壳聚糖对镉胁迫下玉米幼苗叶片AsA-GSH循环的调控效应

以玉米（Zea mays L.）品种‘郑单958’为实验材料,分析外施壳聚糖对镉胁迫下玉米幼苗生物量、叶片镉含量、叶片超氧阴离子（O₂^·-）产生速率和过氧化氢（H₂O₂）的含量,以及抗坏血酸-谷胱甘肽（AsA-GSH）循环中抗氧化酶的活性及抗氧化物含量的影响。结果显示,随着镉胁迫时间的延长,玉米幼苗发生氧化胁迫,叶片抗氧化酶（APX、GR、DHAR、MDHAR）活性和抗氧化物（AsA、GSH）的含量降低,镉积累过量会抑制玉米幼苗的生长。施加壳聚糖可以降低镉胁迫下玉米幼苗叶片O₂^·-的产生速率和H₂O₂含量,提高上述抗氧化酶活性和抗氧化物的含量,促进AsA和GSH的再生,维持细胞的氧化还原状态,促进玉米幼苗的生长。研究结果表明壳聚糖处理后玉米幼苗保持了较高的AsA-GSH循环运作效率,提高了抗氧化能力,可有效缓解镉胁迫对玉米幼苗生长的抑制。     

Isolation and characterization of Azotobacter chroococcum from the roots of Zea mays

Abstract Bacteria showing rapid growth on a nitrogenfree medium and acetylene-reducing activity were isolated from maize roots collected from agricultural soils in Spain. The isolates were Gram-negative motile rods and were identified as Azotobacter chroococcum . Acetylene-reducing activity and microbial counts were determined on root segments from 7- and 30-day-old plants. Rates obtained were in the range of 0.0053–0.848 nmol C₂H₂· g⁻¹· h⁻¹. Root populations were 1.4–6.0 × 10⁴ micro-organisms · g⁻¹. These results showed that there was an association between A. chroococcum strains and roots of maize planted in some Spanish soils.     

Maximal biomass production can occur in corn (Zea mays) in the absence of NO3 accumulation in either leaves or roots

In order to investigate effects of limited NO₃ availability in corn ( Zea mays L. cv. Brulouis) 17-day-old plants were grown for a further 25 days on sand in a growth chamber. The plants received frequent irrigation with a complete nutrient solution containing 0.2, 0.6, 1.5 or 3.0 mM NO₃. With 0.2 mM NO; nitrate levels in both roots and leaves diminished rapidly and were almost zero after 10 days treatment. Concurrently, as signs of nitrogen deficiency appeared, shoot growth was restricted, whereas root growth was enhanced. In addition, the concentration of reduced nitrogen and malate in the leaves declined, and in vitro nitrate reductase activity (NRA. EC 1.6.6.1), soluble protein and chlorophyll levels of leaf tissue were depressed and starch concentration was enhanced. With 0.6 mM NO₃ in the nutrient solution, the decrease in NO₃ levels in the tissues and the increase in root development were similar to those observed with 0.2 mM NO₃. However, shoot growth, reduced nitrogen concentration in leaves, and the above-mentioned biochemical characteristics were almost identical to those obtained at 1.5 and 3.0 mM NO₃. This indicates that when supplied with 0.6 mM NO₃, corn plants were able to absorb sufficient NO3 to support maximal biomass production without appreciable NO₃ accumulation in roots or shoot. It is, thus, suggested that the plants responded to low NO₃, availability in medium by enhancing root growth and by maximizing NO₃ reduction relative to NO₃ accumulation.     

Tissue distribution and developmental patterns of NADH-dependent and ferredoxin-dependent glutamate synthase activities in maize (Zea mays) kernels

Both NADH-dependent glutamate synthase (NADH-GOGAT, EC 1.4.1.14) and ferredoxin-dependent glutamate synthase (Fd-GOGAT, EC 1.4.7.1) activities were present in the endosperm, embryo, pedicel and pericarp of maize ( Zea mays L. var. W64A × A619) kernels. The endosperm contained the highest proportions of each activity on a per tissue basis. In the endosperm, NADH-GOGAT and Fd-GOGAT activities increased 12- and 2.5-fold, respectively, during early zein accumulation. NADH-GOGAT and Fd-GOGAT activities were expressed in the upper, middle and lower portions of the endosperm in a manner that paralleled but preceded zein accumulation. Maize endosperm NADH-GOGAT was purified 159-fold using ammonium sulfate fractionation, anion exchange chromatography and dye-ligand chromatography. Apparent K_m values for glutamine, α-ketoglutarate and NADH were 850, 19 and 1 μM, respectively. The results are consistent with endosperm GOGAT functioning to redistribute nitrogen from glutamine, the predominant nitrogenous compound delivered to the endosperm, into other amino acids needed for storage protein synthesis.     

Calmodulin and the Regulation of Succinate Dehydrogenase Activity in the Mitochondria of Zea mays

Succinate dehydrogenase (SDH) was purified by DEAE C-32 chromatography from the mitochondrial fraction of corn ( Zea mays L. ). Free calmodulin (CAM) could not be detected in the purified SDH with the method based on the ability of SDH to stimulate NAD kinase, but it still contained some CAM when measured with the ELISA method. Purified SDH could stimulate NAD kinase only after heating to release free CAM. Plain polyacrylamide gel electrophoresis (PAGE) of the pufffled SDH revealed only one peptide band, but three peptide bands were shown on SDS-PAGE, their molecular weight being 67.0 kD, 30.0 kD, 16.7 kD respectively. The 67.0 kD and 30.0 kD peptides corresponded to the large and small molecular subunit of SDH respectively. The Rf value of the 16.7 kD peptide band was identical to the standard CAM in the SDS-PAGE. From all the above evidence, the authors suggested that CAM might exert its function of SDH regulation in a binding state with the SDH molecule.     

Response of Zea mays to the Inoculation with Azospirillum on Nitrogen Metabolism under Greenhouse Conditions

The maize (Zea mays L.) plants inoculated by N₂-fixing bacterium Azospirillum showed increased activity of glutamate dehydrogenase (GDH) and glutamine synthetase (GS) in root cells free extracts over uninoculated control plants. Maximum differences in NADH-GDH activity were observed during the second and third weeks after sowing. The specific activity of GS showed a greater increase at the end of the assay. The percentage of nitrogen in leaves, root and foliage length, total fresh mass and nitrogenase activity were higher in inoculated plants than in the control ones.     

Relationship between iron chlorosis and alkalinity in Zea mays

Mengel, K. and Geurtzen, G. 1988. Relationship between iron chlorosis and alkalinity in Zea mays . - Physiol. Plant. 72: 460–465.
Maize ( Zea mays L. cv. Anjou 21) grown in nutrient solution with Fe-EDTA and with nitrate as the sole nitrogen source showed typical Fe-chlorosis symptoms after a growth period of 14–21 days. Alkalinity in roots, stems and leaves of the chlorotic plants was high. Transferring the chlorotic plants from the nitrate-containing nutrient solution to a solution of (NH₄)₂SO₄ resulted in a regreening of leaves within 2–3 days which was associated with a decrease in solution pH, a decrease in alkalinity of plant parts, a translocation of Fe from roots to tops and a release of Fe into the outer solution. Similar effects were obtained when Fe chlorotic plants were transferred to a dilute HO solution with pH 3.5.
Spraying chlorotic leaves with indoleacetic acid or with fusicoccin led also to a regreening of leaves without having a major effect on leaf alkalinity.
Interpretation of the experimental results is based on the assumption that nitrate as sole N source leads to a high pH level in the apoplast resulting in the precipitation of Fe compounds, probably Fe oxide hydrate. Ammonium nutrition has the reverse effect since it lowers the apoplast pH and this can result in the dissolution of Fe compounds. Application of indoleacetic acid as well as fusicoccin supposedly stimulates the proton pumps in the plasmalemma of the leaf tissue. The resulting decrease in apoplast leaf pH in the microenvironment also leads to a dissolution of Fe compounds in the apoplast and thus promotes the uptake of Fe by the symplasm.     

Glutamine synthetase activity of the endosperm, embryo and pedicel-placento-chalazal regions of developing maize (Zea mays) kernels

Glutamine synthetase (GS, EC 6.3.1.2) activity in homogenates of the maize ( Zea mays L. hybrid A619 X W64A) kernel pedicel-placento-chalazal (PPCh), endosperm regions was characterized in order to optimize assay (hydroxylamine-dependent γ-glutamyl hydroxymate formation) conditions for quantitating maize kernel GS in crude extracts. The GS activities of all three tissue extracts exhibited optima at pH 7.0 with ATP:Mg²⁺ of 1:1.6. Assays of kernel tissue GS activity required relatively high concentrations of substrates to achieve saturation compared to GS from other plant tissue sources, a point which has not been considered in previous reports of maize kernel GS activity. When measured under optimal assay conditions. PPCh-GS increased to a peak of 51 nmol γ-glutamyl hydroxymate kernel⁻¹ min⁻¹ at 25 days after pollination and then declined throughout the remainder of kernel development. Embryo GS activity increased steadily throughout development to a maximum of 24 nmol γ-glutamyl hydroxymate embryo⁻¹ min⁻¹ by 50 days after pollination. In contrast, endosperm GS activity, which was 25 nmol γ-glutamyl hydroxymate endosperm⁻¹ min⁻¹ at 25 days after pollination, exhibited no discernable pattern of change during kernel development. These findings are discussed with respect to the possible roles PPCh, endosperm and embryo GS play in kernel development.     

Modelling postsilking nitrogen fluxes in maize (Zea mays) using 15N-labelling field experiments

In maize (Zea mays), nitrogen (N) remobilization and postflowering N uptake are two processes that provide amino acids for grain protein synthesis. To study the way in which N is allocated to the grain and to the stover, two different 15N-labelling techniques were developed. 15NO(3-) was provided to the soil either at the beginning of stem elongation or after silking. The distribution of 15N in the stover and in the grain was monitored by calculating relative 15N-specific allocation (RSA). A nearly linear relationship between the RSA of the kernels and the RSA of the stover was found as a result of two simultaneous N fluxes: N remobilization from the stover to the grain, and N allocation to the stover and to the grain originating from N uptake. By modelling the 15N fluxes, it was possible to demonstrate that, as a consequence of protein turnover, a large proportion of the amino acids synthesized from the N taken up after silking were integrated into the proteins of the stover, and these proteins were further hydrolysed to provide N to the grain.     

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.     

A study of the impaired growth of roots of Zea mays seedlings at low oxygen concentrations

Abstract. Seedlings of Zea mays L. were grown in the dark at 27°C. Four-day-old seedlings were then exposed for 3 days to solutions equilibrated with gas mixtures to give O₂ concentrations between 0.02 and 0.25 mol m^?3. Root growth was impaired just as severely at 0.06 as 0.02 mol O₂ m^?3 while growth at 0.16 mol O₂ m^?3 was about the same as in solutions in equilibrium with air (0.25 mol O₂ m^?3). Growth of young seedlings at low O₂ concentrations was inhibited to the same extent in nutrient solution and 0.5 ml m^?3 CaCl₂, showing that the adverse effect of O₂ deficits on growth was not due to less uptake of inorganic nutrients. Furthermore, at low O₂ concentrations neither exposure of the shoots to a relative humidity of 100% (26.0 g H₂O m^?3) nor excision of the entire shoot enhanced root growth relative to that in plants with shoots at a relative humidity of 50% (13.0 g H₂O m^?3). Therefore, for these seedlings growing in the dark, impairment of root growth at low O₂ concentrations was not a consequence of water deficits in the shoot or of other shoot-root interactions. Total soluble sugars and amino acid concentrations were generally greater at low (0.02–0.06 mol O₂m^?3) than at high O₂ concentrations (0.16–0.25 mol O₂ m ^?3). This applied specifically to the root apices (0–2 mm) and expanding (2–15 mm) tissue except in some experiments where sugar concentrations in expanding tissue were slightly greater at high than at low O₂ concentrations. Critical O₂ pressures for respiration of excised root segments were approximately 0.117 and 0.065 mol O₂ m^?3 in the expanding and expanded zones of the roots, respectively. In contrast, the critical O₂ pressure exceeded 0.20 mol O₂ m^?3 in the apex, suggesting that O₂ supply for metabolic processes is most likely to be sub-optimal in this zone. Our results show clearly that the adverse effects of low O₂ concentrations are unlikely to be a consequence of substrate shortage for either respiration or synthesis of macromolecules; low rates of ATP regeneration in growing root tissues are the logical cause for impaired growth in young seedlings while they are being sustained by seed reserves.     

Chilling stress and oxygen metabolizing enzymes in Zea mays and Zea diploperennis*

Abstract. The activities of five active-oxygen scavenging enzymes were compared for cold-lability and three were compared for chilling induction in two Zea genotypes of contrasting susceptibility to photoinhibition during chilling. Activities of superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), dehydroascorbate reductase (DHAR, EC 1.8.5.1), and glutathione reductase (GTR, EC 1.6.4.2) in leaf extracts from plants grown without chilling stress were assayed at 19°C and 5°C. Enzymes from the chilling-susceptible Z. Mays cv. LG11 had lower specific activities at 5°C than did enzymes from the chilling-tolerant Z. diploperennis, except for MDHAR where no significant differences were observed. The activities of SOD and APX from Z. diploperennis were double those of Z. mays at both assay temperatures. Monodehydroa-scrobate reductase and glutathione reductase activities in both species were reduced by 63–78% at a 5°C assay temperature. The dehydroascorbate reductase (DHAR) showed the greatest low-temperature lability losing 96% (Z. diploperennis) and 100% (Z. mays) of its activity at 5°C. To examine possible chilling-induced changes in levels of enzyme activity, plants of both Zea genotypes were transferred to growth chambers at 10°C at moderate light intensities. Glutathione reductase activity was found to increase within 24h in Z. diploperennis, but it decreased slightly in Z. mays. MDHAR activity decreased by 50% in Z. diploperennis but showed only a transient increase in activity in Z. mays.     

Exogenous application of abscisic acid or triadimefon affects the recovery of Zea mays seedlings from heat shock

Maize seedlings ( Zea mays L. cv. DK 246) grown for 1–4 days in the presence of abscisic acid (ABA) or triadimefon (a fungicide) demonstrated an enhanced ability to withstand the effects of a 3-h sub-lethal (40°C) or lethal (45°C) heat shock. Both the ABA and triadimefon treatments were applied solely to the roots of seedlings; however, the ability to withstand a heat shock was induced in both the root and the shoot. The level of protection provided by these agents was dependent upon the time that plants were exposed to them; prolonged exposure reduced tolerance to subsequent stress.     

Transpiration, and nitrogen uptake and flow in two maize (Zea mays L.) inbred lines as affected by nitrogen supply

BACKGROUND AND AIMS: The influence of two nitrogen (N) levels on growth, water relations, and N uptake and flow was investigated in two different inbred lines of maize (N-efficient Zi330 and N-inefficient Chen94-11) to analyse the differences in N uptake and cycling within a plant. METHODS: Xylem sap from different leaves of the inbred lines cultured in quartz sand was collected by application of pressure to the root system. Plant transpiration was measured on a daily basis by weighing five pots of each of the treatments. KEY RESULTS: N-efficient Zi330 had a higher relative growth rate and water-use efficiency at both high (4 mm) and low (0.08 mm) N levels. At a high N level, the amount of N taken up was similar for the two inbred lines; the amount of N transported in the xylem and retranslocated in the phloem was slight greater in Chen94-11 than in Zi330. At a low N level, however, the total amount of N taken up, transported in the xylem and retranslocated in the phloem of Zi330 was 2.2, 2.7 and 2.7 times more, respectively, than that of Chen94-11. Independent of inbred line and N level, the amounts of N transported in the xylem and cycled in the phloem were far more than that taken up by roots at the same time. Low N supply shifted NO(3)(-1) reduction towards the roots. The major nitrogenous compound in the xylem sap was NO(3)(-1), when plants grew at the high N level, while amino acid-N was predominant when plants grew at the low N level. CONCLUSIONS: The N-efficient maize inbred line Zi330 had a higher ability to take up N and cycle N within the plant than N-inefficient Chen94-11 when grown under N-deficiency.     

Influence of elevated CO2 and nitrogen nutrition on photosynthesis and nitrate photo-assimilation in maize (Zea mays L.)

Measurements of CO₂ and O₂ gas exchange and chlorophyll a fluorescence were used to test the hypothesis that elevated atmospheric CO₂ inhibits nitrate (NO₃ ^– ) photo‐assimilation in the C₄ plant, maize (Zea mays L.). The assimilatory quotient (AQ), the ratio of net CO₂ assimilation to net O₂ evolution, decreases as NO₃ ^– photo‐assimilation increases so that the difference in AQ between the ammonium‐ and nitrate‐fed plants (ΔAQ) provided an in planta estimate of NO₃ ^– photo‐assimilation. In fully expanded maize leaves, NO₃ ^– photo‐assimilation was detectable only under high light and was not affected by CO₂ treatments. Furthermore, CO₂ assimilation and O₂ evolution were higher under NO₃ ^– than ammonia (NH₄⁺) regardless of CO₂ levels. In conclusion, NO₃ ^– photo‐assimilation in maize primarily occurred at high light when reducing equivalents were presumably not limiting. Nitrate photo‐assimilation enhanced C₄ photosynthesis, and in contrast to C₃ plants, elevated CO₂ did not inhibit foliar NO₃^– photo‐assimilation.     

The pathway of nitrogen assimilation in plants

The major route of nitrogen assimilation has been considered for many years to occur via the reductive amination of α-oxoglutarate, catalysed by glutamate dehydrogenase. However, recent work has shown that in most bacteria an alternative route via glutamine synthetase and glutamine: 2-oxoglutarate aminotransferase (glutamate synthase) operates under conditions of ammonia limitation. Subsequently the presence of a ferredoxin-dependent glutamate synthase in green leaves and green and blue-green algae, and a NAD(P)H and ferredoxin-dependent enzyme in roots and other non-green plant tissues, has suggested that this route may also function in most members of the plant kingdom. The only exceptions are probably the majority of the fungi, where so far most organisms studied do not appear to contain glutamate synthase. Besides the presence of the necessary enzymes there is other evidence to support the contention that the assimilation of ammonia into amino acids occurs via glutamine synthetase and glutamate synthase, and that it is unlikely that glutamate dehydrogenase plays a major role in nitrogen assimilation in bacteria or higher plants except in circumstances of ammonia excess.     

Localization of the genes controlling B chromosome transmission rate in maize (Zea mays ssp. mays, Poaceae)

In previous papers we found that the frequency of B chromosomes in native races of maize varies considerably in different populations. Moreover, we found genotypes that control high and low transmission rates (TR) of B chromosomes in the Pisingallo race. In the present work crosses were made to determine whether the genes controlling B-TR are located on the normal chromosome set (As) or on the B chromosomes (Bs). We made female f.0B × male m.2B crosses between and within high (H) and low (L) B-TR groups. The Bs were transmitted on the male side in all cases. The mean B-TR from the progeny of f.0B (H) × m.2B (H) and f.0B (H) × m.2B (L) crosses was significantly higher than that from f.0B (L) × m.2B (L) and f.0B (L) × m.2B (H) crosses. The results show that the B-TR of the crosses corresponds to the H or L B-TR of the 0B female parents irrespective of the Bs of the male parent. This indicates that B-TR is genetically controlled by the 0B female parent and that these genes are located on the A chromosomes.     

Oxygen-dependent stomatal opening in Zea mays leaves. Effect of Light and carbon dioxide

The oxygen requirement for stomatal opening in maize plants ( Zea mays L. hybrid INRA 508) was studied at different CO₂ concentrations and light intensities. In the absence of CO₂, stomatal opening always required O₂, but this requirement decreased with increasing light intensity. In darkness, the lowest O₂ partial pressure needed to obtain a weak stomatal movement was about 50 Pa. This value was lowered to ca 10 Pa in light (320 μmol m⁻² s⁻¹).
On the other hand. in the absence of O₂, CO₂enabled stomatal opening to occur in the light, presumably due to the evolved photosynthetic O₂. Thus, CO₂, which generally reduced stomatal aperture, could induce stomatal movement in anoxia and light. The effect of CO₂ on stomatal opening was closely dependent on O₂ concentration and light intensity. Stomatal aperture appeared CO₂-independent at an O₂ partial pressure which was dependent on light intensity and was about 25 Pa at 320 umol m⁻² s⁻¹.
The presence of a plasmalemma oxidase, in addition to mitochondrial oxidase, might explain the differences in the O² requirement at various light intensities. The possible involvement of such a system in relation to the effect of CO₂ is discussed.