Spermine delays leaf senescence in Lactuca sativa and prevents the decay of chloroplast photosystems

Aliphatic polyamines (PAs) are involved in the delay or prevention of plant senescence, but the molecular mechanism is not clarified. The hypothesis is put forward that one of the mechanisms by which PAs modulate leaf senescence and chlorophyll stabilisation could be due to their modification of chlorophyll-bound proteins, catalysed by transglutaminase (TGase, R-glutaminylpeptide-amine γ-glutamyltransferase; E.C. 2.3.2.13). The retardation of leaf senescence of Lactuca sativa L. by spermine (Spm) was examined during induced cell death using leaf discs, or during the normal developmental senescence of leaves. Over 3 days, in leaf discs, Spm caused a delay of chlorophyll (Chl) decay, an increase of endogenous TGase activity, and a three-fold increase in chlorophyll content when supplied together with exogenous TGase. Spm was conjugated, via TGase, mainly to 22–30 kDa proteins. Long-term experiments over 5 days showed a general decrease in all three parameters with or without Spm. When leaves remained on the plants, Spm-sprayed leaves showed an increase in free Spm 1 h after spraying, mainly in the young leaves, whereas over longer periods (15 days) there was an increase in perchloric acid-soluble and -insoluble Spm metabolites. In senescing leaves, Spm prevented degradation of chlorophyll b and some proteins, and increased TGase activity, producing more PA-protein conjugates. Spm was translocated to chloroplasts and bound mainly onto fractions enriched in PSII, but also those enriched in PSI, whose light-harvesting complexes (LHC) sub-fractions contained TGase. Spm was conjugated by TGase mainly to LHCII, more markedly in the light. Immunodetection of TGase revealed multiple proteins in young leaves, possibly representing different TGase isoforms when TGase activity was high, whereas in already senescent leaves, when its activity decreased, one high-molecular-mass band was found, possibly because of enzyme polymerisation. Spm thus protected senescing Lactuca leaves from the decay of their chloroplast photosystem complexes. The senescence-delaying effects of Spm could be mediated by TGase, as TGase was re-activated to the level in young leaves following Spm treatment.     

Seasonal changes in light and temperature affect the balance between light harvesting and light utilisation components of photosynthesis in an evergreen understory shrub

In a temperate climate, evergreen species in the understory are exposed to large changes in photosynthetic photon flux density (PPFD) and temperature over the year. We determined the photosynthetic traits of leaves of an evergreen understory shrub Aucuba japonica at three sites at monthly intervals: understorys of a deciduous forest; an evergreen forest; and a gap in a mixed forest. This set up enabled us to separate the effects of seasonal change in PPFD and temperature on photosynthetic acclimation under natural conditions. The effects of PPFD and temperature were analysed by simple and multiple regression analyses. The amounts of light utilisation components (LU), represented by nitrogen and rubisco contents per area, were higher in winter, when temperature was low and PPFD was high. The LU relative to the amount of light harvesting components (LH), represented by chlorophyll a/b and rubisco/chlorophyll ratios, and the inverse of chlorophyll/nitrogen ratio were also higher in winter. We quantified the effects of PPFD and temperature on the LU and LH components. Across sites PPFD had stronger effects than air temperature, while within a site temperature had stronger effects on photosynthetic acclimation. We concluded that the photosynthetic apparatus is strongly affected by the prevailing PPFD at the time of leaf development. Within a given light regime, however, plants acclimated by increasing LU relative to LH primarily in response to temperature and to a lesser extent to PPFD.     

Polyamines and transglutaminase activity are involved in compatible and self-incompatible pollination of Citrus grandis

Pollination of pummelo (Citrus grandis L. Osbeck) pistils has been studied in planta by adding compatible and self-incompatible (SI) pollen to the stigma surface. The pollen germination has been monitored inside the pistil by fluorescent microscopy showing SI altered morphologies with irregular depositions of callose in the tube walls, and heavy callose depositions in enlarged tips. The polyamine (PA) content as free, perchloric acid (PCA)-soluble and -insoluble fractions and transglutaminase (TGase) activity have been analyzed in order to deepen their possible involvement in the progamic phase of plant reproduction. The conjugated PAs in PCA-soluble fraction were definitely higher than the free and the PCA-insoluble forms, in both compatible and SI pollinated pistils. In pistils, pollination caused an early decrease of free PAs and increase of the bound forms. The SI pollination, showed highest values of PCA-soluble and -insoluble PAs with a maximum in concomitance with the pollen tube arrest. As TGase mediates some of the effects of PAs by covalently binding them to proteins, its activity, never checked before in Citrus, was examined with two different assays. In addition, the presence of glutamyl-PAs confirmed the enzyme assay data and excluded the possibility of a misinterpretation. The SI pollination caused an increase in TGase activity, whereas the compatible pollination caused its decrease. Similarly to bound PAs, the glutamyl-PAs and the enzyme activity peaked in the SI pollinated pistils in concomitance with the observed block of the pollen tube growth, suggesting an involvement of TGase in SI response.     

Transglutaminase activity changes during the hypersensitive reaction, a typical defense response of tobacco NN plants to TMV

The occurrence of glutamyl polyamines (PAs) and changes in activity and levels of transglutaminase (TGase, EC 2.3.2.13), the enzyme responsible for their synthesis, are reported during the progression of the hypersensitive reaction (HR) of resistant NN tobacco plants ( Nicotiana tabacum L. cv. Samsun) to tobacco mosaic virus (TMV). Mature leaves of tobacco were collected over 0–72 h after inoculation with TMV or phosphate buffer (mock). In vivo synthesis of polyamine glutamyl derivatives (glutamyl PAs), catalyzed by TGase activity, was evaluated after supplying labeled putrescine (Pu, a physiological substrate of TGase) to leaves. Results show that, starting from 24 h, mono-( γ -glutamyl)-Pu and bis-( γ -glutamyl)-Sd were recovered in TMV-inoculated samples but not in mock-inoculated ones; 2 days later, in the former, the amount of glutamyl derivatives further increased. An in vitro radiometric assay showed that, in TMV-inoculated leaves, TGase activity increased from 24 h onwards relative to mock controls. An immunoblot analysis with AtPng1p polyclonal antibody detected a 72-kDa protein whose amount increased at 72 h in TMV-inoculated leaves and in the lesion-enriched areas. A biotin-labeled cadaverine incorporation assay showed that TGase activity occurred in S1 (containing soluble proteins), S2 (proteins released by both cell walls and membranes) and S3 (membrane intrinsic proteins) fractions. In S3 fraction, where changes were the most relevant, TGase activity was enhanced in both mock-inoculated and TMV-inoculated samples, but the stimulation persisted only in the latter case. These data are discussed in the light of a possible role of TGase activity and glutamyl PAs in the defense against a viral plant pathogen.     

香果树实生苗的光合特性及其与环境因子的关系

为确定香果树实生苗的适生环境并为其自然更新提出有针对性策略,研究了不同生境(冠下、冠缘、林窗和林缘)中2年生香果树实生苗的净光合速率、水分利用效率、叶绿素含量、苗高、基径、生物量等的变化及其与生态因子之间的关系.结果表明: 4种生境中的光合有效辐射最大值为50～1380 μmol·m^-2·s^-1,冠下和冠缘中香果树实生苗的净光合速率日变化呈单峰型,而林窗和林缘实生苗的净光合速率日变化呈双峰型;香果树实生苗为耐阴植物,但耐阴能力较弱,其功能叶的光饱和点、补偿点和暗呼吸在4种生境中大小顺序为: 林缘>林窗>冠缘>冠下,表观量子效率的变化规律与之相反;林窗和冠缘2种生境中香果树实生苗的适应能力较强,叶片的蒸腾速率、气孔导度、水分利用效率和净光合速率较高;林窗中香果树实生苗叶片的叶绿素含量较低,但实生苗的生长速度最快,生物量最大;香果树实生苗的净光合速率与光合有效辐射和气孔导度呈显著正相关.对于冠下生境,需降低林冠层密度,增加透光率,以利于香果树实生苗的光合作用;对于林缘生境,则需要增加植被盖度,降低光照强度,以利于其快速生长.     

红松阔叶混交林林隙光量子通量密度的时空分布格局

以小兴安岭原始红松阔叶混交林林隙为对象,采用网格法布点,对生长季林隙内各样点光量子通量密度(photosynthetic photon flux density,PPFD)进行连续观测,利用基本统计学和地统计学方法分析其时空分布格局.结果表明:红松阔叶混交林林隙的PPFD高值区日变化明显,最大值出现在12:00,位于林隙北侧.林隙的PPFD 6月最高,7、8、9月依次递减,其中7月PPFD不同位置间的变异系数最大;各月均为中等变异.不同月份林隙PPFD空间异质性的强度和尺度不同,6月变程最大,7月基台值和结构比最大;各月林隙PPFD斑块复杂程度不同,最大值均位于林隙东北侧.郁闭林分和空旷地的月平均PPFD变化次序与林隙一致.各样点月平均PPFD为空旷地最高,林隙次之,郁闭林分最低.     

辽东山区次生林不同大小林窗光照特征比较

以辽东山区天然次生林中3种不同大小林窗(G₁,670 m²;G₂,290 m²和G₃,90 m²)为对象,通过对林窗内光强进行连续观测,比较光量子通量密度(PPFD)的时空分布.结果表明：3种林窗的PPFD日变化均呈现北高南低,且面积越大,PPFD高值区范围越广,异质性越明显;3种林窗的PPFD月变化规律为：林窗内各方位PPFD最大值集中在生长季初期(4—5月),最小值出现的月份则有所差异;3种林窗东部和西部的PPFD出现极值的时间基本一致,且春季光强均明显高于夏、秋季(P<0.05);G₁、G₂、G₃中心点的月平均PPFD分别占全光照的66.59%、49.05%和30.37%,在生长旺盛期,中心点光强分别是林内的37.8倍、27.9倍和10.3倍.受林窗面积不同,以及地形、边缘木高度(林窗形状)等因素的影响,不同大小林窗接收的光强及其分布格局不同,这是导致林窗内更新格局、物种组成发生变化的关键因素.     

Compatible and self-incompatible pollination in Pyrus communis displays different polyamine levels and transglutaminase activity

The polyamine (PA) content and the transglutaminase (TGase) activity have been investigated in Pyrus communis pollination with compatible and self-incompatible (SI) pollen in order to deepen their possible involvement in the progamic phase of plant reproduction. The PA distribution as free, perchloric acid (PCA)-soluble and PCA-insoluble fractions in ungerminated (UGP), germinating pollen (GP), styles and pollinated styles with compatible and SI pollens is discussed in the light of a possible role during pollination. Generally, the conjugated PAs both in PCA-soluble and PCA-insoluble fractions were higher than the free form. Within the conjugated PAs, the PCA-insoluble ones were the highest with the exception of the not pollinated styles. As TGase mediates some of the effects of PAs by covalently binding them to proteins, the activity of this enzyme, never checked before in styles and pollinated styles, was examined. In the SI styles, the TGase activity is higher in comparison to style-pollinated with compatible pollen, and high molecular mass cross-linked products were formed, suggesting an involvement of TGase in SI response. This is the first evidence on the presence of this enzyme activity in not pollinated and pollinated styles.     

Basal leaf senescence in a sunflower (Helianthus annuus) canopy: responses to increased R/FR ratio

Senescence of lower leaves (LS) begins before anthesis in sunflower crop canopies. Using isolated field-grown sunflower plants, it has previously been shown that pre-anthesis LS is dependent on photosynthetic photon flux density (PPFD) and is hastened by increases in far-red light. We tested the hypothesis that increasing the red/far-red ratio (R/FR) perceived by basal leaves within canopies delays LS. To do this, light impinging on the lower surface of north-oriented 8th leaves (cotyledons=0) of crops with maximum leaf area indexes of 3.3 (Experiment 1) and 2.4 (Experiment 2) was enriched (+8.33 μmol m⁻² s⁻¹) with red light using light emitting diode (LED) panels. LED panels constructed with unlit LED or with green LED (PPFD slightly greater than the red LED panels, to compensate for lower efficiency) were used as controls. Compared with controls, additional R significantly ( P <0.05) increased R/FR perceived by the lower surface and significantly ( P <0.01) delayed LS. On average, leaf duration, as time between full expansion and a 70% diminution of chlorophyll content, was 5 days greater for leaves receiving extra red light (maximum observed LD=27 days). We conclude that an increase in the R/FR ratio can delay LS in crop canopies.     

盐适应过程中香根草内源游离态、结合态、束缚态多胺含量的变化

研究了不同浓度NaCl胁迫下,香根草（Vetiteria zizanioides）根、叶中的游离态、结合态、束缚态多胺（PAs）[包括腐胺（Put）,尸胺（Cad）,亚精胺（Sod）和精胺（Spm）]含量的变化。在中度盐胁迫（100,200mmol L^-1NaCl）9天时,香根草基本能够正常生长,但在重度盐胁迫（300mmol L^-1NaCl）下,其生长受到严重抑制。在上述3个不同浓度的NaCl胁迫下,香根草根、叶中游离态Put,Cad,spd,Stma和总的游离态PAs含量明显下降,在高盐浓度下下降幅更大;结合态Put,Cad,Sod,Spm和总的结合态PAs含量显著上升,但在重度盐胁迫下升幅较小或与对照相当;束缚态Put,Cad和总的束缚态PAs含量均减少,而束缚态Spd和Spm含量在叶中是下降的,在根中则增加,且在中度盐胁迫下更明显。对根和叶片而言,除游离态（Spd＋Spm）,Put比值在重度盐胁迫下较对照显著下降外,其它游离态、结合态、束缚态和总的（Spd＋Spm）／Put比值在不同盐胁迫下均上升,在中度盐胁迫下更明显。这表明,维持多胺总量的稳态和较高的（Spd＋Spm）／Put比值是香根草适应中度盐胁迫的一个重要机制。     

Microsite variation in light availability and photosynthesis in a cool-temperate deciduous broadleaf forest in central Japan

An empirical light simulation model was applied to estimate stand scale photosynthesis in a deciduous broadleaved forest in central Japan. Based on diurnal courses of photosynthetically active photon flux density (PPFD), we characterized the components of incoming light within the forest canopy, and found that the instantaneous relative PPFD (PPFD under the canopy relative to that above the canopy) under diffuse light condition was a reliable estimator of the intensity and duration of PPFD. We calculated the daily photosynthesis (A_day) for each PPFD class using photosynthesis–light response curves. Model simulated A_day were corroborated with the estimates obtained from the nearby CO₂ flux tower. The result demonstrated the potential of the light simulation model. The light use efficiency of two dominant species, Betula ermanii as overstory and Sasa senanensis as understory species, were then evaluated. At the forest understory, PPFD under 50 mol m^–2 s^–1 contributed to 77% of the sunshine duration on a completely clear day. Therefore, a higher apparent quantum yield for S. senanensis enhanced the utilization of low PPFD for photosynthesis. On the other hand, at the upper forest canopies, B. ermanii with a higher light-saturated photosynthetic rate used high PPFD efficiently. Consequently, potential of daily net photosynthesis for both B. ermanii and S. senanensis was high under each light condition. Such interspecific difference in the patterns of light utilization was suggested as one of factors allowing coexistence of the two species in the study forest.     

The consequences of delayed greening during leaf development for light absorption and light use efficiency

Many species of rainforest plants have an unusual form of leaf development such that leaves delay greening until after full leaf expansion. Chlorophyll accumulation was measured during leaf development in five woody rainforest species, three with white young leaves, and two with ‘normal’ greening. In the three species with white leaves, the chlorophyll content of the expanding leaves was about 0.4mg dm^?2, whereas in the two species with green young leaves, chlorophyll content was about 2.1 mg dm^?2. Chlorophyll accumulation in greenhouse and field experiments was independent of light level. During leaf expansion, species with delayed chloroplast development only absorb 18–25% of the maximum possible light, compared with 80% for species with normal greening. Furthermore, species with delayed greening have low chlorophyll contents and reduced absorption for at least 30 d after full expansion. At a PPFD typical of the forest under story, the photosynthetic light use efficiency based upon incident radiation was 0.030–0.036 for species with delayed chloroplast development and 0.068–0.085 for the two species with normal greening. The lower light use efficiency of white species was primarily due to decreased light absorption. However, they also had a slightly lower light use efficiency based upon absorbed radiation, suggesting that development of other components of the photo-synthetic apparatus also may be delayed. Despite the fact that delayed greening decreases light absorption and light use efficiency during leaf development, it is extremely common in shade-tolerant species. We suggest that an advantage of delayed greening is that resources are not invested in the leaf until it is fully expanded and better defended from herbivores.     

光强对烟草幼苗形态和生理指标的影响

通过白纱布遮荫模拟不同光生境条件(透光率分别为100%、68.2%、35.4%和16.7%),研究了光强因子对烟草幼苗形态和生理指标的影响.结果表明:随相对光强的减弱,幼苗高度增加,茎粗、干鲜比、叶片厚度和单位叶面积质量均呈降低趋势,幼苗干物质积累减少,但其对叶数的影响不大.弱光条件下,叶片自由水、叶绿素、总氮和蛋白质含量增加,束缚水含量降低,叶绿素a/b值减小,转化酶活性降低;烟草幼苗根系相对不发达,根冠比和根生物量减小,根系活力降低.表明弱光条件不利于培育烟草壮苗,生产中应尽可能改善苗床的光照条件.     

LIGHT EFFECTS ON LEAF MORPHOLOGY IN WATER HYACINTH (EICHHORNIA CRASSIPES)

Water hyacinth leaves in natural populations vary from being long and thin-petioled to being short with inflated petioles. A variety of factors has been used experimentally to alter water hyacinth leaf shape, but what controls the development of leaf morphology in the field has not been established. We measured photosynthetic photon flux density (PPFD) and spectral distribution of radiation in a natural water hyacinth population. PPFD in the center of the water hyacinth mat was reduced to 2.7% of full sunlight, and the red to far red (R:FR) ratio was reduced to 0.28. When shoot tips of plants were exposed to artificial light environments, only plants in the treatment with a R:FR ratio comparable to that in the natural population produced leaves with long, thin petioles. Shoot tips in full sun or covered with clear plastic bags or bags that reduced light quantity without greatly altering light quality produced shorter leaves with inflated petioles. We hypothesize that the altered light quality inside a mat is a major environmental control of water hyacinth leaf morphology.     

Changes in content of free,conjugated and bound polyamines and osmotic adjustment in adaptation of vetiver grass to water deficit

Osmotic adjustment and alteration of polyamines (PAs) have been suggested to play roles in plant adaptation to water deficit/drought stress. In this study, the changes in cell intactness, photosynthesis, compatible solutes and PAs [including putrescine (Put), spermidine (Spd) and spermine (Spm) each in free, conjugated and bound forms] were investigated in leaves of vetiver grass exposed to different intensity of water deficit stress and subsequent rewatering. The results showed that, when vetiver grass was exposed to the moderate (20% and 40% PEG-6000 solutions) and severe (60% PEG solution) water deficit for 6 days, the plant injury degree (expressed as the parameters of plant growth, cell membrane integrity, water relations and photosynthesis) increased and contents of free and conjugated Put decreased with the rise of PEG concentration. Under the moderate water deficit, the plants could survive by the reduced osmotic potential (ψ_s), increased free and conjugated Spd and Spm in leaves. After subsequent rewatering, the osmotic balance was re-established, most of the above investigated physiological parameters were fully or partly recovered to the control levels. However, it was not the case for the severely-stressed and rewatering plants. It indicates that, vetiver grass can cope well with the moderate water deficit/drought stress by using the strategies of osmotic adjustment and maintenance of total contents of free, conjugated and bound PAs in leaves.     

Transglutaminase activity decrease during acclimation to hyposaline conditions in marine seaweed Grateloupia doryphora (Rhodophyta, Halymeniaceae)

Polyamines (PAs), such as diamine putrescine (PUT), triamine spermidine (SPD) and tetraamine spermine (SPM) have been related to environmental stress, including salt stress. A marine red macrophyte alga Grateloupia doryphora (Montagne) Howe was used to investigate the role of PAs during acclimation to moderate hyposaline conditions (incubation 24h in 18 psu seawater as compared to 36 psu of natural seawater). The results obtained showed that a moderate hyposaline shock caused an increase in the free fraction of PUT, SPD and SPM, mainly due to a decrease in TGase activity, together with an apparent increase in the l-arginine dependent PAs synthesis (ODC and arginase decreased, and ADC slightly increased). The photosynthetic rate increased in thalli when exposed to free SPD at 18 psu, but it did not increase at 36 psu.     

Phosphorylation of the D1 photosystem II reaction center protein is controlled by an endogenous circadian rhythm

The light dependence of D1 phosphorylation is unique to higher plants, being constitutive in cyanobacteria and algae. In a photoautotrophic higher plant, Spirodela oligorrhiza, grown in greenhouse conditions under natural diurnal cycles of solar irradiation, the ratio of phosphorylated versus total D1 protein (D1-P index: [D1-P]/[D1] + [D1-P]) of photosystem II is shown to undergo reproducible diurnal oscillation. These oscillations were clearly out of phase with the period of maximum in light intensity. The timing of the D1-P index maximum was not affected by changes in temperature, the amount of D1 kinase activity present in the thylakoid membranes, the rate of D1 protein synthesis, or photoinhibition. However, when the dark period in a normal diurnal cycle was cut short artificially by transferring plants to continuous light conditions, the D1-P index timing shifted and reached a maximum within 4 to 5 h of light illumination. The resultant diurnal oscillation persisted for at least two cycles in continuous light, suggesting that the rhythm is endogenous (circadian) and is entrained by an external signal.     

Changes in photosynthetic activity,pigment composition,electrolyte leakage,lipid peroxidation,and antioxidant enzymes during ex vitro establishment of micropropagated <Emphasis Type="Italic">Rauvolfia tetraphylla</Emphasis> plantlets

The effects of photosynthetic photon flux density (PPFD) on antioxidant metabolism and photosynthetic properties in leaves during ex vitro establishment of micropropagated Rauvolfia tetraphylla plantlets were investigated. In vitro-propagated plantlets were acclimatized at either 50 (Low-light = LL) or 300 (High-light = HL) μmol m⁻²s⁻¹ photosynthetic PPFD for 4 weeks under controlled conditions. Increases in chlorophyll (Chl) a, b and carotenoid levels were observed in plantlets acclimatized at both light intensities. At transplantation, micropropagated plantlets were not photosynthetically active, but the net photosynthetic rate increased in newly formed leaves over time during acclimatization. The observed differences in pigment contents and photosynthetic rates suggested adaptation of plantlets from heterotrophic to autotrophic mode of nutrition during acclimatization. Changes in activities of antioxidant enzymes were also observed during acclimatization. Superoxide dismutase activity increased in plantlets acclimatized at HL intensities. Likewise, changes in activity of catalase and ascorbate peroxidase were also detected. These observed changes reflected the ability of plants in developing an antioxidant enzymatic defense system aiding in survival against oxidative stress and in reducing release of free radicals.     

红松阔叶混交林林隙光量子通量密度、气温和空气相对湿度的时空分布格局

以小兴安岭原始红松阔叶混交林林隙为研究对象,通过对林隙内光量子通量密度(PPFD)、气温和空气相对湿度进行连续观测,比较其间的时空分布格局.结果表明：晴天和阴天阔叶红松林林隙的PPFD日最大值均出现在11:00—13:00,晴天林隙内各个时段最大值出现位置不同,日最大值出现在林隙北侧林冠边缘处;而阴天各个时段最大值均处于林隙的中心.林隙内月平均PPFD 为6月最高、9月最低,极差7月最大.林隙内晴天气温的峰值出现在9:00—15:00,而阴天气温峰值在15：00—19：00,均位于林隙中心南8 m.5:00—9:00林隙各点阴天的气温都高于晴天,9:00—19:00则相反.月平均气温为6月最高、9月最低.晴天和阴天空气相对湿度的峰值均出现在5:00—9:00,日最大值在林隙西侧林冠边缘处,且阴天的相对湿度始终大于晴天.月平均相对湿度为7月最高、6月最低.晴天PPFD的异质性大于阴天,而气温和相对湿度的异质性则不明显.生长季内不同月份PPFD、气温和空气相对湿度的最大值所处位置不同.PPFD和气温的月均值在林隙中心及附近变化梯度较大,而相对湿度的月均值则在林隙边缘变化梯度较大.     

Salinity-induced tissue-specific diurnal changes in nitrogen assimilatory enzymes in tomato seedlings grown under high or low nitrate medium.

We studied the salt stress (100 mM NaCl) effects on the diurnal changes in N metabolism enzymes in tomato seedlings (Lycopersicon esculentum Mill. cv. Chibli F1) that were grown under high nitrogen (HN, 5 mM NO(3)(-)) or low nitrogen (LN, 0.1 mM NO(3)(-)). NaCl stress led to a decrease in plant DW production and leaf surface to higher extent in HN than in LN plants. Total leaf chlorophyll (Chl) content was decreased by salinity in HN plants, but unchanged in LN plants. Soluble protein content was decreased by salt in the leaves from HN and LN plants, but increased in the stems-petioles from LN plants. Nitrate reductase (NR, EC 1.6.1.6) showed an activity peak during first part of the light period, but no diurnal changes were observed for the nitrite reductase (NiR, EC 1.7.7.1) activity. Glutamine synthetase (GS, EC 6.3.1.2) and glutamate synthase (Fd-GOGAT, EC 1.4.7.1) activities increased in HN plant leaves during the second part of the light period, probably when enough ammonium is produced by nitrate reduction. NR and NiR activities in the leaves were more decreased by NaCl in LN than in HN plants, whereas the opposite response was obtained for the GS activity. Fd-GOGAT activity was inhibited by NaCl in HN plant leaves, while salinity did not shift the peak of the NR and Fd-GOGAT activities during a diurnal cycle. The induction by NaCl stress occurred for the NR and GS activities in the roots of both HN and LN plants. Glutamate dehydrogenase (GDH, EC 1.4.1.2) activity shifted from the deaminating activity to the aminating activity in all tissues of HN plants. In LN plants, both aminating and deaminating activities were increased by salinity in the leaves and roots. The differences in the sensitivity to NaCl between HN and LN plants are discussed in relation to the N metabolism status brought on by salt stress.