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。     

不同耐盐性水稻幼苗根氨同化酶对盐胁迫的反应

在盐胁迫下,检测了耐盐性不同的水稻(Oryza sativa L.)品种根部氨同化酶及其相关参数的变化。结果表明,根的可溶性蛋白、谷氨酰胺合成酶(GS)及依赖于NADH的谷氨酸合酶(NADH-GOGAT)活性在高盐浓度下不同程度地降低,其影响大小依次为早花二号(盐敏感品种)、金珠一号(正常栽培品种)、津稻779(耐盐品种),与其耐盐性相一致。在盐胁迫条件下,在耐盐性较高的水稻品种中, GS和GOGAT活性比盐敏感品种高,NH4 浓度维持在较低的水平。Native-PAGE和活性染色结果表明,GSrb更容易受到外界环境的影响。在高浓度盐的胁迫下,早花二号、金珠一号的依赖于NADH的谷氨酸脱氢酶(NADH-GDH)活性都有较显著的升高,津稻779却无明显的变化,这和NH4 含量的变化相一致。盐不同程度地导致可溶性糖(TSS)在金珠一号和津稻779根部积累,而在早花2号的根部,可溶性糖的水平则随盐浓度的不同而表现出不同的变化。在所检测的品种中,脯氨酸的含量均有不同程度的升高,但在高盐浓度下,盐敏感品种的含量较低。这些结果提示,不同的水稻品种对盐胁迫的敏感程度与该品种GS以及GOGAT活性的高低有关。     

外源钙离子对小麦幼苗氮素代谢的影响

以普通小麦豫麦34为材料,研究了不同浓度的外源Ca2 对小麦幼苗氮素代谢的影响.在小麦第一片叶完全展开后,开始外源Ca2 处理,设0 (对照)、2、4 mmol · L-1 和8 mmol · L-1 4个Ca2 浓度梯度.处理5d后,测定氮同化酶活性、氮同化量及其它相关代谢物含量.结果表明,小麦幼苗叶片中硝酸还原酶(NR)和谷氨酰胺合成酶(GS)在2 mmol · L-1 Ca2 处理下活性比对照有显著增加,4 mmol · L-1 Ca2 处理的NR活性增加明显,但GS活性增加不显著;8 mmol · L-1 Ca2 处理下NR和GS活性比对照均明显降低.谷氨酸脱氢酶(NADH-GDH)活性在2 mmol · L-1 Ca2 处理下活性增加不明显,而在4、8 mmol · L-1 Ca2 处理下活性显著增加.小麦幼苗氮同化量以4 mmol · L-1处理最大,2 mmol · L-1处理与4 mmol · L-1之间差异不显著;Ca2 浓度为8 mmol · L-1时,氮素同化量明显降低.结果揭示了小麦幼苗不同氮同化途径对Ca2 的响应不同,GS途径比GDH途径对小麦氮素同化量的增加作用更大;4 mmol · L-1对小麦幼苗的氮素利用可能是最有效的Ca2 浓度.     

干旱胁迫与小麦幼苗亚精胺代谢变化的关系

通过对亚精胺（spermidine,Spd）在抗旱性不同的小麦品种幼苗中的作用的研究,发现抗旱品种周麦18号在渗透胁迫处理时,其叶片中的Spd含量明显大于不抗旱的豫麦51号。用Spd合成的抑制剂甲基乙二醛-双（鸟嘌呤腙MGBG）处理周麦18号,则导致Spd含量下降和抗性的降低,外源Spd又可逆转MGBG对周麦18号在渗透胁迫下的伤害。外源Spd可以明显提高豫麦51号的叶片内Spd含量,并相应提高其抗性。以上结果表明,Spd可以提高小麦幼苗的抗渗透胁迫能力。     

钙离子对盐胁迫小麦幼苗氮代谢的影响

为探讨增强小麦抗盐能力的调控途径,以普通小麦豫麦34为材料,研究了Ca2+对盐胁迫下小麦幼苗氮代谢及生长的影响.采用全营养液培养小麦幼苗至第一片叶完全展开,更换无钙营养液,并开始不同处理.处理分别为低盐胁迫(150 mmol · L-1 NaCl)、低盐胁迫+4 mmol · L-1 Ca2+、高盐胁迫(300 mmol · L-1 NaCl)、高盐胁迫+4mmol · L-1 Ca2+,以无NaCl胁迫的小麦为对照.5 d后取样,测定了氮同化酶活性、代谢物含量、积累量及幼苗生长状况.结果表明,Ca2+明显缓解了低盐胁迫对小麦幼苗的生长抑制,表现在鲜重、叶绿素及可溶性蛋白含量的增加,而对高盐胁迫下小麦幼苗的生长无明显改善效果;Ca2+改善了低盐胁迫下小麦幼苗的氮营养状况,表现在氮积累量的增加,这一效应主要是通过硝酸还原酶(NR)、谷氨酰胺合成酶(GS)以及异柠檬酸脱氢酶(NADP-ICDH)活性的增强而实现的.Ca2+未能改善高盐胁迫下小麦幼苗氮营养状况的主要限制因子在于NADP-ICDH活性未明显增加.     

不同形态氮素对专用型小麦花后氮代谢关键酶活性及籽粒蛋白质含量的影响

采用盆栽方法研究了氮素形态对不同专用型小麦开花后氮素同化关键酶活性及籽粒蛋白质含量的影响。结果表明:不同专用型小麦氮素同化关键酶硝酸还原酶、谷氨酰胺合成酶和谷氨酸合酶对氮素形态的反应不同。强筋小麦豫麦34施用酰胺态氮对旗叶硝酸还原酶和谷氨酰胺合成酶活性、籽粒谷氨酰胺合成酶和谷氨酸合酶活性具有明显的促进作用,最终籽粒蛋白质含量较高;中筋小麦豫麦4 9在施用铵态氮时,3种氮素同化关键酶活性均有较大增强,籽粒蛋白质含量最高;弱筋小麦豫麦5 0硝酸还原酶活性以铵态氮处理最高,而籽粒和旗叶谷氨酰胺合成酶和谷氨酸合酶活性在酰胺态氮处理下明显增强,酰胺态氮对籽粒中蛋白质含量的增加具有明显的促进作用。相关性分析表明,籽粒蛋白质含量与旗叶GS活性和籽粒GOGAT活性呈显著或极显著正相关,与旗叶NR活性和GS活性、籽粒GOGAT活性相关性不显著     

渗透胁迫下小麦根系渗透调节与根冠淀粉水解的研究

用不同浓度的PEG—600对抗旱性不同的小麦幼苗进行渗透胁迫处理,研究了小麦幼苗根系的淀粉酶活性、可溶性糖含量、渗透势、渗透调节能力和根冠淀粉的水解状况。结果表明,随着渗透胁迫程度的加重,抗旱性强的小麦品种昌乐5号和北农2号根系渗透势和饱和渗透势的降低程度大于抗旱性弱的小麦品种鲁麦5号和921842,并且抗旱性强的小麦品种根系的渗透调节能力大于抗旱性弱的小麦品种。随着渗透胁迫程度的加重．各品种小麦根冠淀粉粒均有不同程度的减少。而抗旱性强的品种根冠淀粉粒的减少程度小于抗旱性弱的品种;抗旱性强的小麦品种根系淀粉酶活性显著高于抗旱性弱的小麦品种,但是,随着渗透胁迫程度的加重,抗旱性弱的品种淀粉酶活性增加的幅度远高于抗旱性强的品种。可溶性糖含量的变化趋势与淀粉酶活性的变化趋势一致．即渗透胁迫下根冠淀粉水解程度大的小麦品种,可溶性糖的含量高。但根冠淀粉水解在根系的渗透调节以及在小麦适应水分胁迫中的作用还有待于进一步探讨。     

水分胁迫对小麦幼苗叶片多胺含量的影响

水分胁迫下小麦幼苗叶片多胺含量变化的研究表明,聚乙二醇(PEG)的渗透胁迫明显提高了抗旱性强的周麦系列幼苗叶片游离态Put、Spd和Spm的含量和抗旱性弱的温麦6号的Put含量。外源Spd显著提高了水分胁迫下温麦6号的Spd的含量,对其抗性也有所改善。外源MGBG(Spd和Spm生物合成抑制剂)可提高水分胁迫下周麦12号Put的含量,但降低了Spd和Spm的含量和幼苗的抗性。     

不同耐盐性水稻幼苗根氨同化酶对盐胁迫的反应

在盐胁迫下,检测了耐盐性不同的水稻(Oryza sativa L.)品种根部氨同化酶及其相关参数的变化.结果表明,根的可溶性蛋白、谷氨酰胺合成酶(GS)及依赖于NADH的谷氨酸合酶(NADH-GOGAT)活性在高盐浓度下不同程度地降低,其影响大小依次为早花二号(盐敏感品种)、金珠一号(正常栽培品种)、津稻779(耐盐品种),与其耐盐性相一致.在盐胁迫条件下,在耐盐性较高的水稻品种中,GS和GOGAT活性比盐敏感品种高,NH4 浓度维持在较低的水平.Native-PAGE和活性染色结果表明,GSrb更容易受到外界环境的影响.在高浓度盐的胁迫下,早花二号、金珠一号的依赖于NADH的谷氨酸脱氢酶(AADH-GDH)活性都有较显著的升高,津稻779却无明显的变化,这和NH4 含量的变化相一致.盐不同程度地导致可溶性糖(TSS)在金珠一号和津稻779根部积累,而在早花2号的根部,可溶性糖的水平则随盐浓度的不同而表现出不同的变化.在所检测的品种中,脯氨酸的含量均有不同程度的升高,但在高盐浓度下,盐敏感品种的含量较低.这些结果提示,不同的水稻品种对盐胁迫的敏感程度与该品种GS以及GOGAT活性的高低有关.     

盐旱复合胁迫对小麦幼苗生长和水分吸收的影响

为明确盐害、干旱及盐旱复合胁迫对小麦幼苗生长和水分吸收的影响,从而为盐害和干旱胁迫下栽培调控提供理论依据。以2个抗旱性不同的小麦品种(扬麦16和耐旱型洛旱7号)为材料,采用水培试验,以NaCl和PEG模拟盐旱复合胁迫,研究了盐旱复合胁迫下小麦幼苗生长、根系形态、光合特性及水分吸收特性的变化。结果表明,盐、旱及复合胁迫下小麦幼苗的生物量、叶面积、总根长与根系表面积、叶绿素荧光和净光合速率均显著下降,但是复合胁迫处理的降幅却显著低于单一胁迫。盐旱复合胁迫下根系水导速率和根系伤流液强度显著大于单一胁迫,从而提高了小麦幼苗叶片水势和相对含水量。盐胁迫下小麦幼苗Na~+/K~+显著大于复合胁迫,但复合胁迫下ABA含量却显著小于单一的盐害和干旱胁迫。因此,盐旱复合胁迫可以通过增强根系水分吸收及降低根叶中ABA含量以维持较高光合能力,这是盐旱复合胁迫提高小麦适应性的重要原因。洛旱7号和扬麦16对盐及盐旱复合胁迫的响应基本一致,但在干旱胁迫下洛旱7号表现出明显的耐性。     

Regulation of ammonia assimilation in an obligate methylotroph Methylobacillus flagellatum under steady-state and transient growth conditions

The obligate methylotroph Methylobacillus flagellatum was grown in the presence of different ammonium concentrations and the regulation of the enzymes associated with ammonium assimilation was investigated in steady-state and transient growth regimes. As the medium changed from C-limitation to dual C/N- and finally to N-limitation, the culture passed through three definite growth phases. The NADP⁺-dependent glutamate dehydrogenase (GDH) was present under ammonium limitation of the culture growth (at 2 mmol l^-1 of ammonium in the growth medium) and increased in response to an increase in nitrogen availability. Glutamine synthetase (GS) and glutamate synthase (GOGAT) activities were negligible during C- and C/N-limitation. In N-limited cells the GOGAT activity increased as the dilution rate increased up to 0.35 h^-1, and then sharply dropped. In the N-sufficient cultures both NAD⁺- and NADP⁺-dependent isocitrate dehydrogenase (NAD-ICDH and NADP-ICDH) activities were up-regulated as dilution rate increased, but in the N-limited culture the NAD-ICDH activity was up-regulated whereas NADP-ICDH one was down-regulated. Pulse additions of ammonium and methanol demonstrated the coordinate regulation of the GDH and ICDHs activities. When pulses were added to the C/N-limited cultures, there was an immediate utilization of the nutrients, resulting in an increase in biomass; at the same time the GDH and ICDH activities increased and the GS and GOGAT activities decreased. When the same ammonium/methanol pulse was added into the N-limited culture, there was a 3-hours delay in the culture response, after which the substrates were utilized at rates close to the ones shown by the C/N-limited culture after the analogous pulse.     

A study of the role of glutamate dehydrogenase in the nitrogen metabolism of Arabidopsis thaliana

Glutamate-dehydrogenase (GDH, EC 1.4.1.2) activity and isoenzyme patterns were investigated in Arabidopsis thaliana plantlets, and parallel studies were carried out on glutamine synthetase (GS, EC 6.3.1.2). Both NADH-GDH and NAD-GDH activities increased during plant development whereas GS activity declined. Leaves deprived of light showed a considerable enhancement of NADH-GDH activity. In roots, both GDH activities were induced by ammonia whereas in leaves nitrogen assimilation was less important. It was demonstrated that the increase in GDH activity was the result of de-novo protein synthesis. High nitrogen levels were first assimilated by NADH-GDH, while GS was actively involved in nitrogen metabolism only when the enzyme was stimulated by a supply of energy, generated by NAD-GDH or by feeding sucrose. When methionine sulfoximine, an inhibitor of GS, was added to the feeding solution, NADH-GDH activity remained unaffected in leaves whereas NAD-GDH was induced. In roots, however, there was a marked activation of GDH and no inactivation of GS. It was concluded that NADH-GDH was involved in the detoxification of high nitrogen levels while NAD-GDH was mainly responsible for the supply of energy to the cell during active assimilation. Glutamine synthetase, on the other hand was involved in the assimilation of physiological amounts of nitrogen. A study of the isoenzyme pattern of GDH indicated that a good correlation existed between the relative activity of the isoenzymes and the ratio of aminating to deaminating enzyme activities. The NADH-GDH activity corresponded to the more anodal isoenzymes while the NAD-GDH activity corresponded to the cathodal ones. The results indicate that the two genes involved in the formation of GDH control the expression of enzymes with different metabolic functions.Abbreviations GDH glutamate dehydrogenase - GS glutamine synthetase - MSO methionine sulfoximine     

Nickel and Al-excess inhibit nitrate reductase but upregulate activities of aminating glutamate dehydrogenase and aminotransferases in growing rice seedlings

The present study was undertaken to examine the influence of toxic levels of Ni and Al, on the activities of key nitrogen assimilatory enzymes in roots and shoots of growing rice seedlings. When seedlings of two inbred rice (Oryza sativa L.) cvs. Malviya-36 and Pant-12, sensitive to both Ni and Al, were raised in sand cultures containing 200 and 400 μM NiSO₄ or 80 and 160 μM Al₂(SO₄)₃, a marked inhibition in the activities of NO₃ ⁻ assimilatory enzymes NR and GS was observed in roots as well as shoots during a 5–20 day growth period. Both Ni and Al treatments, in growth medium, stimulated the activity of aminating glutamate dehydrogenase (NADH-GDH) whereas the activity of deaminating GDH (NAD⁺-GDH) decreased under metal toxicities. The activities of the aminotransferases studied; alanine aminotransferase (AlaAT) and aspartate amino transferase (AspAT) increased due to Ni and Al treatments. Results suggest that both Ni and Al treatments impair N assimilation in rice seedlings by inhibiting the activities of NR and GS and that GDH appears to play a role in assimilation of NH₄ ⁺ in metal stress conditions. Further, higher activity of aminotransferases in metal stressed seedlings might be helpful in meeting higher demand of amino acids under stressed conditions.     

Glutamine synthetase and glutamate dehydrogenase contribute differentially to proline accumulation in leaves of wheat (Triticum aestivum) seedlings exposed to different salinity

To investigate the roles of ammonium-assimilating enzymes in proline synthesis under salinity stress, the activities of glutamine synthetase (GS; EC 6.3.1.2) and NADH-dependent glutamate dehydrogenase (NADH-GDH; EC 1.4.1.2) were determined in leaves of wheat (Triticum aestivum) seedlings exposed to salt stress at 150 and 300 mM NaCl for 5d. At the lower salinity, only GS activity increased markedly. At 300 mM NaCl, however, NADH-GDH activity increased while GS activity decreased. A significant accumulation of proline was found only at high-salinity exposure while glutamate, a proline precursor, increased dramatically under both low and high salinity. These data suggests that GS-catalysis might be the main glutamate synthesis pathway under low salinity. At 300 mM NaCl, glutamate seems to be preferentially produced through the process catalyzed by NADH-GDH. The increase of ammonium in salinity-stressed wheat seedlings might have resulted from increased photorespiration, which is responsible for the higher NADH-GDH activity. The activity of Delta(1)-pyrroline-5-carboxylate reductase (P5CR; EC 1.5.1.2) was significantly enhanced at 300 mM NaCl but remained unchanged at 150 mM. Delta(1)-Pyrroline-5-carboxylate synthetase (P5CS) activity did not show a specific response, indicating that P5CR might be the limiting step in proline synthesis from glutamate at high salinity.     

Nitrogen metabolism in leaves of a tank epiphytic bromeliad: characterization of a spatial and functional division

The leaf is considered the most important vegetative organ of tank epiphytic bromeliads due to its ability to absorb and assimilate nutrients. However, little is known about the physiological characteristics of nutrient uptake and assimilation. In order to better understand the mechanisms utilized by some tank epiphytic bromeliads to optimize the nitrogen acquisition and assimilation, a study was proposed to verify the existence of a differential capacity to assimilate nitrogen in different leaf portions. The experiments were conducted using young plants of Vriesea gigantea. A nutrient solution containing NO₃⁻/NH₄⁺ or urea as the sole nitrogen source was supplied to the tank of these plants and the activities of urease, nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (NADH-GDH) were quantified in apical and basal leaf portions after 1, 3, 6, 9, 12, 24 and 48 h. The endogenous ammonium and urea contents were also analyzed. Independent of the nitrogen sources utilized, NR and urease activities were higher in the basal portions of leaves in all the period analyzed. On the contrary, GS and GDH activities were higher in apical part. It was also observed that the endogenous ammonium and urea had the highest contents detected in the basal region. These results suggest that the basal portion was preferentially involved in nitrate reduction and urea hydrolysis, while the apical region could be the main area responsible for ammonium assimilation through the action of GS and GDH activities. Moreover, it was possible to infer that ammonium may be transported from the base, to the apex of the leaves. In conclusion, it was suggested that a spatial and functional division in nitrogen absorption and NH₄⁺ assimilation between basal and apical leaf areas exists, ensuring that the majority of nitrogen available inside the tank is quickly used by bromeliad's leaves.     

High temperature effects on ammonium assimilation in leaves of two Festuca arundinacea cultivars with different heat susceptibility

The aim of this study was to determine the effects of high temperature stress on ammonium assimilation in leaves of two tall fescue cultivars (Festuca arundinacea), Jaguar 3 brand (J3) (heat-tolerant) and TF 66 (T6) (heat-sensitive). High temperature stress for either 10 d or 20 d, and particularly the 20 d stress, produced dramatic changes in ammonium assimilation. After 20 d of stress treatment, the accumulations of total nitrogen, nitrate, soluble protein and total free amino acid (20 amino acids) decreased in both cultivars. Moreover, the activities of main regulatory enzymes, such as nitrate reductase, glutamine synthetase (GS), NADH-dependent glutamate synthase (GOGAT), as well as Δ¹-pyrroline-5-carboxylate reductase (P5CR), also decreased in both cultivars when exposed to 20 d stress. Heat stress had little influence on ammonium accumulation in J3, but this was not the case with T6. The accumulations of nitrate, ammonium, soluble protein, and total free amino acid between the two cultivars were different. This suggests that accumulations of these nitrogen forms were associated with heat tolerance in both tall fescue cultivars. Changes of both NADH-glutamate dehydrogenase (NADH-GDH) activity and Glx (glutamine and glutamic acid) concentration in both cultivars indicated that there is an alternative system for assimilation of nitrogen through glutamate dehydrogenase (GDH) in T6 during longer high temperature stress periods. Our results provide an insight to further selection and breeding of heat-tolerant tall fescue turfgrass cultivars.     

Bottle gourd rootstock-grafting affects nitrogen metabolism in NaCl-stressed watermelon leaves and enhances short-term salt tolerance

The plant growth, nitrogen absorption, and assimilation in watermelon (Citrullus lanatus [Thunb.] Mansf.) were investigated in self-grafted and grafted seedlings using the salt-tolerant bottle gourd rootstock Chaofeng Kangshengwang (Lagenaria siceraria Standl.) exposed to 100 mM NaCl for 3 d. The biomass and NO₃⁻ uptake rate were significantly increased by rootstock while these values were remarkably decreased by salt stress. However, compared with self-grafted plants, rootstock-grafted plants showed higher salt tolerance with higher biomass and NO₃⁻ uptake rate under salt stress. Salinity induced strong accumulation of nitrate, ammonium and protein contents and a significant decrease of nitrogen content and the activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), and glutamate synthase (GOGAT) in leaves of self-grafted seedlings. In contrast, salt stress caused a remarkable decrease in nitrate content and the activities of GS and GOGAT, and a significant increase of ammonium, protein, and nitrogen contents and NR activity, in leaves of rootstock-grafted seedlings. Compared with that of self-grafted seedlings, the ammonium content in leaves of rootstock-grafted seedlings was much lower under salt stress. Glutamate dehydrogenase (GDH) activity was notably enhanced in leaves of rootstock-grafted seedlings, whereas it was significantly inhibited in leaves of self-grafted seedlings, under salinity stress. Three GDH isozymes were isolated by native gel electrophoresis and their expressions were greatly enhanced in leaves of rootstock-grafted seedlings than those of self-grafted seedlings under both normal and salt-stress conditions. These results indicated that the salt tolerance of rootstock-grafted seedlings might (be enhanced) owing to the higher nitrogen absorption and the higher activities of enzymes for nitrogen assimilation induced by the rootstock. Furthermore, the detoxification of ammonium by GDH when the GS/GOGAT pathway was inhibited under salt stress might play an important role in the release of salt stress in rootstock-grafted seedlings.     

Nitrogen Metabolism of the Marine Microalga Chlorella autotrophica

The levels of glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in Chlorella autotrophica (clone 580) are strongly regulated by the nitrogen source and salt concentration of the medium. GS is present at high levels in NO₃⁻-grown cells, and at maximum levels in nitrogen-starved cells. However, the levels of GS in these cells are somewhat decreased by increasing salinity. Cells growing on NH₄⁺ have high NADPH-GDH activity, the levels of which increase with increasing NH₄⁺ supply, while GS decreases to a very low level under these conditions. Salinity intensifies the induction of NADPH-GDH activity in NH₄⁺-grown cells. The levels of NADH-GDH are low in this alga, but present under all growth conditions. Methionine sulfoximine (MSX) has little effect on growth and nitrogen assimilation of the alga in the presence of NH₄⁺.