54mg/kg-d之间,平均1.62mg/kg-d,约有10%的土壤处于N固定化状态。极大多数NO3--N含量特别高的土壤施N量高、pH低于4.0,净硝化速率呈负数,土壤中积累的NO3-是大量施氮的结果。15个茶园土壤总硝化速率在0.75～11.56mg/kg.d之间,平均为4.66mg/kg.d。土壤微生物对NO3--N的同化速率在0.29～7.67mg/kg.d之间,平均为3.01mg/kg.d,占总硝化速率的33.7%～76.7%,平均为61.5%；茶园土壤与pH呈中性的蔬菜土壤差异不大,但显著高于森林土壤。
4.土壤硝化速率的高低主要取决于土地使用类型(土壤植被)及其管理水平,与茶树种植模式和植茶树龄也有较大的关系,与土壤pH的关系则相对较小。蔬菜、茶园和森林三种土壤比较,蔬菜土壤的硝化速率最高,茶园土壤次之,森林土壤最低；不同生产力水平茶园比较：高产>中产>低产茶园。施肥对土壤硝化速率有十分重要的影响。随着施N量的提高,土壤硝化作用随之增强,氮肥是刺激和促进硝化微生物数量和活性的重要因素。加入到蔬菜土壤的NH4+被迅速硝化,使土壤硝化速率随培养时间的变化表现为一级动力学方程,而加入到茶园和森林土壤中的NH4+被逐渐缓慢硝化,在35天的培养期内硝化的NH4+占60%～80%,硝化速率随培养时间的变化呈直线方程。适当降低茶园施N量,平衡供应土壤养分对于提高N素利用率,减少土壤NO3--N积累以及由此产生的环境污染有重要意义。不同种植模式土壤的净硝化速率表现为有机茶园>常规茶园>荒芜茶园,不同植茶年龄土壤的总硝化速率表现为50年>90年>9年生茶园>森林。土壤中添加不同剂量的CaCO3后,总硝化速率没有随pH的提高而增加。
5.导致酸性茶园土壤具有较强硝化活性的主要微生物是硝化古菌,而非硝化细菌。传统培养法没有检测到高生产力水平茶园土壤氨氧化细菌。DNA检测表明茶园土壤中硝化古菌(AO A) amoA基因拷贝数在5.5×104至2.4×107/g之间,平均为1.36×107/g,茶园土壤明显高于森林土壤；土壤硝化细菌(AOB) amoA基因拷贝数在未检出至4.40×106/g之间,平均为1.20×106/g。土壤硝化势与硝化古菌amoA基因拷贝数呈极显著正相关(p<0.001),但与硝化细菌amoA基因拷贝数的相关性不明显;土壤pH与硝化古菌amoA基因拷贝数的相关性不明显,但与硝化细菌amoA基因拷贝数却呈极显著正相关(p<0.001)；AOA与AOB amoA基因拷贝数之比随土壤pH的降低呈指数曲线显著提高,表明AOA是酸性茶园土壤硝化作用的主要微生物。茶园土壤AOA和AOB群落组成随上壤pH和施N量发生变化,不同石灰施用量茶园土壤间AOA群落组成、高N与低N施肥土壤间AOB群落组成有显著区别。不同基因型的AOA和AOB种类对土壤pH和施N量具有不同的要求,说明不同种类的AOA和AOB对土壤生态环境具有不同的要求。
6.茶园土壤具有较高的N20释放速率,施氮量高是最重要的原因之一。浙江142个茶园土壤N20释放速率在0～4960μg/m2.h之间,平均为467μg/m2.h。不同土壤类型间表现为：高产>中产>低产茶园和菜园>森林土壤。土壤N20释放速率的年周期变化与气温(地温)变化一致。随着施N量的提高,N20释放速率显著增加。茶园土壤N20-N占施N量的比例在1.43%-3.44%之间,平均为2.28%。N20释放速率随土壤含水量的提高明显增加,土壤含水量与施N量存在一定的正交互作用,上壤有机质含量高的土壤释放的N20也显著高于有机质含量低的土壤,但pH对土壤N20释放速率的影响不大。土壤中添加杀菌剂会明显降低N20释放速率,N20释放速率与硝酸还原酶活性呈正相关,表明茶园土壤释放的N20主要是生物反硝化产生的。
Tea is a globally important crop and is unusual because it both requires and acidifies the soil in which it grows. In spite of the low pH, high NO3-accumulation in and N2O emission from tea soils have been found, resulting in low N use efficiency and a great potential for diffuse pollution. In this study, represent tea soils with different basic properties, from tea gardens with varied management levels and planting models were collected from Zhejiang province, eastern China. A special attention was focused on tea soils with different production levels and tea stand ages, and their adjacent forest or/and vegetable soils in which tea soil originated. A series of field investigation and laboratory experiments were carried out to study soil microbial properties, biomass C, N and P, microbial quotient, net and gross nitrification, microbial nitrifiers, denitrification and N2O emission rate and their impact factors. The main results are as follows:
1. The microbial biomass C from75tea soils ranged from38.1to607.2mg/kg with an average of208.1mg/kg, microbial quotient C (ratio of microbial biomass C to total organic C) ranged from0.21%to4.55%with a mean of1.25%; the microbial biomass ninhydrin-N ranged from1.76to76.14mg/kg with an average of20.67mg/kg, microbial quotient ninhydrin-N ranged from0.26%to9.09%with a mean of1.80%; the microbial biomass P ranged from undetected to60.07mg/kg with a mean of14.93mg/kg, microbial quotient P ranged from0to10.20%with a mean of3.06%. Compared to other arable, grass and forest soils in literature, the microbial biomass, microbial quotient C and ninhydrin-N in tea soils were lower. However, the microbial biomass P was higher.
2. The microbial biomass content in tea garden soils was strongly influenced by tea cultivation intensity (production level), tea planting duration, soil pH, fertilizer application and tea planting model. Soil pH was highest in the forest soil and decreased with increasing productivity and age of tea stand. Soil total organic C, N, P and available nutrients significantly increased. Microbial biomass C, ninhydrin-N, PLFAs declined in the order forest> low> high> middle production, and biomass C and ATP declined in the order tea stand age50> age9> forest> age90. Microbial biomass ninhydrin-N and microbial quotient N declined in the order forest≥age9> age50> age90. Soil pH had a strong influence on the microbial biomass, demonstrated by positive linear correlations with: microbial biomass C, microbial biomass ninhydrin-N, PLFAs, the microbial quotient C, ninhydrin-N and P, the basal respiration rate and specific respiration rate. Appropriate fertilization could increase microbial biomass P. However, over application of N fertilizers reduced microbial biomass C, microbial biomass ninhydrin-N. With the increase of N application, the ratio of biomass C to total organic C, biomass ninhydrin-N to total N and biomass P to total P all significantly decreased. A principal component (PC) analysis of PLFA data showed consistent shift in the community composition with productivity level and stand age. The Gran positive bacterial PLFA biomarkers were significantly higher than Gran negative bacterial and fungal PLFA biomarkers. The ratio of fungal:bacterial PLFA biomarkers was negatively and linearly correlated with specific respiration rate in the soils (R2=0.93, p<0.05). Converting tea fields from conventional to organic increased soil pH and total organic C, the longer under organic cultivation, the higher pH and total organic C. Organic tea fields had higher microbial biomass C, ninhydrin-N, P and their microbial quotients than conventional fields.
3. In spite of low pH, tea garden soils had strong nitrification. The NO3--N concentrations in130tea field soils ranged from0to286.8mg/kg with an average of41.7mg/kg. About41%and12%of soil samples below20and above100mg/kg, respectively. Soil net nitrification rate ranged from-6.08to6.54mg/kg-d with an average of1.62mg/kg-d. About10%soils were in N immobilization. The NO3--N and net nitrification rate increased with increase of N application rate. The majority of soils with very higher NO3-accumulation had higher N application rate, pH below4.0, and even negative net nitrification rate. The gross nitrification rate from15soils ranged from0.75to11.56mg/kg.d with a mean of4.66mg/kg.d. Soil microbial NO3--N consumption rate ranged from0.29to7.67mg/kg.d with a mean of3.01mg/kg.d, accounting for33.7%-76.7%(mean61.5%) of gross nitrification rate. Nitrificaton rate in the tea soils had no significantly different with the neutral pH vegetable soil, but significant higher than the forest soil.
4. Type of land use and its management, tea planting model and duration were more effective to impact soil nitrification rate than soil pH. The net and gross nitrification rate was highest in vegetable soil, followed by tea soils, and the lowest in the forest soil. Net nitrification rate decreased in the order forest> age50> age90> age9tea stand, and organic> conventional> abandoned soils. Gross nitrification rate declined in the order tea age50> age90> age9> forest. The nitrification rate increased with increasing tea production and N application rates. The NH4+added to the vegetable soil was immediately and completely nitrified, resulted in nitrification pattern in first-order kinetics, but that in the tea and forest soils was nitrified more slowly with nearly linear nitrification pattern. About60%-80%of the added NH4+in the tea and forest soils was nitrified during35days of incubation, indicating nitrifiers do exist in these soils. High nitrogen application rate is the main cause of NO3-accumulation in tea soils and should be reduced to increase N use efficiency and minimize NO3-pollution of water resources. Liming did not increase soil gross nitrification rate.
5. The key microorganism for strong nitrification in tea soils were ammonia-oxidizing archaea (AOA), instead of ammonia-oxidizing bacteria (AOB). No AOB were found by traditional incubation method in the soil of high tea production field. Soil DNA sequence analysis after quantitative polymerase chain reaction (Q-PCR) and terminal-restriction fragment length polymorphism (T-RFLP) showed the abundance of AOA and AOB, and nitrification potentials were significantly higher in the tea soils than adjacent forest soils. The AOA amoA gene copy numbers ranged from5.5×104to2.4×107/g with an average of1.36X107/g, and the AOB amoA gene copy numbers ranged from below detection limits to4.40×106/g with a mean of1.20×106/g. A significant relationship between AOA abundance and nitrification potential (p<0.001) was found, but not between the AOB abundance and nitrification potential. Soil pH had a significantly negative relationship with AOB abundance, but not with AOA abundance. There was an exponential increase in the ratio of AOA to AOB amoA gene copies with decreasing soil pH values in the tea gardens. The AOB copy numbers were below detection limits in some highly acidic tea soils, but these soils still had high nitrification rates. These results suggest that nitrification is mainly driven by AOA and not AOB in highly acidic tea soils. We also found that different genotypes of AOA and AOB adapt to a particular soil pH and N content which suggest that functional microbial populations also follow niche based community assembly.
6. High N2O emission rate was found in tea garden soils, high N application rate was the main cause. The N2O emission rate in142tea soils ranged from0to4960μg/m2.h with an average of467μg/m2.h. It decreased in the order high> middle> low tea production and vegetable> forest soils. Its annual change was in accordance with the soil temperature. N2O emission rate was significantly increased with the increase of N application rate and of soil moisture contents, which had a synergistic effect on N2O emission rate. The annual fertilizer-induced emission factor ranged from1.43%to3.44%with a mean of2.28%. High soil organic C increased N2O emission, but no effect of soil pH on N2O emission was found. There were a significant reduction of N2O emission by bactericides and significant relationship between N2O emission rate and nitrate reductase, indicating that N2O emitted by microbial denitrification.
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