15N2 incorporation by rhizosphere soil influence of rice variety,organic matter and combined nitrogen

Summary Heterotrophic nitrogen fixation by rhizosphere soil samples from 20 rice cultivars grown under uniform field conditions was estimated employing¹⁵N-tracer technique. Rhizosphere soil samples from different rice cultivars showed striking differences with regard to their ability to incorporate¹⁵N₂. Rhizosphere samples from rice straw-amended (3 and 6 tons/ha) soil exhibited more pronounced nitrogen-fixing activity than the samples from unamended soil; while the activity of the rhizosphere samples from soils receiving combined nitrogen (40 and 80 kg N/ha) was relatively low. However, the inhibitory effect of combined nitrogen was not expressed in the presence of rice straw at 6 tons/ha. Results suggest that plant variety, application of combined nitrogen and organic matter influence the rhizosphere nitrogen fixation.     

Effect of Rice Plants on Nitrogenase Activity of Flooded Soils

In samples of flooded soil containing blue-green algae (cyanobacteria), the presence of rice plants did not influence the nitrogenase activity of the algae. Nitrogenase activity of heterotrophic bacteria was enhanced by the presence of rice plants, but this activity was not affected by changes in plant density. The rate of nitrogen fixation in the rhizosphere, however, varied significantly among the 16 rice varieties tested. A simple method was devised to test the nitrogen-fixing activity in the root zone of rice varieties, and data were obtained showing marked differences in the activities of the 16 varieties. In tests of two varieties with dissimilar rates of nitrogen fixation in their rhizospheres, the variety which had the greater root weight and lesser shoot weight and which supported greater methane formation had the greater nitrogenase activity.     

氮肥运筹对免耕水稻根系生长、根际土壤特性及产量的影响

为探索氮肥运筹对免耕条件下水稻根系生长以及对根际土壤特性、产量的影响,以金优253为材料进行试验。结果表明：平衡施肥显著提高单株根系干重、根长、单株生物量、根半径、单株根表面积、根长密度及根系活力,实收单产高于重穗肥和重基肥处理,且与重基肥差异达95％的显著水平,主要是有效穗数、结实率的增加。平衡施肥显著提高0~10 cm土层的0~2 mm根际土壤有机质、碱解氮含量及脲酶、蔗糖酶活性。因此平衡施肥能明显促进免耕水稻根系生长和有效穗数的增加,对提高水稻产量具有促进作用。     

Nitrogen transformations and balance in a constructed wetland for nutrient-polluted river water treatment using forage rice in Japan

A pilot-scale surface-flow wetland planted with a new rice variety (Oryza sativa ’Kusahonami’) developed for livestock feed was constructed for treating nutrient-polluted river water. To calculate the balance between nitrogen removal and rice plant uptake of nitrogen, nitrogen removal from river water and nitrogen interactions among plants, soil water, and soil were investigated for this constructed wetland over two growing seasons in 2004 and 2005. The constructed wetland removed 33% of the total nitrogen entering with the river water. Rice plants were found to constitute the major nitrogen storage, with plant uptake being the major removal mechanism. The total inorganic nitrogen concentration in the rhizosphere changed seasonally because of plant uptake. Most nitrogen taken up by rice plants was contained in the aboveground biomass, with the mean amount being 34.0 g N m⁻². However, the nitrogen balance calculation suggested that rice plants uptake some nitrogen from soil, decreasing the available nitrogen in the soil of the lined impermeable wetland.     

Physiological and genetic characterization of rice nitrogen fixer PGPR isolated from rhizosphere soils of different crops

Aims

We aimed to identify plant growth-promoting rhizobacteria that could be used to develop a biofertilizer for rice.

Methods

To obtain plant growth-promoting rhizobacteria, rhizosphere soils from different crops (rice, wheat, oats, crabgrass, maize, ryegrass, and sweet potato) were inoculated to rice plants. In total, 166 different bacteria were isolated and their plant growth-promoting traits were evaluated in terms of colony morphology, indole-3-acetic acid production, acetylene reduction activity, and phosphate solubilization activity. Moreover, genetic analysis was carried out to evaluate their phylogenetic relationships based on 16S rRNA sequence data.

Results

Strains of Bacillus altitudinis, Pseudomonas monteilii, and Pseudomonas mandelii formed associations with rice plants and fixed nitrogen. A strain of Rhizobium daejeonense showed nitrogen fixation activity in an in vitro assay and in vivo. Strains of B. altitudinis and R. daejeonense derived from rice rhizosphere soil, strains of P. monteilii and Enterobacter cloacae derived from wheat rhizosphere soil, and a strain of Bacillus pumilus derived from maize rhizosphere soil significantly promoted rice plant growth.

Conclusions

These methods are effective to identify candidate species that could be developed as biofertilizers for target crops.     

稻田水氮氧环境因子对水稻生长发育、光合作用和氮利用的调控研究进展

水分和氮素是影响水稻生长发育的两个重要环境因子。适宜的水氮耦合模式可通过“以水调氮、以水控氧”调控稻田根际氮形态和溶氧量等环境因子,促进良好根系形态构建,提高叶片光合速率和光合产物“源-库”分配平衡,提高水稻群体质量和产量形成。同时,稻田水氮氧环境因子驱动的微生物调控机制在水稻-土壤系统氮高效利用方面也发挥重要作用。本文重点阐述了水氮耦合下水分、氮形态和溶氧量对水稻生长发育、光合作用、碳氮代谢、稻田氮转化过程及其微生物调控机制等方面的研究进展,展望并提出了未来亟待加强的研究方向:1)开展水氮耦合下水稻根际溶氧量时空动态分布特征及氧环境调控关键因子研究;2)明确不同基因型水稻根源信号增氧响应特征及其对水稻生长发育的影响调控机制;3)阐明根际氧环境驱动的关键微生物过程对稻田氮转化和氮素利用的影响。     

水分管理调控水稻氮素利用研究进展

水、氮是调控水稻生长发育的两个重要环境因子。通过"以水调氧"增加根际溶氧量(如干湿交替、好氧栽培等)能够提升土壤硝化势和氧化还原电位,刺激土壤氮的矿化作用,使水稻处于NH+4与NO-3混合营养中,并能通过诱导水稻的生理特性及改善根系的吸收功能增强其抗旱性能,提高水稻产量及氮素利用率。光合作用是形成干物质的主要途径,土壤氮水平、氮形态与水稻光合速率紧密相关,提高叶片光合速率将有助于提高水稻的氮素利用率和产量。从稻田水分管理对土壤氮素形态特征、水稻氮吸收利用、光合速率及氮环境效应的影响等方面综述了国内外相关研究进展,并指出进一步的研究方向。     

Influence of some environmental factors on nitrogen fixation in the rhizosphere of rice

Summary Nitrogen fixers make up a large percentage of the total microflora in the rhizosphere of lowland rice. There are more aerobic nitrogen fixers than there are anaerobic ones. When soil crumbs from the root zone were placed on a nitrogen free agar medium and inoculated at 0, 5, 10, and 21 percent oxygen concentration, colonies of aerobic nitrogen fixers reached their greatest diameter at 5 and 10 percent oxygen. In acetylene reduction assays rice plants grown in paddy fields and in solution culture were tested for the nitrogenase activities of their roots at different oxygen tensions. Nitrogenase activity was highest at 3 percent oxygen, lower at 0 percent, and far lower at 21 percent. When rice was grown in solution culture the redox potential of the nutrient solution strongly influenced nitrogenase activity. With declining redox potential, nitrogenase activity increased to a maximum value but dropped sharply as redox potential further decreased. Ten ppm of combined nitrogen as urea depressed nitrogenase activity on excised roots. Combined nitrogen applied to one part of the root system affected, to some extent, nitrogen fixation on other roots kept in a solution without nitrogen. Nitrogenase activity in a fertility trial with lowland rice, examined at several dates, showed no inhibitory effect of fertilizer nitrogen, however, presumably because the nitrogen concentration in the soil solution rapidly decreased. Instead, an overall stimulating effect of nitrogen dressing was noticeable. Diurnal fluctuations of nitrogenase activity in the rhizosphere, with a peak in the afternoon and low fixation rates after low solar radiation, suggest a photosynthetic effect on nitrogen fixation. re]19751208     

Root-induced changes in radial oxygen loss, rhizosphere oxygen profile, and nitrification of two rice cultivars in Chinese red soil regions

Aims

To evaluate the external and internal morphological differences of roots that might influence rice root radial oxygen loss (ROL) and the corresponding rhizosphere nitrification activity, growth characteristics and nitrogen nutrition of rice.

Methods

The root ROL and rhizosphere oxygen profile were determined using a miniaturised Clark-type oxygen microelectrode system, and the rhizosphere nitrification activity was studied with a short-term nitrification activity assay.

Results

The rice biomass, nitrogen accumulation and nitrogen use efficiency (NUE) of ZH (high yield) were significantly higher than those of HS (low yield). The root biomass, number, diameter and porosity of ZH were also much greater than those of HS. The inner and surface oxygen concentrations of the root of ZH were significantly higher than those of HS. The order of paddy soil oxygen penetration depth was ZH?>?HS?>?CK, and the order of the oxygen concentrations detected in the water layer and rhizosphere soil was the same. The rhizosphere nitrification activity and nitrate concentration of ZH were significantly higher than those of HS.

Conclusions

More porous and thicker roots improved the individual root ROL, and more adventitious root numbers enhanced the entire plant ROL and correspondingly improved the rhizosphere nitrification activity, which might influence the growth and nitrogen nutrition of rice.     

Application of the fluorescent antibody technique to the study of an isolate of Beijerinckia in soil.

Fluorescent antibody was prepared against a temperate-soil isolate of Beijerinckia obtained from a rhizosphere of rice growing in Camargue (France). The antibody did not cross-react with any of 6 species of Azotobacter, 4 species of Beijerinckia, or 44 unidentified soil bacteria isolated from a spectrum of rhizospheres, but strongly stained the homologous Beijerinckia isolate. The isolate grew well in autoclave Camargue soil, but increased in numbers only slightly in nonsterile soil during 9 days. Preliminary examination of rice plants grown in the laboratory in soil from which the Beijerinckia was originally isolated did not show detectable Beijerinckia in the rhizosphere. The fluorescent antibody was sufficiently sensitive and specific to permit more extensive study of Beijerinckia in relation to nitrogen fixation in the rhizospher of rice.     

耕作播种方式对稻茬小麦根系发育、土壤水分和硝态氮含量的影响

2016—2018年,以深旋耕播种模式为对照,研究了浅旋耕播种和免耕带旋播种模式对稻茬小麦根系发育、土壤水分和硝态氮含量的影响。结果表明: 孕穗期以前免耕带旋播种和浅旋耕播种处理耕层土壤含水量高于深旋耕播种处理,而硝态氮含量低于深旋耕播种处理。拔节和开花期根重密度和根表面积密度处理间差异不显著。2016—2017年,3种耕播方式的产量和地上部分氮吸收量无显著差异;2017—2018年,免耕带旋播种和浅旋耕播种处理的产量较深旋耕播种分别增加10.9%和10.5%,地上部分氮吸收量分别增加17.5%和12.0%。与深旋耕播种和浅旋耕播种处理相比,免耕带旋播种处理播种效率高、断垄率低。综上,免耕带旋播种处理可提高稻茬小麦的播种质量,增强土壤保墒能力,降低氮淋溶风险,促进产量和环境效益的协同提升。     

Algal and bacterial non-symbiotic nitrogen fixation in paddy soils

Summary The nitrogen fixing activity of three Ivory Coast soils was tested in the laboratory by the acetylene reduction assay and the Kjeldahl method. Nitrogen fixation due to algae was estimated to be of the order of 4 to 8 (acetylene method) and 7 μg N per g soil per day (Kjeldahl method). Nitrogen fixation due to bacterial activity in the rice rhizosphere was estimated to be of the order of 2 to 5 (acetylene method) and 1 to 3 μg N per g soil per day (Kjeldahl method). These results emphasize the importance of the bacterial nitrogen fixation in the rhizosphere which had been hitherto overlooked. Comparison of acetylene method and Kjeldahl method results shows discrepancies the origin of which has been discussed. Time course of acetylene reduction by rhizosphere soils exhibits a lag phase which may be attributed to Postgate's switch off — switch on process.     

翅荚木人工林不同径阶间细根主要功能性状与根际土壤养分的关系

细根能敏感地感知土壤环境变化,对植物生长发育具有重要影响.以6年生翅荚木人工林为对象,对其不同径阶的细根主要功能性状与根际土壤养分特征及两者间关系进行分析.结果表明:细根生物量、根长密度与根体积密度均随径阶增加而增加,比根长与比根面积则随径阶增加呈先升高再下降后升高的趋势,根组织密度则与径阶大小不相关.不同径阶翅荚木根际土壤的pH值及含水率、全碳、全磷、铵态氮、硝态氮和总有效氮含量均存在显著差异,大径阶林木的根际土壤全碳、全氮、硝态氮、总有效氮含量相对较高,小径阶林木的根际土壤含水率、土壤全磷、铵态氮含量相对较高.土壤全氮、全碳、硝态氮和总有效氮含量与林木细根的生物量、根长密度、根体积密度呈显著正相关;土壤全磷与林木细根的根组织密度呈显著正相关,与比根长、比根面积呈显著负相关;土壤含水率与林木细根的生物量和根体积密度均呈显著正相关;根际土壤pH和林木细根的比根长、比根面积呈显著正相关,与根组织密度则呈显著负相关.研究结果可为翅荚木优良种质资源选育提供科学依据.     

Selection of mixed inoculants exhibiting growth-promoting activity in rice plants from undefined consortia obtained by continuous enrichment

Aims

The main objective of this study was to test the hypothesis that isolating and characterizing bacterial suspensions (undefined consortia) from the root/rhizosphere of rice will contribute to the selection of mixtures of rhizobacteria with better growth-promoting activity of rice plants.

Methods

Bacterial consortia were obtained from roots/rhizosphere soil samples of rice plants grown under upland and irrigated production systems. Those undefined consortia were subjected to five consecutive passes every 7 days in NFb (N-free broth) semisolid medium. Thereafter, strains of each growth-promoting consortia were isolated by plating on three different culture media. Then, undefined consortia, as well as mix and single bacterial strains, were characterized in terms of indoleacetic acid production, nitrogen fixation capacity, and growth promotion of rice plants.

Results

Of the 72 consortia analyzed, 41.7 % and 50.0 % increased nitrogenase activity and the production of indolic compounds, respectively, after 5 continuous passes in NFB medium. Three undefined consortia, 11 single strains and 5 strain mixtures, exhibited plant growth promotion in rice plants under greenhouse conditions.

Conclusions

Continuous enrichment in Nfb medium of undefined consortia from root/rhizosphere soil is a good strategy for the selection of plant growth-promoting bacteria for rice plants.     

Nitrogen management in direct sown intermediate deep water rice (15–50 cm)

Summary Field experiments were conducted to assess the efficiency of different forms and methods of urea application for direct sown rice under intermediate deep water situation (15–50 cm). Basal application of N @ 40 kg/ha in the form of prilled urea (PU); urea super granules (USG); and sulphur coated urea (SCU); were tested with a view to improving the early vigour, tillering and grain yield. Deep placement of prilled urea behind a plough; USG placement between rows in moist soil; and also in shallow water were found to increase the recovery of nitrogen. There was no deleterious effect on stand establishment when the seed and fertilizer @ 40, 8.6 and 16.6 kg N, P and K per hectare respectively were drilled in the same furrow in moist soil. Split application of either conditioned urea or application as a foliar spray did not prove effective in an intermediate deep water rice situation.     

稻作系统有机肥替代部分化肥的土壤氮循环特征及增产机制

采用有机肥替代部分化肥是实现化肥使用零增长和作物稳产增产的重要途径。基于近年来的研究进展,探讨了稻作系统有机肥替代部分化肥对水稻产量、氮素利用效率、土壤氮库组分和微生物固氮、氨化、硝化和反硝化等氮循环关键过程的影响。同时,就单施化肥与有机肥替代部分化肥的氮素循环特征进行了比较。有机肥替代部分化肥通过改变稻田土壤氮素循环多个环节(增强氨化过程、协调硝化和反硝化过程、降低氨挥发和减少氮素损失等),改善土壤氮素供给状态(提高小分子有机氮供给、协调无机氮组分与比例、提高土壤微生物量氮和总氮固持),进而促进水稻氮素吸收并协调植株氮素分配过程,最终实现水稻稳产增产。     

Contribution of nitrogen fixation to first year Miscanthus × giganteus

Many characteristics make Miscanthus × giganteus an appealing bioenergy feedstock in temperate North America, but the degree to which this plant species interacts with nitrogen‐fixing bacteria remains understudied. Demonstration of associative nitrogen fixation in Miscanthus would support management with minimal fertilizer inputs that is demanded of long‐term biofuel sustainability. As a first step, we investigate the role of biological nitrogen fixation in nutrition of immature Miscanthus and temporal dynamics of plant‐associated nitrogen fixers. The contribution of biological nitrogen fixation to plant nitrogen acquisition in first year Miscanthus × giganteus was estimated using a yield‐dependent ¹⁵N isotope dilution model. Temporal changes in plant‐associated diazotroph relative abundance and community composition were analyzed with quantitative PCR and terminal restriction fragment length polymorphism of the nifH gene in rhizome and rhizosphere DNA extracts. We estimate 16% of new plant nitrogen was derived by nitrogen fixation during the growing season, despite non‐limiting soil nitrogen. Diazotroph communities from rhizome and rhizosphere changed with plant development and endophytic nitrogen fixers had significantly higher relative abundance and altered community composition at sampling dates in July and August. This study provides evidence for a small, but measurable, benefit of associative nitrogen fixation to first year Miscanthus × giganteus that underscores the potential and need for selection of breeding lines that maximize this trait.     

Deep placement: A method of nitrogen fertilizer application compatible with algal nitrogen fixation in wetland rice soils

Summary The effect of different methods of nitrogen fertilizer application on the algal flora and biological nitrogen fixation (Acetylene-reducing activity) in a wetland rice soil was studied in pot and field experiments. Broadcast application of urea inhibited nitrogen fixation and favored the growth of green algae. In contrast, deep placement of urea supergranules (1–2 g urea granules) did not suppress the growth of N₂-fixing blue-green algae and permitted acetylene-reducing activity on the soil surface to continue virtually uninhibited.     

Field response of rice paddy crop to Azospirillum inoculation: physiology of rhizosphere bacterial communities and the genetic diversity of endophytic bacteria in different parts of the plants

The response of rice plants to the application of inoculant containing two Azospirillum brasilense strains was studied under field conditions. The experiment was performed as three treatments with four replicates in randomized complete blocks arranged as plots of 60 m² in an area on a Vertic Argiudol soil type in the province of Entre Ríos, Argentina. The bacterial rhizosphere community and also the diazotrophic isolates obtained from control and inoculated rice plants were analyzed in relation to their physiology and biological nitrogen fixation (BNF). The MPN of diazotrophs in the rhizosphere varied during the ontogenic cycle. The patterns of distribution of the microbial physiological activities obtained by principal component analysis of community-level physiological profiles (CLPP) showed differences in the utilization of carbon sources by the rhizosphere communities among treatments. Although the analyses of DGGE 16S and nifH profiles have not indicated that the inoculation influenced the genetic diversity of bacterial communities among treatments, they revealed that the banding profiles were altered in different parts of the rice plant by each Azospirillum inoculation treatment. These observations suggest that physiological responses of plant tissues to the inoculation may have occurred. According to agronomic parameters of each treatment, the Azospirillum inoculation increased aerial biomass at the tillering and grain-filling stages. Although the N content accumulated in rice plants increased by 16 and 50 kg ha^?1, the BNF contribution could not be estimated under our experimental conditions by the ¹⁵N balance technique. Based on this field inoculation experiment to rice plants, it is noteworthy that our data suggest that due to Azospirillum inoculation the increase of total N accumulated in rice plants could be a tool to help farmers to improve production and maintain high input of plant residues, providing more organic matter to the soil and guaranteeing sustainability of the system.     

红壤区不同产量籼稻品种苗期根表和根际土壤硝化特征

以不同产量籼稻品种中旱22(ZH,高产品种)和禾盛10号(HS,低产品种)为材料,采用根际培养箱(三室)--速冻切片技术研究了红壤水稻土种植条件下,水稻苗期生长、氮素积累和氮素利用率(NUE)、根孔隙度(POR)、根际土壤矿质态氮含量和硝化强度.结果表明,ZH苗期生长、氮素吸收及NUE均显著优于HS,且ZH单株不定根数量及根系通气组织发育程度(用POR表示)均显著高于HS.ZH根际和土体土壤中铵(NH+4)含量始终低于HS,而硝(NO-3)含量则始终高于HS,但二者根表土壤NH+4和NO-3含量均无显著差异.ZH和HS硝化强度最大发生部位均是在距根表2 mm的根际土壤,分别为:0.48 μmol kg-1h-1和0.31 μmol kg-1h-1.随着距根表越远,硝化强度就越弱,直至距根表10～20 mm处土壤硝化强度就接近于土体土壤.ZH根际土壤硝化强度始终显著高于HS,但二者根表和土体土壤硝化强度均无显著差异.与不种水稻的CK相比,根际土壤硝化强度提高了约2～3.5倍.尽管红壤水稻土硝化作用很弱,但红壤区水稻根际硝化作用与水稻苗期生长和氮素营养密切相关.