红壤稻田不同生育期土壤氨氧化微生物群落结构

以福建省红壤稻田土壤为对象,通过提取土壤总DNA,利用特异引物进行PCR(聚合酶链反应)扩增和DGGE(变性梯度凝胶电泳)并结合DNA克隆测序,研究了水稻生长过程中稻田土壤氨氧化细菌和氨氧化古菌群落结构的变化。结果显示:稻田土壤具有丰富的氨氧化细菌和氨氧化古菌资源。水稻生长过程中土壤氨氧化细菌群落组成较为稳定,只表现出水稻生长前期(苗期、分蘖期)和中后期(孕穗期、成熟期)间存在一定差异。而土壤氨氧化古菌群落组成变化较大,在水稻生长的苗期、分蘖期、孕穗期和成熟期4个时期间均存在一定差异。在水稻生长过程中,土壤氨氧化细菌群落多样性指数无显著性变化,但氨氧化古菌群落多样性指数随水稻生长明显提高,孕穗期后才达到平稳。水稻生长前期土壤硝化势也具有显著上升趋势,孕穗期时达到最高,而后有所下降。土壤硝化势与氨氧化古菌群落多样性指数具有显著正相关性,与氨氧化细菌没有相关性。研究表明,氨氧化古菌对红壤稻田土壤硝化作用的影响程度较大,证实了氨氧化微生物尤其是氨氧化古菌在稻田土壤微生物组成及其生态系统功能中的重要性。     

脲酶抑制剂与硝化抑制剂对稻田土壤硝化、反硝化功能菌的影响

[目的]在农业生产中,脲酶抑制剂(urease inhibitor,UI)与硝化抑制剂(nitrification inhibitor,NI)常作为氮肥增效剂来提高肥料利用率。本文研究了在我国南方红壤稻田施用脲酶抑制剂与硝化抑制剂后,土壤中氨氧化细菌(ammonia oxidizing bacteria,AOB)、氨氧化古菌(ammonia-oxidizing archaea,AOA)以及反硝化细菌的丰度以及群落结构的变化特征,旨在揭示抑制剂的作用机理及其对土壤环境的影响。[方法]试验在我国南方红壤稻田进行,共设5个处理:1)不施氮肥(CK);2)尿素(U);3)尿素+脲酶抑制剂(U+UI);4)尿素+硝化抑制剂(U+NI);5)尿素+脲酶抑制剂+硝化抑制剂(U+UI+NI),3次重复。脲酶抑制剂与硝化抑制剂分别为NBPT[N-(n-butyl)thiophosphrictriamide,N-丁基硫代磷酰三胺]和DMPP(3,4-dimethylpyrazole phosphate,3,4-二甲基吡唑磷酸盐)。通过荧光定量PCR(Real-time PCR)研究水稻分蘖期与孕穗期抑制剂对三类微生物标记基因拷贝数的影响,并分析土壤铵态氮、硝态氮与三种菌群丰度的相关性;利用变性梯度凝胶电泳(DenaturingGradient Gel Electrophoresis,DGGE)分析抑制剂对土壤AOB、AOA以及反硝化细菌群落结构的影响,并对优势菌群进行系统发育分析。[结果]1)荧光定量PCR结果表明,施用氮肥对两个时期土壤中AOB的amoA基因与反硝化细菌nirK基因的拷贝数均有显著提高,而对AOA的amoA基因始终没有明显影响;AOB与nirK反硝化细菌的丰度与两个时期的铵态氮含量、分蘖期的硝态氮含量呈极显著正相关,与孕穗期的硝态氮含量相关性不显著;DMPP仅在分蘖期显著减少了AOB的amoA基因拷贝数,表明DMPP主要通过限制AOB的生长来抑制稻田土壤硝化过程;NBPT对三类微生物的丰度无明显影响;2)DGGE图谱表明,在分蘖期与孕穗期,施用氮肥均明显增加了图谱中AOB的条带数,而对AOA却没有明显影响;氮肥明显增加了孕穗期反硝化细菌的条带数;与氮肥的影响相比,抑制剂NBPT与DMPP对AOA、AOB以及反硝化菌的群落结构影响甚微;系统发育分析结果表明,与土壤中AOB的优势菌群序列较为接近的有亚硝化单胞菌和亚硝化螺菌。[结论]在南方红壤稻田中,施入氮肥可显著提高AOB与反硝化细菌的丰度,明显影响两种菌群的群落结构,而AOA较为稳定;NBPT对三类微生物的群落结构丰度无明显影响;硝化抑制剂DMPP可抑制AOB的生长但仅表现在分蘖期,这可能是其缓解硝化反应的主要途径;这也说明二者对土壤生态环境均安全可靠。     

Temporal shifts in diversity and quantity of nirS and nirK in a rice paddy field soil

Denitrification is an important part of the nitrogen cycle in the environment, and diverse bacteria, archaea, and fungi are known to have denitrifying ability. Rice paddy field soils have been known to have strong denitrifying activity, but the microbes responsible for denitrification in rice paddy field soils are not well known. Present study analyzed the diversity and quantity of the nitrite reductase genes (nirS and nirK) in a rice paddy field soil, sampled four times in one rice-growing season. Clone library analyses suggested that the denitrifier community composition varied over sampling time. Although many clones were distantly related to the known NirS or NirK, some clones were related to the NirS from Burkholderiales and Rhodocyclales bacteria, and some were related to the NirK from Rhizobiales bacteria. These denitrifiers may play an important role in denitrification in the rice paddy field soil. The quantitative PCR results showed that nirK was more abundant than nirS in all soil samples, but the nirK/nirS ratio decreased after water logging. These results suggest that both diversity and quantity changed over time in the rice paddy field soil, in response to the soil condition.     

Diversity and Characterization of Potential H2-Dependent Fe(Ⅲ)-Reducing Bacteria in Paddy Soils

Microbial ferric iron reduction, with organic carbon or hydrogen as the electron donor, is one of the most important biogeochemical processes in anoxic paddy soils; however, the diversity and community structure of hydrogen-dependent dissimilatory iron-reducers remain unknown. Potential H2-dependent Fe(III)-reducing bacteria in paddy soils were explored using enrichment cultures with ferrihydrite or goethite as the electron acceptor and hydrogen as the electron donor. Terminal restriction fragment length polymorphism (T-RFLP) analysis and cloning/sequencing were conducted to reveal bacterial community structure. Results showed that Geobacter and Clostridium were the dominant bacteria in the enrichment cultures. Fe(III) oxide mineral phases showed a strong effect on the community structure; Geobacter and Clostridium were dominant in the ferrihydrite treatment, while Clostridium spp. were dominant in the goethite treatment. These suggested that H2-dependent Fe(III)-reducing bacteria might be widely distributed in paddy soils and that besides Geobacter, Clostridium spp. might also be an important group of H2-dependent Fe(III)-reducing microorganisms.     

Biosurfactant in Membrane Separation of Atrazine from Water

This study examines the effects of land use change on nitrate concentration and the abundances of ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), and narG-containing denitrifiers in shallow groundwater. The results reveal a general increase of nitrate in shallow groundwater following the change of land use from paddy fields to vegetable patches. Furthermore, a significant relationship between NO₃ ^?-N concentrations was observed both in groundwater and in soil at soil depths of 0–20, 20–40, 40–60, 60–80, and 80–100 cm. With regard to gene abundance in groundwater, the AOB amoA gene was most abundant and the AOA amoA gene copy numbers were lowest from the field with long-term paddy cultivation compared with the field under vegetable cultivation. The narG gene copy numbers were higher from the field under short-term vegetable cultivation compared with fields under long-term vegetable cultivation. The NO₃ ^?-N concentrations in groundwater correlated positively with AOA amoA gene copy numbers, negatively with the AOB amoA gene, but with no significant relationship with the narG gene. In conclusion, land use change from paddy fields to vegetable patches increases nitrate in groundwater, which is correlated significantly with nitrate in soil and the abundance of the amoA gene, but is not related to the narG gene in groundwater. This study also suggests that the removal of groundwater nitrate pollution is not feasible through biological denitrification without additional denitrifiers and that it might even become more aggravated because of the AOA.     

南方典型母质发育稻田土壤氨氧化细菌和古菌群落结构特征

依托中国科学院桃源农业生态试验站水稻土易地置土长期定位试验,以广东英利(YL,发育于玄武岩风化物)、江西鹰潭(YT,发育于第四纪红黏土)和湖南桃源古市(TYG,发育于河流冲积物)3种母质发育稻田土壤为供试土壤,采用常规土壤农化分析方法测得3种土壤的理化性质、硝化速率,并结合实时荧光定量PCR技术和Illumina MiSeq测序技术,分析了土壤氨氧化古菌(AOA)和氨氧化细菌(AOB)种群丰度及群落结构的异同。结果表明：土壤硝化速率除分蘖期外,均表现为：YT>YL>TYG。YL与YT土壤中AOA数量占主导地位,AOA/AOB比值比值分别在0.72 ~ 3.05、0.98 ~ 1.52;TYG土壤中AOB数量占主导地位,AOA/AOB比值在0.21 ~ 0.75。群落组成方面,不同母质土壤间AOA、AOB群落结构差异显著,3种土壤AOA优势菌属各不相同,AOB优势菌属均为Nitrosospira。冗余分析表明,pH和可溶性有机碳是造成AOA、AOB群落结构差异的核心因子。综上,在环境背景均统一的前提下,不同母质发育稻田土壤理化性质之间依旧存在差异,这种差异影响着氨氧化微生物AOA、AOB的丰度以及群落结构。     

Effect of agricultural land use change on community composition of bacteria and ammonia oxidizers

Purpose

Soil microbial communities can be strongly influenced by agricultural practices, but little is known about bacterial community successions as land use changes. The objective of this study was to determine microbial community shifts following major land use changes in order to improve our understanding of land use impacts on microbial community composition and functions.

Materials and methods

Four agricultural land use patterns were selected for the study, including old rice paddy fields (ORP), Magnolia nursery planting (MNP), short-term vegetable (STV), and long-term vegetable (LTV) cultivation. All four systems are located in the same region with same soil parent material (alluvium), and the MNP, STV, and LTV systems had been converted from ORP for 10, 3, and 30 years, respectively. Soil bacteria and ammonia oxidizer community compositions were analyzed by 454 pyrosequencing and terminal restriction fragment length polymorphism, respectively. Quantitative PCR was used to determine 16S rRNA and amoA gene copy numbers.

Results and discussion

The results showed that when land use was changed from rice paddy to upland systems, the relative abundance of Chloroflexi increased whereas Acidobacteria decreased significantly. While LTV induced significant shifts of bacterial composition, MNP had the highest relative abundance of genera GP1, GP2, and GP3, which were mainly related to the development of soil acidity. The community composition of ammonia-oxidizing bacteria (AOB) but not ammonia-oxidizing archaea was strongly impacted by the agricultural land use patterns, with LTV inducing the growth of a single super predominant AOB group. The land use changes also induced significant shifts in the abundance of 16S rRNA and bacterial amoA genes, but no significant differences in the abundance of archaea amoA was detected among the four land use patterns. Soil total phosphorous, available phosphorous, NO₃ ^?, and soil organic carbon contents and pH were the main determinants in driving the composition of both bacteria and AOB communities.

Conclusions

These results clearly show the significant impact of land use change on soil microbial community composition and abundance and this will have major implications on the microbial ecology and nutrient cycling in these systems, some of which is unknown. Further research should be directed to studying the impacts of these microbial community shifts on nutrient dynamics in these agroecosystems so that improved nutrient management systems can be developed.     

Distributions and environmental drivers of archaea and bacteria in paddy soils

Purpose

The aim of this study is to investigate the abundance, diversity, and distribution of archaea and bacteria as affected by environment parameters in paddy soils, with focus on putative functional microbial groups related to redox processes. Because there is generally a high iron content in the soil, we also want to test a hypothesis that soil iron concentration significantly affects microbial diversity and distribution.

Materials and methods

Quantitative PCR and barcoded pyrosequencing of 16S ribosomal RNA genes were employed to investigate the abundance and community composition of archaeal and bacterial communities in 27 surface paddy soil samples. Pearson’s correlation, analysis of variance, partial least squares regression, principal coordinates analysis, and structural equation models were performed for the analyses of gene copy numbers, α-diversity, β-diversity, and relative abundances of archaea and bacteria and their relationships with environmental factors.

Results and discussion

Archaeal abundance was correlated greatest with temperature, but bacterial abundance was affected mainly by soil organic matter and total nitrogen content. Soil pH and concentrations of different ions were associated with archaeal and bacterial β-diversity. The relative abundances of Euryarchaeota and Thaumarchaeota were 61.3 and 13.1% of archaea and correlated with soil pH, which may affect the availability of substrates to methanogens and ammonia oxidizers. Dominant bacterial phyla were Proteobacteria (32.4%), Acidobacteria (17.8%), Bacteroidetes (9.3%), and Verrucomicrobia (6.0%). The relative abundances of putative bacterial reducers of nitrate, Fe(III), sulfate, and sulfur, and oxidizers of ammonia, nitrite, reduced sulfur, and C1 compounds had positive, negative, or non-significant correlations with the concentrations of their substrates. Soil iron concentration was correlated only with the distributions of some putative iron-reducing bacteria.

Conclusions

In paddy soils characterized by dynamic redox processes, archaea and bacteria differ in relationships of abundance, diversity, and distribution with environmental factors. Especially, the concentrations of electron donors or acceptors can explain the distributions of some but not all the putative functional microbial groups related to redox processes. Depending on pH range, soil pH has a strong impact on microbial ecology in paddy soils.

     

稻蟹共作对稻田水体底栖动物多样性的影响

稻蟹共作是一种生态型种养新模式,而底栖动物是该复合生态系统的重要组成部分。为研究稻蟹共作对稻田底栖动物多样性的影响,在水稻分蘖期、拔节期、扬花期和成熟期,采集不同放养密度的稻蟹共作稻田(仔蟹放养密度10 ind.m 2,低密度,LD;仔蟹放养密度30 ind.m 2,高密度,HD)和常规稻田(不放蟹,常规栽培,CK)水体的底栖动物,分析不同稻田生态系统底栖动物的种类构成、密度和多样性。结果表明:水稻分蘖期,3种放蟹密度处理稻田底栖动物的种类数、密度、多样性指数和Pielou均匀度指数均无显著差异(P>0.05);在拔节期和扬花期,养蟹稻田底栖动物的种类数和密度均小于常规稻田,且与放蟹密度呈反比;常规稻田底栖动物的Shannon-Wiener多样性指数在拔节期和扬花期均显著高于高密度养蟹稻田(P<0.05),而只在拔节期显著高于低密度养蟹稻田(P<0.05),且Pielou均匀度指数也在拔节期显著高于低密度养蟹稻田(P<0.05);成熟期,3种处理底栖动物的种类数和密度均达到最低点,Shannon-Wiener多样性指数无显著差异(P>0.05),高密度养蟹稻田的Pielou均匀度指数显著高于常规稻田(P<0.05)。研究结果可为稻蟹共作稻田底栖动物的多样性保护和稻蟹共作技术的可持续发展提供理论指导,促进稻蟹共作技术的发展和推广。     

Long-term submergence of non-methanogenic oxic upland field soils helps to develop the methanogenic archaeal community as revealed by pot and field experiments

The community structure of methanogenic archaea is relatively stable,i.e.,it is sustained at a high abundance with minimal changes in composition,in paddy field soils irrespective of submergence and drainage.In contrast,the abundance in non-methanogenic oxic soils is much lower than that in paddy field soils.This study aimed to describe methanogenic archaeal community development following the long-term submergence of non-methanogenic oxic upland field soils in pot and field experiments.In the pot experiment,a soil sample obtained from an upland field was incubated under submerged conditions for 275 d.Soil samples periodically collected were subjected to culture-dependent most probable number(MPN)enumeration,polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE)analysis of archaeal 16 S r RNA gene,and quantitative PCR analysis of the methyl-coenzyme M reductase alpha subunit gene(mcr A)of methanogenic archaea.The abundance of methanogenic archaea increased from 102 to 103 cells g-1 dry soil and 104 to 107 copies of mcr A gene g-1 dry soil after submergence.Although no methanogenic archaeon was detected prior to incubation by the DGGE analysis,members from Methanocellales,Methanosarcinaceae,and Methanosaetaceae proliferated in the soils,and the community structure was relatively stable once established.In the field experiment,the number of viable methanogenic archaea in a rice paddy field converted from meadow(reclaimed paddy field)was monitored by MPN enumeration over five annual cycles of field operations.Viability was also determined simultaneously in a paddy field where the plow layer soil from a farmer’s paddy field was dressed onto the meadow(dressed paddy field)and an upland crop field converted from the meadow(reclaimed upland field).The number of viable methanogenic archaea in the reclaimed paddy field was below the detection limit before the first cultivation of rice and in the reclaimed upland field.Then,the number gradually increased over five years and finally reached 103–104 cells g-1 dry soil,which was comparable to that in the dressed paddy field.These findings showed that the low abundance of autochthonous methanogenic archaea in the non-methanogenic oxic upland field soils steadily proliferated,and the community structure was developed following repeated and long-term submergence.These results suggest that habitats suitable for methanogenic archaea were established in soil following repeated and long-term submergence.     

Growth,morphological and yield responses of irrigated wheat (Triticum aestivum L.) genotypes to water stress

Water shortages is a major constraint in wheat production in South Africa. It is important therefore to assist irrigated wheat farmers to identify water stress tolerant growth stages in irrigated wheat genotypes. This study evaluated new wheat genotypes for water stress at different growth stages. An 8 (genotypes) × 2 (water treatments) × 3 (growth stages) factorial experiment was laid out in a randomised complete block design with three replicates. The results indicated that plant height was not affected (p > .05) by water stress at tillering and grain filling. Water stress imposed at the tillering stage reduced the number of fertile tillers (p < .05) in susceptible genotypes while at the flowering and grain filling stages all genotypes were tolerant (p > .05). Aboveground biomass was only affected (p < .05) by water stress imposed at the tillering stage. Water stress reduced grain yield on the genotypes where stress was imposed at the tillering stage (p < .05); whereas when stress was imposed at flowering and grain filling the grain yield was not reduced (p > .05). This study provided evidence to suggest that most genotypes were tolerant to water stress at the flowering and grain filling stages.     

氮肥对稻田土壤反硝化细菌群落结构和丰度的影响

以氮肥田间定位试验为研究对象,利用PCR-DGGE(聚合酶链反应变性梯度凝胶电泳)和荧光定量PCR(real-time PCR)技术,通过对反硝化细菌nirS基因的检测,分析了定位试验第2年稻田反硝化细菌群落结构和丰度的变化。DGGE图谱及依据其条带位置和亮度数字化数值进行的主成分分析(PCA)结果均显示:在氮肥定位试验第2年,与不施肥对照(CK)比较,在水稻各个生育期(分蘖期、齐穗期和成熟期)内,施用氮肥[150kg(N)·hm-2]的稻田根层土或表土中的反硝化细菌群落结构均无明显变化;且稻田根层土或表土中的反硝化细菌群落结构在水稻各个生育期间也均无明显差异。荧光定量PCR结果显示,在水稻生长发育过程中,施用氮肥的稻田根层土或表土中的反硝化细菌nirS基因拷贝数始终显著(P<0.05)高于其对应的不施肥对照。此外,无论施用氮肥与否,根层土中的反硝化细菌nirS基因拷贝数在水稻成熟期时都会显著(P<0.05)降低;但表土中的nirS基因拷贝数在水稻各生育期间无明显变化;且水稻成熟期时施用氮肥和不施肥的稻田表土中nirS基因拷贝数都显著(P<0.05)高于根层土。同时,与对照比较施用氮肥可促进水稻增产44%。研究表明,短期定位试验中施用氮肥能够显著提高稻田土壤反硝化细菌的丰度,但对其群落结构没有明显影响。     

稻田厌氧氨氧化菌群落结构对氮肥的响应

为探明稻田厌氧氨氧化菌多样性及其对氮肥用量的响应状况,利用厌氧氨氧化菌16S rRNA基因特异引物对定位试验稻田土壤DNA进行PCR-DGGE(聚合酶链反应变性梯度凝胶电泳)并结合DNA克隆测序,研究了氮肥供应量对厌氧氨氧化菌群落结构的影响。DGGE图谱及依据其条带位置和亮度数值计算的多样性指数均显示:高氮处理[N3:225 kg(N).hm 2]的厌氧氨氧化菌群落结构多样性在表层或根层土壤中均显著(P<0.05)高于中、低氮[N2:150 kg(N).hm 2;N1:75 kg(N).hm 2]处理和不施肥对照(CK);同时,高氮处理下表层土壤厌氧氨氧化菌群落多样性指数显著高于根层土壤(P<0.05)。冗余分析(RDA)结果表明,表层土壤中厌氧氨氧化菌群落结构组成与不同氮肥水平处理存在显著相关性(P=0.006)。此外,本试验获得厌氧氨氧化菌DGGE条带DNA序列18条,登录GenBank并获得登录号。研究表明稻田厌氧氨氧化菌群落结构对高氮水平具有较强的响应,尤其是在表层土壤中。     

Diversity of methanogenic archaeal communities in Japanese paddy field ecosystem, estimated by denaturing gradient gel electrophoresis

Diversity of methanogenic archaeal communities in Japanese paddy field ecosystem was evaluated by the denaturing gradient gel electrophoresis (DGGE) after PCR amplification of the 16S rRNA genes (16S rDNAs), sequencing analysis and data evaluation by principal component analysis. Data were obtained from samples collected from the plowed soil layer, rice roots, rice straws incorporated in soil, plant residues (mixture of weeds, rice litters, rice roots, and rice stubbles) in soil, and composing rice straw. The number of bands of DGGE profiles ranged from 12 to 26 with the highest numbers in rice roots and rice straws incorporated in soil. However, the diversity indices based on both the numbers and intensity of bands indicated that the community of the plowed soil layer was the most diverse, even, and stable. Sequencing of the main DGGE bands showed the presence of Methanomicrobiales, Methanosarcinales, Methanobacteriaceae, and Methanocellales. The plowed soil layer included all phylogenetic groups of the methanogenic archaea of the other studied habitats, with prevalence of the members of Methanomicrobiales and Methanocellales. The phylogenetic diversity was compared with that of paddy soils collected in Italy, China, and the Philippines and that of 12 anaerobic environments (fen, waste, coast, permafrost, natural gas field, bovine rumen, riparian soil, termite, ciliate endosymboints, lake sediment, landfill, and seep rumen). The phylogenetic diversity was more similar among paddy soils than with the other anaerobic environments. Probably, the methanogenic archaeal communities of the paddy field soils were characterized by indigenous members and some of the members of the community of the plowed soil layer colonized rice roots, rice straws, and plant residues.     

How different or similar are nematode communities between a paddy and an upland rice fields across a flooding-drainage cycle?

The paddy field is being recognised as a biodiversity hotspot fostering a variety of organisms. However, there are few studies on the ecology of paddy field nematodes. We characterised nematode communities in rice paddy fields by comparing them with upland fields of rice or soybean. We examined nematode communities of the top (0-15 mm) and second (15-50 mm) soil layers before flooding (March or April), during flooding (June or July) and during the draining period (October) 2007-2009. We found that the nematode community in the paddy was different than that in the upland fields during all periods. Rhabdolaimus, Tobrilus, Mesodorylaims and Monhysteridae characterised the top of the paddy and Hirschmanniella characterised the second layer of the paddy. Total nematode density was generally lower in the paddy than in the upland field. However, the density in the paddy top layer increased with time from the flooding period to the draining period, during which time it was about the same as (or even greater than) the peak density in the upland fields. The density in the second layer of the paddy remained lower than that in the top layer of the paddy throughout the time course. Community diversity values were generally greater in the paddy top layer than in the paddy second layer across the six sampling periods, but periodic differences between the paddy and upland fields or between soil layers were not significant. During the flooding period, the F/(F + B) (13-37) and Enrichment Index (17-38) values were lower in the paddy than in the upland fields (32-47, 37-74, respectively) to reflect that bacteria dominate over fungi with slow decomposition due to anaerobic conditions in the flooded paddy field. In addition, particularly in the top layer, the Maturity (2.0-2.4) and Structure Index (23-72) values were greater in the paddy than in the upland fields (1.7-2.1, 9-15, respectively), indicating a well-developed ecosystem under water. These unique nematode communities persisted during the draining period, but there was a rapid increase in opportunistic bacterivores, which increased the EI values. We suggest that bactivorous nematodes in the families Cephalobidae and Chronogasteridae, herbivores in the genus Hirschmanniella, and fungivores in the genus Filenchus may be specific to paddy field soil rather than to pond and lake sediments.     

Prey-predator dynamics in communities of culturable soil bacteria and protozoa: differential effects of mercury

We investigated whether the prey-predator dynamics of bacteria and protozoa were affected by inorganic mercury at concentrations of 0, 3.5 and 15 mg Hg(II) kg soil⁻¹. The amount of bioavailable Hg was estimated using a biosensor-assay based on the mer-lux gene fusion. The numbers of bacterial CFUs on the general medium 1/100 tryptic soy agar (TSA) were significantly decreased when the soil had been amended with Hg. In contrast, no effect was seen on the number of CFUs on the Pseudomonas-specific medium Gould's S1 agar. Protozoan numbers estimated by the most probable number (MPN) method with 1/100 TSB as growth medium were also negatively affected by Hg. The different fractions of protozoa were affected to different degrees suggesting that amoebae were less sensitive than slow-growing flagellates, which again were less sensitive than the fast-growing flagellates. In contrast, Hg did not induce any detectable changes in the diversity of flagellate morphotypes. In the treatment with 15 mg Hg kg⁻¹ a transiently increased number of bacteria was seen at day 6 probably concomitant with a decrease in the numbers of protozoa. This might indicate that Hg affected the prey-predator dynamics in communities of culturable bacteria and protozoa in soil. Furthermore, we showed that the number of Pseudomonas spp. was not affected by Hg whereas the number of bacteria growing on a general medium was.     

Tillage and residue management practices affect soil biological indicators in a rice–wheat cropping system in north-western India

Agricultural productivity relies on a wide range of ecosystem services provided by the soil biota. Sustainable management practices, such as tillage and residue management, can influence structure and function of the soil microbiota, with direct consequences for the associated ecosystem services. Although there is increasing evidence that different tillage regimes alter the soil biological indices, we only have a limited understanding of their temporal changes in a rice (Oryza sativa L.)–wheat (Triticum aestivum L.) cropping system. We evaluated the effects of combinations of tillage, crop residue management and green manuring on soil biological indicators after 5 years of the practising rice–wheat system (RWS). Four main plot treatments in rice included the following: (a) PTR_W0, puddled transplanted rice with no wheat straw retained; (b) PTR_W25, puddled transplanted rice with 25% anchored wheat stubbles retained; (c) PTR_W0 + Sesbania aculeate L. green manure (GM); and (d) PTR_W25+GM, puddled transplanted rice with 25% anchored wheat stubbles retained+ GM. There were three subplot treatments in the subsequent wheat crop: (a) CTW_R0, conventional tillage wheat with rice residue removed; (b) ZTW_R0, zero tillage wheat with rice residue removed; and (c) ZTW_R100, ZTW with 100% rice residue retained as mulch. The PTR_W25+GM treatment, followed by ZTW_R100, significantly increased soil microbial biomass carbon, basal soil respiration, microbial quotient and mineralization quotient measured during wheat-growing season. These biological indicators were higher at vigorous vegetative wheat growth stage than at flowering stage and decreased at maturity. The principal component analysis of the assayed variables showed that all the variables significantly contributed to the variability in parameters examined and were more related to maximum tillering stage of wheat growth than to maturity or at sowing of wheat. Three highly effective biological indicators were microbial biomass carbon, microbial quotient and mineralization quotient, which responded significantly to changes in tillage and residue management practices in the RWS. We conclude that crop residues and green manure have significant to improve soil biochemical processes by improving soil organic carbon and soil biological indicators in rice–wheat cropping system.     

不同施肥处理稻田系统磷素输移特征研究

磷是水体富营养化限制性元素,近年来由于磷肥的过量施用,农田迁移的磷素已成为水体磷素的主要来源。本研究通过野外测坑定位试验,研究有机肥处理(OT)、混施肥处理(MT)和化肥处理(CT)3种施肥处理下,稻田中磷素的迁移流失特征及这3种处理对水稻产量和磷素利用率的影响,以探求稻田系统的最佳施磷方式。结果表明,CT、MT和OT 3种施肥方式的磷径流流失负荷分别为0.56 kg(P)·hm-2、1.13 kg(P)·hm-2和4.20 kg(P)·hm-2,渗漏流失负荷分别为0.42 kg(P)·hm-2、0.44 kg(P)·hm-2和0.45 kg(P)·hm-2;磷的径流流失占流失总量的56.86%~90.38%,是水稻田磷素流失的主要途径。磷的径流流失主要受施肥和降雨的影响,50%左右磷的流失发生在第1次径流过程;磷素渗漏流失特征不受施磷处理的影响,80%以上的流失发生在施肥后的前30 d。在磷素流失形态上,坑面水、渗漏水和径流水中磷素的主要形态均为可溶性磷;在土壤方面,MT处理和OT处理能保证土壤磷营养,CT处理的土壤有效磷和有机质含量则显著下降。3种施肥处理的水稻产量显著高于空白对照,且MT最高,为6 728.84 kg·hm-2;磷肥利用率CT和MT处理显著高于OT,CT和MT间差异不显著。综合比较,混施肥处理在磷素流失、土壤养分利用和水稻产量等方面更符合我国生态农业发展的要求。     

Community structure of methanogenic archaea in paddy field soil under double cropping (rice-wheat)

Community structure of methanogenic archaea in paddy field soil under double cropping (rice [Oryza sativa L.] and wheat [Triticum aestivum L.]) was studied by the denaturing gradient gel electrophoresis (DGGE) method. Soil samples under flooded and upland conditions were collected 7 and 6 times, respectively, from two paddy fields throughout a year, and two primer sets, 0357F-GC/0691R and newly designed 1106F-GC/1378R, were used for DGGE analysis. The 25 and 29 different bands were observed on the DGGE gels with the primers 0357F-GC/0691R and 1106F-GC/1378R, respectively. DGGE band patterns of the methanogenic archaeal community were stable throughout a year including the cultivation periods of rice under flooded conditions and of wheat under upland conditions. Cluster analysis and principal component analysis suggested that the difference in the soil type (sampling region) largely influenced the community structures of methanogenic archaea in paddy field soil, while the effects of sampling period and different fertilizer treatments on them were small. Most of the sequences obtained from the DGGE bands were closely related to Methanomicrobiales, Methanosarcinaceae, Methanosaetaceae and Rice cluster-I.