Cell wall peroxidase activity,hydrogen peroxide level and NaCl-inhibited root growth of rice seedlings

The changes in cell-wall peroxidase (POD) activity and H₂O₂ level in roots of NaCl-stressed rice seedlings and their correlation with root growth were investigated. Increasing concentrations of NaCl from 50 to 150 mM progressively reduced root growth and increased ionically bound cell-wall POD activity. NaCl had no effect on covalently bound cell-wall POD activities. The reduction of root growth by NaCl is closely correlated with the increase in H₂O₂ level. Exogenous H₂O₂ was found to inhibit root growth of rice seedlings. Since ammonium and proline accumulation are associated with root growth inhibition caused by NaCl, we determined the effects of NH₄Cl or proline on root growth, cell-wall POD activity and H₂O₂level in roots. External application of NH₄Cl or proline markedly inhibited root growth, increased cell-wall POD activity and increased H₂O₂ level in roots of rice seedlings in the absence of NaCl. An increase in cell-wall POD activity and H₂O₂ level preceded inhibition of root growth caused by NaCl, NH₄Cl or proline. NaCl or proline treatment also increased NADH-POD and diamine oxidase (DAO) activities in roots of rice seedlings, suggesting that NADH-POD and DAO contribute to the H₂O₂ generation in the cell wall of NaCl- or proline-treated roots. NH₄Cl treatment increased NADH-POD activity but had no effect on DAO activity, suggesting that NADH-POD but not DAO is responsible for H₂O₂ generation in cell wall of NH₄Cl-treated roots.     

Cell wall peroxidase against ferulic acid, lignin, and NaCl-reduced root growth of rice seedlings

The changes in cell wall peroxidase activity against ferulic acid (FPOD) and lignin level in roots of NaCl-stressed rice seedlings and their correlation with root growth were investigated. Increasing concentrations of NaCl from 50 to 150 mmol L⁻¹ progressively decreases root growth. The reduction of root growth by NaCl is closely correlated with the increase in FPOD activity extracted from the cell wall. In contrast, lignin level was reduced by NaCl. Since proline and ammonium accumulations are associated with root growth inhibition caused by NaCl, we determined the effect of proline or NH₄Cl on root growth and FPOD in roots. Exogenous application of NH₄Cl or proline markedly inhibited root growth and increased FPOD activity in rice seedlings in the absence of NaCl. An increase in FPOD activity in roots preceded inhibition of root growth caused either by NaCl, NH₄Cl, or proline. Our results suggest that cell-wall stiffening catalyzed by FPOD may participate in the regulation of root growth reduction of rice seedlings caused by NaCl.     

NaCl induced changes in ionically bound peroxidase activity in roots of rice seedlings

The changes in ionically bound peroxidase activity in roots of NaCl-stressed rice seedlings and their correlation with root growth were investigated. Increasing concentrations of NaCl from 50 to 150 mM progressively decreases root growth. The reduction of root growth by NaCl is closely correlated with the increase in ionically bound peroxidase activity. Since proline and ammonium accumulations are associated with root growth inhibition caused by NaCl, we determined the effects of proline or NH4Cl on root growth and ionically bound peroxidase activity in roots. External application of proline or NH4Cl markedly inhibited root growth and increased ionically bound peroxidase activity in roots of rice seedlings in the absence of NaCl. An increase in ionically bound peroxidase activity in roots preceded inhibition of root growth caused by NaCl, NH4Cl or proline. Mannitol inhibited root growth, but decreased rather than increased ionically bound peroxidase activity at the concentration iso-osmotic with NaCl. The inhibition of root growth and the increase in ionically bound peroxidase activity in roots by NaClis reversible and is associated with ionic rather than osmotic component. This revised version was published online in June 2006 with corrections to the Cover Date.     

Glutathione reduces the inhibition of rice seedling root growth caused by cadmium

We investigated the effects of agents known to affect cellular glutathione (reduced form, GSH) levels on the growth of rice seedlings treated with Cd. CdCl₂ was more effective than CdSO₄ in inhibiting root growth. However, CdCl₂ had no effect on shoot growth. GSH, a substrate for phytochelatin synthesis, was effective in counteracting growth inhibition of roots by CdCl₂. Root growth in the CdCl₂ medium was found also to be enhanced by the addition of L-glutamic acid and L-cysteine, both of which are substrates for GSH formation. Buthionine sulfoximine, an inhibitor of GSH synthesis, rendered the roots susceptible to growth inhibition by Cd. Our results suggest that GSH level may play a role in regulating Cd-inhibited growth of rice roots.Abbreviations BSO buthionine sulfoximine - GSH reduced form glutathione     

Arbuscular mycorrhizas alter root system architecture of Citrus tangerine through regulating metabolism of endogenous polyamines

Cadmium (Cd) is readily taken up by the roots of rice seedlings, leading to growth reduction. H₂O₂ is a constituent of oxidative metabolism and is itself a reactive oxygen species. In this study, the participation of H₂O₂ in CdCl₂-inhibited growth of rice roots was investigated. CdCl₂ treatment increased H₂O₂ production in rice roots. CdCl₂ treatment had no effect on the activities of superoxide dismutase, ascorbate peroxidase, and glutathione reductase, but inhibited the activity of catalase (CAT) in rice roots. CdCl₂-inhibited root growth and -increased H₂O₂ content were lessened in the presence of diphenyleneiodonium chloride, an inhibitor of H₂O₂ generating NADPH oxidase. However, this stimulation of root growth in CdCl₂-treated seedlings is small (about 5%). Calcium (Ca) is important in many physiological processes in plants. Attempts were also made to determine whether the action of Ca on CdCl₂-inhibited growth of rice roots is associated with H₂O₂. CaCl₂ application reduced the production of H₂O₂, the decrease in CAT activity, and the inhibition of root growth caused by CdCl₂. The effects of CaCl₂ application could be reversed by exogenous H₂O₂. Our results indicate that the Cd causes a decline in CAT and to a lower extent a stimulation of NADPH oxidase in rice roots, with the subsequent generation of H₂O₂, an agent responsible for growth inhibition.     

Photosynthetic pigments content, <Emphasis Type="Italic">δ</Emphasis>-aminolevulinic acid dehydratase and acid phosphatase activities and mineral nutrients concentration in cadmium-exposed <Emphasis Type="Italic">Cucumis sativus</Emphasis> L.

In this study, the effects of cadmium chloride (CdCl₂) on plant growth, histology of roots, photosynthetic pigments content, δ-aminolevulinic acid dehydratase (ALA-D; E.C. 4.2.1.24) and acid phosphatase activities (AP; E.C. 3.1.3.2), soluble phosphorus (Pi) measurement and mineral nutrients content in cucumber seedlings (Cucumis sativus L.) were investigated. Cucumber seedlings were grown in vitro in an agar-solidified substrate containing four CdCl₂ treatments (0, 100, 400, and 1000 μM) for ten days. Cd was readily absorbed by seedlings and its content was greater in the roots than in the shoot. Cd reduced shoot and root length, and fresh and dry biomass of seedlings. Inhibition of root cell elongation in Cd-treated seedlings was observed by the increase of the mean radial size of cells belonging to three zones of the root tip. The highest level of Cd reduced in a similar manner chlorophyll a, chlorophyll b and total chlorophyll contents. Increasing concentrations of Cd resulted in a linear decrease in carotenoids levels of cotyledons. Interestingly, the ALA-D activity in cotyledons was inhibited only at the highest level of Cd. Root and shoot AP activities were, respectively, activated and inhibited at all CdCl₂ concentrations. Root Pi concentration was increased in all Cd treatments and it was not altered in the shoot tissues. Moreover, in general, the nutrient contents were increased in the root and decreased in the shoot. Therefore, we suggest that Cd affects negatively growth, photosynthetic pigments, ALA-D and AP activities and partition of mineral nutrients in cucumber seedlings.     

Microalgae Improve the Photosynthetic Performance of Rice Seedlings (<i>Oryza sativa</i> L.) under Physiological Conditions and Cadmium Stress

The aim of this study was to assess the impact of the microalgae Chlorella vulgaris on the rice seedlings at physiological conditions and under cadmium (Cd) stress. We examined the effects of C. vulgaris in the nutrient solution on rice seedlings grown hydroponically in the presence and the absence of 150 μM CdCl₂, using the low (77 K) temperature and pulse amplitude modulated (PAM) chlorophyll fluorescence, P700 photooxidation measurements, photochemical activities of both photosystems, kinetic parameters of oxygen evolution, oxidative stress markers (MDA, H₂O₂ and proline), pigment content, growth parameters and Cd accumulation. Data revealed that the application C. vulgaris not only stimulates growth and improves the functions of photosynthetic apparatus under physiological conditions, but also reduces the toxic effect of Cd on rice seedlings. Furthermore, the presence of the green microalgae in the nutrient solution of the rice seedlings during Cd exposure, significantly improved the growth, photochemical activities of both photosystems, the kinetic parameters of the oxygen-evolving reactions, pigment content and decreased lipid peroxidation, H₂O₂ and proline content. Data showed that the alleviation of Cd-induced effects in rice seedlings is a result of the Cd sorption by microalgae, as well as the reduced Cd accumulation in the roots and its translocation from the roots to the shoots.     

Antioxidative response to cadmium in roots and leaves of tomato plants

Treatment of tomato seedlings (Lycopersicon esculentum Mill. cv. 63/5 F1) with increasing CdCl₂ concentrations in the culture medium resulted in Cd accumulation more important in roots than in leaves. Biomass production was severely inhibited, even at low Cd concentration. Cd reduced chlorophyll content in leaves and enhanced lipid peroxidation. An increase in antioxidative enzyme (superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, glutathione reductase) activities was more pronounced in leaves than in roots, while catalase activity increased only in roots. In addition, changes in isoenzyme composition were observed using the non-denaturing polyacrylamid gel electrophoresis.     

Effects of exogenous nitric oxide on cadmium toxicity, element contents and antioxidative system in perennial ryegrass

The effects of sodium nitroprusside (SNP, a donor of NO) on cadmium (Cd) toxicity in ryegrass seedlings (Lolium perenne L.) were studied by investigating the symptoms, plant growth, chlorophyll content, lipid peroxidation, H⁺-ATPase enzyme and antioxidative enzymes. Addition of 100???M CdCl₂ caused serious chlorosis and inhibited the growth of ryegrass seedlings, and dramatically increased accumulation of Cd in both shoots and roots, furthermore, the absorption of macro and micronutrients were inhibited. Addition of 50, 100, 200???M SNP significantly decreased the transport of Cd from roots to shoots, alleviated the inhibition of K, Ca, Mg and Fe, Cu, Zn absorption induced by Cd, reduced the toxicity symptoms and promoted the plant growth. The accumulation of reactive oxygen species (ROS) significantly increased in ryegrass seedlings exposed to Cd, and resulted in the lipid peroxidation, which was indicated by accumulated concentration of thiobarbituric acid-reactive substances. Addition of 50, 100, 200???M SNP significantly decreased the level of ROS and lipid peroxidation. Activities of antioxidant enzymes also showed the same changes. Addition of 50, 100, 200???M SNP increased activities of superoxide dismutase, peroxidase, catalase and ascorbate peroxidase in ryegrass seedlings exposed to Cd. Addition of 100???M SNP had the most significant alleviating effect against Cd toxicity while the addition of 400???M SNP had no significant effect with Cd treatment.     

Cadmium induced oxidative stress and changes in soluble and ionically bound cell wall peroxidase activities in roots of seedling and 3–4 leaf stage plants of <Emphasis Type="Italic">Brassica juncea</Emphasis> (L.) czern

Metabolic adaptations to heavy metal toxicity in plants are thought to be related with developmental growth stage and the type of metal by which plant is affected. In the present study, changes in ionically bound CWP, soluble peroxidase activity, H(2)O(2) level and Malonaldehyde content in roots of cadmium and copper stressed seedlings and cadmium stressed 3-4 leaf stage plants of Brassica juncea were investigated. Cadmium inhibits root growth and reduces fresh biomass. The reduction in root growth and fresh biomass is correlated with increased lipid peroxidation and reduced tolerance. Treatment with cadmium resulted in an increase in ionically bound CWP activity in roots of seedlings but no significant change in its activity was found in roots of 3-4 leaf stage plants. Increased level of H(2)O(2) in roots of cadmium and copper treated seedlings, show a direct correlation with increased activity of ionically bound CWP. H(2)O(2) level in 3-4 leaf stage plant roots was found to be very low. Soluble peroxidase activity decreased in cadmium (50 and 100 mu-icroM) treated seedlings but it was ineffective to cause any change in its activity in 3-4 leaf stage plants. Copper treated seedlings showed an increase in ionically bound CWP activity, H(2)O(2) level and MDA content. Ascorbic acid (50 mM) pretreated seedlings shows significant decrease in ionically bound CWP activity when exposed to 50 muM cadmium. Hence, it is concluded that inhibition of root growth in Brassica juncea seedlings by cadmium, is associated with CWP catalyzed H(2)O(2) dependent reactions which are involved in metabolic adaptations to heavy-metal stress.     

Relative importance of Na+, Cl−, and abscisic acid in NaCl induced inhibition of root growth of rice seedlings

The relative importance of endogenous abscisic acid (ABA), as well as Na⁺ and Cl^– in NaCl-induced responses related to growth in roots of rice seedlings were investigated. The increase in ammonium, proline and H₂O₂ levels, and cell wall peroxidase (POD) activity has been shown to be related to NaCl-inhibited root growth of rice seedlings. Increasing concentrations of NaCl from 50 to 150 mM progressively decreased root growth and increased both Na⁺ and Cl^–. Treatment with NaCl in the presence of 4,4-diisothiocyano-2,2-disulfonic acid (DIDS, a nonpermeating amino-reactive disulfonic acid known to inhibit the uptake of Cl^–) had less Cl^– level in roots than that in the absence of DIDS, but did not affect the levels of Na⁺, and responses related to growth in roots. Treatment with 50 mM Na-gluconate (the anion of which is not permeable to membrane) had similar Na⁺ level in roots as that with 100 mM NaCl. It was found that treatment with 50 mM Na-gluconate effected growth reduction and growth-related responses in roots in the same way as 100 mM NaCl. All these results suggest that Cl^– is not required for NaCl-induced responses in root of rice seedlings. Endogenous ABA level showed no increase in roots of rice seedlings exposed to 150 mM NaCl. It is unlikely that ABA is associated with NaCl-inhibited root growth of rice seedlings.     

Disturbed ammonium assimilation is associated with growth inhibition of roots in rice seedlings caused by NaCl

The effects of NaCl on changes in ammonium level and enzyme activities of ammonium assimilation in roots growth of rice (Oryza sativa L.) seedlings were investigated. NaCl was effective in inhibiting root growth and stimulated the accumulation of ammonium in roots. Accumulation of ammonium in roots preceded inhibition of root growth caused by NaCl. Both effects caused by NaCl are reversible. Exogenous ammonium chloride and methionine sulfoximine (MSO), which caused ammonium accumulation in roots, inhibited root growth of rice seedlings. NaCl decreased glutamine synthetase and glutamate synthase activities in roots, but increased glutamate dehydrogenase activity. The growth inhibition of roots by NaCl or MSO could be reversed by the addition of L-glutamic acid or L-glutamine. The current results suggest that disturbance of ammonium assimilation in roots may be involved in regulating root growth reduction caused by NaCl.Abbreviations GDH glutamate dehydrogenase - GOGAT glutamate synthase - GS glutamine synthetase - MSO methionine sulfoximine     

Cadmium accumulation in Atriplex halimus subsp. schweinfurthii and its influence on growth,proline, root hydraulic conductivity and nutrient uptake

Atriplex halimus subsp. schweinfurthii is a newly found cadmium (Cd)-hyperaccumulator, but there have been no detailed studies on its physiological responses when Cd is hyperaccumulated. A. halimus was grown in hydroponic conditions to investigate the effect of cadmium chloride (CdCl₂) on growth, water status, leaf chlorophyll concentration, proline and Cd accumulation. Treatments were prepared by adding 0, 50, 100, 200 and 400 μM CdCl₂ to the nutrient medium. Plant growth was significantly affected at high-Cd treatments. Increased CdCl₂ decreased chlorophyll concentration, transpiration and root hydraulic conductivity (L₀). Hence water flux had only a little effect on the uptake of Cd in A. halimus seedlings. In contrast, proline content increased with increasing CdCl₂ concentration. Plants accumulated substantial amount of Cd in different plant parts (shoot and root). Most of the Cd taken up was retained in roots (606.51 μg g⁻¹DW after 15 d at 400 μM CdCl₂). The addition of Cd in the culture medium affected calcium (Ca) and potassium (K) nutrition in both shoot and root. A. halimus provides a new plant resource for exploring the mechanism of Cd hyperaccumulation and has potential for use in the phytostabilization of Cd-contaminated salt soils.     

Osmotic stress-induced changes in cell wall peroxidase activity and hydrogen peroxide level in roots of rice seedlings

The changes in activity of peroxidase (POD) extracted from the cellwalls and the level of H₂O₂ in rice seedling rootstreatedwith mannitol and their correlation with root growth were investigated.Increasing concentrations of mannitol from 92 to 276 mM, which isiso-osmotic with 50 to 150 mM NaCl, progressively reduced rootgrowth and increased POD activities extracted from the cell walls of riceroots.The reduction of growth was also correlated with an increase inH₂O₂ level. Both diamine oxidase (DAO) and NADHperoxidase(NADH-POD) are known to be responsible for the generation ofH₂O₂. Mannitol treatment increased DAO but not NADH-PODactivities in roots of rice seedlings, suggesting that DAO contributes to thegeneration of H₂O₂ in the cell walls of mannitol-treatedroots. An increase in the level of H₂O₂ and the activityof POD extracted from the cell walls of rice roots preceded root growthreduction caused by mannitol. An increase in DAO activity coincided with anincrease in H₂O₂ in roots caused by mannitol. Since DAOcatalyses the oxidation of putrescine, the demonstration that mannitolincreasesthe activity of DAO in roots is consistent with those that mannitol decreasesthe level of putrescine. In conclusion, cell-wall stiffening catalysed by PODispossibly involved in the regulation of root growth reduction caused bymannitol.     

A comparative study of the effects of abscisic acid and methyl jasmonate on seedling growth of rice

The effects of abscisic acid (ABA) and methyl jasmonate (MJ) on growth of rice seedlings were compared. The lowest tested concentration of ABA and MJ that inhibited seedling growth was found to be 4.5 and 0.9 µM, respectively. Growth inhibition by ABA is reversible, whereas that by MJ is irreversible. GA₃ was found to be more effective in reversing inhibition of shoot growth by ABA than by MJ. KCl partially relieved MJ-inhibited, but not ABA-inhibited, growth of rice seedlings. The beneficial effect of K⁺ on growth of rice seedlings in MJ medium could not be replaced by Li⁺, Na⁺ or Cs⁺. MJ treatment caused a marked release of K⁺ into the medium. In order to understand whether cell wall-bound peroxidase activity was inversely related to rice seedling growth, effects of ABA and MJ on cell wall-bound peroxidase activity were also examined. Results indicated that both ABA and MJ increased cell wall-bound peroxidase activity in roots and shoots of rice seedlings. Although MJ (4.5 µM) was less effective in inhibiting root growth than ABA (9 µM), MJ was found to increase more cell wall-bound peroxidase activity in roots than ABA.     

The decline in potassium concentration is associated with cadmium toxicity of rice seedlings

Cadmium (Cd) is one of the most dangerous environmental pollutants, among other things, affecting plant mineral composition. Thus, in this study, we investigated the changes in potassium (K) concentration in Cd-treated rice (Oryza sativa L.) seedlings of two cultivars. On treatment with 5 μM CdCl₂, the Cd concentration increased in the shoot and roots of Cd-sensitive cultivar (cv. Taichung Native 1, TN1) but not or slightly in the Cd-tolerant cultivar (cv. Tainung 67, TNG67). The decrease in K concentration in the shoot and roots of TN1 caused by Cd was more pronounced than that of TNG67. Exogenous addition of KCl decreased Cd concentration and reduced Cd toxicity of TN1 seedlings. Evidence presented in this study suggests that the improvement of K status is able to reduce toxicity of rice seedlings to CdCl₂.     

Effect of nitrogen deficiency on antioxidant status and Cd toxicity in rice seedlings

Effect of nitrogen (N) deficiency on antioxidant status and Cd toxicity in rice seedlings was investigated. N deficiency resulted in a reduction of shoot growth but not root growth. The contents of N-containing compounds such as nitrate, chlorophyll, and protein decreased in leaves of rice seedlings grown under N deficiency. Accumulation of abscisic acid and H₂O₂ in leaves was induced by N deficiency. The content of ascorbate and the activities of ascorbate peroxidase, glutathione reductase, and catalase in N-deficient leaves were lower than their respective control leaves. However, glutathione content was not affected and superoxide dismutase activity was increased by N deficiency. Cd toxicity in N-deficient seedlings was more pronounced than that in N-sufficient ones. Pretreatment with ascorbate or L-galactono-1,4-lactone, a biosynthetic precursor of ascorbate resulted in a reduction of Cd toxicity enhanced by N deficiency. N deficiency also resulted in an enhancement of Cd uptake in rice seedlings. The possible mechanism of Cd toxicity enhanced by N deficiency is discussed.     

Effects of Melatonin on Growth,Physiology and Gene Expression in Rice Seedlings Under Cadmium Stress

Melatonin (MLT) is a hormonal substance found in many organisms and can improve plant stress resistance. In this study, the japonica rice variety Y32 and indica rice variety NJ6 were cultivated in hydroponics under different concentrations of CdCl₂ at the two-leaf stage. The growth, physiological and biochemical responses of the seedlings and the expression of cadmium (Cd)-related genes under exogenous melatonin (MLT) treatment were assessed. The results indicated that Cd stress destroyed the dynamic balance between reactive oxygen species (ROS) production and removal, resulting in ROS accumulation, membrane lipid peroxidation, and impaired growth and development. Following the application of exogenous MLT to rice seedlings, increases in plant biomass including both underground and above-ground areas were observed. MLT also scavenged the inhibition of superoxide dismutase (SOD) and peroxidase (POD) in a concentration dependent manner in response to Cd stress. Catalase (CAT) activity and malondialdehyde (MDA) expression also decreased following MLT treatment. Amongst the six Cd-related genes assessed, five genes were down-regulated and one was up-regulated in response to MLT treatment. Taken together, these data demonstrate that MLT improves the resilience of rice seedlings at the biochemical, physiological, and molecular levels, and diminishes the damage caused by Cd stress.     

Toxicity in leaves of rice exposed to cadmium is due to hydrogen peroxide accumulation

The production of H₂O₂ in detached rice leaves of Taichung Native 1 (TN1) caused by CdCl₂ was investigated. CdCl₂ treatment resulted in H₂O₂ production in detached rice leaves. Diphenyleneiodonium chloride (DPI) and imidazole (IMD), inhibitors of NADPH oxidase (NOX), prevented CdCl₂-induced H₂O₂ production, suggesting that NOX is a H₂O₂-genearating enzyme in CdCl₂-treated detached rice leaves. Phosphatidylinositol 3-kinase inhibitors wortmanin (WM) or LY294002 (LY) inhibited CdCl₂-inducted H₂O₂ production in detached rice leaves. Exogenous H₂O₂ reversed the inhibitory effect of WM or LY, suggesting that phosphatidylinositol 3-phosphate is required for Cd-induced H₂O₂ production in detached rice leaves. Nitric oxide donor sodium nitroprusside (SNP) was also effective in reducing CdCl₂-inducing accumulation of H₂O₂ in detached rice leaves. Cd toxicity was judged by the decrease in chlorophyll content. The results indicated that DPI, IMD, WM, LY, and SNP were able to reduce Cd-induced toxicity of detached rice leaves. Twelve-day-old TN1 and Tainung 67 (TNG67) rice seedlings were treated with or without CdCl₂. In terms of Cd toxicity (leaf chlorosis), it was observed that rice seedlings of cultivar TN1 are Cd-sensitive and those of cultivar TNG67 are Cd-tolerant. On treatment with CdCl₂, H₂O₂ accumulated in the leaves of TN1 seedlings but not in the leaves of TNG67. Prior exposure of TN1 seedlings to 45^oC for 3 h resulted in a reduction of H₂O₂ accumulation, as well as Cd tolerance of TN1 seedlings treated with CdCl₂. The results strongly suggest that Cd toxicity of detached leaves and leaves attached to rice seedlings are due to H₂O₂ accumulation.     

Cadmium decreases crown root number by decreasing endogenous nitric oxide,which is indispensable for crown root primordia initiation in rice seedlings

Cadmium (Cd) is toxic to crown roots (CR), which are essential for maintaining normal growth and development in rice seedlings. Nitric oxide (NO) is an important signaling molecule that plays a pivotal role in plant root organogenesis. Here, the effects of Cd on endogenous NO content and root growth conditions were studied in rice seedlings. Results showed that similar to the NO scavenger, cPTIO, Cd significantly decreased endogenous NO content and CR number in rice seedlings, and these decreases were recoverable with the application of sodium nitroprusside (SNP, a NO donor). Microscopic analysis of root collars revealed that treatment with Cd and cPTIO inhibited CR primordia initiation. In contrast, although SNP partially recovered Cd-caused inhibition of CR elongation, treatment with cPTIO had no effect on CR elongation. l-NMMA, a widely used nitric oxide synthase (NOS) inhibitor, decreased endogenous NO content and CR number significantly, while tungstate, a nitrate reductase (NR) inhibitor, had no effect on endogenous NO content and CR number. Moreover, enzyme activity assays indicated that treatment with SNP inhibited NOS activity significantly, but had no effect on NR activity. All these results support the conclusions that a critical endogenous NO concentration is indispensable for rice CR primordia initiation rather than elongation, NOS is the main source for endogenous NO generation, and Cd decreases CR number by inhibiting NOS activity and thus decreasing endogenous NO content in rice seedlings.