The miRNAs and their regulatory networks responsible for pollen abortion in Ogura-CMS Chinese cabbage revealed by high-throughput sequencing of miRNAs, degradomes, and transcriptomes.

Chinese cabbage (Brassica rapa ssp. pekinensis) is one of the most important vegetables in Asia and is cultivated across the world. Ogura-type cytoplasmic male sterility (Ogura-CMS) has been widely used in the hybrid breeding industry for Chinese cabbage and many other cruciferous vegetables. Although, the cause of Ogura-CMS has been localized to the orf138 locus in the mitochondrial genome, however, the mechanism by which nuclear genes respond to the mutation of the mitochondrial orf138 locus is unclear. In this study, a series of whole genome small RNA, degradome and transcriptome analyses were performed on both Ogura-CMS and its maintainer Chinese cabbage buds using deep sequencing technology. A total of 289 known miRNAs derived from 69 families (including 23 new families first reported in B. rapa) and 426 novel miRNAs were identified. Among these novel miRNAs, both 3-p and 5-p miRNAs were detected on the hairpin arms of 138 precursors. Ten known and 49 novel miRNAs were down-regulated, while one known and 27 novel miRNAs were up-regulated in Ogura-CMS buds compared to the fertile plants. Using degradome analysis, a total of 376 mRNAs were identified as targets of 30 known miRNA families and 100 novel miRNAs. A large fraction of the targets were annotated as reproductive development related. Our transcriptome profiling revealed that the expression of the targets was finely tuned by the miRNAs. Two novel miRNAs were identified that were specifically highly expressed in Ogura-CMS buds and sufficiently suppressed two pollen development essential genes: sucrose transporter SUC1 and H (+) -ATPase 6. These findings provide clues for the contribution of a potential miRNA regulatory network to bud development and pollen engenderation. This study contributes new insights to the communication between the mitochondria and chromosome and takes one step toward filling the gap in the regulatory network from the orf138 locus to pollen abortion in Ogura-CMS plants from a miRNA perspective.

Accumulation and cellular toxicity of aluminum in seedling of Pinus massoniana.

BACKGROUND: Masson pine (Pinus massoniana) is one of the most important timber species with adaptable, fast growing, versatile advantages in southern China. Despite considerable research efforts, the cellular and molecular mechanisms of A1 toxicity and resistance in P. massoniana are still poorly understood. The effects of Al on uptake and translocation of Al and other minerals, cell division and nucleolus in P. massoniana were investigated. RESULTS: The results indicated that Al accumulated mainly in the roots, and small amounts were transported to aboveground organs. In the presence of Al, the contents of Mg and Fe in stems increased and decreased in roots. Accumulation of Mn in the organs was inhibited significantly. Evidence from cellular experiments showed that Al had an inhibitory effect on the root growth at all concentrations (10⁻5 - 10⁻² M) used. Chromosome fragments, chromosome bridges, C-mitosis and chromosome stickiness were induced during mitosis in the root tip cells. Al induced the formation of abnormal microtubule (MT) arrays, consisting of discontinuous wavy MTs or short MT fragments at the cell periphery. MT organization and function of the mitotic spindle and phragmoplast were severely disturbed. The nucleolus did not disaggregate normally and still remained its characteristic structure during metaphase. Nucleolar particles containing argyrophilic proteins were accumulated and leached out from the nucleus to the cytoplasm. Evidence confirmed that these proteins contained nucleophosmin (B23), nucleolin (C23) and fibrillarin. Western immunoblot analysis revealed that the contents of three nucleolar proteins increased significantly. CONCLUSION: Based on the information provided in this article, it is concluded that root tips of plants are the most sensitive organ to environmental stresses and the accumulation of Al ions primarily is in roots of P. massoniana, and small amounts of Al are transported to aboveground. Root apical meristems play a key role in the immediate reaction to stress factors by activating signal cascades to the other plant organs. Al induces a series of the cellular toxic changes concerning with cell division and nucleolus. The data presented above can be also used as valuable and early markers in cellular changes induced by metals for the evaluation of metal contamination.

Effects of lead on the morphology and structure of the nucleolus in the root tip meristematic cells of Allium cepa L.

To study the toxic mechanisms of lead (Pb) in plants, the effects of Pb on the morphology and structure of the nucleolus in root tip meristematic cells of Allium cepa var. agrogarum L. were investigated. Fluorescence labeling, silver-stained indirect immunofluorescent microscopy and western blotting were used. Fluorescence labeling showed that Pb ions were localized in the meristematic cells and the uptake and accumulation of Pb increased with treatment time. At low concentrations of Pb (1-10 µM) there were persistent nucleoli in some cells during mitosis, and at high concentration (100 µM) many of the nucleolar organizing regions were localized on sticky chromosomes in metaphase and anaphase cells. Pb induced the release of particles containing argyrophilic proteins to be released from the nucleus into the cytoplasm. These proteins contained nucleophosmin and nucleolin. Pb also caused the extrusion of fibrillarin from the nucleus into the cytoplasm. Western blotting demonstrated the increased expression of these three major nucleolar proteins under Pb stress.

Three major nucleolar proteins migrate from nucleolus to nucleoplasm and cytoplasm in root tip cells of Vicia faba L. exposed to aluminum.

Results from our previous investigation indicated that Al could affect the nucleolus and induce extrusion of silver-staining nucleolar particles containing argyrophilic proteins from the nucleolus into the cytoplasm in root tip cells of Vicia faba L. So far, the nucleolar proteins involved have not been identified. It is well known that nucleophosmin (B23), nucleolin (C23), and fibrillarin are three major and multifunctional nucleolar proteins. Therefore, effects of Al on B23, C23, and fibrillarin in root tip cells of V. faba exposed to 100 µM Al for 48 h were observed and analyzed using indirect immunofluorescence microscopy and Western blotting. The results from this work demonstrated that after 100 µM of Al treatment for 48 h, B23 and C23 migrated from the nucleolus to the cytoplasm and fibrillarin from the nucleolus to the nucleoplasm. In some cells, fibrillarin was present only in the cytoplasm. Western blotting data revealed higher expression of the three major nucleolar proteins in Al-treated roots compared with the control and that the B23 content increased markedly. These findings confirmed our previous observations.

Cadmium accumulation, activities of antioxidant enzymes, and malondialdehyde (MDA) content in Pistia stratiotes L.

The aquatic plant Pistia stratiotes L. (water lettuce) was studied due to its capability of absorption of contaminants in water and its subsequent use in wetlands constructed for wastewater treatment. The effects of Cd on root growth, accumulation of Cd, antioxidant enzymes, and malondialdehyde (MDA) content in P. stratiotes were investigated. The results indicated that P. stratiotes has considerable ability to accumulate Cd. Cadmium induced higher superoxide dismutase (SOD) and peroxidase (POD) activities than catalase activity, suggesting that SOD and POD provided a better defense mechanism against Cd-induced oxidative damage. The accumulation of Cd promoted MDA production.

Aluminum can induce alterations in the cellular localization and expression of three major nucleolar proteins in root tip cells of Allium cepa var. agrogarum L.

A 50 µM aluminum (Al) could induce nucleolar materials containing the argyrophilic proteins scattered in the nuclei and extruded from the nuclei into the cytoplasm in the root tip cells of Allium cepa. Unfortunately, what kinds of nucleolar proteins are affected has not been reported till now. In order to go deeper into the understanding of the cytological effects of Al on nucleolus and nucleolar proteins, alterations in the cellular localization and expression of three major nucleolar proteins: nucleophosmin, nucleolin, and fibrillarin were further examined under the treatment with Al in the root tip cells of A. cepa in the present study. Cytological effects of Al on nucleolus were observed by silver-staining method and three major nucleolar proteins: nucleophosmin, nucleolin, and fibrillarin were examined by western blotting. The results indicated that in the presence of 50 µM Al for 48 h the nucleolar proteins were translocated from nucleolus to nucleoplasm and cytoplasm. Western blotting data demonstrated the relatively higher expression of the three major nucleolar proteins when compared with control. Evidence from the present investigation indicated that Al had toxic effects on Ag-NOR proteins, nucleophosmin and nucleolin, and other kinds of nucleolar proteins, fibrillarin.

Effects of aluminum on nucleoli in root tip cells and selected physiological and biochemical characters in Allium cepa var. agrogarum L.

BACKGROUND: Increased Al concentration causes reduction of mitotic activity, induction of nucleolar alteration, increase of the production of ROS and alteration of several antioxidant enzyme activities in plant cells. Allium cepa is an excellent plant and a useful biomarker for environmental monitoring. Limited information is available about the effects of Al on nucleoli, antioxidant enzyme system, contents of MDA and soluble protein in A. cepa. Therefore, we carried out the investigation in order to better understand the effects of Al on the growth, nucleoli in root tip cells and selected physiological and biochemical characters. RESULTS: The results showed that the root growth exposed to 50 µM Al was inhibited significantly. 50 µM Al could induce some particles of argyrophilic proteins scattered in the nuclei and extruded from the nucleoli into the cytoplasm. The nucleolus did not disaggregate normally and still remained its characteristic structure during metaphase. Nucleolar reconstruction was inhibited. 50 µM Al induced high activities of SOD and POD in leaves and roots significantly (P < 0.05) when compared with control, whereas the level of CAT was low significantly (P < 0.05). At 50 µM Al the content of MDA in leaves was high significantly (P < 0.05) at 9(th) day and in roots increased (P < 0.05) with prolonging the treatment time during 6-12 days. The soluble protein content in leaves treated with 50 µM Al was high significantly (P < 0.05) at 6(th) day and increased with prolonging the treatment time. CONCLUSIONS: We suggest that variations in nucleoli and the alterations of antioxidant enzyme activities, MDA and soluble protein contents in Allium cepa can serve as useful biomarkers, which can provide valuable information for monitoring and forecasting effects of exposure to Al in real scenarios conditions. Among the antioxidant enzymes SOD and POD appear to play a key role in the antioxidant defense mechanism under Al toxicity condition. Data from MDA concentration show that Al indirectly produces superoxide radicals, resulting in increased lipid peroxidative products and oxidative stress.

Pb-induced cellular defense system in the root meristematic cells of Allium sativum L.

BACKGROUND: Electron microscopy (EM) techniques enable identification of the main accumulations of lead (Pb) in cells and cellular organelles and observations of changes in cell ultrastructure. Although there is extensive literature relating to studies on the influence of heavy metals on plants, Pb tolerance strategies of plants have not yet been fully explained. Allium sativum L. is a potential plant for absorption and accumulation of heavy metals. In previous investigations the effects of different concentrations (10(-5) to 10(-3) M) of Pb were investigated in A. sativum, indicating a significant inhibitory effect on shoot and root growth at 10(-3) to 10(-4) M Pb. In the present study, we used EM and cytochemistry to investigate ultrastructural alterations, identify the synthesis and distribution of cysteine-rich proteins induced by Pb and explain the possible mechanisms of the Pb-induced cellular defense system in A. sativum. RESULTS: After 1 h of Pb treatment, dictyosomes were accompanied by numerous vesicles within cytoplasm. The endoplasm reticulum (ER) with swollen cisternae was arranged along the cell wall after 2 h. Some flattened cisternae were broken up into small closed vesicles and the nuclear envelope was generally more dilated after 4 h. During 24-36 h, phenomena appeared such as high vacuolization of cytoplasm and electron-dense granules in cell walls, vacuoles, cytoplasm and mitochondrial membranes. Other changes included mitochondrial swelling and loss of cristae, and vacuolization of ER and dictyosomes during 48-72 h. In the Pb-treatment groups, silver grains were observed in cell walls and in cytoplasm, suggesting the Gomori-Swift reaction can indirectly evaluate the Pb effects on plant cells. CONCLUSIONS: Cell walls can immobilize some Pb ions. Cysteine-rich proteins in cell walls were confirmed by the Gomori-Swift reaction. The morphological alterations in plasma membrane, dictyosomes and ER reflect the features of detoxification and tolerance under Pb stress. Vacuoles are ultimately one of main storage sites of Pb. Root meristematic cells of A. sativum exposed to lower Pb have a rapid and effective defense system, but with the increased level of Pb in the cytosol, cells were seriously injured.

Cytogenetical and ultrastructural effects of copper on root meristem cells of Allium sativum L.

Different copper concentrations, as well as different exposure times, were applied to investigate both cytogenetical and ultrastructural alterations in garlic (Allium sativum L.) meristem cells. Results showed that the mitotic index decreased progressively when either copper concentration or exposure time increased. C-mitosis, anaphase bridges, chromosome stickiness and broken nuclei were observed in the copper treated root tip cells. Some particulates containing the argyrophilic NOR-associated proteins were distributed in the nucleus of the root-tip cells and the amount of this particulate material progressively increased with increasing exposure time. Finally, the nucleolar material was extruded from the nucleus into the cytoplasm. Also, increased dictyosome vesicles in number, formation of cytoplasmic vesicles containing electron dense granules, altered mitochondrial shape, disruption of nuclear membranes, condensation of chromatin material, disintegration of organelles were observed. The mechanisms of detoxification and tolerance of copper are briefly discussed.

Cadmium induction of lipid peroxidation and effects on root tip cells and antioxidant enzyme activities in Vicia faba L.

The effects of different concentrations (1-50 microM) of Cd on root growth, cell division and nucleoli in root tip cells, protective enzyme activities and lipid peroxidation in Vicia faba were investigated in order to better understand the processes of Cd-induced senescence. The results indicated that lower concentration of Cd (1 microM) had no obviously influence on the root growth during 24-48 h treatment, but higher concentrations (5-50 microM) inhibited significantly after 48 and 72 h. The mitotic index decreased with increasing of Cd concentration and duration of treatment except for the group exposed to 1 microM Cd. Cd induced c-mitosis, chromosome bridges, chromosome stickiness and lagging chromosomes. The rate of aberrant dividing cells increased with prolonging duration of treatment and increasing of Cd concentration. On nucleolus, some particulates containing the argyrophilic proteins were extruded from the nucleus into the cytoplasm in the cells stressed by Cd and some were scattered in the nucleus. After the treatment with Cd (10 microM Cd, 48 h), the nucleolus did not disaggregate normally and still remain its characteristic structure during metaphase and the particles of similar silver-stained materials were localized on chromosomes. In leaves, Catalase (CAT) activity declined but Peroxidase (POD) activity increased with increasing of the duration of treatment. In roots, CAT activity increased with increasing of the duration of treatment, POD activity increased during early days and then declined. Superoxide dismutase (SOD) activity showed an upward trend with increasing of the duration of treatment after 3 and 6 days, then declined both in leaves and roots (9 days). SOD and POD had highest activities at 50 microM Cd in leaves. CAT activity was lowest at 50 microM Cd. Malondialdehyde (MDA) content increased with the increasing of Cd concentrations and duration of treatment in leaves. In roots, MDA content showed an upward trend with increasing of the duration of treatment at early time and then declined.

Cytogenetical and ultrastructural effects of copper on root meristem cells of Allium sativum L

Different copper concentrations, as well as different exposure times, were applied to investigate both cytogenetical and ultrastructural alterations in garlic (Allium sativum L.) meristem cells. Results showed that the mitotic index decreased progressively when either copper concentration or exposure time increased. C-mitosis, anaphase bridges, chromosome stickiness and broken nuclei were observed in the copper treated root tip cells. Some particulates containing the argyrophilic NOR-associated proteins were distributed in the nucleus of the root-tip cells and the amount of this particulate material progressively increased with increasing exposure time. Finally, the nucleolar material was extruded from the nucleus into the cytoplasm. Also, increased dictyosome vesicles in number, formation of cytoplasmic vesicles containing electron dense granules, altered mitochondrial shape, disruption of nuclear membranes, condensation of chromatin material, disintegration of organelles were observed. The mechanisms of detoxification and tolerance of copper are briefly discussed.(AU)

Cytogenetical and ultrastructural effects of copper on root meristem cells of Allium sativum L

Different copper concentrations, as well as different exposure times, were applied to investigate both cytogenetical and ultrastructural alterations in garlic (Allium sativum L.) meristem cells. Results showed that the mitotic index decreased progressively when either copper concentration or exposure time increased. C-mitosis, anaphase bridges, chromosome stickiness and broken nuclei were observed in the copper treated root tip cells. Some particulates containing the argyrophilic NOR-associated proteins were distributed in the nucleus of the root-tip cells and the amount of this particulate material progressively increased with increasing exposure time. Finally, the nucleolar material was extruded from the nucleus into the cytoplasm. Also, increased dictyosome vesicles in number, formation of cytoplasmic vesicles containing electron dense granules, altered mitochondrial shape, disruption of nuclear membranes, condensation of chromatin material, disintegration of organelles were observed. The mechanisms of detoxification and tolerance of copper are briefly discussed.

Pb/Cu effects on the organization of microtubule cytoskeleton in interphase and mitotic cells of Allium sativum L.

The effects of lead and copper on the arrangement of microtubule (MT) cytoskeleton in root tip cells of Allium sativum L. were investigated. Batch cultures of garlic were carried out under defined conditions in the presence 10(-4) M Pb/Cu of various duration treatments. With tubulin immunolabelling and transmission electron microscopy (TEM), we found four different types of MT structures depending on the cell cycle stage: the interphase array, preprophase band, mitotic spindle and phragmoplast were typical for the control cells. Pb/Cu affected the mechanisms controlling the organization of MT cytoskeleton, and induces the following aberrations in interphase and mitotic cells. (1) Pb/Cu induced the formation of atypical MT arrays in the cortical cytoplasm of the interphase cells, consisting of skewed, wavy MT bundles, MT fragments and ring-like tubulin aggregations. (2) Pb/Cu disordered the chromosome movements carried out by the mitotic spindle. The outcome was chromosome aberrations, for example, chromosome bridges and chromosome stickiness, as well as inhibition of cells from entering mitosis. (3) Depending on the time of exposure, MTs disintegrated into shorter fragments or they completely disappeared, indicating MT depolymerization. (4) Different metals had different effects on MT organization. MTs were more sensitive to the pressure of Cu ions than Pb. Moreover, TEM observations showed that the MTs were relatively short and in some places wavy when exposed to 10(-4) M Pb/Cu solutions for 1-2 h. In many sections MTs were no longer visible with increasing duration of treatment (>4 h). Based on these results, we suggested that MT cytoskeleton is primarily responsible for Pb/Cu-associated toxicity and tolerance in plants.

Uptake and accumulation and oxidative stress in garlic (Allium sativum L.) under lead phytotoxicity.

The effects of different concentrations of Pb on growth of Allium sativum L, Pb uptake and accumulation, antioxidant enzyme activity and malondialdehyde content were investigated. The results indicated that shoot growth at high concentration of Pb (10(-3) M) and roots growth at 10(-3) M and 10(-4) M Pb were significantly inhibited. Lead ions were accumulated mainly in the roots and only small amounts were translocated to bulbs and shoots. SOD activities in shoot and roots exposed to 10(-3) M Pb were observed to be high. Plants exposed to 10(-3) M Pb showed a significant increase in POD activity in roots versus the control and other Pb treatments. In roots, CAT activity and MDA concentration at 10(-3) M Pb is high significantly. The mechanisms of Pb toxicity and tolerance in garlic are briefly discussed.

Effects of Cd(2+) on seedling growth of garlic (Allium sativum L.) and selected physiological and biochemical characters.

In this study, we determined the effects of Cd(2+) (from 10(-5) to 10(-3)M) on the growth of leaves and roots of garlic seedlings in Hoagland's nutrient solution. We also characterized the influences of Cd(2+) on a few key cellular activities, e.g., antioxidant enzymes, content of malondialdehyde (MDA), composition of photosynthetic pigments and fluorescence properties of chlorophyll on day 5, 10 and 15. The results indicated that the presence of 10(-3)M Cd(2+), but not other tested Cd(2+) concentrations, significantly decreased the development of leaves and roots of the seedlings. This level of Cd(2+) increased the activities of antioxidant enzymes and the level of MDA. With the exception of carotenoid on day 5, the presence of 10(-3)M Cd(2+) showed no significant effects on the contents of photosynthetic pigments. Intermediate concentrations of Cd(2+) caused variable effects, ranging from lowering to increasing the pigments in garlic seedlings. The presence of 10(-3)M Cd(2+), but not other tested concentrations, decreased efficiency of energy harvesting associated with photosystem II as measured by the ratio of variable to maximum chlorophyll fluorescence of intact leaves. However, Cd(2+) generated no definitive trends on photochemical quenching. Possible significance of experimental findings in relation to Cd(2+) level in garlic seedling is discussed.

Hexavalent chromium uptake and its effects on mineral uptake, antioxidant defence system and photosynthesis in Amaranthus viridis L.

The effects of different concentrations (10(-6)M, 10(-5)M and 10(-4)M) of K2Cr2O7Cr(VI) on some minerals (Mn, Fe, Cu and Zn), lipid peroxidation, activities of antioxidant enzymes, photosynthetic function, and chlorophyll fluorescence characteristics were investigated in hydroponically grown Amaranthus viridis L. Results indicated that chromium was accumulated primarily in roots. In the roots and shoots, the Cr content increased with the increasing Cr(VI) concentrations, and induced decrease of Mn, Fe, Cu and Zn. Chromium Cr(VI) induced oxidation stress and lipid peroxidation in A. viridis L. shown by the increased concentration of MDA. The increased activities of POD and SOD indicated that they could serve as important components of antioxidant defense mechanisms to minimize Cr induced oxidative injury. The net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO2 concentration were reduced only by high Cr(VI) treatments (10(-5)M and 10(-4)M). The chlorophyll fluorescence parameters Fv/Fm, Fv(')/Fm('), Phi PSII and qP, decreased in Cr(VI)-treated, but qN and NPQ showed an increase in Cr(VI) treated plants.

Cadmium accumulation and its effects on metal uptake in maize (Zea mays L.).

The effects of different concentrations of Cd on growth of maize (Zea mays L.) and metal uptake were investigated. Cd accumulations in roots and shoots and the interactions among other metals (Mn, Fe, Cu and Zn) were analyzed using inductively coupled plasma atomic emission spectrometry (ICP-AES). The concentrations of cadmium chloride (CdCl(2).2.5H2O) used ranged from 10(-4) M to 10(-6) M. Cd had stimulatory effects during the first 5 days on root length of Nongda No. 108 at 10(-6) M and 10(-5) M Cd concentrations. Seedlings exposed to 10(-4) M Cd solution exhibited substantial growth reduction, and root growth even stopped. Root growth of Liyu No. 6 was stimulated at concentrations of 10(-5) M and 10(-6) M Cd during the entire experiment (15 days). Cadmium inhibited root growth of Liyu No. 6 at 10(-4) M Cd after 10 days of treatment. The Cd accumulation in roots and shoots of the two cultivars increased significantly (P < 0.05) with increasing Cd concentration and duration of treatment. Cadmium concentrated mainly in the roots, and small amounts were transferred to shoots. The proportion of Cd in the roots of Nongda No. 108 decreased with increases in Cd concentrations and duration of treatment, except for the group exposed to 10(-4) M Cd. In Liyu No. 6, the proportion of Cd in the root decreased progressively with an increase in Cd concentrations. Liyu No. 6 has a greater ability to remove Cd from solution and accumulate it when compared with Nongda No. 108. Liyu No. 6 can be considered a Cd-hyperaccumulator, according to the current accepted shoot concentration that defines hyperaccumulation as 0.01% (w/w) for cadmium. This cultivar, producing many roots and a high biomass and with great ability to accumulate Cd can play an important role in the treatment of soils stressed by Cd.

Accumulation of copper by roots, hypocotyls, cotyledons and leaves of sunflower (Helianthus annuus L.).

The effects of different concentrations of copper sulfate on the growth of and the accumulation of Cu2+ by root, hypocotyl, cotyledon and leaf growth of sunflower (Helianthus annuus L.) were examined in this study. The concentrations of copper sulfate (CuSO4 x 5H2O) used were in the range from 10(-5) to 10(-3) M. Seedlings exposed to 10(-5) M Cu2+ solution exhibited a 33% increase in growth (P < 0.005) when compared with the root length of the control. The seedlings treated with 10(-3) M Cu2+ were significantly inhibited in shoot growth (P < 0.005). The Cu2+ content in roots, hypocotyls, cotyledons and leaves increased with increasing solution Cu2+ concentration. The roots of plants exposed to 10(-3) M Cu2+ accumulated a large amount of Cu (1070 microgram/g DW), and the Cu2+ level was approximately 25 fold higher than that of control. The Cu2+ contents in sunflower roots treated with 10(-4) and 10(-5) M Cu2+ were about 3.3 and 2.6 fold higher than the control, respectively. Also, the Cu2- level of the roots exposed to 10(-3) M Cu2+ was approximately 7.7 and 9.8 fold respectively, in comparison with the roots of plants grown in 10(-4) and 10(-5) M Cu2+. At 10(-3) M Cu2+, the Cu accumulated mainly in the roots (about 73%), and small amounts of Cu2+ (27%) were translocated to the hypocotyls, cotyledons and leaves. The Cu2+ concentration in the roots was less than that of the above parts of seedlings in treated groups with 10(-5) - 10(-4) M Cu2+. H. annuus has potential ability to accumulate Cu without being overly sensitive to Cu toxicity.