不同生育期花生叶片蛋白质含量及氮代谢相关酶活性分析

以5个珍珠豆型花生(Arachis hypogaea Linn.)品种(系)‘汕E’(‘Shan E’)、‘汕G’(‘Shan G’)、‘TH’、‘TJ’和‘泉花7号’(‘Quanhua No.7’)为研究对象,分析了花针期、结荚期和饱果期花生叶片中可溶性蛋白质含量及硝酸还原酶(NR)、谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)活性的变化趋势,并比较了5个品种(系)荚果和秆产量的差异。结果表明:在3个生育期内,5个花生品种(系)叶片可溶性蛋白质含量和GDH活性的变化趋势基本一致,而NR和GS活性的变化趋势则有差异。其中,可溶性蛋白质含量均呈"低—高—低"的变化趋势,在结荚期最高;GDH活性均逐渐升高,至饱果期达最高;‘泉花7号’叶片NR活性呈"高—低—高"的变化趋势,而其他4个品种(系)叶片NR活性均逐渐降低;‘汕E’、‘TJ’和‘泉花7号’叶片GS活性呈逐渐降低趋势,而‘汕G’和‘TH’叶片GS活性呈"低—高—低"的变化趋势。总体上看,5个品种(系)中,‘汕G’和‘泉花7号’叶片的可溶性蛋白质含量及NR和GDH活性、‘汕E’叶片的NR和GS活性以及‘TH’叶片的GDH活性均较高。5个品种(系)的2个产量指标(单株荚果鲜质量和单株秆鲜质量)均有明显差异,总体上看,‘汕G’、‘泉花7号’和‘TH’的2个产量指标均较高,而‘汕E’和‘TJ’的2个产量指标均较低。综合分析结果显示:‘汕G’和‘泉花7号’叶片可溶性蛋白质含量及NR和GDH活性均相对较高,其荚果和秆产量也均较高,表明花生荚果和秆产量与不同生育期叶片氮代谢水平有一定关系。     

亚低温条件下外源褪黑素对甜瓜幼苗氮代谢及渗透调节物质的影响

以甜瓜品种‘羊角酥瓜’幼苗为试材,利用人工气候室进行亚低温处理(昼/夜18℃/12℃)6 d,研究外源褪黑素(MT)对亚低温条件下甜瓜幼苗叶片氮代谢酶[硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)和谷氨酸脱氢酶(GDH)]活性、叶片总氮、铵态氮、硝态氮和渗透调节物质含量的影响.结果表明:与对照相比,亚低温处理降低了甜瓜总氮、硝态氮及硝酸还原酶活性,增加了铵态氮含量,抑制了甜瓜幼苗的生长.外源MT可显著提高亚低温下甜瓜幼苗氮代谢酶的活性,尤其可显著提高叶片GS和GOGAT活性,从而有效降低铵态氮含量;外源MT还可以提高叶片脯氨酸、可溶性蛋白质和可溶性糖的含量,进而降低了亚低温对细胞膜的伤害,表现为甜瓜叶片相对电导率和丙二醛(MDA)含量较低.总之,在亚低温条件下,外源MT在一定程度上可通过降低甜瓜叶片铵态氮含量和促进渗透调节物质的积累,降低膜质过氧化水平,从而增加其对亚低温的适应性.     

亚低温条件下外源褪黑素对甜瓜幼苗氮代谢及渗透调节物质的影响

以甜瓜品种‘羊角酥瓜’幼苗为试材,利用人工气候室进行亚低温处理(昼/夜18 ℃/ 12 ℃)6 d,研究外源褪黑素（MT）对亚低温条件下甜瓜幼苗叶片氮代谢酶\[硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)和谷氨酸脱氢酶(GDH)\]活性、叶片总氮、铵态氮、硝态氮和渗透调节物质含量的影响.结果表明：与对照相比,亚低温处理降低了甜瓜总氮、硝态氮及硝酸还原酶活性,增加了铵态氮含量,抑制了甜瓜幼苗的生长.外源MT可显著提高亚低温下甜瓜幼苗氮代谢酶的活性,尤其可显著提高叶片GS和GOGAT活性,从而有效降低铵态氮含量;外源MT还可以提高叶片脯氨酸、可溶性蛋白质和可溶性糖的含量,进而降低了亚低温对细胞膜的伤害,表现为甜瓜叶片相对电导率和丙二醛(MDA)含量较低.总之,在亚低温条件下,外源MT在一定程度上可通过降低甜瓜叶片铵态氮含量和促进渗透调节物质的积累,降低膜质过氧化水平,从而增加其对亚低温的适应性.     

Ni胁迫对不同基因型谷子幼苗生长及氮素代谢的影响

采用盆栽土培法,研究了不同浓度Ni2+(0、25、50、100、150、200 mg/kg)对4种基因型谷子(13-36、B-7、晋谷51号、晋谷52号)幼苗生长,Ni2+富集与转运能力,叶片中硝态氮、氨态氮、可溶性蛋白质、脯氨酸含量及氮代谢相关酶硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、谷氨酸脱氢酶(GDH)活性的影响。结果表明:Ni2+胁迫下,4种基因型谷子幼苗的根长、苗长、生物量随Ni2+浓度增加逐渐降低,体内Ni2+含量逐渐增加,与对照组差异显著(P0.05)。在所试浓度范围内,4种基因型谷子幼苗叶片中的硝态氮含量、NR、GS、GOGAT活性表现为低浓度(25—50mg/kg)增高和高浓度(50—200 mg/kg)降低,而GDH活性在Ni2+浓度为100mg/kg以上时下降,氨态氮含量在50—150 mg/kg处理组中为对照的1.14—3.02倍。不同浓度Ni2+处理后,4种基因型谷子幼苗叶片中的脯氨酸含量均有不同程度的提高,而可溶性蛋白质含量呈明显下降趋势。实验结果证明,Ni2+胁迫抑制了谷子幼苗对硝态氮的吸收,降低了叶片中NR、GS、GOGAT活性,影响了氨的同化作用,使谷子幼苗的氮素代谢发生紊乱,不同基因型谷子对Ni2+胁迫的毒性效应存在差异。4种基因型谷子对Ni2+的耐性顺序为13-36B-7晋谷51晋谷52。     

氮素水平对花生氮素代谢及相关酶活性的影响

 在大田高产条件下研究了氮素水平对花生(Arachis hypogaea)可溶性蛋白质、游离氨基酸含量及氮代谢相关酶活性的影响, 结果表明, 适当提高氮素水平既能增加花生各器官中可溶性蛋白质和游离氨基酸的含量, 又能提高硝酸还原酶、谷氨酰胺合成酶和谷氨酸脱氢酶等氮素同化酶的活性, 使其达到同步增加; 氮素水平过高虽能提高硝酸还原酶和籽仁蛋白质含量, 但谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)的活性下降; N素施肥水平不改变花生植株各器官中可溶性蛋白质、游离氨基酸含量以及硝酸还原酶(NR)、谷氨酰胺合成酶、谷氨酸脱氢酶活性的变化趋势, 但适量施N (A2和A3处理)使花生各营养器官中GS、GDH活性提高; 氮素水平对花生各叶片和籽仁中GS、GDH活性的高低影响较大, 但对茎和根中GDH活性大小的影响较小。     

盐碱胁迫下外源硫化氢对裸燕麦叶片氮代谢和产量构成因素的影响

为探讨外源信号硫化氢(H₂S)对盐碱胁迫下裸燕麦(Avena nuda)氮代谢的调控机制及产量构成因素的影响,采用盆栽土培试验,以‘定莜9号’品种为材料,设置盆土不添加盐碱和添加3.00 g·kg^-1盐碱(摩尔比NaCl∶Na₂SO₄∶Na₂CO₃∶NaHCO₃=12∶8∶1∶9)与抽穗期喷水和喷50μmol·L^-1 H₂S供体硫氢化钠(NaHS)溶液,共4个处理。喷施3 d后分别在第7天和第14天取叶片检测氮代谢相关物质含量和关键酶活性,成熟期调查产量构成因素及产量。结果表明：(1)喷施后第7天,盐碱胁迫显著降低裸燕麦叶片氨态氮、硝态氮、可溶性蛋白质含量和蛋白酶、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、谷氨酸脱氢酶(GDH)、谷草转氨酶(GOT)、谷丙转氨酶(GPT)活性,对游离氨基酸含量和硝酸还原酶(NR)活性无显著影响;喷施NaHS溶液使盐碱胁迫下裸燕麦叶片氨态氮、游离氨基酸...     

施氮和去除子叶对大叶桃花心木幼苗叶片硝酸还原酶活性的影响

研究了热带落叶乔木大叶桃花心木(Swietenia macrophylla)在施氮和去除子叶后幼苗叶片的硝酸还原酶活性(NRA)变化。结果表明,在非施氮(对照)条件下,NRA随着幼苗叶片的发育先升高后降低;施氮后幼苗叶片NRA在各取样时期(除35 d外)均显著高于非施氮处理(P<0.05),并随着取样时期的延续,叶片NRA逐渐降低。在幼苗发育的不同时期去除子叶,4周后,叶片NRA均显著升高(P<0.05)。     

浑球红假单胞菌氨同化途径的研究

本文测定了浑球红假单胞菌(Rhodobacter sphaeroides)菌株601谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、谷氨酸脱氢酶(GDH)和丙氨酸脱氢酶(ADH)的活性。低氨时,GS/GOGAT活力高,GDH活力低,高氨时,GS/GOGAT活力低,GDH活力高。在以分子氮或低浓度氨为氮源的培养条件下,加入GS抑制刑MSX(L—methionine—DL—sulphoximine),细菌生长受到抑制。但是,生长在以谷氨酸为氮源的细菌则不受影响。上述结果表明,浑球红假单胞菌菌株601氨同化是通过GS/GOGAT途径和GDH途径。     

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.     

根际低氧胁迫对网纹甜瓜果期根系氮代谢的影响

采用气雾法栽培系统,研究了根际低氧(10%和5% O2)胁迫对网纹甜瓜(Cucumis melo)果实发育期间根系氮代谢的影响。结果表明:在30 d试验期内,与对照(21% O2)相比,低氧胁迫网纹甜瓜根系硝态氮(NO3--N)含量、铵态氮(NH4+-N)含量、硝酸还原酶(NR)活性、谷氨酰氨合成酶(GS)活性先增后降;在低氧处理20 d时,网纹甜瓜上述指标显著高于对照,低氧处理30 d时,显著低于对照,且5% O2处理变化的幅度大于10% O2处理;谷氨酸脱氢酶(GDH)活性在低氧处理10 d时显著低于对照,处理20 d时,与对照差异不显著;谷草转氨酶(GOT)活性在低氧处理20 d时与对照差异不显著,处理30 d时,显著低于对照;在5% O2处理期间谷丙转氨酶(GPT)活性显著低于对照,10% O2处理20 d时显著低于对照;在低氧处理期间,根系中可溶性蛋白质含量、根系伤流液中氨基酸的总量和大部分氨基酸的含量均随着根际O2浓度的降低而减少;网纹甜瓜果实发育期间根际O2浓度长期处于10% O2以下时,根系对氮的吸收、代谢能力下降,植株氮循环水平降低。     

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     

Changes in the levels of enzymes involved in ammonia assimilation during the development of Phaseolus vulgaris seedlings.Effects of exogenous ammonia

Seeds of Phaseolus vulgaris L. cv. White Kidney were germinated and grown either in a nitrogen-free or in an ammonia-supplied medium. The changes in the soluble protein concentration and in the levels of glutamine synthetase (GS, EC 6.3.1.2), NADH–glutamate synthase (NADH-GOGAT, EC 1.4.1.14), ferredoxin-glutamate synthase (Fd-GOGAT, EC 1.4.7.1) and glutamate dehydrogenase (GDH, EC 1.4.1.2), both NADH- and NAD⁺-dependent, were examined in cotyledons and roots during the first 10 days after sowing. Soluble protein declined rapidly in the cotyledons and increased slightly in the roots. GS activity was initially high both in cotyledons and roots but subsequently decreased during seedling growth. Exogenous ammonia hardly affected GS activity. High levels of NADH-GOGAT were present both in cotyledons and roots during the first days of germination. The activity then gradually declined in both organs. In contrast, Fd-GOGAT in cotyledons was initially low and progressively increased with seedling development. In roots, the levels of Fd-GOGAT were higher in young than in old seedlings. Supply of ammonia to the seedlings increased the levels of NADH-GOGAT and Fd-GOGAT both in cotyledons and roots. NADH-GDH (aminating) activity gradually increased during germination. In contrast, the levels of NAD⁺-GDH (deaminating) activity were highest during the first days of germination. Exogenous ammonia did not significantly affect the activities of GDH.     

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。     

Effect of selected compounds on the activity of glutamate dehydrogenase from triticale roots

The influence of selected factors on the activity of highly purified GDH in triticale roots was investigated in vitro. In the presence of 2-ME, NADH-GDH activity increased by 400 %, while NADPH-GDH activity rose by 500 %. No effect of reducing factors on NAD(P)⁺-GDH reaction was detected. The sulphydryl groups inhibitors, such as p-chloromercuribenzoate (p-CMB) and iodoacetamide, proved the strongest inhibitors of the aminative reaction. Metal-binding compounds: ethylenediaminetetraacetic acid disodium salt (EDTA) and Zinkov also considerably inhibited NAD(P)H-GDH activity. Diisopropylfluorophosphate (DFP) and pepstatin A, the inhibitors specific for -OH serine and COO⁻ aspartic acid groups respectively, caused a non-significant NAD(P)H-GDH activity decrease. Cd²⁺, Co²⁺, Hg²⁺, Mg²⁺, Pb²⁺ and Zn²⁺ ions strongly inhibited the amination reaction, whereas their inhibiting effect upon NAD⁺-GDH activity was negligible. Among the applied ions, only Ca²⁺ activated NADH-GDH.     

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.     

Regulation of glutamate dehydrogenase activity by amino acids in maize seedlings

Root or secondary leaf segments from maize ( Zea mays L. cv. Ganga safed-2) seedlings were incubated with 9-amino acids and two amides separately, each at 5 m M for 24 h, to study their effects on glutamate dehydrogenase (GDH) activity. Most of the compounds tested inhibited the specific activity of NADH-GDH and increased that of NAD⁺-GDH in the roots in the presence as well as in the absence of ammonium. In the leaves, such effects were recorded only with a few amino acids. Total soluble protein in the root and leaf tissues increased with the supply of most of the amino compounds. The effect of glutamate on enzyme activity and protein was concentration dependent in both tissues. When the enzyme extracts from root or leaf tissues were incubated with some of the amino acids, NADH-GDH declined while NAD⁺-GDH increased in most cases. The inhibition of NADH-GDH increased with increasing concentration of cysteine from 1 to 5 m M . The experiments demonstrate that most of the amino acids regulated GDH activity, possibly through some physicochemical modulation of the enzyme molecule.     

低氧胁迫下外源硝态氮对樱桃根系功能及氮代谢相关酶活性的影响

以樱桃组培苗‘吉塞拉5号’（Prunun cerasus × P. canescens）为试材,采用营养液水培控制溶氧浓度的方法,研究了短期低氧胁迫下外源硝态氮对其根系功能及氮代谢相关酶活性的影响.结果表明：与对照（7.5 mmol NO₃^-·L^-1）相比,低氧加氮处理（15和22.5 mmol NO₃^-·L^-1）使樱桃体内代谢原料充足,保证了各类酶蛋白的合成,使植株根系活力升高,根系呼吸未受到明显抑制,与氮代谢相关的硝酸还原酶（NR）、谷氨酰胺合成酶（GS）及谷氨酸脱氢酶（NADH-GDH）活性升高,从而为低氧逆境下樱桃根系的吸收作用提供了足够的能量,保证了糖酵解和电子传递的顺利进行,并及时同化了NO₃^-还原生成NH₄⁺,避免了铵毒害,缓解了樱桃的低氧伤害,且22.5 mmol NO₃^-·L^-1处理的缓解效果优于15 mmol·L^-1处理;低氧缺氮处理（0 mmol NO₃^-·L^-1）的樱桃植株根系活力下降,根系呼吸受到抑制,NR、GS及NADH-GDH活性降低.这说明低氧胁迫下,适当提高生长介质中的NO₃^-浓度可调控樱桃的根系功能及氮代谢,缓解低氧胁迫对樱桃根系的伤害.     

An approach to the genetics of nitrogen use efficiency in maize

To study the genetic variability and the genetic basis of nitrogen (N) use efficiency in maize, a set of recombinant inbred lines crossed with a tester was studied at low input (N-) and high input (N+) for grain yield and its components, grain protein content, and post-anthesis nitrogen uptake and remobilization. Other physiological traits, such as nitrate content, nitrate reductase, glutamine synthetase (GS), and glutamate dehydrogenase activities were studied at the level of the lines. Genotypexnitrogen (GxN) interaction was significant for yield and explained by variation in kernel number. In N-, N-uptake, the nitrogen nutrition index, and GS activity in the vegetative stage were positively correlated with grain yield, whereas leaf senescence was negatively correlated. Whatever N-input, post-anthesis N-uptake was highly negatively related to N-remobilization. As a whole, genetic variability was expressed differently in N+ and N-. This was confirmed by the detection of QTLs. More QTLs were detected in N+ than in N- for traits of vegetative development, N-uptake, and grain yield and its components, whereas it was the reverse for grain protein content and N-utilization efficiency. Several coincidences between genes encoding for enzymes of N metabolism and QTLs for the traits studied were observed. In particular, coincidences in three chromosome regions of QTLs for yield and N-remobilization, QTLs for GS activity and a gene encoding cytosolic GS were observed. This may have a physiological meaning. The GS locus on chromosome 5 appears to be a good candidate gene which can, at least partially, explain the variation in nitrogen use efficiency.