The germination process is critical for plant growth and development and it is largely affected by environmental stress, especially salinity. Recently, hydrogen sulfide (H2S) is well known to act as a signaling molecule in a defense mechanism against stress conditions but poorly understood regulating seed germination. In this study, the effects of NaHS (the H2S donor) pretreatment on various biochemical (hydrogen peroxide (H2O2) content and amylase and protease activity) and physiological properties (germination rate) during seed germination of oilseed rape (Brassica napus L. cv. Mosa) were examined under salt stress. The seed germination and seedling growth of oilseed rape were inhibited by NaCl treatment but it was alleviated by NaHS pretreatment. The NaCl treatment increased H2O2 content leading to oxidative stress, but NaHS pre-treatments maintained much lower levels of H2O2 in germinating seeds under salt stress. Amylase activity, a starch degradation enzyme, significantly increased over 2-fold in control, NaHS pretreatment, and NaHS pretreatment under NaCl during seed germination compared to NaCl treatment. Protease activity was highly induced in NaHS-pretreated seeds compared to NaCl treatment, accompanied by a decrease in protein content. These results indicate that NaHS pretreatment could improve seed germination under salt stress conditions by decreasing H2O2 accumulation and activating the degradation of protein and starch to support seedling growth.

Sulfur is an essential element in plants, including amino acids, vitamin synthesis, and acting as an antioxidant. However, the interaction between endogenous sulfur and proline synthesis has not been yet fully documented. White clover (Trifolium repens L.) is known as a species highly sensitive to sulfate supply. Therefore, this study aimed to elucidate the role of sulfur in regulating proline metabolism in relation to ammonia detoxification and hydrogen peroxide (H2O2) accumulation in white clover. The detached leaves of white clover were immersed in solution containing different concentration of sulfate (0, 10, 100, and 1000 mM MgSO4). As MgSO4 concentrations were increased, the concentration of H2O2 increased up to 2.5-fold compared to control, accompanied with H2O2 detection in leaves. Amino acid concentrations significantly increased only at higher levels (100 and 1000 mM MgSO4). No significant difference was observed in protein concentration. Proline and Δ1-pyrroline-5-carboxylate (P5C) concentrations slightly decreased at 10 and 100 mM MgSO4 treatments, whereas it rapidly increased over 1.9-fold at 1000 mM MgSO4 treatment. Ammonia concentrations gradually increased up to 8.6-fold. These results indicate that exogenous sulfur levels are closely related to H2O2 and ammonia synthesis but affect proline biosynthesis only at a higher level.

The objective of this study was to determine effects of phosphorus on lignification and carbohydrate metabolism in Kentucky bluegrass under drought stress. Drought stress was induced by reducing of water to plants in pots. Two types of phosphorus were applied as potassium phosphate (PO4 3-; P) or potassium phosphonate (PO3 3-; PA) in drought-stressed plants. Drought had significant negative effects on plant growth, as revealed by reduced biomass of shoot. Drought-induced increase of lignin content was concomitant with the increase of phenylalanine ammonia-lyase (PAL). Soluble sugar content was highly increased but fructan content was largely decreased by drought stress. However, the application of phosphorus was efficient to ameliorate the adverse effects of drought. PA application improved reduced shoot growth and relative water content, and inhibited lignification synthesis with a reduction of PAL activity. P or PA application maintained soluble sugar and fructan content at similar levels to controls under drought stress. These results indicate that phosphorus application may mitigate the drought stress by inhibiting the lignification and promoting the fructan assimilation.

The objective of this study was to determine phosphorus effects on drought stress-induced oxidative stress in Kentucky bluegrass. Drought stress was induced by reducing of water to plants in pots. Two types of phosphorus were applied as potassium phosphate (P) or potassium phosphonate (PA). Application of phosphorus was efficient to ameliorate the adverse effects of drought. Osmotic potential, total chlorophyll and carotenoid content were significantly decreased by drought stress, but was relieved by P or PA application. Superoxide (O2 •−) concentration was significantly increased more than 14-fold under drought-stressed plants, was accompanied with increase of hydrogen peroxide (H2O2) and lipid peroxidation (MDA). However, malondialdehyde (MDA) was much less in P or PA applied plants under drought stress condition. Activities of catalase (CAT), ascorbate peroxidase (APX) and guaiacol-peroxidase (GPX) were largely increased by drought stress and its increase rate was much higher in P or PA applied plants except APX. These results indicate that drought stress-induced oxidative stress is alleviated by P or PA application due to the increase of activities of antioxidant enzymes.

Oilseed rape is known to crop having low nitrogen use efficiency (NUE) but requires high levels of N fertilizer. NUE is associated with N remobilization from source to sink organ, consequently affects seed yield. Remobilization of leaf N is also related to transport of C/N metabolites in phloem. However, interaction between seed yield and phloem transport was not fully documented. In response to seed yield, N and C metabolites and their transport into seed from bolting to pod filling stage investigated in two contrasting genotypes (Capitol and Pollen) cultivated under ample (HN) or limiting nitrate (LN) supply. Seed yield was significantly reduced in N limitation and its reduction rate was much lower in Capitol than in Pollen compared to HN treated plants. Amino acid and protein content was higher in Capitol than in Pollen at bolting stage. They gradually decreased during plant development but not significant between two cultivars and/or two treatments. Glucose, fructose and sucrose content were 1.8-,1.6- or 1.25-fold higher in LN condition than in HN condition, respectively. Amino acid and sucrose content in phloem were largely higher in Capitol than in Pollen under LN condition. These results indicate that the higher seed yield might be related to greater transport ability of amino acid and sucrose in phloem under LN condition.

Soil is the main nitrogen (N) provider for plants but N in soil is not all available to advanced plants. Mineralization is a critical biological process for transferring organic N to inorganic N that can be used by plants directly. To investigate the effect of different levels of soil temperature and water content to soil mineralization, a field experiment was established on three different sites (A, B and C). We measured soil temperature, moisture and electrical conductivity once daily after swine slurry application. Average soil moisture and temperature in site A is the highest among three sites (40.9% and 9.7°C, respectively). Following is in site C (37.3% and 9.6°C) and the lowest is in site B (28.0% and 9.0°C). Ammonium N (NH4+-N) and nitrate N (NO3--N) were determined on the first and fifth day after treatment. Compared with site B and C, site A always had the highest soil total N content (1.54 g N kg-1 on day one; 1.22 g N kg-1 on day five) and highest NO3-- N content (93.18 mg N kg-1 on day one; 16.22 mg N kg-1 on day five) and a significant decrease on day five. Content of NH4+-N in site B and C reduced while in site A, it increased by 6.7%. Results revealed that net N mineralization positively correlated with soil temperature (P<0.5, r=0.675*) and moisture (P<0.01, r=0.770**), suggesting that to some extent, higher soil moisture and temperature contribute more to inorganic N that can be used by plants.

The objective of this study was to determine the effect of injection application of pig slurry on ammonia (NH3) and nitrous oxide (N2O) emission from timothy (Phleum pretense L.) sward. The three treatments were applied: 1) only water as a control, 2) pig slurry application by broadcasting, 3) pig slurry application by injection. The pig slurry was applied at a rate of 200 kg N ha-1. Total NH3 and N2O emission, expressed as a cumulative amount throughout the measurement time (40 days), was 2.68 kg NH3-N ha-1 and 6.58 g N2O-N ha-1, respectively, in the control. The injection application of pig slurry decreased total NH3 and N2O emission by 39.8% and 33.3%, respectively, compared to broadcasting application of pig slurry. The present study clearly showed that injection application exhibited positive roles in reducing N losses through NH3 and N2O emission.

To investigate the impact of nitrogen (N) mineral on reproductive potential of Brassica napus L, plants were treated with different levels of N treatment (N0; N100; N500). The half of N content for each treatment were applied at the beginning of the early vegetative stage and the rest was applied at the late vegetative stage. Nitrogen content in plant tissues such as root, stem and branch, leaf, pod and seed was analyzed and harvest index (HI) was calculated as percentage of seed yield to total plant weight. Biomass and nitrogen content were significantly affected by different levels of N supply. Biomass was significantly decreased by 59.2% in nitrogen deficiency (N0) but significantly increased by 50.3% in N excess (N500), compared to control (N100). Nitrogen content in all organs was remarkably increased with nitrogen levels. N distribution to stem and branches, and dead leaves was higher in N-deficient (N0) and N excessive plants (N500) than in control (N100). However, nitrogen allocated to seed was higher in control (N100) than in other treatments (N0 or N500), accompanied by higher HI. These results indicate that the optimum level of N supply (N100) improve HI and N distribution to seed and excessive N input is unnecessary.

In order to compare greenhouse gases emission from different animal manures and to explore how different animal manures effect on soil mineralization, three kinds of materials, cattle, goat and chicken manure were amended to soil for 14 days incubation as CtS (cattle manure-amended soil), GS (goat manure-amended soil) and ChS (chicken manure-amended soil). Cumulative NH3 emissions in all treatments were rapidly increased until day 7 and then it was slightly increased in three manure-amended soils but maintained in control until day 14. GS had the highest NH3 emission at 0.14 mg kg-1 during the entire experimental period. Emissions of CO2 were highly increased by 7.8-, 9.0- and 12.4-fold in CtS, GS and ChS, respectively, compared to control at day 14. A significant increase of N2O emission in all treatments occurred within 5 days and then it was slightly increased until day 14. N2O emission was 2-fold higher in all manure-amended soils than that of control. Compared to day 1, inorganic N (NH4 + plus NO3 --N) content was highly increased in all four treatments at day 14. The increase rate was the highest in CtS treatment. Net N mineralization was increased by 4.0-, 2.4- and 2.9-fold in CtS, GS and ChS, respectively, compared to control. These results indicate that increase of NH3, CO2 and N2O gas emissions was positively related to high N mineralization.

Mineralization is an important biological process for conversion of organic nitrogen (N) to inorganic N which can be used by plants directly. To investigate the effect of different manures on soil mineralization, the soil amended with cattle (CtM), goat (GM), chicken manure (ChM) and pig slurry (PS) were incubated under in vitro condition and ammonium N (NH4 +-N), ammonification rate and ammonia emission were determined for eighty-four days. NH4 +-N was the highest in PS-amended soil for the whole experimental period. NH4 +-N in PS-amended soil was gradually decreased until day 84, whereas it was rapidly decreased for the first 14 days and then slightly increased until 84 days in ChM-, CtM- and GM-amended soil. The ammonification rate showed negative value for the first 14 days in all treatments. From day 14, ammonification rate started to increase in CtM- and ChM-amended soil, whereas it was maintained in GM- and PS-amended soil until day 84. The daily ammonia emission was the highest in PS-amended soil (41mg kg-1 d-1), followed by CtM-, ChM-, and GM-amended soil at day 1. It was gradually decreased until day 84 in all treatments. The total NH3 emission was the highest in PS-amended soil with 0.6 mg kg-1 for 84 days, while less than 0.1 mg kg-1 in three other plots. These results indicate that different manures showed different soil ammonification rate and NH3 emission.

The objective of this study was to determine the effect of nitrification inhibitor dicyandiamide (DCD) and urease inhibitor hydroquinone (HQ) on ammonia (NH3) and nitrous oxide (N2O) emission from pig slurry applied to Timothy (Phleum pretense L.) sward. The daily emission of ammonia (NH3) and nitrous oxide (N2O) was monitored for 9 days in three different treatments; 1) control (only pig slurry application), 2) DCD treatment (pig slurry + DCD), and 3) HQ treatment (pig slurry + HQ). Most NH3 emission occurred after 4~5 days in three treatments. Total NH3 emission, expressed as a cumulative amount throughout the measurement time, was 1.33 kg N ha 1 in the control. The DCD and HQ treatment decreased total NH3 emission by 16.3% and 25.1%, respectively, compared to the control. Total N2O emission in the control was 47.1 g N ha 1. The DCD and HQ treatment resulted in a reduction of 67.9% and 41.8% in total N2O emission, respectively, compared to the control. The present study clearly indicated that nitrification and urease inhibitor exhibited positive roles in reducing N losses through NH3 and N2O emission.

This study was conducted to assess the efficacy of slurry acidification in reducing ammonia emission from manure storage and application. The non-fermented cattle manure (NFC) and swine slurry (SS) were acidified by sulfuric acid and stored in an acryl chamber for 168 and 96 hours, respectively. Ammonia emitted from the chamber was collected using an acid trap system. The amount of ammonia emission was significantly reduced when the livestock manures were treated with sulfuric acid. The absolute amount of ammonia in NFC increased rapidly starting from 48 h and 72 h in the control (pH 8.6) and acidified NFC (pH 6.5), respectively. The absolute amount of ammonia was the highest at 96 h (3.65 g kg ¯¹ h ¯¹) in the control and at 144 h (2.34 g kg ¯¹ h ¯¹) in pH 6.5 NFC. The cumulative ammonia content in the control continuously increased until 96 h and was maintained until 168 h, whereas the increase rate of emission gas accumulation in acidified NFC was much less throughout the experimental period. Acidification of SS mitigated ammonia emission as proven in NFC. The cumulative amount of ammonia emission was decreased by 49.4% and 92.3% in the acidified SS at pH 6.5 and pH 5.5, respectively, compared to the control at 96 h after treatment. These results indicate that ammonia emission can be significantly reduced by sulfuric acid treatment of livestock manure during processing and the subsequent land application.

To investigate lignification process and its physiological significance under water-deficit condition, the responses of peroxidases, polyphenol oxidase(PPO) and phenylalanine ammonia-lyase(PAL) in relation to leaf water status to the short term of water deficit treatment in the leaves with different maturities in forage rape were measured. The significant decrease in relative water content(RWC) and leaf osmotic potential(Ψπ) were apparent after 5 d of water-deficit treatment. The activity of guaiacol peroxidase(GPOD), ascorbate peroxidase(APOD), coniferyl alcohol peroxidase(CPOD), and syringaldazine peroxidase(SPOD) was depressed especially in middle and old leaves when compared with that of control leaves. On the other hand, in young leaves, a significant increase in CPOD(+34%) and SPOD(+24%) activity as affected by water-deficit treatment was apparent. The activation of PAL and PPO was observed in middle and old leaves for PAL and in young and middle leaves for PPO. These results suggest that peroxidases in middle and old leaves did not involve in lignification under mild water-deficit stress, whereas CPOD and SPOD in young leaves participate in lignification by a coordination with PAL and PPO to incorporate phenol and lignin into the cell walls.

Molybdenum (Mo) in rhizosphere influences sulfate assimilation as well as a number of other physiological aspects. In this study, the activity of key enzymes in sulfate assimilatory pathways, such as ATP sulfurylase (ATPs), adenosine 5'-phosphosulphate reductase (APR), as well as the responses of reactive oxygen species (ROS), were analyzed to elucidate the metabolic and physiological effects of external Mo supply to detached leaves of Trifolium repens L. Mo supply with a range from 1 mM to 40 mM depressed the activity of ATPs throughout the entire time course. In the leaves exposed to 1 mM Mo, a continuous decrease in the activity of ATPs was confirmed by Native-PAGE. The APR activity was also declined by Mo treatment. The accumulation of H2O2 and O2 were not significant up to 10 mM Mo, whereas a remarked accumulation was detected under 40 mM Mo supply. The data suggest that an external supply of Mo has an inhibitory effect on sulfate assimilation, and induces oxidative stress only at an extremely high concentration.

Sulphur deficiency has become widespread over the past several decades in most of the agricultural area. Oilseed rape (Brassica napus L.) is a very sensitive to S limitation which is becoming reduction of quality and productivity of forage. Few studies have assessed the sulphur mobilization in the source-sink relationship, very little is known about the regulatory mechanism in interaction between sulphur and nitrogen during the short-term sulphur deficiency. In this study, therefore, amount of sulphur and nitrogen incorporated into amino acids and proteins as affected by different S-supplied level (Control: 1 mM SO4 2 , S-deficiency: 0.1 mM SO4 2 , and S-deprivation: 0 mM SO4 2 ) were examined. The amount of sulphur in sulphate (S-sulphate) was significantly decreased by 25.8% in S-deprivation condition, compare to control, but not nitrogen in nitrate (N-nitrate). The markedly increase of sulphur and nitrogen incorporated amino acids (S-amino acids and N-amino acids) was observed in both S-deficiency and S-deprivation treatments. The amount of nitrogen incorporated proteins (N-protein) was strongly decreased as sulphur availability while the amount of sulphur incorporated into proteins (S-protein) was not affected. A highly significant (p<0.001) relationship between S-sulphate and S-amino acid was observed whereas the increase of N-amino acids is closely associated with decrease of N-proteins. These data indicate that increase of sulphur and nitrogen incorporated into amino acids was from different nitrogen and sulphur metabolites, respectively.