不同浓度的NO^—3和NH^＋4对小麦根谷氨酰胺合成酶及其相关酶的影响

利用酶活性测定和 Northern分子杂交等技术 ,研究了小麦幼苗根在不同浓度的 Na NO3 和(NH4) 2 SO4的供应下 ,其谷氨酰胺合成酶 (GS)、天冬酰胺合成酶 (AS)、谷氨酸脱氢酶 (GDH)、硝酸还原酶 (NR)以及 GS- m RNA的变化。结果表明 :NH 4 处理的小麦 ,其根部 GS活性比 NO-3 处理的高 ;高浓度处理的比低浓度处理的高 ;Northern杂交结果说明 GS- m RNA转录量与 GS活性一致 ;3mmol/ L NO-3促进了 AS的活性。AS酶活性变化与 GS酶活性变化无明显依赖关系。在实验的条件下 ,没能测出 GDH的活性 ,不同浓度的 NO-3 和 NH 4 处理对 NR活性没有明显的规律。     

不同NH4^＋／NO3^-比例的氮素营养对棉花氮素代谢的影响

The influence of different NH4^+／NO3^- ratios on nitrogen metabolism of cotton was studied under controlled hydroponics, The results showed that compared with single nitrate nutrition, solutions with 25/75, 50/50, 75/25 and 100/0 of NH4^+／NO3^- significantly increased the soluble protein accumulation in leaves and roots of cotton, and the maximum content of soluble protein in leaves and roots appeared respectively in the solution with 50/50 and 75/25 of NH4^+／NO3^- The soluble protein content in roots was increased with the increase of NH4+ percentage, but was slightly less in the solution of 100/0 than 75/25, which was probably related to the excess NH4^+ limiting boot metabolism. With the increase of NH4+ percentage, the nitrate content in petiole and the nitrate reductase activity in functional blade declined, but ammoniac nitrogen content increased in every organ of cotton. These results showed that foreign nitrogen affected the nitrogen metabolism of cotton in a different way, and the nitrogen absorption by cotton was probably related to different forms of foreign nitrogen.     

南瓜种子萌发及子叶发育时谷氨酰胺合成酶和其它氨同化酶的变化

在发育的新生组织中 ,来自种子胚乳储存蛋白的降解和氨基酸分解代谢产生的氨由谷氨酰胺合成酶 ( Glutamine synthetase,GS)重新同化 ,生成的谷氨酰胺 ( Gln)被转运到正在生长着的部分。GS是高等植物氮素代谢的关键酶 [1] ,这个酶能同化不同来源的氨。 GS有多种同工酶 ,存在于植物的各种组织和器官中。它们是由一小的同源但分离的核基因家族编码的 [2 3 ] ,这些不同的 GS在植物氮素同化中起着非重叠的作用 [4] ,它们的表达受到环境、发育进程以及组织或细胞类型等许多因素的影响。在大多数已研究过的植物叶片中存在两种 GS,即胞液型GS(…     

Growth and Major Nutrient Concentrations in Brassica campestris Supplied with Different NH4+/NO3- Ratios

In the present study, we investigated whether growth and main nutrient ion concentrations of cabbage (Brassica campestris L.) could be increased when plants were subjected to different NH4+/NO3- ratios. Cabbage seedlings The results showed that cabbage growth was reduced by 87% when the proportion of NH4+-N in the nutrient solution was more than 75% compared with a ratio NH4+/NO3- of 0.5:0.5 35 d after transplanting, suggesting a possible toxicity seedling weight, root length, and H2PO4- (P), K+, Ca2+, and Mg2+ concentrations were all higher than those in plants 0.5 NH4+/NO3-. The present results indicate that an appropriate NH4+/NO3- ratio improves the absorption of other nutrients and maintains a suitable proportion of N assimilation and storage that should benefit plant growth and the quality of cabbage as a vegetable.     

蔗糖对不同氮源培养下水稻根部氨同化相关酶活性的影响

糖、有机酸以及氨基酸影响碳-氮代谢过程中的相关酶的基因表达和活性。将蔗糖分别加入到含有相同氮素浓度的（NH4）2SO4（NH4^＋）或丙氨酸（Ala）作为氮源的营养液中培养水稻,测定幼苗根的谷氨酰胺合成酶（GS）、依赖于NADH的谷氨酸合酶（NADH-GOGAT）、磷酸烯醇式丙酮酸羧化酶（PEPC）和依赖于NADP的异柠檬酸脱氢酶（NADP-ICDH）的活性。结果显示,蔗糖诱导NH4^＋氮源中幼苗根的GS、NADH-GOGAT活性,抑制Ala氮源中幼苗根的这两种酶活性,蔗糖对PEPC和NADP-ICDH活性的影响也不同;未加蔗糖时,以Ala作为氮源的幼苗根的GS、NADH-GOGAT、PEPC和NADP-ICDH的活性明显高于以NH4^＋为氮源时的活性;生物量和蛋白质水平的变化与上述参数的变化基本一致。基于Ala碳骨架的存在,这些结果表明,碳／氮平衡是影响这些酶活性差别表达的主要原因。     

Effect of Nucleotides and Related Compounds on Glutamine Synthetase Activity in Chick Embryo Retina: A Biochemical and Immunohistochemical Study

Abstract: The effect of a number of nucleotides and related compounds on glutamine synthetase (GS) induction in retina from 12-day chick embryo was studied with both biochemical and immunohistochemical techniques. A number of these compounds gave rise to GS activity comparable to that induced by treatment with cortisol, which is known to give rise to precocious induction of the enzyme in this system. Of the cyclic nucleotides examined, cAMP (0.5-1.2 × 10^?3 M) gave essentially no increase in GS activity. In contrast, dibutyryl cAMP (0.8 × 10^?3 M) had a more significant effect on GS activity, as did 8-bromo-cAMP and cGMP at the same concentration. The activity elicited by these nucleotides was generally half that obtained by treatment with 2.8 × 10^?7 M-cortisol for the same length of time, 8-Bromo-cGMP (0.8 × 10^?3 M) had an effect comparable to the aforementioned concentration of cortisol. Since phosphodiesterase activity is minimal in the chick retina at 12 days of development, addition of isobutylmethylxanthine (1 × 10^?5 M) to this system had, as would be expected, little effect on GS activity. Of the noncyclic compounds, 8-bromoguanosine often gave rise to GS activity comparable to that obtained with cortisol. The other compounds (8-bromo-5′-GMP, guanosine, adenosine, and 5′-AMP) generally had less of an effect on GS. In general, the degree of staining in the immunohistochemical localization of GS corresponded well with the biochemical results and showed the enzyme to be present in regions consistent with the distribution of Muller cells and their processes. Thin-layer chromatography and radioimmunoassay of the nucleotides did not show any steroid impurity in any of the compounds used in the study, even when determinations were carried out at five times the concentration of nucleotide used in the experiments.     

高糖环境对Mu ller细胞谷氨酸转运体及谷氨酰胺合成酶表达的影响

目的：研究高糖环境对原代培养新生7天SD乳鼠视网膜Muller细胞谷氨酸转运合成系统的影响及其可能机制。方法：新生7天SD乳鼠视网膜Muller细胞原代培养并模拟高糖环境构建乳鼠视网膜muller细胞体外高糖环境模型。处理分为3组：对照组,高糖组,高糖＋白藜芦醇干预组。培养时间为24h,通过westernblot等检测方法,对照观察各组Muller细胞谷氨酸转运体（GLAST）、谷氨酰胺合成酶（GS）的表达情况。结果：模拟高糖环境可以造成新生SD乳鼠视网膜Muller细胞谷氨酸转运体（GLAST）表达的降低（0．225foldVScontrol,P〈0．05）,并导致其表达的谷氨酰胺合成酶（GS）表达水平的显著降低（0．653foldVScontrol,P〈0．05）;而干预药物白藜芦醇作用后可明显逆转新生SD乳鼠Mu ller细胞谷氨酸转运体（GLAST）（1．133foldvSHGgroup,P〈0．05）、谷氨酰胺合成酶（GS）（1．720foldVSHGgroup,P〈0．05）等蛋白的表达水平。结论：模拟高糖环境可以影响视网膜M0ller细胞谷氨酸转运体（GLAST）、谷氨酰胺合成酶的表达,其结局可能导致视神经细胞因谷氨酸堆积而导致的兴奋性毒性,白藜芦醇能提高Mcjller细胞谷氨酸转运体（GLAST）、谷氨酰胺合成酶表达,从而保护视神经细胞。     

不同光强下氮素形态对番茄谷氨酰胺合成酶和光呼吸的影响

强光(800μmol·m-2·s-1)下供应铵态氮的番茄植株与供应硝态氮的相比,其生长受到显著抑制,谷氨酰胺合成酶(GS)活性和光呼吸速率显著下降,同时净光合速率(Pn)和叶绿素荧光参数(Fv/Fm、Fv/F.)值下降;而弱光(200μmol·m-2·s-1)下供应铵态氮与硝态氮植株之间的这些参数差异不显著.     

The Influence of NO-3 and NH+4 Nutrition on the Gas Exchange Characteristics of the Roots of Wheat (Triticum aestivum) and Maize (Zea mays) Plants

Respiratory oxygen consumption by roots was 1·4- and1·6-fold larger in NH⁺₄-fed than in NO^-₃-fed wheat (Triticumaestivum L.) and maize (Zea mays L.) plants respectively. Higherroot oxygen consumption in NH⁺₄-fed plants than in NO^-₃-fedplants was associated with higher total nitrogen contents inNH⁺4-fed plants. Root oxygen consumption was, however, not correlatedwith growth rates or shoot:root ratios. Carbon dioxide releasewas 1·4- and 1·2-fold larger in NO⁺3-fed thanin NH⁺₄-fed wheat and maize plants respectively. Differencesin oxygen and carbon dioxide gas exchange rates resulted inthe gas exchange quotients of NH^-₄-fed plants (wheat, 0·5;maize, 0·6) being greatly reduced compared with thoseof NO^-₃-fed plants (wheat, 1·0; maize, 1·1). Measuredrates of HCO^-₃ assimilation by PEPc in roots were considerablylarger in 4 mM NH⁺₄-fed than in 4 NO^-₃ plants (wheat, 2·6-fold;maize, 8·3-fold). These differences were, however, insufficientto account for the observed differences in root carbon dioxideflux and it is probable that HCO^-₃ uptake is also importantin determining carbon dioxide fluxes. Thus reduced root extension in NH⁺₄-fed compared with NO^-₃-fedwheat plants could not be ascribed to differences in carbondioxide losses from roots.Copyright 1993, 1999 Academic Press Triticum aestivum, wheat, Zea mays, maize assimilation, ammonium assimilation, root respiration     

有机酸去除污泥重金属前后硝态氮和铵态氮浓度变化

研究了柠檬酸、草酸和乙酸溶液对污泥中重金属(Cd、Pb、Cu和Zn)的去除效果,以及处理前后析出液和污泥中硝态氮和铵态氮的浓度变化.结果表明,0.8mol.L-1柠檬酸溶液可去除污泥中76.0%的Pb和92.5%的Zn,是较好的重金属去除剂.污泥经有机酸处理后,有大量的硝态氮和铵态氮溶解于析出液中,与加入蒸馏水的对照处理相比,有机酸可大幅度增加析出液中铵态氮的含量,减少硝态氮含量.由于污泥处理过程中有其他形态的氮的转化,处理后污泥中仍含有较高浓度的硝态氮和铵态氮.0.5mol.L-1草酸处理的析出液中硝态氮和铵态氮浓度分别为2.8和888.1mg.L-1,且重金属含量不高,可作为较好的液体肥料进行回收利用.     

Effects of Different Nitrogen and Phosphorus Synergistic Fertilizer on Enzymes and Genes Related to Nitrogen Metabolism in Wheat

In recent years, in order to improve nutrient use efficiency, especially nitrogen use efficiency, fertilizer value-added technology has been developed rapidly. However, the mechanism of the effect of synergistic fertilizer on plant nitrogen utilization is not clear. A study was, therefore, conducted to explore the activities and gene expression of key enzymes for nitrogen assimilation and the gene expression of nitrogen transporters in wheat after the application of synergistic fertilizer. Soil column experiment was set up in Qingdao Agricultural University experimental base from October 2018 to June 2019. Maleic acid and itaconic acid were copolymerized with acrylic acid as cross-linking monomer to make a fluid gel, which was sprayed on the fertilizer surface to make nitrogen and phosphorus synergistic fertilizer. A total of 6 treatments was set according to different nitrogen and phosphorus fertilizer ratios: (1) 100% common nitrogen fertilizer+100% common phosphate fertilizer (2) 70% nitrogen synergistic fertilizer+100% phosphorus synergistic fertilizer; (3) 100% nitrogen synergistic fertilizer+70% phosphorus synergistic fertilizer; (4) 100% nitrogen synergistic fertilizer+100% phosphorus synergistic fertilizer; (5) 70% nitrogen synergistic fertilizer+70% phosphorus synergistic fertilizer; (6) 100% commercial nitrogen synergistic fertilizer+100% commercial phosphorus synergistic fertilizer. The results are as follows: (1) the enzyme activities of wheat plants under synergistic fertilizer condition were higher than those under ordinary fertilizer, except under the treatment that nitrogen and phosphorus synergistic fertilizer were both reduced; (2) the expression level of the genes under the treatment “100% nitrogen synergistic fertilizer+100% phosphorus synergistic fertilizer” was significantly higher than those in other treatments. Combined with the higher performance of nitrogen concentration in various parts of the plant under the condition of applying synergistic fertilizer, this study indicated that the application of synergistic fertilizer can improve the nitrogen metabolism of the plant by increasing the nitrogen level in the rhizosphere soil, inducing the expression of nitrogen transporter genes and key assimilation enzymes genes.

     

NaHSO3对盐胁迫下小麦幼苗氮同化酶及脯氨酸含量的影响

以普通小麦(Triticum aestivumL.)为材料,研究了NaHSO3对不同盐度胁迫下小麦幼苗氮素同化酶和脯氨酸含量的调节。结果表明,盐胁迫降低了叶片中硝酸还原酸(NR)的活性,加入NaHSO3之后,NR活性表现出进一步的降低。谷氨酰胺合成酶(GS)在低浓度盐胁迫下活性增加,在高浓度盐胁迫下活性降低;NaHSO3加入时,即便在低盐浓度下GS活性也降低。依赖于NADH的谷氨酸脱氢酶(NADH-GDH)和依赖于NADP的异柠檬酸脱氢酶(NADP-ICDH)的变化趋势一致,在盐胁迫下它们的活性都明显增加;NaHSO3加入促进了它们活性的进一步增加,尤其对NADH-GDH活性的促进更为明显。游离脯氨酸在高浓度盐胁迫下大量积累,在低浓度盐胁迫下含量增加不明显;NaHSO3促进了盐胁迫下脯氨酸的积累,提示了NaHSO3促进了盐胁迫下小麦幼苗碳氮营养元素的贮存。     

聚球藻7942高效泌氨突变种的获得及其泌氨、谷氨酰胺合成酶活性、光合和生长

将含有Ａｎａｂａｅｎａｓｐ．ＰＣＣ７１２０反义ｇｌｎＡ基因片段的穿梭表达质粒ｐＤＣ－ＡＴＧＳ转化单细胞蓝藻聚球藻Ｓｙｎｅ－ｃｈｏｃｏｃｃｕｓ　ｓｐ．ＰＣＣ７９４２,通过同源重组,外源ＤＮＡ定位整合到染色体上。经过抗菌素筛选,获得一种高效泌氨的Ｓｙｎｅｃhｏｃｏｃｃｕｓ　ｓｐ．７９４２突变种。将此突变种固定化在聚氨脂泡沫中后,定量测定其谷氨酰胺合成酶（ＧＳ）活性。结果表明,固定化后的突变藻培养９ｄ后泌氨活性比自由生活的野生藻高１５６倍,ＧＳ活性降低７３．６％;其生长速度与同条件下野生藻相近,７７Ｋ荧光光谱表明突变种固定化后光系统Ⅱ活性提高４４％。     

Establishment of a Highly Efficient Ammonia Secreting Mutant of Synechococcus sp. PCC 7942 and the Glutamine Synthetase Activity,Photosynthesis and Growth in Its Immobilized Cells

A recombinate plasmid pDC-ATGS was constructed, which contained the antisense fragment of glnA gene from Anabaena sp. PCC 7120 and transformed the unicellular cyanobactefium Synechococcus sp. PCC 7942. The foreign DNA was inserted into the site of glnA locus of the chromosome through the homologous recombination. By using neomyisin, a highly efficient ammonia secretion mutant was selected. After immobilized, the cells of the mutant within polyurethane (PU) foams, glutamine synthetase (GS) and NIt4+ secretory activity of GS, and its growth and photosynthesis were measured. It was shown that NH4+ secretion of the immobilized mutant was enhanced 156 folds which was much higher than that of free-living cells of the wild type. The activity of GS was decreased by 73.6%. Growth of the mutant was the same as that of the wild type. The activity of photosystem Ⅱ in the immobilized mutant cells increased by 44% with 77 K fluorescence spectrum measurement.     

Effect of Cortico-Striate Pathway Lesion on the Activities of Enzymes Involved in Synthesis and Metabolism of Amino Acid Neurotransmitters in the Striatum

The activities of several enzymes involved in the metabolism of aspartate and glutamate were measured in striatal (nucleus caudatus and putamen) homogenates 2-3, 6-7, and 35-40 days following frontoparietal and frontal cortical ablation. The activity of glutamine synthetase (GS) was substantially increased (46-48%) on the operated side 6-7 days following the lesion whereas smaller changes were observed at 2-3 and 35-40 days after lesion. In contrast, decreased levels of glutaminase and malate dehydrogenase (MDH) were observed by 6-7 days while no significant change was found at either 2-3 or 35-40 after the lesion. The activities of glutamate dehydrogenase (GDH) and glutamate decarboxylase (GAD) were elevated after 35-40 days whereas no changes in the levels of either GDH or aspartate aminotransferase (ASAT) were found at 2-3 or 6-7 days after the fronto-parietal decortication. When only the frontal cortex was removed quantitatively similar changes were observed in striatal GS and glutaminase activity. The content of glutamate and glutamine in the denervated striatum followed qualitatively the changes in glutaminase and GS. The results indicate that the degeneration of cortico-striatal terminals causes a profound glial reaction in the striatum, and both glutaminase and MDH are present in relatively high concentrations in the corticostriatal terminals.     

Effect of divalent cations on the short-term NH 4 + inhibition of nitrogen fixation in Azotobacter chroococcum

Incubation of Azotobacter chroococcum in the presence of micromolar concentrations of MnCl₂, but not MgCl₂, prevented nitrogenase activity from NH ₄ ⁺ inhibition. Mg(II), at a 100-fold concentration with respect to Mn(II), counteracted the protective effect of Mn(II) on nitrogenase activity. When Mn(II) was added to cells that had been given NH₄Cl, stopping of NH ₄ ⁺ uptake and recovery of nitrogenase activity took place, and a raise of NH ₄ ⁺ concentration in medium developed. Furthermore, incubation of A. chroococcum cells with 20 M Mn(II) under air, but not under an argon: oxygen (79%:21%) gas mixture, resulted in NH ₄ ⁺ excretion to the external medium. The Mn(II)-mediated uncoupling of nitrogen fixation from ammonium assimilation leads us to conclude that Mn(II) may act as a physiological inhibitor of glutamine synthetase.Abbreviations Hepes N-2-Hydroxyethylpiperazine-N-ethanesulfonic acid - Mops 3-(N-Morpholino)propanesulfonic acid     

Alteration of cyanobacterial glutamine synthetase activity in vivo in response to light and NH 4 +

Extractable glutamine synthetase activity of the cyanobacterium Anabaena cylindrica was reduced by approximately 50% when N₂-fixing cultures were treated with 10 mM NH ₄ ⁺ or were placed in darkness. The deactivated enzyme could be rapidly reactivated (within 5 min) by adding 40 mM 2-mercaptoethanol to the biosynthetic reaction mixture. The enzyme could also be reactivated in vivo by replacing the culture in light or by removing NH ₄ ⁺ . When the enzyme was deactivated by simultaneously adding NH ₄ ⁺ and placing the culture in darkness, reactivation occurred on reillumination and removal of NH ₄ ⁺ . The removal of NH ₄ ⁺ in darkness did not result in reactivation. On in vitro reactivation of glutamine synthetase from dark or NH ₄ ⁺ -treated cultures the maximum glutamine synthetase activity observed frequently exceeded that of glutamine synthetase extracted from untreated cultures. Anacystis nidulans showed a similar type of reversible dark deactivation to A. cylindrica but Plectonema boryanum and a Nostoc did not. With A. cylindrica, a direct positive correlation between the size of the intracellular pool of glutamate and biosynthetic glutamine synthetase activity occurred during light/dark shifts, and on treatment with NH ₄ ⁺ . The changes in activity of glutamine synthetase in A. cylindrica in response to light resemble in some respects the light modulation of enzymes of the oxidative and reductive pentose phosphate pathways noted in cyanobacteria by others.     

Influences of different nitrogen sources on nitrogen- and water-use efficiency, and carbon isotope discrimination, in C3 Triticum aestivum L. and C4 Zea mays L. plants

The impacts of various nitrogen sources, i.e. NO⁻ ₃, NH₄ ⁺ or NH₄NO₃ in combination with gaseous NH₃, on nitrogen-, carbon- and water-use efficiency and ¹³C discrimination (δ¹³C) by plants of the C₃ species Triticum aestivum L. (wheat) and the C₄ species Zea mays L. (maize) were studied. Triticum aestivum and Z. mays were hydroponically grown with 2 mol · m⁻³ of N supplied as NO⁻ ₃, NH₄ ⁺ or NH₄NO₃ for 21 and 18 d, respectively, and thereafter exposed to gaseous NH₃ at 320 μg · m⁻³ or to ambient air for 7 d. In T. aestivum and Z. mays over a 7-d growth period, nitrogen-use efficiency (NUE) values were influenced by N-sources in the decreasing order NH₄NO₃-N > NO⁻ ₃-N > NH₄ ⁺-N and NO⁻ ₃-N > NH₄NO₃-N > NH₄ ⁺-N, respectively. Fumigation with NH₃ decreased the NUE values of plants grown with any of the N-forms. During 28- and 7-d growth periods, N-sources affected water-use efficiency (WUE) values in the decreasing order of NH₄ ⁺-N > NO⁻ ₃-N≈NH₄NO₃-N in non-fumigated T. aestivum, while fumigation with NH₃ increased the WUE of NO⁻ ₃-grown plants. There were insignificant effects of N-sources on WUE values of Z. mays over 25- and 7-d growth periods. Furthermore, δ¹³C values in plant tissues (leaves, stubble and roots) were higher (less negative) in NH₄ ⁺-grown plants of T. aestivum and Z. mays than in those supplied with NH₄NO₃ or NO⁻ ₃. Regardless of the N-form supplied to the roots of the plant species, exposure to NH₃ caused more-positive δ¹³C values in the plant tissues. These results indicate that the variations in N-source were associated with small but significant variations in δ¹³C values in plants of T. aestivum and Z. mays. These differences in δ¹³C values are in the direction expected from differences in WUE values over long or short growth periods and with differences in the extent of non-Rubisco (ribulose-1,5-bisphosphate carboxylase-oxygenase, EC 4.1.1.39) carboxylate contribution to net C acquisition, as a function of N-source. Received: 12 September 1997 / Accepted: 13 January 1998