• Title/Summary/Keyword: nitrate reductase (NR) activity

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Cloning and Sequence Analysis of Spinach (Spinacia oleracea L. cv Ace) Nitrate Reductase cDNA (시금치 nitrate reductase cDNA 클로닝 및 염기서열 분석)

  • Park, Nu-Ri;Chung, Jong-Bae;Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.45 no.3
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    • pp.129-133
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    • 2002
  • Suppression of nitrate accumulation in spinach and lettuce through foliar application of chitosan formula containing micronutrients is related with the increase of the nitrate reductase (NR) activity. If NR in spinach were highly expressed to increase the assimilatory activity, nitrate content could be reduced. For this, NR cDNA was cloned from the isolated mRNAs of spinach using reverse transcriptase-PCR. Nucleotide sequence of cloned spinach NR cDNA showed highly deduced amino acid sequence identity ($71{\sim}82%$) with other known plant NR genes. Only two nucleotide-base differences were observed in the cloned NR cDNA compared with that of the published spinach NR cDNA.

Production and Characterization of Nitrate Reductase Deficient Mutants in Petunia parviflora

  • Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.6
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    • pp.706-715
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    • 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.

Analysis of Nitrate Reductase Activity for Dominant Tree Leaves in the Northern Aspect Forest of Changbai Mountain, China (중국 장백산 북사면 산림에서 우세목의 잎 내 질소 환원 효소 활성도 분석)

  • wen, Li-Yu;Kim, Ji-Hong
    • Journal of Korea Foresty Energy
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    • v.22 no.3
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    • pp.29-36
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    • 2003
  • This paper concerned the application of improvement in Vivo of Traditional Method for determination of nitrate reductase (NR) activity of leaves to dominant tree species in five forest communities of northern aspect of Changbai Mountain. The results indicated that the NR activity of tree species was related to shade tolerance, and the intolerant tree species had higher NR activity. The NR of a species was also related to the vertical structure and ecological site condition. The tree species, which have higher NR activities should be selected for fast growing and high yield tree species.

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Verticillium dahliae toxins-induced nitric oxide production in Arabidopsis is major dependent on nitrate reductase

  • Shi, Fu-Mei;Li, Ying-Zhang
    • BMB Reports
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    • v.41 no.1
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    • pp.79-85
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    • 2008
  • The source of nitric oxide (NO) in plants is unclear and it has been reported NO can be produced by nitric oxide synthase (NOS) like enzymes and by nitrate reductase (NR). Here we used wild-type, Atnos1 mutant and nia1, nia2 NR-deficient mutant plants of Arabidopsis thaliana to investigate the potential source of NO production in response to Verticillium dahliae toxins (VD-toxins). The results revealed that NO production is much higher in wild-type and Atnos1 mutant than in nia1, nia2 NR-deficient mutants. The NR inhibitor had a significant effect on VD-toxins-induced NO production; whereas NOS inhibitor had a slight effect. NR activity was significantly implicated in NO production. The results indicated that as NO was induced in response to VD-toxins in Arabidopsis, the major source was the NR pathway. The production of NOS-system appeared to be secondary.

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
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    • v.34 no.1
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    • pp.49-57
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    • 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.

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Some Aspects to the in vivo Nitrate Reductase Activity in Carex species (사초속 식물의 질산환원효소 활성의 특징)

  • 추연식
    • Journal of Life Science
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    • v.10 no.1
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    • pp.52-60
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    • 2000
  • Up to now, there have been done much efforts in regard to nitrate reductase activity (NRA) of dicotyledonous herbs and important crop monocotyledons, but few to wild plants having canopy structure such as Carex. The objective of the present study are to determine: a) the optimum in vivo NR assay conditions for leaf samples of Carex species, b) changes of NRA according to section within leaf and leaf ages, c) diurnal variations. Optimized assay media of each Carex species were determined. NRA of C. rostrata adapted to oligotrophic habitats is readily saturated at lower substrate concentration than those of C. distans and C. gracilis, adapted to meso- and eutrophic habitats, respectively. All Carex species investigated have higher NRA in leaves than in roots. NRA of all species showed maximal values at the middle section of each leaf and in the youngest fully expanded leaves. Compared to C. gracilis, NR in leaves of C. distans was adapted readily to the light period. On the whole, Carex showed rather delayed diurnal variation. Even if the in vivo nitrate reductase assay based on nitrite estimation does not give an accurate estimation of total nitrate reduced, it still serves as a useful tool to find out relative differences in varying environmental conditions. Additionally, in vivo RNA measurements are helpful to understand nitrate reduction and basic nitrogen metabolism of Carex species having different canopy structure.

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Effects of Nrogen Form and Light Conditions on the Nitrate Reductase Activity of Ulva pertusa (Chlorophyta) and Ecklonia cava (Phaeophyta) (구멍갈파래 (Ulva pertusa Kjellman)와 감태 (Ecklonia cava Kjellman)의 질산환원요소 활성에 미치는 질소원 형태와 빛의 효과)

  • Hwang, Jae-Ran;Kang, Yun-Hee;Oak, Jung-Hyun;Lee, Sang-Rae;Chung, Ik-Kyo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.1
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    • pp.64-70
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    • 2011
  • Nitrate reductase (NR) is activated by nitrogen sources (${NO_3}^-$ and ${NH_4}^+$) and irradiance. This study investigated the effects of these factors on the NR activity of Ulva pertusa (Chlorophyta) and Ecklonia cava (Phaeophyta). In addition, the ammonium (${NH_4}^+$) and nitrate (${NO_3}^-$) uptake rates of the two species were examined. U. pertusa took up most of the ${NO_3}^-$ and ${NH_4}^+$ in the medium during a 3hour incubation, while E. cava had a relatively high uptake rate after 3 hours. The NR activities of the two species were affected by the nitrogen source and irradiance and were highest when they were exposed to ${NO_3}^-$-rich medium and high irradiance. However, the patterns of NR activity differed between the two species. In ${NO_3}^-$-rich medium and high irradiance, U. pertusa achieved the highest NR activity ($2.01{\pm}0.07\;{\mu}mol$ ${NO_2}^-$ $g^{-1}$ DW $h^{-1}$) within the first 3 hours and then this activity decreased drastically. By contrast, the NR activity of E. cava ($0.36{\pm}0.04\;{\mu}mol$ ${NO_2}^-$ $g^{-1}$ DW $h^{-1}$) was constant for 12 hours. When exposed to darkness, the NR activity of U. pertusa decreased dramatically, while that of E. cava increased gradually for 12 hours. Therefore, E. cava is able to maintain NR activity during the dark because of its adequate carbohydrate reserves and substrate.

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
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    • v.6 no.4
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    • pp.259-264
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    • 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
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    • v.52 no.2
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    • pp.153-161
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    • 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.

Effects of added chlorides and their concentrations on growth and nitrate content in leaf lettuce(Lactuca sativa L.) in hydroponics.

  • Lee, Guang-Jae;Shin, Hyun-Man;Kim, Ki-Sik;Lee, Cheol-Hee;Kim, Jin-Han
    • Plant Resources
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    • v.2 no.1
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    • pp.26-30
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    • 1999
  • Nitrate in edible vegetables is converted to nitrite by nitrate reductase(NR) and/or bacteria in intestines. Nitrite and amino, in the intestine of some animals and human, bind to form nitrosamine, which is toxic and known as carcinogen. This study was carried out to examine the effect of added chlorides and their concentrations on growth, yield and nitrate content in leaf lettuce plants in hydroponics. Seeds of lettuce cv, "Samsunjokchukmyon" were planted on April 29, and seedlings were planted on June 2, and were cultured until July 5 in 1998. KCI and CaCl₂ were used as chloride source and their concentrations were 1, 2 and 4 me/L, respectively, in the lettuce standard nutrient solution for National Horticultural Research Institute(NHRI). Completely randomized design with 3 replications was used. Nitrate content and NR activity were measured 2 and 5 weeks after planting(WAP). The obtained results were summarized as the follows : Leaf weight per plant was difference from harvest dates and treatments, but total leaf weight was not significantly different among treatments. Number of leaves was higher in KCI 2 me/L, CaCl₂1 me/L and control at 2 WAP than the others, and was higher in KCI 1 me/L, and control at 3 WAP than others, and was higher in control at 5 WAP. Total number of harvested leaves was the highest in control with 14, which followed by KCI 2 me/L and CaCl₂1 me/L. Nitrate content was decreased by addition of chloride in nutrient solution. Nitrate content in the 3rd and 9th leaves was significantly decreased. NR activity was higher in control and CaCl₂ addition treatments, while KCI addition treatments reduced NR activity. However, no direct relationship with nitrate was observed. Growth characteristics such leaf length and leaf width were not significantly influenced by chloride addition.

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