Effect of Cadmium on gamma-Glutamylcysteine Synthesis in Maize Seedlings

Cysteine, γ-glutamylcysteine, and glutathione and the extractable activity of the enzymes of glutathione biosynthesis, γ-glutamylcysteine synthetase (EC 6.3.2.2) and glutathione synthetase (EC 6.3.2.3), were measured in roots and leaves of maize seedlings (Zea mays L. cv LG 9) exposed to CdCl₂ concentrations up to 200 micromolar. At 50 micromolar Cd²⁺, γ-glutamylcysteine contents increased continuously during 4 days up to 21-fold and eightfold of the control in roots and leaves, respectively. Even at 0.5 micromolar Cd²⁺, the concentration of γ-glutamylcysteine in the roots was significantly higher than in the control. At 5 micromolar and higher Cd²⁺ concentrations, a significant increase in γ-glutamylcysteine synthetase activity was measured in the roots, whereas in the leaves this enzyme activity was enhanced only at 200 micromolar Cd²⁺. Labeling of isolated roots with [³⁵S]sulfate showed that both sulfate assimilation and glutathione synthesis were increased by Cd. The accumulation of γ-glutamylcysteine in the roots did not affect the root exudation rate of this compound. Our results indicate that maize roots are at least in part autonomous in providing the additional thiols required for phytochelatin synthesis induced by Cd.     

谷胱甘肽还原酶和超氧化物歧化酶在玉米幼苗热激诱导的交叉适应中的作用

玉米幼苗经过热激处理后,在随后的高温、低温、干旱和盐胁迫环境中,其存活率明显高于未热激的幼苗,表明热激能提高植物的抗热性、抗冷性、抗旱性和抗盐性,证实了玉米幼苗交叉适应现象的存在;热激还可提高谷胱甘肽还原酶(GR)和超氧化物歧化酶(SOD)的活性,且在上述4种胁迫过程中GR、SOD活性水平与玉米幼苗的存活率呈正相关,表明GR和SOD参与玉米幼苗交叉适应的形成.     

IAA Oxidase Inhibitors from Normal and Mutant Maize Plants

Extracts of maize (Zea mays L.) plants contain substances which, in vitro, inhibit an indoleacetic acid (IAA) oxidase enzyme from maize. The extracts can be freed of inhibitors by dialysis or by passage through columns of polyvinylpyrrolidone powder. Inhibitor-free extracts contain an IAA oxidase enzyme which requires a phenolic co-factor and is stimulated by Mn²⁺.     

Localization of [gamma]-Glutamylcysteine Synthetase and Glutathione Synthetase Activity in Maize Seedlings

Fresh weight, protein, cysteine, [gamma]-glutamylcysteine, glutathione, and the extractable activity of the enzymes of glutathione biosynthesis, [gamma]-glutamylcysteine synthetase (EC 6.3.2.2) and glutathione synthetase (EC 6.3.2.3), were measured in roots, scutella, endosperms, and shoots of 3-, 7-, and 11-d-old maize (Zea mays L. cv LG 9) seedlings. In 3-d-old seedlings, the scutella represented 14% of the seedling fresh weight, containing 43% of total protein and 63 and 55% of the activity of [gamma]-glutamylcysteine synthetase and glutathione synthetase, respectively; in 11-d-old seedlings, the corresponding values were 4.5% for fresh weight, 8.0% for protein content, and 14 and 20% for the enzyme activities. The highest concentrations of thiols were found for cysteine (0.27 mM) in the roots, for glutathione (4.4 mM) in the shoots, and for [gamma]-glutamylcysteine (13 [mu]M) in the scutella of 3-d-old seedlings. The enzyme activities of roots were localized in subcellular fractions after sucrose density gradient centrifugation. Nearly half of the [gamma]-glutamylcysteine synthetase activity was detected in the root proplastids of 4-d-old seedlings, whereas <10% of the glutathione synthetase activity was localized in this organelle. Our results demonstrate the importance of scutella in glutathione synthesis in the early stage of seedling development. Unlike chloroplasts, root plastids show only a small proportion of glutathione synthetase activity.     

In Situ Microdialysis for Monitoring of Extracellular Glutathione Levels in Normal,Ischemic and Post-Ischemic Skeletal Muscle

Microdialysis probes were inserted into the tibialis anterior muscle and into the femoral vein of anaesthetised Sprague-Dawley rats for monitoring of reduced (GSH) and oxidized (GSSG) extracellular glutathione. The dialysates were analysed using HPLC. The levels of GSH and GSSG were high immediately after implantation in the skeletal muscle and declined to steady state levels after 90 minutes into the same range as that found in the venous dialysate. Total ischemia was induced two hours after implantation of the dialysis probe after steady state levels had been reached. The extracellular levels of GSH increased during total ischemia and had doubled at the end of the ischemic period compared to preischemic values. During the following initial 30 minutes of reperfusion the levels increased further to four-fold the preischemic levels. The levels of GSSG also increased (100%) during the initial 30 minutes of reperfusion. The extracellular GSH levels remained elevated for 1 hour of reperfusion, but the GSSG levels returned to preischemic levels. The results indicate that intermittent hypoxia or anoxia in muscle tissue through hypoperfusion or ischemia decreases intracellular GSH stores by leakage, reducing the intracellular antioxidative capacity and increasing the risk for oxidative reperfusion injury upon final normalization of tissue blood supply.     

In Situ Microdialysis for Monitoring of Extracellular Glutathione Levels in Normal, Ischemic and Post-Ischemic Skeletal Muscle

Microdialysis probes were inserted into the tibialis anterior muscle and into the femoral vein of anaesthetised Sprague-Dawley rats for monitoring of reduced (GSH) and oxidized (GSSG) extracellular glutathione. The dialysates were analysed using HPLC. The levels of GSH and GSSG were high immediately after implantation in the skeletal muscle and declined to steady state levels after 90 minutes into the same range as that found in the venous dialysate. Total ischemia was induced two hours after implantation of the dialysis probe after steady state levels had been reached. The extracellular levels of GSH increased during total ischemia and had doubled at the end of the ischemic period compared to preischemic values. During the following initial 30 minutes of reperfusion the levels increased further to four-fold the preischemic levels. The levels of GSSG also increased (100%) during the initial 30 minutes of reperfusion. The extracellular GSH levels remained elevated for 1 hour of reperfusion, but the GSSG levels returned to preischemic levels. The results indicate that intermittent hypoxia or anoxia in muscle tissue through hypoperfusion or ischemia decreases intracellular GSH stores by leakage, reducing the intracellular antioxidative capacity and increasing the risk for oxidative reperfusion injury upon final normalization of tissue blood supply.     

Changes in Isozyme Profiles of Catalase,Peroxidase, and Glutathione Reductase during Acclimation to Chilling in Mesocotyls of Maize Seedlings

The response of antioxidants to acclimation and chilling in various tissues of dark-grown maize (Zea mays L.) seedlings was examined in relation to chilling tolerance and protection from chilling-induced oxidative stress. Chilling caused an accumulation of H2O2 in both the coleoptile + leaf and the mesocotyl (but not roots), and acclimation prevented this accumulation. None of the antioxidant enzymes were significantly affected by acclimation or chilling in the coleoptile + leaf or root. However, elevated levels of glutathione in acclimated seedlings may contribute to an enhanced ability to scavenge H2O2 in the coleoptile + leaf. In the mesocotyl (visibly most susceptible to chilling), catalase3 was elevated in acclimated seedlings and may represent the first line of defense from mitochondria-generated H2O2. Nine of the most prominent peroxidase isozymes were induced by acclimation, two of which were located in the cell wall, suggesting a role in lignification. Lignin content was elevated in mesocotyls of acclimated seedlings, likely improving the mechanical strength of the mesocotyl. One cytosolic glutathione reductase isozyme was greatly decreased in acclimated seedlings, whereas two others were elevated, possibly resulting in improved effectiveness of the enzyme at low temperature. When taken together, these responses to acclimation illustrate the potential ways in which chilling tolerance may be improved in preemergent maize seedlings.     

植物铁吸收、转运和调控的分子机制研究进展

铁是植物正常生命活动所必需的微量矿质元素,铁离子的吸收、转运和利用是一个复杂的过程,很多基因参与了这一过程。本文对近10年来发现和分离的参与植物铁吸收、转运及调控的基因研究进展进行了综述。根据最近的研究结果,提出了植物控制铁吸收的分子调控模式(机理I)。     

植物铁吸收、转运和调控的分子机制研究进展

铁是植物正常生命活动所必需的微量矿质元素, 铁离子的吸收、转运和利用是一个复杂的过程, 很多基因参与了这一过程。本文对近10年来发现和分离的参与植物铁吸收、转运及调控的基因研究进展进行了综述。根据最近的研究结果, 提出了植物控制铁吸收的分子调控模式(机理I)。     

Different Ways of Formation of Root System in Plants,Maize and Flax Seedlings Taken as Examples

Comparison of the appearance and development of lateral roots in the flax and maize seedlings has shown the way of root branching in the flax, as distinct from that in most plants. Some primordia in the flax main root did not develop immediately into lateral roots, but remained quiescent, which determines different reactions of the maize and flax root systems to experimental influences. Decapitation of the main root in the maize did not leads to a significant increase in the number of lateral roots, while in the flax, their number noticeably increased due to the development of previously quiescent primordia into lateral roots. The treatment with synthetic auxin did not induce the formation of additional primordia and lateral roots in the maize roots. In the flax, the number of primordia increased significantly and that of lateral roots increased to a somewhat lesser extent. Apparently, the development of a primordium into a lateral root proceeds in two stages and they have different regulation.     

Polyamine Uptake and Translocation in Plants

Recently, evidence has increased for both long- and short-distance transport of polyamines (PAs) in living organisms, but the mechanisms involved and physiological significance of PAs translocation are still not well understood. This review deals with various aspects of polyamine uptake and transport in higher plant tissues.     

Sulfate Assimilation in C(4) Plants: Intercellular and Intracellular Location of ATP Sulfurylase, Cysteine Synthase, and Cystathionine beta-Lyase in Maize Leaves

The activity of ATP sulfurylase, cysteine synthase, and cystathionine β-lyase was measured in crude leaf extracts, bundle sheath strands, and mesophyll and bundle sheath chloroplasts to determine the location of sulfate assimilation of C₄ plant leaves. Almost all the ATP sulfurylase activity was located in the bundle sheath chloroplasts while cysteine synthase and cystathionine β-lyase activity was located, in different proportions, in both chloroplast types.

A new spectrophotometric assay for measuring ATP sulfurylase activity is also described.

     

Absorption,Translocation and Metabolism of Nicotinamide in Some Plants

The seedlings of rice, eggplant and tomato at the 5th leaf stage of growth readily absorbed exogenous ¹⁴C-nicotinamide through the root and the foliage in water culture. Within the 24 hr period after the bigining of cultivation, the radioactivity gradually translocated from the part treated with ¹⁴C-nicotinamide to the whole plant body. This compound was rapidly metabolised in the plants to at least six metabolites, in which three compounds were identified as nicotinic acid, NAD and NADP. ¹⁴C-Nicotinic acid was also taken up quickly through the root of rice and its metabolism showed a similar pattern to that of ¹⁴C-nicotinamide. The incorporation of radioactivity into NAD and NADP from ¹⁴C-nicotinamide added to cultivating solution at a concentration of 0.21 ppm was decreased to 10~20% by the simultaneous addition of unlabeled nicotinic acid at a concentration about 1000 times higher than that of the labeled one. It was concluded that the biosynthesis of these pyridine nucleotides from nicotinamide was chiefly via nicotinic acid. The formation of ¹⁴C-nicotinamide in the ¹⁴C-nicotinic acid metabolism suggested a breakdown of NAD. Three unknown compounds observed in both the metabolisms described above were not intermediates in the pyridine nucleotide biosynthesis.     

亚硒酸钠对小麦幼苗中谷胱甘肽过氧化物酶和谷胱甘肽转硫酶活性以及谷胱甘肽含量的影响

用1.0 mg·L-1的亚硒酸钠根施小麦幼苗,测定亚硒酸钠对谷胱甘肽过氧化物酶和谷胱甘肽转硫酶活性以及还原性谷胱甘肽含量的结果表明,外源亚硒酸钠对麦苗地上部的谷胱甘肽过氧化物酶和谷胱甘肽转硫酶活性均有诱导作用,使麦苗体内的谷胱甘肽含量水平增加.     

Induction of Nitrate Reductase and Peroxidase Activity in the Seedlings of Normal and High Lysine Maize

Steady state levels of in vivo nitrate reductase activity in the endosperm, scutella, roots and shoots of maize seedlings were higher in normal maize than those in high lysine maize. Activity of peroxidase in the roots, however, was higher in the high lysine cultivar. The nitrate reductase activity increased with the supply of nitrate in all parts of the seedlings of both cultivars, although the rates of increment in the endosperm were lower than those in scutella, roots and shoots. In relation to substrate concentration, a saturation was achieved at 5 to 10 mM of nitrate except in the endosperm, where activity increased slowly up to 100 mM at least. Final levels of enzyme activity were significantly higher in the scutella of normal than in that of high lysine seedlings. In vitro enzyme activity in the roots also increased with the supply of nitrate in both cultivars, reaching maximum at 5 to 10 mM nitrate.     

Some Aspects of Nitrate Assimilation in the Seedlings of Normal and Opaque-2 Mutant of Maize

The effects of 10 mM nitrate on the growth and nitrogenous componentsof Zea mays L. var. W64A wild type (normal) were compared tothose on its opaque-2 (high lysine) mutant during the first10 d of seedling growth at a constant temperature of 26 °Cand with a 16 h photoperiod. Nitrate supply had no effect onthe growth of embryonic axes in both lines till day 6. Growthof both lines was enhanced slightly after that time, however.Increases in 80% (v/v) ethanolsoluble and protein nitrogen werealso observed only after day 4 when the supply of nitrogen fromthe storage proteins in the endosperm was limiting. Nitratehad no effect on the synthesis of chlorophyll during leaf developmentbut it did increase the total chlorophyll in mature and senescingprimary leaves. The increase in nitrogenous components or chlorophyllin opaque-2 was more pronounced than in the normal type. Itmight be related to the lower proline or higher lysine in themutant.     

The Redox State of Glutathione,Cysteine and Homocysteine in the Extracellular Fluid in the Skin

Glutathione, the most abundant low-molecular weight thiol in the skin, has been shown to protect the skin from both photobiological and chemical injury. The thiols, glutathione in particular, have also been shown to be crucially involved in defence against contact allergens. Since the levels of extracellular thiol concentrations are important determinants of intracellular thiol status, we have compared the normal concentrations and the redox status of the main low-molecular weight thiol components in the extracellular fluid at the dermo-epidermal junction with the corresponding plasma levels. In their sulfhydryl form, all three thiols, i.e. glutathione, cysteine and homocysteine, were more abundant in experimental skin blister fluid than in plasma, as were the free disulfides of glutathione and homocysteine, whereas the free disulfides of cysteine were about the same in blister fluid and in plasma. Protein mixed disulfide levels were higher in plasma than in blister fluid. The present results provide information concerning the extracellular defence in the skin.     

The Production, Storage, and Translocation of Carbohydrates in Developing Potato Plants

¹⁴C-tracer experiments showed that the export of assimilateslabelled with a short pulse of ¹⁴CO₂ continued for 5 weeks.There appeared to be an approximately exponential loss of ethanol-insoluble¹⁴C from the haulm with a half-time of about 3.5 weeks. Initiallythere was a poor correlation between the fresh weight of thetubers and their ¹⁴C content but the correlation became goodafter 5 weeks. Field data showed that, in many instances, the rate of tubergrowth was greater than the rate of growth of the whole plant.This suggests the transfer of large amounts of dry matter fromthe haulm to the tubers. Dry-matter loss from the haulm couldaccount for the discrepancy and the amount transferred in thisway was about 10 per cent of the final dry weight of the tuber. There was an increase in net assimilation rate when the leafarea of the plants started to decline. It is suggested thatthis increase was due to an increase in the rate of photosynthesisbrought about by the influence of the rapidly growing tubers.