Enhancing saline stress tolerance in soybean seedlings through optimal NH4+/NO3- ratios: a coordinated regulation of ions, hormones, and antioxidant potential.

BACKGROUND: Nitrogen (N) availability is crucial in regulating plants' abiotic stress resistance, particularly at the seedling stage. Nevertheless, plant responses to N under salinity conditions may vary depending on the soil's NH4+ to NO3- ratio. METHODS: In this study, we investigated the effects of different NH4+:NO3- ratios (100/0, 0/100, 25/75, 50/50, and 75/25) on the growth and physio-biochemical responses of soybean seedlings grown under controlled and saline stress conditions (0-, 50-, and 100-mM L- 1 NaCl and Na2SO4, at a 1:1 molar ratio). RESULTS: We observed that shoot length, root length, and leaf-stem-root dry weight decreased significantly with increased saline stress levels compared to control. Moreover, there was a significant accumulation of Na+, Cl-, hydrogen peroxide (H2O2), and malondialdehyde (MDA) but impaired ascorbate-glutathione pools (AsA-GSH). They also displayed lower photosynthetic pigments (chlorophyll-a and chlorophyll-b), K+ ion, K+/Na+ ratio, and weakened O2•--H2O2-scavenging enzymes such as superoxide dismutase, catalase, peroxidase, monodehydroascorbate reductase, glutathione reductase under both saline stress levels, while reduced ascorbate peroxidase, and dehydroascorbate reductase under 100-mM stress, demonstrating their sensitivity to a saline environment. Moreover, the concentrations of proline, glycine betaine, total phenolic, flavonoids, and abscisic acid increased under both stresses compared to the control. They also exhibited lower indole acetic acid, gibberellic acid, cytokinins, and zeatine riboside, which may account for their reduced biomass. However, NH4+:NO3- ratios caused a differential response to alleviate saline stress toxicity. Soybean seedlings supplemented with optimal ratios of NH4+:NO3- (T3 = 25:75 and T = 4 50:50) displayed lower Na+ and Cl- and ABA but improved K+ and K+/Na+, pigments, growth hormones, and biomass compared to higher NH4+:NO3- ratios. They also exhibited higher O2•--H2O2-scavenging enzymes and optimized H2O2, MDA, and AsA-GSH pools status in favor of the higher biomass of seedlings. CONCLUSIONS: In summary, the NH4+ and NO3- ratios followed the order of 50:50 > 25:75 > 0:100 > 75:25 > 100:0 for regulating the morpho-physio-biochemical responses in seedlings under SS conditions. Accordingly, we suggest that applying optimal ratios of NH4+ and NO3- (25/75 and 50:50) can improve the resistance of soybean seedlings grown in saline conditions.

Peculiarity of the early metabolomic response in tomato after urea, ammonium or nitrate supply.

Nitrogen (N) is the nutrient most applied in agriculture as fertilizer (as nitrate, Nit; ammonium, A; and/or urea, U, forms) and its availability strongly constrains the crop growth and yield. To investigate the early response (24 h) of N-deficient tomato plants to these three N forms, a physiological and molecular study was performed. In comparison to N-deficient plants, significant changes in the transcriptional, metabolomic and ionomic profiles were observed. As a probable consequence of N mobility in plants, a wide metabolic modulation occurred in old leaves rather than in young leaves. The metabolic profile of U and A-treated plants was more similar than Nit-treated plant profile, which in turn presented the lowest metabolic modulation with respect to N-deficient condition. Urea and A forms induced some changes at the biosynthesis of secondary metabolites, amino acids and phytohormones. Interestingly, a specific up-regulation by U and down-regulation by A of carbon synthesis occurred in roots. Along with the gene expression, data suggest that the specific N form influences the activation of metabolic pathways for its assimilation (cytosolic GS/AS and/or plastidial GS/GOGAT cycle). Urea induced an up-concentration of Cu and Mn in leaves and Zn in whole plant. This study highlights a metabolic reprogramming depending on the N form applied, and it also provide evidence of a direct relationship between urea nutrition and Zn concentration. The understanding of the metabolic pathways activated by the different N forms represents a milestone in improving the efficiency of urea fertilization in crops.

Elevated CO2 and ammonium nitrogen promoted the plasticity of two maple in great lakes region by adjusting photosynthetic adaptation.

Introduction: Climate change-related CO2 increases and different forms of nitrogen deposition are thought to affect the performance of plants, but their interactions have been poorly studied. Methods: This study investigated the responses of photosynthesis and growth in two invasive maple species, amur maple (Acer ginnala Maxim.) and boxelder maple (Acer negundo L.), to elevated CO2 (400 µmol mol-1 (aCO2) vs. 800 µmol mol-1 (eCO2) and different forms of nitrogen fertilization (100% nitrate, 100% ammonium, and an equal mix of the two) with pot experiment under controlled conditions. Results and discussion: The results showed that eCO2 significantly promoted photosynthesis, biomass, and stomatal conductance in both species. The biochemical limitation of photosynthesis was switched to RuBP regeneration (related to Jmax) under eCO2 from the Rubisco carboxylation limitation (related to Vcmax) under aCO2. Both species maximized carbon gain by lower specific leaf area and higher N concentration than control treatment, indicating robust morphological plasticity. Ammonium was not conducive to growth under aCO2, but it significantly promoted biomass and photosynthesis under eCO2. When nitrate was the sole nitrogen source, eCO2 significantly reduced N assimilation and growth. The total leaf N per tree was significantly higher in boxelder maple than in amur maple, while the carbon and nitrogen ratio was significantly lower in boxelder maple than in amur maple, suggesting that boxelder maple leaf litter may be more favorable for faster nutrient cycling. The results suggest that increases in ammonium under future elevated CO2 will enhance the plasticity and adaptation of the two maple species.

Nitrate reduces copper toxicity by preventing oxidative stress and inhibiting copper translocation from roots to shoots in Liriodendron Chinense.

Nitrogen forms can affect metal accumulation in plants and tolerance to metals, but a few published studies on the effects on Cu toxicity and Cu accumulation in plants are scarce. Thus, the objective of this study was to evaluate the responses of Liriodendron chinense to different nitrogen forms, by the oxidative stress, antioxidant enzymes system, GSH-AsA cycle, Cu uptake, translocation, and accumulation under Cu stress. We found that Cu-induced growth inhibiting was alleviated by added exclusive NO3--N. Adding N as NH4+-N with or without NO3--N was aggravated as evidenced by significantly elevated malonaldehyde (MDA) and hydrogen peroxide (H2O2) compared to N-Null. Cu exposure and adding NH4+-N inhibited superoxide dismutase activity, but remarkably stimulated the activities of catalase and peroxidase, the efficiency of glutathione-ascorbate (GSH-AsA) cycle, and the activity of glutathione reductase and nitrate reductase, with respect to the control. However, adding exclusive NO3--N progressively restored the alteration of antioxidant to prevent Cu-induced oxidative stress. Additionally, adding exclusive NO3--N significantly promoted the Cu uptake and accumulation in roots, but reduced Cu concentration in leaves, accompanied by the inhibited Cu translocation factor from roots to shoots by 36.7%, when compared with N-Null. Overall, adding NO3--N alleviated its Cu toxicity by preventing Cu-induced oxidative stress and inhibiting Cu translocation from roots to shoots, which provides an effective strategy for phytostabilization in Cu-contaminated lands.

Effects of hypobaria, hyperoxia, and nitrogen form on the growth and nutritional quality of lettuce.

The objectives of this research were to investigate the impact of hypobaria, hyperoxia, and nitrogen form on the growth and nutritional quality of plants. Pre-culture 20-day-old lettuce (Lactuca sativa L. var. Rome) seedlings grew for 25 days under three levels of total atmospheric pressure (101, 54, and 30 kPa), two levels of oxygen partial pressure (21 and 28 kPa), and two forms of nitrogen (NO3N and NH4N). The ratios of NO3N to NH4N included 3: 1, 4: 0, 2: 2, and 0: 4. The nitrogen quantity included two levels, i.e. N1, 0.1 g N kg-1 dry matrix and N2, 0.2 g N kg-1 dry matrix. The growth status of lettuce plants in different treatments differentiated markedly. Regardless of the nitrogen factor, the growth status of lettuce plants treated with total atmospheric pressure/oxygen partial pressure at 54/21 was equivalent to the treatment of 101/21. Under the hypobaric condition (54 kPa), compared with 21 kPa oxygen partial pressure, hyperoxia (28 kPa) significantly inhibited the growth of lettuce plants and the biomass (fresh weight) decreased by 60.9%-69.9% compared with that under 101/21 treatment. At the N1 level, the sequence of the biomass of lettuce plants supplied with different ratios of NO3N to NH4N was 3: 1 > 4: 0 > 2: 2 > 0: 4, and there were higher concentrations of chlorophyll and carotenoid of lettuce plants supplied with the higher ratio of NO3 to NH4. At the N2 level, the effects of different ratios of NO3N to NH4N on lettuce plants were similar to those at the N1 level. The high nitrogen (N2) promoted the growth of lettuce plants such as 54/21/N2 treatments. Both form and nitrogen level did not affect the stress resistance of lettuce plants. Hypobaria (54 kPa) increased the contents of N, P, and K and hyperoxia (28 kPa) decreased the content of organic carbon in lettuce plants. The high nitrogen (N2) improved the content of total N and the N uptake. The ratios of NO3N to NH4N were 4: 0 and 3: 1, lettuce could absorb and utilize N effectively. This study demonstrated that hyperoxia (28 kPa) inhibited the growth of lettuce plants under the hypobaric condition (54 kPa), and high level of nitrogen (0.2 g N kg-1 dry matrix) and NO3N: NH4N at 3: 1 markedly enhanced the growth, the contents of mineral elements and the nutritional quality of lettuce plants.

Differential Nitrous oxide emission and microbiota succession in constructed wetlands induced by nitrogen forms.

Nitrous oxide (N2O) emission during the sewage treatment process is a serious environmental issue that requires attention. However, the N2O emission in constructed wetlands (CWs) as affected by different nitrogen forms in influents remain largely unknown. This study investigated the N2O emission profiles driven by microorganisms in CWs when exposed to two typical nitrogen sources (NH4+-N or NO3--N) along with different carbon source supply (COD/N ratios: 3, 6, and 9). The results showed that CWs receiving NO3--N caused a slight increase in total nitrogen removal (by up to 11.8 %). This increase was accomplished by an enrichment of key bacteria groups, including denitrifiers, dissimilatory nitrate reducers, and assimilatory nitrate reducers, which enhanced the stability of microbial interaction. Additionally, it led to a greater abundance of denitrification genes (e.g., nirK, norB, norC, and nosZ) as inferred from the database. Consequently, this led to a gradual increase in N2O emission from 66.51 to 486.77 ug-N/(m2·h) as the COD/N ratio increased in CWs. Conversely, in CWs receiving NH4+-N, an increasing influent COD/N ratio had a negative impact on nitrogen biotransformation. This resulted in fluctuating trend of N2O emissions, which decreased initially, followed by an increase at later stage (with values of 122.87, 44.00, and 148.59 ug-N/(m2·h)). Furthermore, NH4+-N in the aquatic improved the nitrogen uptake by plants and promoted the production of more root exudates. As a result, it adjusted the nitrogen-transforming function, ultimately reducing N2O emissions in CWs. This study highlights the divergence in microbiota succession and nitrogen transformation in CWs induced by nitrogen form and COD/N ratio, contributing to a better understanding of the microbial mechanisms of N2O emission in CWs with NH4+-N or NO3--N at different COD/N ratios.

Appropriate Nitrogen form Ratio and UV-A Supplementation Increased Quality and Production in Purple Lettuce (Lactuca sativa L.).

Purple lettuce (Lactuca sativa L. cv. Zhongshu Purple Lettuce) was chosen as the trial material, and LED intelligent light control consoles were used as the light sources. The purpose was to increase the yield and quality of purple lettuce while lowering its nitrate level. By adding various ratios of NO3--N and NH4+-N to the nutrient solution and 20 µmol m-2 s-1 UV-A based on white, red, and blue light (130, 120, 30 µmol m-2 s-1), the effects of different NO3--N/NH4+-N ratios (NO3--N, NO3--N/NH4+-N = 3/1, NH4+-N) and UV-A interaction on yield, quality, photosynthetic characteristics, anthocyanins, and nitrogen assimilation of purple lettuce were studied. In order to produce purple lettuce hydroponically under controlled environmental conditions, a theoretical foundation and technological specifications were developed, taking into account an appropriate UV-A dose and NO3--N/NH4+-N ratio. Results demonstrate that adding a 20 µmol m-2 s-1 UV-A, and a NO3--N/NH4+-N treatment of 3/1, significantly reduced the nitrate level while increasing the growth, photosynthetic rate, chlorophyll, carotenoid, and anthocyanin content of purple lettuce. The purple leaf lettuce leaves have an enhanced capacity to absorb nitrogen. Furthermore, plants have an acceleration of nitrogen metabolism, which raises the concentration of free amino acids and soluble proteins and promotes biomass synthesis. Thus, based on the NO3--N/NH4+-N (3/1) treatment, adding 20 µmol m-2 s-1 UV-A will be helpful in boosting purple lettuce production and decreasing its nitrate content.

Effects of Different Forms and Proportions of Nitrogen on the Growth, Photosynthetic Characteristics, and Carbon and Nitrogen Metabolism in Tomato.

Optimal plant growth in many species is achieved when the two major forms of N are supplied at a particular ratio. This study investigated optimal nitrogen forms and ratios for tomato growth using the 'Jingfan 502' tomato variety. Thirteen treatments were applied with varying proportions of nitrate nitrogen (NN), ammonium nitrogen (AN), and urea nitrogen (UN). Results revealed that the combination of AN and UN inhibited tomato growth and photosynthetic capacity. Conversely, the joint application of NN and UN or NN and AN led to a significant enhancement in tomato plant growth. Notably, the T12 (75%UN:25%NN) and T4 (75%NN:25%AN) treatments significantly increased the gas exchange and chlorophyll fluorescence parameters, thereby promoting the accumulation of photosynthetic products. The contents of fructose, glucose, and sucrose were significantly increased by 121.07%, 206.26%, and 94.64% and by 104.39%, 156.42%, and 61.40%, respectively, compared with those in the control. Additionally, AN favored starch accumulation, while NN and UN favored fructose, sucrose, and glucose accumulation. Gene expression related to nitrogen and sugar metabolism increased significantly in T12 and T4, with T12 showing greater upregulation. Key enzyme activity in metabolism also increased notably. In summary, T12 enhanced tomato growth by upregulating gene expression, increasing enzyme activity, and boosting photosynthesis and sugar accumulation. Growers should consider using NN and UN to reduce AN application in tomato fertilization.

Effects of nitrogen stress and nitrogen form ratios on the bacterial community and diversity in the root surface and rhizosphere of Cunninghamia lanceolata and Schima superba.

Background: The bacterial communities of the root surface and rhizosphere play a crucial role in the decomposition and transformation of soil nitrogen (N) and are also affected by soil N levels and distribution, especially the composition and diversity, which are sensitive to changes in the environment with high spatial and temporal heterogeneity of ammonium N (NH4 +-N) and nitrate N (NO3 --N). Methods: One-year-old seedlings of Cunninghamia lanceolata and Schima superba were subjected to N stress (0.5 mmol L-1) and normal N supply (2 mmol L-1), and five different N form ratios (NH4 +-N to NO3 --N ratio of 10:0, 0:10, 8:2, 2:8, and 5:5) were created. We analyze the changes in composition and diversity of bacteria in the root surface and rhizosphere of two tree species by high-throughput sequencing. Results: Differences in the composition of the major bacteria in the root surface and rhizosphere of C.lanceolata and S. superba under N stress and N form ratios were not significant. The dominant bacterial phyla shared by two tree species included Proteobacteria and Bacteroidota. Compared to normal N supply, the patterns of diversity in the root surface and rhizosphere of two tree species under N stress were distinct for each at five N form ratios. Under N stress, the bacterial diversity in the root surface was highest at NH4 +-N to NO3 --N ratio of 10:0 of C. lanceolata, whereas in the root surface, it was highest at the NH4 +-N to NO3 --N ratio of 0:10 of S. superba. The NH4 +-N to NO3 --N ratio of 5:5 reduced the bacterial diversity in the rhizosphere of two tree species, and the stability of the bacterial community in the rhizosphere was decreased in C. lanceolata. In addition, the bacterial diversity in the root surface was higher than in the rhizosphere under the N stress of two tree species. Conclusion: The bacterial compositions were relatively conserved, but abundance and diversity changed in the root surface and rhizosphere of C. lanceolata and S. superba under N stress and different N form ratios. The heterogeneity of ammonium and nitrate N addition should be considered for N-stressed environments to improve bacterial diversity in the rhizosphere of two tree species.

Nitrogen acquisition strategy and its effects on invasiveness of a subtropical invasive plant.

Introduction: Preference and plasticity in nitrogen (N) form uptake are the main strategies with which plants absorb soil N. However, little effort has been made to explore effects of N form acquisition strategies, especially the plasticity, on invasiveness of exotic plants, although many studies have determined the effects of N levels (e.g. N deposition). Methods: To address this problem, we studied the differences in N form acquisition strategies between the invasive plant Solidago canadensis and its co-occurring native plant Artemisia lavandulaefolia, effects of soil N environments, and the relationship between N form acquisition strategy of S. canadensis and its invasiveness using a 15N-labeling technique in three habitats at four field sites. Results: Total biomass, root biomass, and the uptakes of soil dissolved inorganic N (DIN) per quadrat were higher for the invasive relative to the native species in all three habitats. The invader always preferred dominant soil N forms: NH4 + in habitats with NH4 + as the dominant DIN and NO3 - in habitats with NO3 - as the dominant DIN, while A. lavandulaefolia consistently preferred NO3 - in all habitats. Plasticity in N form uptake was higher in the invasive relative to the native species, especially in the farmland. Plant N form acquisition strategy was influenced by both DIN levels and the proportions of different N forms (NO3 -/NH4 +) as judged by their negative effects on the proportional contributions of NH4 + to plant N (f NH4 +) and the preference for NH4 + (ß NH4 +). In addition, total biomass was positively associated with f NH4 + or ß NH4 + for S. canadensis, while negatively for A. lavandulaefolia. Interestingly, the species may prefer to absorb NH4 + when soil DIN and/or NO3 -/NH4 + ratio were low, and root to shoot ratio may be affected by plant nutrient status per se, rather than by soil nutrient availability. Discussion: Our results indicate that the superior N form acquisition strategy of the invader contributes to its higher N uptake, and therefore to its invasiveness in different habitats, improving our understanding of invasiveness of exotic plants in diverse habitats in terms of utilization of different N forms.

Effects of different soil water holding capacities on vegetable residue return and its microbiological mechanism.

With the gradual expansion of the protected vegetable planting area, dense planting stubbles and increasing labor cost, the treatment of vegetable residues has become an urgent problem to be solved. Soil bacterial community structure plays an important role in vegetable residue return and is susceptible to environmental changes. Therefore, understanding the influences of different soil water holding capacities on plant residue decomposition and soil bacterial communities is important for biodegradation. During the whole incubation period, the weight loss ratio of plant residue with 100% water holding capacity was 69.60 to 75.27%, which was significantly higher than that with 60% water holding capacity in clay and sandy soil, indicating that high water holding capacity promoted the decomposition of plant residue. The degradation of lignin and cellulose was also promoted within 14 days. Furthermore, with the increase in soil water holding capacity, the contents of NH4+ increased to 5.36 and 4.54 times the initial value in the clay and sandy soil, respectively. The increase in napA and nrfA resulted in the conversion of NO3- into NH4+. The increase in water holding capacity made the bacterial network structure more compact and changed the keystone bacteria. The increase in water holding capacity also increased the relative abundance of Firmicutes at the phylum level and Symbiobacterium, Clostridium at the genus level, which are all involved in lignin and cellulose degradation and might promote their degradation. Overall, these findings provide new insight into the effects of different soil water holding capacities on the degradation of plant residues in situ and the corresponding bacterial mechanisms.

Nitrogen addition increases topsoil carbon stock in an alpine meadow of the Qinghai-Tibet Plateau.

Soil carbon (C) sequestration plays a critical role in mitigating climate change. Nitrogen (N) deposition greatly affects soil C dynamics by altering C input and output. However, how soil C stocks respond to various forms of N input is not well clear. This study aimed to explore the impact of N addition on soil C stock and to elucidate the underlying mechanisms in an alpine meadow on the eastern Qinghai-Tibet Plateau. The field experiment involved three N application rates and three N forms, using a non-N treatment as a control. After six years of N addition, the total C (TC) stocks in the topsoil (0-15 cm) were markedly increased by an average of 12.1 %, with a mean annual rate of 20.1 ‰, and no difference was found between the N forms. Irrespective of rate or form, N addition significantly increased the topsoil microbial biomass C (MBC) content, which was positively correlated with mineral-associated and particulate organic C content and was identified as the most important factor that affecting the topsoil TC. Meanwhile, N addition significantly increased the aboveground biomass in the years with moderate precipitation and relatively high temperature, which leads to higher C input into soils. Owing to decreased pH and/or activities of ß-1,4-glucosidase (ßG) and cellobiohydrolase (CBH) in the topsoil, organic matter decomposition was most likely inhibited by N addition, and this inhibiting effect varied under different N forms. Additionally, TC content in the topsoil and subsoil (15-30 cm) exhibited parabolic and positive linear relationship with the topsoil dissolved organic C (DOC) content, respectively, indicating that DOC leaching might be an important influencing factor for soil C accumulation. These findings improve our understanding of how N enrichment affects C cycles in alpine grassland ecosystems and suggest that soil C sequestration in alpine meadows probably increases with N deposition.

Ammonium improved cell wall and cell membrane P reutilization and external P uptake in a putrescine and ethylene dependent pathway.

Ammonium promotes rice P uptake and reutilization better than nitrate, under P starvation conditions; however, the underlying mechanism remains unclear. In this study, ammonium treatment significantly increased putrescine and ethylene content in rice roots under P deficient conditions, by increasing the protein content of ornithine decarboxylase and 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase compared with nitrate treatment. Ammonium treatment increased rice root cell wall P release by increasing pectin content and pectin methyl esterase (PME) activity, increased rice shoot cell membrane P release by decreasing phosphorus-containing lipid components, and maintained internal P homeostasis by increasing OsPT2/6/8 expression compared with nitrate treatment. Ammonium also improved external P uptake by regulating root morphology and increased rice grain yield by increasing the panicle number compared with nitrate treatment. The application of putrescine and ethylene synthesis precursor ACC further improved the above process. Our results demonstrate for the first time that ammonium increases rice P acquisition, reutilization, and homeostasis, and rice grain yield, in a putrescine- and ethylene-dependent manner, better than nitrate, under P starvation conditions.

Leaf cytokinin accumulation promotes potato growth in mixed nitrogen supply by coordination of nitrogen and carbon metabolism.

The source and sink balance determines crop growth, which is largely modulated by nitrogen (N) supplies. The use of mixed ammonium and nitrate as N supply can improve plant growth, however mechanisms involving the coordination of carbon and N metabolism are not well understood. Here, we investigated potato plants responding to N forms and confirmed that, compared with sole nitrate supply, mixed N (75 %/25 % nitrate/ammonium) enhanced leaf area, photosynthetic activity and N metabolism and accordingly resulted in outgrowth of stolons and shoot axillary buds. Cytokinin transportation in xylem sap and local cytokinin synthesis in leaves were up-regulated in mixed-N-treated potato plants relative to sole nitrate provision; and exogenous application of 6-benzylaminopurine in addition to sole nitrate restored leaf area, photosynthetic capacity and N content in leaves to the similar as those under mixed-N treatment. Partial defoliation, an effective method to enhance the sink strength, induced more cytokinin content in leaflets under two treatments relative to their respective controls and ultimately resulted in larger photosynthesis capacity and leaf area. These results suggest that mixed-N-enhanced plant growth through the coordination of carbon and N metabolism largely depends on the signal molecule cytokinin modulated by N supplies.

Interactive effects of aquatic nitrogen and plant biomass on nitrous oxide emission from constructed wetlands.

Understanding of mechanisms in nitrous oxide (N2O) emission from constructed wetland (CW) is particularly important for the establishment of related strategies to reduce greenhouse gas (GHG) production during its wastewater treatment. However, plant biomass accumulation, microbial communities and nitrogen transformation genes distribution and their effects on N2O emission from CW as affected by different nitrogen forms in aquatic environment have not been reported. This study investigated the interactive effects of aquatic nitrogen and plant biomass on N2O emission from subsurface CW with NH4+-N (CW-A) or NO3--N (CW-B) wastewater. The experimental results show that NH4+-N and NO3--N removal efficiencies from CW mesocosms were 49.4% and 87.6%, which indirectly lead to N2O emission fluxes of CW-A and CW-B maintained at 213 ± 67 and 462 ± 71 µg-N/(m2·h), respectively. Correlation analysis of nitrogen conversion dynamic indicated that NO2--N accumulation closely related to N2O emission from CW. Aquatic NH4+-N could up-regulate plant biomass accumulation by intensifying citric acid cycle, glycine-serine-threonine metabolism etc., resulting in more nitrogen uptake and lower N2O emission/total nitrogen (TN) removal ratio of CW-A compared to CW-B. Although the abundance of denitrifying bacteria and N2O reductase nosZ in CW-B were significantly higher than that of CW-A, after fed with mixed NH4+-N and NO3--N influent, N2O fluxes and N2O emission/TN removal ratio in CW-A were extremely close to that of CW-B, suggesting that nitrogen form rather than nitrogen transformation microbial communities and N2O reductase nosZ determines N2O emission from CW. Hence, the selection of nitrate-loving plants will play an important role in inhibiting N2O emission from CW.

Identification of Quantitative Trait Loci Associated With Iron Deficiency Tolerance in Maize.

Iron (Fe) is a limiting factor in crop growth and nutritional quality because of its low solubility. However, the current understanding of how major crops respond to Fe deficiency and the genetic basis remains limited. In the present study, Fe-efficient inbred line Ye478 and Fe-inefficient inbred line Wu312 and their recombinant inbred line (RIL) population were utilized to reveal the physiological and genetic responses of maize to low Fe stress. Compared with the Fe-sufficient conditions (+Fe: 200 µM), Fe-deficient supply (-Fe: 30 µM) significantly reduced shoot and root dry weights, leaf SPAD of Fe-efficient inbred line Ye478 by 31.4, 31.8, and 46.0%, respectively; decreased Fe-inefficient inbred line Wu312 by 72.0, 45.1, and 84.1%, respectively. Under Fe deficiency, compared with the supply of calcium nitrate (N1), supplying ammonium nitrate (N2) significantly increased the shoot and root dry weights of Wu312 by 37.5 and 51.6%, respectively; and enhanced Ye478 by 23.9 and 45.1%, respectively. Compared with N1, N2 resulted in a 70.0% decrease of the root Fe concentration for Wu312 in the -Fe treatment, N2 treatment reduced the root Fe concentration of Ye478 by 55.8% in the -Fe treatment. These findings indicated that, compared with only supplying nitrate nitrogen, combined supply of ammonium nitrogen and nitrate nitrogen not only contributed to better growth in maize but also significantly reduced Fe concentration in roots. In linkage analysis, ten quantitative trait loci (QTLs) associated with Fe deficiency tolerance were detected, explaining 6.2-12.0% of phenotypic variation. Candidate genes considered to be associated with the mechanisms underlying Fe deficiency tolerance were identified within a single locus or QTL co-localization, including ZmYS3, ZmPYE, ZmEIL3, ZmMYB153, ZmILR3 and ZmNAS4, which may form a sophisticated network to regulate the uptake, transport and redistribution of Fe. Furthermore, ZmYS3 was highly induced by Fe deficiency in the roots; ZmPYE and ZmEIL3, which may be involved in Fe homeostasis in strategy I plants, were significantly upregulated in the shoots and roots under low Fe stress; ZmMYB153 was Fe-deficiency inducible in the shoots. Our findings will provide a comprehensive insight into the physiological and genetic basis of Fe deficiency tolerance.

[Effects of nitrogen deposition and biochar application on soil N2O fluxes in a Moso bamboo plantation]. / 氮沉降和施生物质炭对毛竹林土壤N2O通量的影响.

In July 2019-July 2020, we conducted a field trial to examine the effects of nitrogen addition (60 kg N·hm-2·a-1), biochar application (10 t·hm-2), and their combination on soil N2O emission and the relationship between soil N2O emission and environmental factors in a typical Moso bamboo (Phyllostachys edulis) plantation in Hangzhou City of Zhejiang Province. Soil N2O flux of Moso bamboo plantation was measured by the static chamber-gas chromatography technique. The results showed that nitrogen addition treatment increased the annual cumulative N2O emission by 14.6%, while biochar application and the combination treatment reduced it by 20.8% and 10.6%, respectively. Soil N2O flux rate was significantly correlated with soil temperature, NO3--N concentration, urease and protease activities, and soil NH4+-N concentration across all treatments. In conclusion, under the background of nitrogen deposition, the application of biochar would have a significant reduction effect on soil N2O fluxes in Moso bamboo plantations.

Ammonium regulates redox homeostasis and photosynthetic ability to mitigate copper toxicity in wheat seedlings.

As an essential plant micronutrient, copper (Cu) is required as a component of several enzymes, but it can be highly toxic to plants when present in excess quantities. Nitrogen (N) application can help to alleviate the phytotoxic effects of heavy metals, including Cu, and different N forms significantly affect the uptake and accumulation of heavy metals in plants. The aim of this study was to determine the effects of different N forms, i.e., ammonium (NH4+) and nitrate (NO3-), on Cu detoxification in wheat seedlings. The inhibition of seedling growth under excess Cu was more obvious in wheat plants supplied with NO3- than in those supplied with NH4+. This growth inhibition was directly induced by excess Cu accumulation and reduced absorption of other mineral nutrients by the plants. Compared with seedlings treated with NO3-, those treated with NH4+ showed a decrease in Cu-induced toxicity as a result of increased antioxidant capacity in the leaves and a lower redox potential in the rhizosphere. Furthermore, treatment with NH4+ decreased the loss of mineral nutrients in wheat seedlings exposed to excess Cu. In conclusion, compared with supplying NO3-, supplying NH4+ to wheat seedlings under Cu stress improved their ability to maintain their nutritional and redox balance and increased their antioxidant capacity, thereby preventing a decline in photosynthesis. According to our results, NH4+ is more effective than NO3- in reducing Cu phytotoxicity in wheat seedlings.

Organic nitrogen sources promote andrographolide biosynthesis by reducing nitrogen metabolism and increasing carbon accumulation in Andrographis paniculata.

Nitrogen (N) form affects secondary metabolites of medicinal plants, but the physiological and molecular mechanisms remain largely unknown. To fully understand the response of andrographolide biosynthesis to different N forms in Andrographis paniculata, the plants were fed with nutritional solution containing sole N source of nitrate (NO3-), ammonium (NH4+), urea or glycine (Gly), and the growth, carbon (C) and N metabolisms and andrographolide biosynthesis were analyzed. We found that plants grown in urea and Gly performed greater photosynthetic rate and photosynthetic N use efficiency (PNUE) than those grown in NO3- and NH4+. Organic N sources reduced the activities of enzymes involving in C and N metabolisms such as glutamine synthase (GS), glutamate synthase (GOGAT) and NADH-dependent glutamate dehydrogenase (NADH-GDH), invertase (INV), isocitrate dehydrogenase (ICDH) and glycolate oxidase (GO), resulting in reduced depletion of carbohydrates and increased starch accumulation. However, they enhanced andrographolide content by up-regulating the key genes in its biosynthetic pathway including HMGR, DXS, GGPS and ApCPS. Besides, NH4+ decreased leaf SPAD value, contents of soluble protein and amino acids and GO activity, but increased photosynthetic rate and contents of soluble sugar and starch in comparison to NO3-. Andrographolide biosynthesis was also up-regulated. The results revealed that increasing accumulation of carbohydrates, especially starch, was beneficial to the biosynthesis of andrographolide; organic N sources decreased carbohydrate depletion by reducing N metabolism, and promoted plant growth and andrographolide biosynthesis synergistically.

Converting rice husk to biochar reduces bamboo soil N2O emissions under different forms and rates of nitrogen additions.

The effects of biochar application combined with different forms and rates of inorganic nitrogen (N) addition on nitrous oxide (N2O) emissions from forest soils have not been well documented. A microcosm experiment was conducted to study the effects of rice husk and its biochar in combination with the addition of N fertilizers in different forms (ammonium [NH4+] and nitrate [NO3-]) and rates (equivalent to 150 and 300 kg N ha-1 yr-1) on N2O emissions from Lei bamboo (Phyllostachys praecox) soils. The application of rice husk significantly increased cumulative N2O emissions under the addition of both NO3--N and NH4+-N. Biochar significantly reduced cumulative N2O emissions by 15.2 and 5.8 µg N kg-1 when co-applied with the low and high rates of NO3--N, respectively, compared with the respective NO3--N addition rate without biochar. There was no significant difference in soil N2O emissions between the two NH4+-N addition rates, and cumulative N2O emission decreased with increasing soil NH4+-N concentration, mainly due to the toxic effect caused by the excessive NH4+-N on soil N2O production from the nitrification process. Cumulative N2O emissions recorded 18.74 and 14.04 µg N kg-1 under low and high rates of NO3--N addition, respectively, which were higher than those produced by NH4+-N addition. Our study demonstrated that the conversion of rice husk to biochar could reduce N2O emissions under the addition of different N forms and rates. Moreover, rice husk or its biochar in combination with NH4+-N fertilizer produced less N2O in Lei bamboo soil, compared with NO3--N fertilizer.