Differential expression of vacuolar and defective cell wall invertase genes in roots and seeds of metalliferous and non-metalliferous populations of Rumex dentatus under copper stress.

Acid invertase activities in roots and young seeds of a metalliferous population (MP) of Rumex dentatus were previously observed to be significantly higher than those of a non-metalliferous population (NMP) under Cu stress. To date, no acid invertase gene has been cloned from R. dentatus. Here, we isolated four full-length cDNAs from the two populations of R. dentatus, presumably encoding cell wall (RdnCIN1 and RdmCIN1 from the NMP and MP, respectively) and vacuolar invertases (RdnVIN1 and RdmVIN1 from the NMP and MP, respectively). Unexpectedly, RdnCIN1 and RdmCIN1 most likely encode special defective invertases with highly attenuated sucrose-hydrolyzing capacity. The transcript levels of RdmCIN1 were significantly higher than those of RdnCIN1 in roots and young seeds under Cu stress, whereas under control conditions, the former was initially lower than the latter. Unexpected high correlations were observed between the transcript levels of RdnCIN1 and RdmCIN1 and the activity of cell wall invertase, even though RdnCIN1 and RdmCIN1 do not encode catalytically active invertases. Similarly, the transcript levels of RdmVIN1 in roots and young seeds were increased under Cu stress, whereas those of RdnVIN1 were decreased. The high correlations between the transcript levels of RdnVIN1 and RdmVIN1 and the activity of vacuolar invertase indicate that RdnVIN1 and RdmVIN1 might control distinct vacuolar invertase activities in the two populations. Moreover, a possible indirect role for acid invertases in Cu tolerance, mediated by generating a range of sugars used as nutrients and signaling molecules, is discussed.

[Effect of Nano Zeolites on pH, CEC in Soil and Cd Fractions in Plant and Soil at Different Cadmium Levels].

Soil incubation experiment and pot experiment were carried out to investigate the influence of nano zeolite (NZ) and ordinary zeolite (OZ) on the soil pH, cation exchange capacity, concentration of soil Cd, soil Cd fraction and Cd uptake by Chinese cabbage when exposed to different Cd pollution levels(1, 5, 10 and 15 mg·kg-1). The results of soil incubation experiment showed that the nano zeolite and ordinary zeolite dose(5, 10 and 20 g·kg-1) increased the soil pH and cation exchange capacity, and decreased the concentration of soil exchangeable Cd, while increased the concentration of Cd in carbonate, Fe-Mn oxide, organic matter and residual fraction. The lowest EX-Cd was observed in the high nano zeolite (20 g·kg-1) treatment. The soil pH and cation exchange capacity was extremely negatively correlated with the concentration of soil exchangeable Cd(P<0.01),and extremely positively correlated with the concentration of soil Fe-Mn oxide Cd(P<0.01). The results of pot experiment showed that the FDC of exchangeable Cd in soil decreased by 6.4%-63.2%, and the FDC of water-extractable and ethnol-extractable Cd in Chinese cabbage decreased by 2.1%-56% and 11.8%-100% with zeolite application, respectively. Moreover, the reduction effect of nano zeolite on Cd concentration in soil and plant was better than that of ordinary zeolite. The FDC of Cd fraction in shoot of Chinese cabbage showed correlation with the FDC of carbonate Cd and organic matter Cd in soil(P<0.05) when exposed to 1 mg·kg-1 Cd pollution. Moreover, correlation was also found in the FDC of organic matter Cd and residual Cd in soil(P<0.05) under 5 mg·kg-1 Cd pollution.

[Effect of Nano Zeolite on Growth, Activity of Antioxidant Enzyme, and Chemical Fractions and Concentration of Cd in Chinese Cabbage].

Pot experiment was carried out to investigate the influence of nano zeolite (NZ) and ordinary zeolite (OZ) on the growth, activities of antioxidant enzyme, Cd fraction and Cd concentration in two varieties of Chinese cabbage. The results showed that the activities of SOD, CAT and POD in the shoot and root of two varieties increased, and the dry weight of shoot and the total dry weight of Chinese cabbage increased by 4.5%-96.5% and 3.4%-88.4%, respectively. The application of zeolite effectively reduced the Cd concentration of shoot and root in the two varieties of Chinese cabbage, and the reduction range increased with the increase of the application amount of zeolite. Under the low Cd condition (1mg·kg-1 Cd), the Cd concentration of edible parts (shoots) in SD4 variety and XJC3 variety decreased by 1.0%-75.0% and 19.5%-68.9% (except the Cd1+OZ5/OZ10 processing), respectively. Under the high Cd condition (5 mg·kg-1 Cd), the Cd concentration of the edible parts (shoots) in SD4 variety and XJC3 variety decreased by 7.2%-53.2%(except Cd5+OZ10 processing) and 0.7%-63.0%, respectively. The Cd concentrations of the edible parts (shoots) in tow varieties of Chinese cabbage in the nano zeolite treatments decreased by 10.5%-65.7% compared with the ordinary zeolite treatments. In Cd contaminated soils (1 mg·kg-1 and 5 mg·kg-1 Cd),) the total extraction amount of Cd in the edible parts (shoots) of SD4 variety reduced by 12.4%-68.8% and 13.2%-55.6% (except of Cd5+OZ5 processing), and the total extraction amount of Cd in the edible parts (shoots) of XJC3 variety reduced by 9.4%-71.5% and 3.1%-38.7% (except of Cd1+OZ5 processing), respectively. The application amount of nano zeolite (NZ) and ordinary zeolite (OZ) was more, the reduction range of the total extraction amount of Cd in the edible parts (shoots) of two varieties of Chinese cabbage was greater.

Copper-induced alteration in sucrose partitioning and its relationship to the root growth of two Elsholtzia haichowensis Sun populations.

Hydroponic culture was used to comparatively investigate the copper (Cu)-induced alteration to sucrose metabolism and biomass allocation in two Elsholtzia haichowensis Sun populations with one from a Cu-contaminated site (CS) and the other from a non-contaminated site (NCS). Experimental results revealed that biomass allocation preferred roots over shoots in CS population, and shoots over roots in NCS population under Cu exposure. The difference in biomass allocation was correlated with the difference in sucrose partitioning between the two populations. Cu treatment (45 µM) significantly decreased leaf sucrose content and increased root sucrose content in CS population as a result of the increased activities of leaf sucrose synthesis enzymes (sucrose phosphate synthetase and sucrose synthase) and root sucrose cleavage enzyme (vacuolar invertase), which led to increased sucrose transport from leaves to roots. In contrast, higher Cu treatment increased sucrose content in leaves and decreased sucrose content in roots in NCS population as a result of the decreased activities of root sucrose cleavage enzymes (vacuolar and cell wall invertases) that led to less sucrose transport from leaves to roots. These results provide important insights into carbon resource partitioning and biomass allocation strategies in metallophytes and are beneficial for the implementation of phytoremediation techniques.

[Effect of Nano Zeolite on Chemical Fractions of Cd in Soil and Uptake by Chinese Cabbage at Different Soil pH and Cadmium Levels].

Incubation experiments were carried out to investigate the influence of different nano zeolite(NZ) and ordinary zeolite(OZ) levels(0, 5, 10 and 20 g·kg-1) on the fraction distribution coefficient (FDC) of Cd and soil CEC at different soil pH (4, 5, 6, 7 and 8) when exposed to different cadmium(Cd) levels(1, 5, 10 and 15 mg·kg-1), and pot experiment were carried out to investigate the effects of nano zeolite(NZ) and ordinary zeolite(OZ) on the growth, Cd concentration and Cd accumulation of Chinese cabbage. The results showed that nano zeolite and ordinary zeolite decreased the concentration and FDC of exchangeable Cd (EX-F), and increased the concentration and FDC of carbonate(CAB-F), Fe-Mn oxide(FMO-F), organic matter (OM-F) and residual fraction(RES-F) in incubation experiments. At the end of incubation, the FDC of soil exchangeable Cd decreased from 72.0%-88.0% to 2.4%-10.7%. The decreasing effect of zeolite on the concentration and FDC of exchangeable Cd (EX-F) increased with the increase of zeolite, and the decreasing effect of nano zeolite (NZ) was better than that of ordinary zeolite (OZ). During the culture of 28 d, the concentration of different Cd fractions in soil was in order of EX-F>RES-F>FMO-F>CAB-F>OM-F under different pH conditions. Exchangeable fraction Cd was the dominant fraction of Cd in soil during the whole incubation. Soil CEC had significant negative correlation with soil exchangeable Cd (EX-F) (P<0.01), and significant positive correlation with the concentrations of Fe-Mn oxide(FMO-F) and organic matter (OM-F) in soil(P<0.01). Nano zeolite and ordinary zeolite effectively increased soil CEC, and soil CEC increased with the increase of the pH value of soil in the zeolite treatments. Significant negative correlation was found in soil pH with soil exchangeable Cd (EX-F)(P<0.01). The dry weight of plant tissues in Chinese cabbage increased by 14.3%-131.4% in the presence of nano zeolite(NZ) and ordinary zeolite(OZ), and Cd concentration of shoot and root decreased by 1.0%-75.0% and 3.8%-53.2%, respectively. Higher concentration and accumulation of Cd were observed in XJC3 variety than those in SD 4 variety. Compared with ordinary zeolite (OZ), nano zeolite (NZ) was better in increasing the biomass of Chinese cabbage as well as decreasing accumulation of Cd in Chinese cabbage.

[Effect of exogenous selenium on accumulation and chemical forms of cadmium in cucumber (Cucumis satiuus L.)].

Pot experiments were carried out to investigate the influence of different selenium (Se) levels (0, 0.5 and 1.0 mg x L(-1)) on the plant growth, concentration of malonaldehyde (MDA), activities of antioxidant enzymes, accumulation and chemical forms of cadmium (Cd) in cucumber when exposed to Cd (20 mg x kg(-1)). The results showed that dry weights of leaf, stem, root, fruit and plant, and concentrations and accumulation of Cd significantly differed between two varieties of cucumber Yanbai and Jinyou 1. With increasing levels of Se, the contents of MDA in the leaves of Yanbai increased, but the contents of MDA in the leaves of Jinyou 1 decreased. The contents of MDA in the roots of Yanbai first increased and then decreased, while the contents of MDA in Jinyou 1 first decreased and then increased. The variation trends of CAT, SOD and POD in the leaves and roots of both varieties were different with increasing levels of Se. The concentrations of different chemical forms of Cd in the fruit decreased after spraying of Se, compared with the control. Cadmium concentrations in the leaves, stem, roots and fruit of both varieties decreased by 3.2%-17.9%, 14.6%-28.2%, 5.1%-18.5% and 60.6%-75.8% in the presence of Se when exposed to Cd. Accumulation of Cd in the plant of both varieties was in order of Jinyou 1 > Yanbai in the presence or absence of Na2SeO3.

[Effect of Nano Zeolite on Chemical Fractions of Cd in Soil and Its Uptake by Cabbage].

Incubation experiments were carried out to investigate the influence of different nano zeolite (NZ) and ordinary zeolite (OZ) levels(0, 5, 10 and 20 g · kg⁻¹) on the change trends in fraction distribution coefficient (FDC) of Cd when exposed to different Cadmium (Cd) levels (1, 5, 10 and 15 mg · kg⁻¹), and pot experiments were carried out to investigate their influence on soil Cd fraction and Cd uptake by cabbage. The results in incubation experiments showed that the application of nano zeolite as well as ordinary zeolite effectively decreased the FDC of exchangeable Cd and increased the FDC of Fe-Mn oxide fraction. The FDC of soil Cd from 0 d to 28 d was deceased at first, then increased and tended to be stable, and finally increased. At the end of incubation, the FDC of soil exchangeable Cd decreased from 72.0%-88.0% to 30.0%-66.4%. Exchangeable fraction Cd was the most dominant Cd fraction in soil during the whole incubation. The results in pot experiment indicated that the application of nano zeolite and ordinary zeolite decreased the concentration and FDC of soil exchangeable Cd, and concurrently the concentration and FDC of Cd in carbonate, Fe-Mn oxide, organic matter and residual fraction were increased. The lowest EX-Cd was observed in the treatment with high dose of nano zeolite (20 g · kg⁻¹). The FDC of exchangeable Cd showed significant negative relationship with the soil pH (P < 0.05), and was concurrently extremely positively correlated with Cd concentration in shoot and root of cabbage (P < 0.01). Soil pH increased by 1.8%-45.5% and 6.1%-54.3% in the presence of zeolite when exposed to 5 mg · kg⁻¹ 1 and Cd, respectively; FDC of exchangeable Cd decreased by 16.3%-47.7% and 16.2%-46.7%; Cd concentration in each tissues of cabbage decreased by 1.0%-75.0% and 3.8%-53.2%, respectively. Moreover, the reduction effect of nano zeolite on soil and plant Cd was better than that of ordinary zeolite. The growth of cabbage was stimulated by low and medium zeolite doses (≤ 10 g · kg⁻¹), while inhibited by high zeolite doses (20 g · kg⁻¹). Compared to ordinary zeolite, the biomass of Chinese cabbage was significantly increased by Nano zeolite, while the exchangeable Cd in soil as well as Cd concentration and Cd accumulation of cabbage were significantly reduced.

[Effect of Ryegrass and Arbuscular Mycorrhizal on Cd Absorption by Varieties of Tomatoes and Cadmium Forms in Soil].

Field trial was carried out to investigate the effects of ryegrass and arbuscular mycorrhizal single or compound treatment to two varieties of tomato ("Defu mm-8" and "Luobeiqi") on the plant growth, concentrations and accumulations of Cd as well as the impact on microorganisms, enzyme activities, pH and Cd forms in soil when exposed to Cd (5.943 mg · kg⁻¹). The results showed that dry weights of fruit, root, stem, leaf and plant significantly increased by single or compound treatment of ryegrass and arbuscular mycorrhizal by 14.1%-38.4% and 4.2%-18.3%, 20.9%-31.5% and 8.4%-10.3%, 13.0%-16.8% and 3.0%-9.5%, 10.7%- 16.8% and 2.7%-7.6%, 14.3%-36.6% and 4.5%-16.8%, respectively. The amounts of bacteria, fungi, actinomycetes of soil and the activities of urease, invertase, acid phosphatase, catalase in soil were increased by single or compound treatment of ryegrass and arbuscular mycorrhizal, and the soil microorganism amounts and enzyme activities significantly differed between the two varieties of tomato and treatments (P < 0.05). Soil pH was increased by single or compound treatment of ryegrass and arbuscular mycorrhizal, while the concentrations of EXC-Cd, CAB-Cd, Fe-Mn-Cd and total Cd in soil were decreased, and the total Cd content was decreased by 16.9%-27.8%. Cadmium concentrations in fruit, leaf, stem and root of both varieties were significantly decreased by 6.9%-40.9%, 5.7%-40.1%, 4.6%-34.7% and 9.8%-42.4%, respectively. Cadmium accumulations in tomato were in order of leaf > stem > root > fruit. Comparing the two tomato varieties, Cd concentrations and Cd accumulations in fruit and plant were in order of "Luobeiqi" < "Defu mm-8" in the presence or absence of single or compound treatment of ryegrass and arbuscular mycorrhizal.

[Effects of ryegrass and arbuscular mycorrhiza on activities of antioxidant enzymes, accumulation and chemical forms of cadmium in different varieties of tomato].

Pot experiments were carried out to investigate the effects of ryegrass and arbuscular mycorrhiza on the plant growth, malondialdehyde (MDA), antioxidant enzyme activities of leaf and root, accumulation and chemical forms of cadmium (Cd) in tow varieties of tomato when exposed to Cd (20 mg x kg(-1)). The results showed that dry weights of fruit and plant, and contents of malondialdehyde (MDA) and antioxidant enzyme activities of leaf and root, and concentrations and accumulations of Cd significantly differed between two varieties of tomato. Dry weights of fruit, roots, stem, leaf and plant were increased by single or combined remediation of ryegrass and arbuscular mycorrhiza, while MDA contents and antioxidant enzyme activities of leaf and root reduced. The total extractable Cd, F(E), F(W), F(NaCl), F(HAc), F(HCl), and F(R) in fruit of two varieties of tomato reduced by 19.4% - 52.4%, 31.0% - 75.2%, 19.7% - 59.1%, 3.1% - 48.2%, 20.0% - 65.0%, 40.7% - 100.0% and 15.2% - 50.0%, respectively. Cadmium accumulations in tomato were in the order of leaf > stem > fruit > root. Cadmium concentrations in leaf, stem, root and fruit of both varieties decreased by single or combined remediation of ryegrass and arbuscular mycorrhiza, and Cd accumulations of stem and plant of two varieties also reduced. Cd accumulations in fruit of two varieties decreased by 42.9% and 43.7% in the combined remediation treatments, respectively. Tolerance and resistance of 'LUO BEI QI' on Cd was more than 'De Fu mm-8', and Cd concentrations and Cd accumulations in fruit and plant were in the order of 'LUO BEI QI' < 'De Fu mm-8' in the presence or absence of single or combined remediation of ryegrass and arbuscular mycorrhiza.

Physiological responses of biomass allocation, root architecture, and invertase activity to copper stress in young seedlings from two populations of Kummerowia stipulacea (maxim.) Makino.

In the current study, we hypothesize that mine (metallicolous) populations of metallophytes form a trade-off between the roots and shoots when under copper (Cu) stress to adapt themselves to heavy metal contaminated habitats, and thus, differ from normal (non-metallicolous) populations in biomass allocation. To test the hypothesis, two populations of the metallophyte Kummerowia stipulacea, one from an ancient Cu mine (MP) and the other from a non-contaminated site (NMP), were treated with Cu(2+) in hydroponic conditions. The results showed that MP plants had higher root/shoot biomass allocation and more complicated root system architecture compared to those of the NMP plants when under Cu stress. The net photosynthetic capacity was more inhibited in the NMP plants than in the MP plants when under Cu stress. The sugar (sucrose and hexose) contents and acid invertase activities of MP plants were elevated while those in NMP plants were inhibited after Cu treatment. The neutral/alkaline invertase activities and sucrose synthase level showed no significant differences between the two populations when under Cu stress. The results showed that acid invertase played an important role in biomass allocation and that the physiological responses were beneficial for the high root/shoot biomass allocation, which were advantageous during adaptive evolution to Cu-enriched mine soils.

Cloning and characterization of acid invertase genes in the roots of the metallophyte Kummerowia stipulacea (Maxim.) Makino from two populations: Differential expression under copper stress.

The roots of metallophytes serve as the key interface between plants and heavy metal-contaminated underground environments. It is known that the roots of metallicolous plants show a higher activity of acid invertase enzymes than those of non-metallicolous plants when under copper stress. To test whether the higher activity of acid invertases is the result of increased expression of acid invertase genes or variations in the amino acid sequences between the two population types, we isolated full cDNAs for acid invertases from two populations of Kummerowia stipulacea (from metalliferous and non-metalliferous soils), determined their nucleotide sequences, expressed them in Pichia pastoris, and conducted real-time PCR to determine differences in transcript levels during Cu stress. Heterologous expression of acid invertase cDNAs in P. pastoris indicated that variations in the amino acid sequences of acid invertases between the two populations played no significant role in determining enzyme characteristics. Seedlings of K. stipulacea were exposed to 0.3µM Cu(2+) (control) and 10µM Cu(2+) for 7 days under hydroponics׳ conditions. The transcript levels of acid invertases in metallicolous plants were significantly higher than in non-metallicolous plants when under copper stress. The results suggest that the expression of acid invertase genes in metallicolous plants of K. stipulacea differed from those in non-metallicolous plants under such conditions. In addition, the sugars may play an important role in regulating the transcript level of acid invertase genes and acid invertase genes may also be involved in root/shoot biomass allocation.

Comparative study of root growth and sucrose-cleaving enzymes in metallicolous and non-metallicolous populations of Rumex dentatus under copper stress.

Sucrose metabolism in roots of metallophytes is very important for root growth and maintenance of heavy metal tolerance. However, rare researches have been carried out on this topic so far. We tested here a hypothesis that roots of copper-tolerant plants should manifest higher activities of sucrose-cleaving enzymes than non-tolerant plants for maintaining root growth under Cu stress. Plants of two contrasting populations of metallophyte Rumex dentatus, one from an ancient Cu mine (MP) and the other from a non-mine site (NMP), were treated with Cu in controlled experiments. Cu treatment resulted in a higher root biomass and root/shoot biomass ratio in MP compared to NMP. More complicated root system architecture was showed in MP under Cu stress. Activities and transcript levels of acid invertase as well as contents of sucrose and reducing sugar in MP were elevated under Cu treatment, while activities of neutral/alkaline invertase and sucrose synthase showed no significant differences between two populations. The results indicate important roles of acid invertase in governing root growth under Cu stress.

[Effect of exogenous iron on accumulation and chemical forms of cadmium, and physiological characterization in different varieties of tomato].

Pot experiments were carried out to investigate the influence of different iron (Fe) levels (0, 50, 100, 200 and 400 micromol.L-1) on the plant growth, activities of antioxidant enzymes, accumulation and chemical forms of cadmium (Cd) in tomato when exposed to Cd (10 mg.kg-1). The results showed that dry weights of fruit, roots, stem, leaf, and plant, and concentration and accumulation of Cd significantly differed between the two varieties of tomato tested. Dry weights of fruit, roots, stem, leaf, and plant increased in the presence of Fe. Activities of superoxide dismutase (SOD) of root in both varieties, and activities of catalase (CAT) of root in Yufenl09 deceased at first, and then increased with increasing levels of Fe, but activities of peroxidase (POD) of root in both varieties and activities of CAT of root in 4641 increased at first, and then reduced with increasing levels of Fe. Chemical forms of Cd in fruit of tomato were in the order of FR > FHCl > FE > FNaCl > FHAC > FW. All chemical forms of Cd obviously decreased after the application of adequate Fe compared to the control, while FHcI and FR of 4641, and the total extractable Cd, FE, FNaCl and FR of Yufenl09 increased at 400 micromol.L-1Fe treatment. Cadmium accumulations of tomato were in the order of leaf > stew > fruit > roots. Cadmium concentration in leaf, stem, roots and fruit of both varieties decreased by 7. 1% -25.3% , 30. 8% -50. 4% , 13.0% -45. 1% and 2.8% -11.7% in the presence of Fe when exposed to Cd, and the Cd concentration of fruit and Cd accumulations in plant were in the order of 4641 < Yufenl09 in the presence or absence of Fe.

Differential response to copper stress in the reproductive resources and allocation of metallophyte Kummerowia stipulacea.

Abundant seed production is a key life history trait for plant to maintain the stability of the whole population in adverse environments such as heavy metal contaminated mine area. In the current studies, we hypothesize that mine (metallicolous) populations of metallophytes have formed specialized reproductive strategies to adapt themselves to the heavy metal contaminated habitats, and differ from normal (non-metallicolous) populations in reproductive allocation. To test this hypothesis, the differences in reproductive resources and reproductive allocation between the copper mine and non-copper mine populations of pseudo-metallophyte Kummerowia stipulacea were comparatively examined under controlled Cu exposure experiments. Compared to non-copper mine population, copper mine population shows an increased seed output and larger reproductive effort under Cu stress. The increase of reproductive allocation in metallicolous population depends on not only seed size but also seed number per plant. The plants of metallicolous population increase allocation to the reproductive organs at the expense of a curtailment of allocation to vegetative traits, resulting in plants with shorter height and fewer branch numbers. There is little evidence displaying effect of root nodule on the reproductive resources and allocation. In addition, plants in metallicolous population reduce the transfer of Cu from roots to aboveground parts. These data suggest that plants of metallicolous population tend to invest more resources to reproductive output and increase their reproductive allocation in the adaptive evolution to Cu-enriched mine soils.

[Cd uptake in rice cultivars and Cd fractions in soil treated with organic acids and EDTA].

A pot experiment was conducted to examine the yield, quality and cadmium (Cd) uptake in different rice cultivars, and Cd speciation in soil after exposing to Cd (0, 1 and 5 mg x kg(-1)) in the presence of organic acids and ethylenediamine tetraacetic acid (EDTA). The results showed that general increase in the yield for cultivars Xiushui63 and II you527 was observed. Yield of two rice cultivars were in order of organic acids treatment or organic acids + 1/2EDTA treatment > EDTA treatment. The exchangeable, carbonate related and ferric-manganese oxidation related Cd increased; while organic complexation Cd and residules decreased in the presence of organic acids and EDTA. Cadmium concentrations in grain, straw and roots of both cultivars markedly reduced in the presence of organic acids and EDTA. Grain Cd concentration was the lowest for plants treated with EDTA, followed by organic acids + 1/2EDTA, and the highest Cd concentration in grain was found in the treatment with organic acids. Grain Cd concentration decreased by 9.0% to 49.3% and 16.5% to 30.6% at 1 mg x kg(-1) Cd in the presence of organic acids and EDTA, and by 12.7% to 28.5% and 4.3% to 19.1% at 5 mg x kg(-1) Cd. Cadmium concentration and accumulation in plants and total Cd content in soil were higher in Xiushui63 than in that in II you527. Grain Cd concentration decreased, and yield and quality of two rice cultivars increased at the same time in the presence of organic acids + 1/2EDTA.

[Effect of phosphor on accumulation and chemical forms of cadmium, and physiological characterization in different varieties of Capsicum annuum L].

Pot experiments were carried out to investigate the influence of different Phosphor (P) levels (0, 0.3% and 0.5%) on the plant growth, activities of antioxidant enzymes, accumulation and chemical forms of cadmium (Cd) in Capsicum annuum L. when exposed to Cd (10 mg x kg(-1)). The results showed that dry weights of leaf, fruit, roots and total dry weights of plant, and concentration and accumulation of Cd significantly differed between two varieties of Capsicum annuum L. Dry weights of fruit and total plant of Chaotianjiao increased by P (0.3% and 0.5%), while that of Yanjiao425 was inhibited. Activities of catalase (CAT) were increased at first, and then reduced in the presence of P; Activities of superoxide dismutase (SOD) and peroxidase (POD) of Chaotianjiao increased with increasing levels of P, but activities of SOD and POD of Yanjiao425 decreased with increasing levels of P. Chemical forms of Cd in fruit of Capsicum annuum L. were in order of F(NaCl) > F(HAC) > F(E) > Fr > F(HC) > F(W). The total extractable Cd, ethanol-extractable Cd, hydrochloric acid-extractable Cd and residual Cd in fruit of Ynajiao425 obviously decreased in the presence of P compared to the control, while the total extractable Cd, water-extractable Cd, acetic acid-extractable Cd and residual Cd in fruit of Chaotianjiao increased. Cadmium accumulations of Capsicum annuum L. were in order of roots > stew > leaf > fruit. Cadmium accumulations in fruit and plant of Yanjiao425 were decreased by 47.7% and 58.5% , 5.5% and 13. 1% in the presence of 0.3% and 0.5% P when exposed to Cd, and Cd accumulations in fruit and plant of Chaotianjiao were decreased by 23.6% in the presence of 0.3% P.

Differential expression of acid invertase genes in roots of metallicolous and non-metallicolous populations of Rumex japonicus under copper stress.

Recent evidence indicates that during copper (Cu) stress, the roots of metallicolous plants manifest a higher activity of acid invertase enzymes, which are rate-limiting in sucrose catabolism, than non-metallicolous plants. To test whether the higher activity of acid invertases is the result of higher expression of acid invertase genes, we isolated partial cDNAs for acid invertases from two populations of Rumex japonicus (from metalliferous and non-metalliferous soils), determined their nucleotide sequences, and designed primers to measure changes in transcript levels during Cu stress. We also determined the growth of the plants' roots, Cu accumulation, and acid invertase activities. The seedlings of R. japonicus were exposed to control or 20 µM Cu(2+) for 6d under hydroponic conditions. The transcript level and enzyme activity of acid invertases in metallicolous plants were both significantly higher than those in non-metallicolous plants when treated with 20 µM. Under Cu stress, the root length and root biomass of metallicolous plants were also significantly higher than those of non-metallicolous plants. The results suggested that under Cu stress, the expression of acid invertase genes in metallicolous plants of R. japonicus differed from those in non-metallicolous plants. Furthermore, the higher acid invertase activities of metallicolous plants under Cu stress could be due in part to elevated expression of acid invertase genes.

Effects of copper on phenology and reproduction in Rumex dentatus from metalliferous and non-metalliferous sites.

The responses of phenology and reproductive traits to copper stress in two populations of Rumex dentatus were comparatively studied with pot culture experiments. Seeds used for the experiments were, respectively, collected from metalliferous and normal soils. It was found that the responses of phenology and reproductive traits to Cu treatment between the two populations were significantly different. Compared to the non-metallicolous population, the metallicolous population of R. dentatus had a short life cycle, large reproductive effort, and high fertility under Cu stress. In addition, the reproductive effort in metallicolous population of R. dentatus was maintained at the expense of a curtailment of vegetative development. The results suggested that change in phenological traits and more resources allocation to reproduction might play an important role in the adaptation of metallicolous population of R. dentatus to the Cu-enriched mine soils.

[Effect of different zinc levels on accumulation and chemical forms of cadmium, and physiological characterization in Capsicum annuum L].

Pot experiments were carried out to investigate the influence of different zinc (Zn) levels (0, 100, 200, 400 and 600 micromol x L(-1)) on the plant growth,activities of antioxidant enzymes, contents of chlorophyll a and b, accumulation and chemical forms of cadmium (Cd) in Capsicum annuum L. when exposed to Cd (20 mg x kg(-1)). The results showed that dry weights of leaf, stem, fruit and root, and contents of chlorophyll a and b in Capsicum annuum L. were increased by Zn ( < or = 400 micromol x L(-1)), while inhibited by high Zn (600 micromol x L(-1)). Activities of superoxide dismutase (SOD) and catalase (CAT) were reduced by Zn ( < or =400 micromol x L(-1)), the lowest activities of SOD and CAT were recorded in 400 micromol x L(-1) Zn, but activities of SOD and CAT were increased when Zn >400 micromol x L(-1). Cadmium concentrations in stem, fruit and root of Capsicum annuum L. were decreased by 2.7%-5.4%, 7.5%-28.1% and 7.6%-21.8% in the presence of Zn when exposed to Cd. The total extractable Cd, NaCl- extractable Cd, water-extractable Cd and ethanol-extractable Cd in fruit were reduced by 7.7%-21.8%, 4.11%-23.6%, 54.5%-66.8% and 4.8%-86.7% in the presence of Zn,while acetic acid- extractable Cd and residual Cd were increased by 28.0%-68.0% and 12.6%-25.0%.

Effect of Cu stress on the invertase activity and root growth in two populations of Rumex dentatus L. with different Cu tolerance.

There has been no study on key enzymes in sucrose cleavage in metallophyte plants so far, which may be crucial for the plants' root growth and heavy metal tolerance maintenance. Acid invertases are rate-limiting enzymes in sucrose metabolism. Here, we tested the hypothesis that the roots of copper-tolerant plants should manifest a higher activity of acid invertases than nontolerant plants both for supporting growth and for their maintaining tolerance under Cu stress. Two populations of Rumex dentatus L., one from an ancient waste heap at a Cu mine (Cu-tolerant population), and the other from a noncontaminated site (Cu nontolerant population), were used in the experiments. The seedlings of Rumex dentatus L. were exposed to 0, 10, and 40 microM CuCl(2) for 14 days. Cu exposure had a stronger inhibition on root growth and thus resulted in a lower root/shoot ratio in the plants of nontolerant population compared with the Cu-tolerant population. Cu exposure showed a stronger inhibition of acid invertase activity of Cu nontolerant plants than Cu tolerant plants, whereas neutral/alkaline invertase was insensitive to Cu. A positive correlation between the activity of acid invertases and the root growth and root/shoot ratio was observed. The results suggested that the higher activities in acid invertases of Cu-tolerant population might at least partly associate with the plants' Cu tolerance, and their higher activities in acid invertases in turn played an role in maintenance of the Cu tolerance by supplying carbon and energy for tolerance mechanisms.