烤烟叶片衰老期氨气挥发特征及其生理调控研究

以烤烟品种K326为试验材料,利用氨气收集装置测定烟叶的氨气挥发量,并利用谷氨酰胺合成酶(GS)抑制剂(Glufosinate)处理叶片和质外体提取等方法,研究了叶片氨气挥发及其与氮代谢相关生理指标的关系。结果表明:(1)随着叶片的衰老,氨气挥发量在叶龄70d时最大(10.96μg.m-2.h-1),与衰老初期(叶龄40d)相比增加了2.15倍;质外体NH4+浓度和pH、氨气补偿点逐渐上升,GS和硝酸还原酶(NR)活性下降,谷氨酸脱氢酶(GDH)活性升高,可溶性蛋白和总氮降解,叶片NH4+浓度升高。(2)GS抑制剂处理后,叶片组织NH4+浓度和氨气补偿点升高,氨气挥发量增大,与对照相比差异显著。(3)氨气挥发量与质外体NH4+浓度、质外体pH和氨气补偿点呈极显著或显著正相关,与GS活性呈显著负相关,与GDH活性呈显著正相关,与叶片组织NH4+浓度等其他指标相关性不显著。研究认为,烤烟叶片衰老期间氨挥发量大幅上升,挥发量的大小受气孔氨气补偿点、GS和GDH活性的直接调控,以及其他氮素代谢相关指标的间接调控,其中GS起主导作用。     

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.     

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。     

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     

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。     

外源Spd对盐碱胁迫下番茄幼苗氮代谢及主要矿质元素含量的影响

采用水培方法,研究了盐碱与Spd处理对两品种番茄(中杂9号和金棚朝冠)幼苗氮代谢及主要矿质元素含量的影响.结果表明:盐碱胁迫下,番茄幼苗干生物量显著减少,植株生长受到抑制;叶片和根系硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT)活性及硝态氮(NO3--N)、全N、全K、Ca2+、Mg2+含量显著降低,铵态氮(NH4+-N)、Na+含量显著增加;两品种叶片及中杂9号根系谷氨酸脱氢酶(GDH)活性显著升高,金棚朝冠根系GDH活性变化不显著;叶片全P含量显著降低,根系全P含量显著升高(金棚朝冠)或无显著变化(中杂9号).Spd处理通过增强NR、GS、GOGAT活性提高了植株对NH4+的同化利用率,有效缓解了盐碱胁迫导致的氮代谢紊乱,进而促进不同器官对P、K、Ca、Mg、Na的吸收、释放或转运,在一定程度上维持了各元素之间的相对平衡,从而增强植株对逆境的适应能力.此外,盐碱对中杂9号的抑制作用及外源Spd对其氮代谢紊乱和营养失衡的缓解作用高于金棚朝冠.     

乙烯对旱后复水玉米某些生理特性的影响

研究了在不同水分条件下盆栽的玉米(Zea mays L)品种'郑单958'幼苗,经乙烯利和AgNO3处理后,其复水前后某些生理特征的变化.结果表明:复水前,在不同程度干旱胁迫下施用乙烯利有利于增加玉米的生物量、叶片相对含水量、总叶绿素含量、可溶性蛋白质含量、叶片硝酸还原酶(NR)活性和谷氨酰胺合成酶(GS)活性;而AgNO3处理明显降低生物量、叶片相对含水量、可溶性蛋白质含量、NR活性和GS活性.复水后,乙烯利处理的玉米生物量、总叶绿素含量、可溶性蛋白质含量明显高于对照,叶片相对含水量、NR和GS活性与对照相近;经轻度与重度干旱后复水,AeNO3处理明显降低玉米生物量和可溶性蛋白含量,对叶片相对含水量、NR、GS活性影响不明显.因此,施用乙烯利有利于提高干旱后复水条件下苗期玉米对干旱的抗性.     

不同氮素形态配比对甘薯前期氮代谢的影响及其生理机制

以3种不同类型的甘薯(Ipomoea batatas (L.) Lam.)为实验材料,根据氮素的3种形态设置5个配比处理(N1～N5),分别在栽秧后15、25和35 d取样测定甘薯不同器官的氮含量、功能叶氮代谢酶活性变化以及酶调控基因表达情况。结果显示:在同一生育期,N4和N5处理铵态氮和硝态氮配施下植株氮素的积累量明显高于其它处理;在甘薯生长发育前期,叶片含氮量先降低后上升,茎、须根和膨大根以及全株含氮量均呈上升趋势; N4处理能够显著提高硝酸还原酶(NR)、谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)活性; N3处理能够明显提高谷氨酸合成酶(GOGAT)的活性; NR活性随肥料中硝态氮比例的增加而提高;增加肥料配比中硝态氮比例可使调控NR活性的基因上调表达,N4和N5处理可使GS调控基因上调表达,但抑制GOGAT调控基因的表达。酰胺态氮在前期对氮代谢相关酶调控基因无显著影响。研究结果表明,在甘薯生长发育前期,硝态氮和铵态氮配施能够显著提高氮素积累量、代谢酶活性和调控基因表达量,铵态氮∶硝态氮∶酰胺态氮=1∶2∶0的配施方案为本实验条件下的最佳配施组合。     

慢性氨暴露对中华鳖幼鳖血浆总氨氮、皮质酮浓度及组织氨代谢酶活性的影响

本文观测了慢性氨暴露对中华鳖(Pelodiscussinensis)幼鳖血浆总氨氮、皮质酮浓度及与氨代谢有关的酶活性的影响。将中华鳖暴露在总氨氮(TAN)浓度为32.4、57.6、83.5mg/L(分别记为1、2、3组)的水环境中饲养42d,以自然晾晒脱氯自来水饲养组为对照(记为0组),pH值控制在7.80-7.85。检测氨暴露2、4、8、24、48h、42d后血浆TAN与8、24、48h、42d后皮质酮浓度,42d后肝、肌肉与脑中谷氨酰胺合成酶(Glutaminesynthetase,GS)、谷氨酸脱氢酶(Glutamatedehydrogenase,GDH)的活性以及特定生长率(Specificgrowthrate,SGR)的变化。结果表明:0、1组血浆TAN随时间没有明显变化,2、3组血浆TAN随时间呈现先增加后降低的趋势,分别在8h和48h达到峰值,42d时各组间血浆TAN没有显著差异。氨暴露显著影响暴露初期血浆皮质酮水平,暴露后24h,1、3组血浆皮质酮水平显著高于对照组;42d后除2组外其他组间无显著差异。4组之间肝、肌肉和脑中GS活性均没有显著差异,肝和肌肉GDH(氨化和去氨化方向)活性也没有显著差异。各暴露组脑GDH活性和对照组相比差异不明显,但各处理组间氨化和去氨化方向脑GDH酶活差异显著。42d饲养期间各处理组SGR没有显著差异。     

谷氨酰胺合成酶基因GS1和GS2的高效表达增强转基因水稻对氮素缺乏的耐性

构建了同时含有胞质谷氨酰胺合成酶(GS1)cDNA和叶绿体谷氨酰胺合成酶(GS2)cDNA的植物表达载体p2GS,通过农杆菌介导法用它们转化了水稻品种"中花10号"的成熟胚愈伤组织,经潮霉素(Hyg)筛选培养及分化再生,获得了抗Hyg的转基因水稻植株.PCR和基因组Southern杂交分析结果证明,GS1和GS2基因均已经整合到转基因水稻的基因组内.Northern杂交实验结果证实,GS1和GS2基因在转基因水稻的转录水平上得到了有效表达.在以0.7 mmol/L的(NH4)2SO4取代了其中氮成分的MS培养基上测试植株生长量,结果表明转基因植株鲜重增长量显著高于对照,证明高效表达GS增强了转基因水稻对土壤氮素缺乏的耐性.     

Influence of Different Concentrations of NO-3 and NH+4 on the Activity of Glutamine Synthetase and Other Relevant Enzymes of Nitrogen Metabolism in Wheat Roots

Influence of different concentrations of NO3 and NH+ on the activity of glutamine synthetase (GS), asparagine synthetase (AS), glutamate dehydrogenase (GDH), nitrate reductase (NR) and the changes of GS-mRNA in wheat roots have been studied with enzymes activity assay and Northern blot. The results showed that the higher GS activity was found in roots of wheat when NH+4-N was the sole nitrogen source than when NO3-N was the sole nitrogen source. GS-mRNA of Northern blot was simillar to GS activity. 3 mmol/L NO3- promoted the activity of AS. The change of AS was independent of the change of GS. GDH activity was not been detected, and change in regulation of NR activity was not found.     

不同氮源对小麦幼苗谷氨酰胺合成酶的影响

利用ＤＥＡＥ－纤维素柱层析、酶活性测定、Ｎｏｒｔｈｅｒｎ 分子杂交等技术,研究了小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．）幼苗的根、叶和离体叶在不同氮源培养条件下谷氨酰胺合成酶（ＧＳ）活性和同工酶变化, 以及不同氮源对ＧＳ基因转录－ＧＳ－ｍ ＲＮＡ 的影响． 同时与硝酸还原酶（ＮＲ）活性进行比较, 结果表明∶当以ＮＨ＋４ 作唯一氮源时,小麦幼苗根谷氨酰胺合成酶（ＧＳｒ）和叶细胞质谷氨酰胺合成酶（ＧＳ１）活性要比以ＮＯ－３ 作唯一氮源的高．当以ＮＯ－３ 为唯一氮源时, ＮＯ－３ 则促进完整叶片和离体叶片叶绿体谷氨酰胺合成酶（ＧＳ２）活性． 从转录水平上看,ＮＨ＋４ 促进根ＧＳ－ｍ ＲＮＡ 的合成,而ＮＯ－３ 促进叶ＧＳ－ｍ ＲＮＡ 的合成     

Development of nitrogen-assimilating enzymes in sunflower cotyledons during germination as affected by the exogenous nitrogen source

Activities of nitrate reductase (NR; EC 1.6.6.1), nitrite reductase (NiR; EC 1.7.7.1), glutamine synthetase (GS; EC 6.3.1.2) and glutamate dehydrogenase (GDH; EC 1.4.1.3) were measured in cotyledons of sunflower (Helianthus annuus L. cv Peredovic) seedlings during germination and early growth under various external nitrogen sources. The presence of NO ₃ ^- in the medium promoted a gradual increase in the levels of NR and NiR activities during the first 7 d of germination. Neither NR nor NiR activities were increased in a nitrogen-free medium or in media with either NH ₄ ⁺ or urea as nitrogen sources. Moreover, the presence of NH ₄ ⁺ did not abolish the NO ₃ ^- -dependent appearance of NR and NiR activities. The increase of NR activity was impaired both by cycloheximide and chloramphenicol, which indicates that both cytoplasmic 80S and plastidic 70S ribosomes are involved in the synthesis of the NR molecule. By contrast, the appearance of NiR activity was only inhibited by cycloheximide, indicating that NiR seems to be exclusively synthesized on the cytoplasmic 80S ribosomes. Glutamine-synthetase activity was also strongly increased by external NO ₃ ^- but not by NH ₄ ⁺ or urea. The appearance of GS activity was more efficiently suppressed by cycloheximide than chloramphenicol. This indicates that GS is mostly synthesized in the cytoplasm. The cotyledons of the dry seed contain high levels of GDH activity which decline during germination independently of the presence or absence of a nitrogen source. Cycloheximide, but not chloramphenicol, greatly prevented the decrease of GDH activity.Abbreviations GDH glutamate dehydrogenase - GS glutamine synthetase - NiR nitrite reductase - NR nitrate reductase     

Glutamine synthetase and arginine inhibition of nitrate reductase activity in Anabaena cycadeae

Wild-type Anabaena cycadeae with normal glutamine synthetase (GS) activity utilized arginine as sole N source whereas a mutant strain lacking GS activity did not. Nitrate reductase (NR) activity, higher in the mutant strain than the wild-type strain, was inhibited by arginine though arginine-dependent NH ₄ ⁺ generation was higher in the mutant strain than in the wild-type. This suggests that (1) NR activity is NO _inf3 ^sup- -inducible and arginine-repressible; and (2) while GS activity is required for the assimilation of arginine as sole N-source, it is not required for arginine inhibition of NR activity.S. Singh was with the Department of Biochemistry, North-Eastern Hill University, Shillong-793014, India, and is now with P.S. Bisen at the Department of Microbiology, Barkatullah University, Bhopal-462026, India     

NaCl stress effects on enzymes involved in nitrogen assimilation pathway in tomato "Lycopersicon esculentum" seedlings

Tomato plants (Lycopersicon esculentum Mill, cv. Chibli F1) grown for 10 days on control medium were exposed to differing concentrations of NaCl (0, 25, 50, and 100mM). Increasing salinity led to a decrease of dry weight (DW) production and protein contents in the leaves and roots. Conversely, the root to shoot (R/S) DW ratio was increased by salinity. Na(+) and Cl(-) accumulation were correlated with a decline of K(+) and NO(3)(-) in the leaves and roots. Under salinity, the activities of nitrate reductase (NR, EC 1.6.6.1) and glutamine synthetase (GS, EC 6.3.1.2) were repressed in the leaves, while they were enhanced in the roots. Nitrite reductase (NiR, EC 1.7.7.1) activity was decreased in both the leaves and roots. Deaminating activity of glutamate dehydrogenase (GDH, EC 1.4.1.2) was inhibited, whereas the aminating function was significantly stimulated by salinity in the leaves and roots. At a high salt concentration, the nicotinamide adenine dinucleotide reduced (NADH)-GDH activity was stimulated concomitantly with the increasing NH(4)(+) contents and proteolysis activity in the leaves and roots. With respect to salt stress, the distinct sensitivity of the enzymes involved in nitrogen assimilation is discussed.     

外源Spd对盐碱胁迫下番茄幼苗氮代谢及主要矿质元素含量的影响

采用水培方法,研究了盐碱与Spd处理对两品种番茄（中杂9号和金棚朝冠）幼苗氮代谢及主要矿质元素含量的影响.结果表明： 盐碱胁迫下,番茄幼苗干生物量显著减少,植株生长受到抑制;叶片和根系硝酸还原酶（NR）、谷氨酰胺合成酶（GS）、谷氨酸合成酶（GOGAT）活性及硝态氮（NO₃^--N）、全N、全K、Ca²⁺、Mg²⁺含量显著降低,铵态氮（NH₄⁺-N）、Na⁺含量显著增加;两品种叶片及中杂9号根系谷氨酸脱氢酶（GDH）活性显著升高,金棚朝冠根系GDH活性变化不显著;叶片全P含量显著降低,根系全P含量显著升高（金棚朝冠）或无显著变化（中杂9号）.Spd处理通过增强NR、GS、GOGAT活性提高了植株对NH₄⁺的同化利用率,有效缓解了盐碱胁迫导致的氮代谢紊乱,进而促进不同器官对P、K、Ca、Mg、Na的吸收、释放或转运,在一定程度上维持了各元素之间的相对平衡,从而增强植株对逆境的适应能力.此外,盐碱对中杂9号的抑制作用及外源Spd对其氮代谢紊乱和营养失衡的缓解作用高于金棚朝冠.     

Effect of Azotobacter strains on sugar beet callus proliferation and nitrogen metabolism enzymes

The effect of five Azotobacter chroococcum strains and nitrogen content in nutrient media on callus growth of two Beta vulgaris L. cultivars were investigated, as well as the activity of nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in inoculated callus tissue. On medium with full nitrogen content (1 N) the inoculation with A. chroococcum strain A2 resulted in the highest calli mass, while strains A8 and A14 maximally increased NR activity. On media with 1/8 N the highest effect on calli growth, GS and GDH activity had the strain A8. The strain A2/1 significantly increased callus proliferation on medium without N. Asymbiotic association between sugar beet calli and Azotobacter depended on genotype/strain interaction and was realised in presence of different nitrogen levels. This revised version was published online in July 2006 with corrections to the Cover Date.