• Title/Summary/Keyword: Nitrate reductase

Search Result 119, Processing Time 0.028 seconds

Production and Characterization of Nitrate Reductase Deficient Mutants in Petunia parviflora

  • Lee, Cheol-Hee
    • Korean Journal of Plant Resources
    • /
    • v.19 no.6
    • /
    • pp.706-715
    • /
    • 2006
  • Nitrate reductase deficient (NR) mutant lines were selected indirectly by their resistance to 100mM chlorate in cell cultures of P. parviflora. A total of 585 chlorate resistant lines were confirmed by a second passage on a high concentration of chlorate. Frequency of spontaneous mutation was $9.7{\times}10^{-7}$ in 3 month old suspension-cultured cells, and in non-selective media containing amino acids as sole nitrogen source. The frequency of mutation could be increased up to 11-fold by culture for 12 months. Out of 40 randomly selected calli, 22 were fully deficient in NR. The rest of the clones contained a decreased level of NR activity. Further characterization was carried out in 13 mutant lines which were fully deficient in NR and in 5 mutant lines containing residual (0-7.0%) NR activity, as compared to wild-type cells cultured on the same medium. The $NR^-$ mutants were tentatively classified as defective in the NR apoenzyme (nia-type; 11 mutant lines including the 5 with residual NR activity) or in the molybdenum cofactor (cnx-type; 7 mutant lines) by the XDH activity. The cnx-type could be further classified into two groups. In one group (5 mutant lines) of these, the NR activity could be partially restored by nonphysiologically high (1.0mM) molybdate in the culture medium. Both types of $NR^-$ mutants were unable to grow on minimal medium containing nitrate as sole nitrogen source, but grew well on amino acids. They also proved to be extremely sensitive to the standard medium ($MSP_1$) containing nitrate and ammonium. Shoot regeneration was obtained only in the $NR^-$ mutants, which contained residual NR activity, but they so far have failed to grow into plants.

Suppression of Nitrate Accumulation in Vegetables by Foliar Application of Micronutrients (미량원소 엽면 처리에 의한 엽채류의 질산태 질소 축적 억제)

  • Eom, Jin-Sup;Park, Nu-Ri;Park, Sang-Gyu;Park, Shin;Chung, Jong-Bae
    • Applied Biological Chemistry
    • /
    • v.44 no.4
    • /
    • pp.240-245
    • /
    • 2001
  • Suppression of nitrate accumulation in vegetables through foliar application of micronutrients was investigated. Spinach and lettuce were grown in pots under greenhouse condition. Micronutrient solutions containing Cu, Mn, Mo, and Zn were used; chitosan was added into one and the other contained chitosan oligomers. The micronutrient solutions were sprayed on the leaves at 3 and 4 weeks after transplanting of 20-day-old seedlings. Plants were harvested at 5-weeks after transplanting. Yield, contents of chlorophyll, Brix value, micronutrient, and nitrate, and nitrate reductase activity were measured. Fresh weights of lettuce and spinach were significantly increased by the foliar application of micronutrients. Contents of chlorophyll and micronutrients were higher in micronutrient-treated plants, while those of nitrate were reduced by about 10 and 14-23% in lettuce and spinach, respectively. Compared to the control plants, nitrate reductase activity was higher in plants treated with micronutrients. Results of this study indicate the effect of micronutrients on the suppression of nitrate accumulation was relatively small in comparison to the contents of nitrate in leaves of spinach and lettuce. To maximize the effect, nutrient composition in solution, application time, and frequency should be further examined, taking into consideration nitrogen level in soil and other environmental factors including light condition.

  • PDF

Amelioration of $Cd^{++}$ Toxicity by $Ca^{++}$ on Germination, Growth and Changes in Anti-Oxidant and Nitrogen Assimilation Enzymes in Mungbean(Vigna mungo) Seedlings

  • Kochhar Sunita;Ahmad Gayas;Kochhar Vinod Kumar
    • Journal of Plant Biotechnology
    • /
    • v.6 no.4
    • /
    • pp.259-264
    • /
    • 2004
  • The present study describes the ameliorating effect of $Ca^{++}\;on\;Cd^{++}$ toxicity on the germination, early growth of mungbean seedlings, nitrogen assimilation enzyme. s-nitrate reductase (NR), nitrite reductase (NIR), anti-oxidant enzymes (POD, CAT and SOD) and on the accumulation of hydrogen peroxide and sulphydryls. $Cd^{++}$ inhibited seed germination and root and shoot length of seedlings. While NR activity was down- regulated, the activities of NIR, POD and SOD were up- regulated with $Cd^{++}$ treatment. $Cd^{++}$ treatment also increased the accumulation of sulphydryls and peroxides, which is reflective of increased thiol rich proteins and oxidative stress. $Ca^{++}$ reversed the toxic effects of $Cd^{++}$ on germination and on early growth of seedlings as well as on the enzyme activities, which were in turn differentially inhibited with a combined treatment with calcium specific chelator EGTA. The results indicate that the external application of $Ca^{++}$ may increase the tolerance capacity of plants to environmental pollutants by both up and down regulating metabolic activities. Abbreviations: $Cd^{++}= cadmium,\;Ca^{++} = calcium$, NR= nitrate reductase, NIR=nitrite reductase, POD = peroxidse, SOD= superoxide dismutase, CAT= catalase, EGTA= ethylene glycol-bis( $\beta-aminoethyl ether$)-N,N,N,N-tetraacetic acid.

Impact of Sulphur and Nitrogen Application on Seed and Xanthotoxin Yield in Ammi majus L.

  • Ahmad, Saif;Jamal, Arshad;Fazili, Inayat Saleem;Alam, Tanweer;Khan, Mather Ali;Kamaluddin, Kamaluddin;Iqbal, Mohd;Abdin, Malik Zainul
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.52 no.2
    • /
    • pp.153-161
    • /
    • 2007
  • Field experiments were conducted to determine the physiological and biochemical basis of the interactive effect of sulphur (S) and nitrogen (N) application on seed and xanthotoxin yield of Ammi majus L. Six treatments were tested ($T_1$ = control-without manure and fertilizers, $T_2$ = manure @ 9 kg $plot^{-1}-10\;t\;ha^{-1},\;T_3=A_0N_{50}K_{25}P_{25},\;T_4=S_{40}N_{50}K_{25}P_{25},\;T_5=S_{40}N_{100}K_{25}P_{25}\;T_6=S_{20+20}N_{50+50}K_{25}P_{25})$). Nitrate reductase (NR) activity and ATP-sulphurylase activity in the leaves were measured at various phonological stages, as the two enzymes catalyze rate-limiting steps of the assimilatory pathways of nitrate and sulphate, respectively. The activities of these two enzymes were strongly correlated with seed and xanthotoxin yield. The highest nitrate reductase activity, ATP-sulphurylase activity and xanthotoxin yield were achieved with the treatment $T_4$. Any variation from this treatment decreased the activity of these enzymes, resulting in a reduction of the seed and xanthotoxin yield in Ammi majus L. The higher seed and xanthotoxin yield achieved in Ammi majus L. at treatment $T_4$ could be due to optimization of leaf soluble protein and photosynthetic rate, as these parameters are Influenced by S and N assimilation.

Effect of Nitrogen and Sulphur Application on Nitrate Reductase and ATP-sulphurylase Activities in Soybean

  • Jamal Arshad;Fazli Inayat Saleem;Ahmad Saif;Abdin Malik Zainul;Yun Song-Joong
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.51 no.4
    • /
    • pp.298-302
    • /
    • 2006
  • A number of field experiments were conducted to assess the role of combined application of nitrogen and sulfur to increase the seed and oil yield of nonnodulating soybean (Glycine max (L) Merr.) cv. PK-416 $(V_1)$ and cv. PK-1024 $(V_2)$. Six combinations of N and S in three replicates each were used for this purpose i.e. $0\;S+23.5kg\;N\;ha^{-1}(T_1);\;0\;S+23.5+20kg\;N \;ha^{-1}(T_2);\;40\;S+23.5kg\;N\;ha^{-1}(T_3);\;40\;S+23.5+20kg\;N\;ha^{-1}(T_4);\;20+20\;S+23.5kg\;N\;ha^{-1}(T_5);\;20+20\;S+23.5+20kg\;N\;ha^l(T_6)$. Nitrate reductase (NR) and ATP-sulphurylase activities in the leaves were measured at various growth stages as the two enzymes catalyze the rate limiting steps of the assimilatory pathways of nitrate and sulphate, respectively. The activities of these enzymes were strongly correlated with seed yield. The higher seed, oil and protein yields were achieved with the treatment $T_6$ in both the cultivars due to optimization of NR activity and ATP-sulphurylase activity, as these parameters were influenced by N and S assimilation. Any variation from this combination was observed to decrease the activity of these enzymes resulting in reductions in the seed, oil and protein yield of soybean.

Characterization of heterotrophic nitrification and aerobic denitrification by Alcaligenes faecalis NS13 (Alcaligenes faecalis NS13에 의한 호기성 종속영양 질산화 및 탈질화)

  • Jung, Taeck-Kyung;Ra, Chang-Six;Joh, Ki-Seong;Song, Hong-Gyu
    • Korean Journal of Microbiology
    • /
    • v.52 no.2
    • /
    • pp.166-174
    • /
    • 2016
  • In order to find an efficient bacterial strain that can carry out nitrification and denitrification simultaneously, we isolated many heterotrophic nitrifying bacteria from wastewater treatment plant. One of isolates NS13 showed high removal rate of ammonium and was identified as Alcaligenes faecalis by analysis of its 16S rDNA sequence, carbon source utilization and fatty acids composition. This bacterium could remove over 99% of ammonium in a heterotrophic medium containing 140 mg/L of ammonium at pH 6-9, $25-37^{\circ}C$ and 0-4% of salt concentrations within 2 days. It showed even higher ammonium removal at higher initial ammonium concentration in the medium. A. faecalis NS13 could also reduce nitrate and nitrous oxide by nitrate reductase and nitrous oxide reductase, respectively, which was confirmed by detection of nitrate reductase gene, napA, and nitrous oxide reducase gene, nosZ, by PCR. One of metabolic intermediate of denitrification, $N_2O$ was detected from headspace of bacterial culture. Based on analysis of all nitrogen compounds in the bacterial culture, 42.8% of initial nitrogen seemed to be lost as nitrogen gas, and 46.4% of nitrogen was assimilated into bacterial biomass which can be removed as sludge in treatment processes. This bacterium was speculated to perform heterotrophic nitrification and aerobic denitrification simultaneously, and may be utilized for N removal in wastewater treatment processes.

A Study on the Source of Reductants for Nitrate Reduction in Rice (Oryza sativa cv. Tongil) Roots (벼(Oryza sativa cv. Tongil) 뿌리에 있어서 Nitrate 환원에 필요한 환원력의 공급원에 관한 연구)

  • Chang, Nam-Kee;Choe, Hong-Gwan
    • The Korean Journal of Ecology
    • /
    • v.7 no.2
    • /
    • pp.85-90
    • /
    • 1983
  • There was a decrease in nitrate reductase activity (NRA) measured in vivo in rice roots (Oryza sativa cv. Tongil) grown in anaerobic culture solution. But it was reversed by addition of malonate to the in vivo nitrate reduction assay medium. Malonate increased the in vivo NRA during 2-5 hours incubation and decreased it in longer incubation hours. In vivo NRA was stimulated by addition of NaHCO3 to the assay medium, but not by Na2CO3. The stimulation of NRA by NaHCO3 was not observed in shoot removed rice roots. It is suggested that CO2 from NaHCO3 is carboxylated by phosphoenol pyruvate carboxylase, results in increasing the malate contents in the roots, and stimulates the in vivo NRA. NADH needed in nitrate reduction is supported by malate oxidation. In rice roots, it seems probable that malate oxidation in the mitochondria is more important to nitrate reduction than malae oxidation in cytoplasm.

  • PDF

Effects of Iron, Chelators and Nitrate Concentration on in vivo Fluorescence and Nitrate Reductase of the Red Tide Organism Amphidinium carterae

  • Yang, Sung-Ryull;Song, Hwan-Seok;Pae, Se-Jin;Huh, Sung-Hoi
    • Journal of the korean society of oceanography
    • /
    • v.34 no.1
    • /
    • pp.49-57
    • /
    • 1999
  • A red tide organism, Amphidinium carterae was incubated under different iron/chelator and nitrate concentrations to investigate the factors controlling the growth. The chelation capacity played a critical role in regulating the nitrate reductase (NR) activity and in vivo fluorescence of this organism. However, there was a significant difference between the NR activity and in vivo fluorescence in response to trace metals and chelator treatments. In vivo fluorescence was the highest in FeEDTA 10 ${\mu}$M treatments and the lowest in DTPA 10 ${\mu}$M treatments. This indicates that the availability of the trace metal is important in regulating the in vivo fluorescence of this photosynthetic microalgae In contrast, NR activity showed the highest values in trace metal enriched treatments, and trace metal + DTPA treatments showed fairly high NR activities. This suggests that DTPA treatment did not hinder the NR activity as much as it did in vivo fluorescence. In vivo fluorescence and NR activity increased with nitrate concentration of up to 50 ${\mu}$M and remained relatively constant or the rate of increase decreased above that concentration, indicating that initial nitrate concentration of higher than a certain level would not accelerate the growth of A. carterae. Further investigation is needed to elucidate the reason for the difference in timing sequence between the NR and in vivo fluorescence in response to different metal treatments and chelation capacity.

  • PDF

Influence of light Regime on Nitrate Reductase Activity and Organic and norganic Solute Composition of Four Sedges (Carex spp.)

  • Choo, Yeon-Sik;Roland-Albert;Song, Seung-Dal
    • Animal cells and systems
    • /
    • v.2 no.4
    • /
    • pp.455-462
    • /
    • 1998
  • A survey was conducted on the inorganic and organic solute patterns of plants in connection with nitrate metabolism according to different light regimes (1.9, 16.0, 91.5 $Wm^{-2}$). Besides measuring in vivo NRA, we also quantitatively analyzed ater-soluble inorganic ions, organic acids, low molecular weight carbohydrates, amino aciss and total N (% DW). Among 4 Carex species, C. pilosa is known as shade-adapted species and the others as half (C. gracilis) to full (C. rostrata & C. distans) light-adapted species. Compared to species adapted to high light intensity, shade-adapted C. pilosa showed reduced productivity under the highest light intensity. In general, nitrate and amino acid levels decreased at higher light intensity, while sugar and organic acid concentrations increased. In C. pilosa osmolality tended to rise with increasing light intensity, while in the other species it tended to fall. Under low light intensity, the drop in soluble carbohydrate contents is osmotically compensated for by an enhanced nitrate concentration. It is concluded that competition between nitrate and $CO_2$reduction for reductants and ATP from photosynthesis may have important ecological consequences for the adaptation of plants to low or high light conditions. Additionally, the patterns of ionic changes due to increased light intensities were essentially the same in all selected species, indicating similar characteristics of heir mineral ion and organic acid metabolism as well as in field-grown Carex species.

  • PDF