Identification of Rice Varieties with High Tolerance or Sensitivity to Copper

ABSTRACT

To study tolerance of rice (Oryza sativa L.) to excessive copper (Cu), 167 varieties were screened for Cu tolerance or sensitivity. Based on the elongation of the root, No. 1139 and No. 1195 were chosen as Cu-tolerant and Cu-sensitive variety. Compared with the control (0.32 μ mol L^{? 1} Cu), treatments with 5 and 8 μ mol L^{? 1} Cu for 10 days had no significant effect on the root and shoot dry masses of Cu-tolerant variety No. 1139, but significantly decreased root and shoot dry masses of variety No. 1195. Copper at 5 and 8 μ mol L^{? 1} also significantly decreased contents of chlorophyll and carotenoid and the ratio of variable to maximal chlorophyll fluorescence (Fv/Fm) in the leaves of the Cu-sensitive variety No. 1195. Roots and shoots of variety No. 1139 contained significantly higher the concentration of Cu than variety No. 1195 exposed to 5 and 8 μ mol L^{? 1} Cu. However, variety No. 1139 had higher Cu proportions on the cell walls in shoots and roots than variety No. 1195.     

Identification and characterization of salt- and thermo-tolerant Leucaena-nodulating Rhizobium strains

 Rhizobium-legume symbioses are important for their nitrogen input, but salinity and elevated temperature in arid and semi-arid areas limit their effectiveness, and therefore plant growth and productivity. Sixteen Rhizobium strains isolated from root nodules of Leucaena trees grown in different geographical areas of Egypt varied in their degree of tolerance to salinity and in their symbiotic effectiveness with Leucaena leucocephala under saline conditions. Three strains were tolerant to >3% NaCl. L. leucocephala grown in the greenhouse at concentrations of NaCl up to 1.0% and inoculated either with strain DS 78 or strain DS 158 displayed significantly better growth than those plants grown at the same levels of salinity and inoculated with reference strain TAL 583. Although nine of the Rhizobium strains grew at 42  °C, their mean generation times were lengthened two- to fourfold. When daylight growth temperatures were elevated from 30  °C to 42  °C, nodule number and mass, nitrogenase activities and shoot top dry weight of plants inoculated with strains DS 78, DS 157 and DS 158 significantly increased, whereas these parameters decreased in plants inoculated with strain TAL 583. Rhizobium strains that effectively nodulate Leucaena under adverse saline conditions and at high temperatures were thus isolated, identified and characterized. Received: 12 September 1997     

Effect of Sodium Chloride on Growth,Nutrient Accumulation,and Nitrogen Fixation of Common Bean Plants in Symbiosis with Isogenic Strains

The effect of sodium chloride (NaCl)-salinity on growth responses and tissues organic solutes and mineral content was investigated in common bean plants inoculated with salt-tolerant Rhizobium tropici wild-type strain CIAT899 and four mutant derivatives having decreased salt-tolerance (DST). Under non-saline conditions two mutants formed partially effective (HB10, HB12) and another two almost ineffective (HB8, HB13) nodules. A great variation of NaCl tolerance in the different symbiosis tested was observed at harvest, 32 day after planting. Common bean plant responded to salinity by decreasing the content of dry plant biomass, nodule number and the nitrogen fixation, and increasing the root to shoot ratio. The salt dose of 25 mM produced an increase of total soluble sugar and free amino acids content. This result suggest that these metabolites might be related with a nodule osmotic adjustment response under saline conditions, however cannot be excluded that the increase of amino acids content could be a consequence of protein degradation. In the other hand, sodium, calcium and phosphorus contents in shoot increased under the saline treatments. Potassium (K) and calcium (Ca) contents, unlike phosphorus (P) content, in shoot were not related with the symbiotic efficiency of mutant, however the decrease of P content suggest that these symbioses have limited their P absorption process independently of the saline treatment. NaCl tolerance associated with a retention of sodium and maintenance of potassium selectivity seem to be a strategy used for the salt stressed common bean plants in symbiosis assayed here.     

Growth and mineral distribution of Sesuvium portulacastrum l., a salt marsh halophyte,under sodium chloride stress

Abstract

Response of Sesuvium portulacastrum L., to the elogenous addition of sodium chloride (NaCl), applied at different concentrations ranging from 100 to 900 mM was evaluated. Leaf area, shoot and root weight of the treated plants were increased significantly. Total nitrogen (N) content of leaves, stem and root was significantly increased up to 600 mM NaCl, and thereafter declined moderately. Accumulation of potassium (K), and calcium (Ca) were exponentially increased with NaCl treatment as also observed in the case of N. Sodium content of shoot and roots of S. portulacastrum increased significantly with increasing NaCl concentrations. Certain essential elements like copper (Cu), iron (Fe), manganese (Mn) and zinc (Zn) were also accumulated in all plant parts significantly, up to 600 mM. However, due to high salinization, growth and development of plants inhibited markedly. Furthermore, elemental concentration decreased beyond 600 mM level of NaCl. Due to aging and senescence, marked changes in leaf area, shoot, root volume and mineral content were observed between sampling time.     

Inoculation with the native Rhizobium gallicum 8a3 improves osmotic stress tolerance in common bean drought-sensitive cultivar

Abstract

Symbiotic nitrogen fixation potential in common bean is considered to be low in comparison with other grain legumes. However, it may be possible to improve the nitrogen fixation potential of common bean using efficient rhizobia. In order to improve osmotic stress tolerance of a drought-sensitive common bean cultivar (COCOT) consumed in Tunisia, plants were inoculated either by the reference strain Rhizobium tropici CIAT 899 or by inoculation with rhizobia isolated from native soils Rhizobium gallicum 8a3. Fifteen days after sowing, osmotic stress was applied by means of 25 mM mannitol (low stress level) or by 75 mM mannitol (high stress level). Fifteen days after treatment plants were harvested and different physiological and biochemical parameters were analysed. Results showed no significant differences between the studied symbioses under control conditions. However after exposure to osmotic stress our results showed better tolerance of COCOT to osmotic stress when inoculated with the native R. gallicum 8a3. This can be partially explained by better water-use efficiency in both leaves and nodules, better relative water content in nodules and better efficiency in utilization of rhizobial symbiosis as compared with COCOT-CIAT 899 symbiosis. Hence, the present study suggested the better use of native soil isolated strains for the inoculation of common bean in order to improve its performance and nitrogen fixation potential under stressful conditions.     

Response of the pigeonpea-Rhizobium symbiosis to salinity stress: variation among Rhizobium strains in symbiotic ability

Summary There were significant differences among pigeonpea [Cajanus cajan (L.) Millsp] Rhizobium sp. strains (IC 3506, IC 3484, IC 3195, and IC 3087) in their ability to nodulate and fix N₂ under saline conditions. Pigeonpea plants inoculated with IC 3087 and IC 3506 were less affected in growth by salinity levels of 6 and 8 dS m^-1 than plants inoculated with the other strains. For IC 3506, IC 3484, and IC 3195, there was a decrease in the number of nodules with increasing salinity, while the average nodule dry weight and the specific nitrogenase activity remained unaffected. However, in IC 3087, the number of nodules increased slightly with increasing salinity. Leaf-P concentrations increased with salinity in the inoculated plants irrespective of the Rhizobium sp. strain, and leaf-N concentrations decreased with increasing salinity in IC 3484 and IC 3195 only. Shoot-Na and-Cl levels were further increased in these salt-sensitive strains only at 8 dS m^-1. Therefore there may be scope for selecting pigeonpea Rhizobium sp. symbioses better adapted to saline conditions. The Rhizobium sp. strains best able to form effective symbioses at high salinity levels are not necessarily derived from saline soils.Submitted as JA No. 919 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

Varietal tolerance to Zinc deficiency in wheat and barley grown in chelatorbuffered nutrient solution and its effect on uptake of Cu,Fe, and Mn

Tolerance to zinc (Zn) deficiency was examined for three wheat (Triticum aestivum L.) and three barley (Hordeum vulgare L.) varieties grown in chelator‐buffered nutrient solution. Four indices were chosen to characterize tolerance to Zn deficiency: (1) relative shoot weight at low compared to high Zn supply (“Zn efficiency index”), (2) relative shoot to root ratio at low compared to high Zn supply, (3) total shoot uptake of Zn under deficient conditions, and (4) shoot dry weight under deficient conditions. Barley and wheat exhibited different tolerance to Zn deficiency, with barley being consistently more tolerant than wheat as assessed by all four indices. The tolerance to Zn deficiency in the barley varieties was in the order Thule=Tyra>Kinnan, and that of wheat in the order Bastian=Avle>Vinjett. The less tolerant varieties of both species accumulated more P in the shoots than the more tolerant varieties. For all varieties, the concentrations of Mn, Fe, Cu, and P in shoot tissue were negatively correlated with Zn supply. This antagonism was more pronounced for Mn and P than for Cu and Fe. Accumulation of Cu in barley roots was extremely high under Zn‐deficient conditions, an effect not so clearly indicated in wheat.     

Improving salt stress responses of the symbiosis in alfalfa using salt-tolerant cultivar and rhizobial strain

Salt stress can affect alfalfa growth directly by adversely affecting metabolism, or indirectly by its effect on Rhizobium capacity for symbiotic N₂ fixation. Growth and carbohydrate metabolism in leaves, roots and nodules of two alfalfa cultivars (Medicago sativa cv Apica and salt-tolerant cv Halo) in association with two rhizobial strains (A2 and salt-tolerant Rm1521) exposed to different levels of NaCl (0, 20, 40, 80 or 160 mM NaCl) were assessed under controlled conditions. For both cultivars, shoot and root biomasses and shoot to root ratio significantly declined with increasing NaCl concentrations. Under 80 mM NaCl, Halo plants yielded 20% more fresh shoot biomass than Apica while plants inoculated with Rm1521 allocated more biomass to the roots than to the shoots compared to A2. Halo plants maintained a steady shoot water content (about 80%) under the entire range of NaCl concentrations. Shoot water content was more variable in Apica. Apica in association with salt-tolerant strain Rm1521 maintained a better water status than with strain A2, as indicated by the higher shoot water content at 80 mM NaCl. Under salt stress, two major compatible sugars involved in plant osmoregulation, sucrose and pinitol, increased in leaves while a large accumulation of starch was observed in roots. In nodules, pinitol, sucrose and starch increased under salt stress and were much more abundant with strain Rm1521 than with A2. This suggests that there could be an active transport from the shoot to the nodules to help maintain nodule activity under NaCl stress and that strain Rm1521 increases the sink strength toward nodules. Our results show that combining cultivars and rhizobial strains with superior salt tolerance is an effective strategy to improve alfalfa productivity in salinity affected areas.     

NODULATION,NITROGENASE ACTIVITY,AND DRY WEIGHT OF CHICKPEA AND PIGEON PEA CULTIVARS USING DIFFERENT BRADYRHIZOBIUM STRAINS

Chickpea [Cicer arietinum (L.)] and pigeon pea [Cajanus cajan (L.) Millsp.] were grown outside in large clay pots from 1992 to 1995 in Edmond, Oklahoma. Plants were studied to evaluate nodulation, nitrogenase activity, and shoot dry weight (DW) of ‘ICCV-2’ and ‘Sarah’ chickpea inoculated with multistrain, TAL 1148, and TAL 480 Bradyrhizobium, as well as ‘Georgia-1’ and ‘ICPL-87’, pigeon pea inoculated with multistrain TAL 1127, and TAL 1132 Bradyrhizobium. Following wheat [Triticum aestivum (L.) emend. Thell.] harvests in the spring, legumes were planted in the summer and harvested at three successive dates during the following months. Leaves and stems from remaining plants were incorporated into the soil after the last harvest. Across year, chickpea measurements were sensitive to temperature and precipitation whereas pigeon pea measurements were sensitive to length of growing season as well as climate. Pigeon pea consistently demonstrated higher nitrogen-fixing capacity and shoot DW compared with chickpea. Nodule and shoot DW of both species increased with plant age whereas nodule count and nitrogenase activity generally increased with plant age and leveled off or decreased at flowering. Sarah chickpea demonstrated higher nodule count and nodule DW than ICCV-2, as did the Georgia-1 pigeon pea compared with ICPL-87. Shoot DW of Georgia-1 pigeon pea was generally higher than that of ICPL-87. Multistrain inoculum improved nodulation and shoot DW of chickpea, and TAL 1127 improved nodulation of pigeon pea compared with other treatments. These results indicate that specific chickpea and pigeon pea cultivars, along with appropriate Bradyrhizobium strains, may improve nitrogen fixation and DW of these species.     

Net fluxes of mineral nutrients,water, and carbohydrate influenced by manganese in root and shoot of Cucumis sativus L. 1

An experiment was conducted with cucumber (Cueumis sativus L., cv. Sumter) plants in the vegetative phase of growth to determine effects of manganese deficiency (0.2 μM Mn) and toxicity (182 μM Mn) on fluxes of several mineral nutrients, water, and carbohydrate in the root and shoot, beginning 43 d after germination. Plants were sampled every three days from 34 to 58 d after germination. First and second derivatives of regression equations were used to estimate fluxes and study source/sink phenomena of dry weight (DW), fresh weight (FW), H₂O, Cu, Fe, Mn, Zn, N, P, and K in root and shoot tissues.

With Mn sufficiency (1.8 μM Mn), both root and shoot acted as sinks for of each of the 10 dependent variables through 58 d. In contrast, Mn deficiency caused net loss of K and N from the root beginning at 53 and 56 d, respectively, and net loss of P and Fe from the shoot beginning at 57 and 58 d, respectively. With Mn toxicity, net loss of Cu, N, and K from the root began at 46, 46, and 51 d, respectively, and net loss of Fe from the shoot began at 55 d. Both Mn deficiency and toxicity increased the shoot:root ratio (S:R) of K and decreased the S:R of Fe, compared to Mn‐sufficiency. Manganese deficiency decreased the S:R of DW and H₂O, and Mn toxicity increased the S:R of FW, DW, H₂0, and N. Under the conditions of this experiment, acute Mn toxicity affected fluxes more severely than did Mn deficiency.     

Fungal flora of soil polluted with copper

The effects of copper pollution on the soil fungal flora was investigated. Soils treated with 100, 200, 400, 800 or 1600 μg Cu g^?1 were used for experiments to study changes in fungal populations, especially the development and dominance of copper-tolerant fungi. Fungi were sampled 1, 3 and 5 months after copper treatment.All the correlation coefficients between the copper contents and the number of fungal colonies plated were positive. The higher the copper concentration in soil, the more 1000 μ Cu ml^?1 tolerant fungi were isolated. The relative number of 1000 μg Cu mr^?1 tolerant fungi from the soil treated with 1600 μg Cu g^?1 was about 30% of those of the control 14 days after treatment. Within the limits of this experiment, the increase in fungal populations was directly correlated with the increase of dominant Cu-tolerant fungi.From control soils, containing low quantities of copper, 1000 μg Cu ml^?1 tolerant fungi were also isolated; whereas, from soils containing high amounts of copper, some Cu-sensitive fungi were isolated. Most of the 1000 μg Cu ml^?1 tolerant fungi were Penicillium spp. It was concluded that the genus Penicillium may be dominant in soils polluted with copper.     

Copper Tolerance and Accumulation of Elsholtzia splendens Nakai in a Pot Environment

ABSTRACT

Elsholtzia splendens Naki has been identified as a copper (Cu) geobotanical indicator. In this study, the effects of Cu supply levels (control, 100, 200, 400, 600, 800, 1000, 1200 mg kg¹) on the growth and Cu accumulation in E. splendens were studied in one pot experiment. The results showed that no reduction in shoot height and dry weight was noted when the plants were grown at Cu supply levels up to 1000 mg kg^?1 in soil. Slight stimulation on shoot growth was noted at Cu levels ≥ 100 mg kg^?1. Copper concentration in shoots and roots increased with increasing Cu levels, and reached a maximum of 1751 and 9.45 mg kg^?1 (DW) at 1200 mg Cu kg^?1. The amount of Cu accumulated in the roots and shoots were 313 and 22 μ g plant^?1 at external Cu levels of 1000 and 800 mg kg^?1, respectively. The shoot/root Cu ratios ranged from 0.005 to 0.008 and more than 92% of the total Cu taken up by E. splendens was accumulated in roots. Furthermore, Cu concentrations in roots and shoots were significant and positively correlated with total soil Cu, water, ammonium nitrate (NH₄NO₃), ammonium (NH₄)-acetate, and ethylenediaminetetraacetic acid (EDTA) extractable Cu. These results indicate that E. splendens can considered as a Cu tolerant and accumulated plant, and root is the major part for accumulation of Cu in E. splendens.     

耐铜苏丹草根内生细菌的分离筛选及其生物学特性研究

从生长在铜矿废弃地土壤中的Cu耐性苏丹草根中分离筛选到二株产1-氨基环丙烷-1-羧酸(ACC)脱氨酶内生细菌K1-6和K3-9菌株,并对菌株生物学特性进行了研究。菌株K1-6和K3-9具有较强的Cu抗性和多种抗生素抗性,菌株K1-6和K3-9能够溶磷和分泌吲哚乙酸(IAA),另外,菌株K3-9还能够产生铁载体和精氨酸脱羧酶,菌株K1-6和K3-9对温度、p H和盐浓度具有一定的耐受性,经16S r DNA序列分析,菌株K1-6和K3-9分别被鉴定为根瘤菌属(Rhizobium sp.K1-6)和肠杆菌属(Enterobacter aerogenes K3-9)。采用平皿培养试验研究了菌株K1-6和K3-9对生长在不同浓度Cu(0、4 mg/L)环境中的苏丹草的生长和吸收Cu的影响。结果表明,接菌处理苏丹草根部和地上部干重分别比对照增加了10.6%~45.5%和13%~40%,差异达显著水平(P0.05);接菌株K1-6处理苏丹草根部和地上部Cu含量比对照增加了46%和85%(P0.05),而接菌株K3-9处理苏丹草根部和地上部Cu含量与对照相比没有显著差异。另外,接菌株K1-6处理苏丹草根部和地上部总Cu吸收量比对照增加了88%和114%(P0.05),接菌株K3-9处理苏丹草根部总Cu吸收量比对照增加了44%(P0.05)。另外,接菌株K1-6和K3-9处理的苏丹草根部吸收的Cu是地上部吸收Cu的16~23倍。研究表明,分离自耐铜苏丹草根部的内生细菌具有多种植物促生特性,能够显著促进苏丹草的生长、提高苏丹草对Cu的耐受性,并强化苏丹草根部对Cu的富集能力。另外,不同的内生细菌对苏丹草的生长、富集和耐受Cu的影响不同。     

Medicago sativa-rhizobia symbiosis under water deficit: Physiological,antioxidant and nutritional responses in nodules and leaves

Biological nitrogen fixation has positive effects in the enhancement of soil nutrition and plants growth, especially under difficult conditions. The study assesses the effect of drought (40% field capacity) on growth, physiological and biochemical parameters of six symbioses involving two Moroccan alfalfa populations Adis-Tata (Ad) and Riche (Rc) and Moapa (Mo) variety and ArfRh2 (Rh2) and AufRh8 (Rh8) rhizobia isolated from Arfoud and Aufous, respectively. Results showed that drought decreased plants growth and nodulation. Physiologically, as response to stress, the nodules membrane stability was negatively affected. Biochemically, activities of peroxidase (POX), catalase (CAT), superoxide dismutase (SOD) and acid phosphatase (APase) increased in leaves and nodules. Ad-Rh2, Rc-Rh2 and Ad-Rh8 have been qualified as more drought tolerant. They showed high antioxidant activities and as consequence less membrane damages especially in their nodules. The increase in nodules health and antioxidant defense could results in the enhancement of plant drought tolerance under stress.     

Selection of Rhizobium strains on their competitive ability for nodulation

The shoot dry weight of alfalfa inoculated with an effective strain of Rhizobium meliloti mixed with an ineffective strain in different ratios was found to be directly proportional to the log of the number of effective nodules. Consequently the comparison of the shoot dry weight of plants inoculated with a mixture of effective and ineffective strains with the shoot dry weight of plants inoculated with the effective strain should allow the estimation of the relative competitiveness of the effective strains. To check this. the competitiveness of 14 antibiotic-resistant strains of R. leguminosarum was evaluated in this way and compared with the ability of the strains to form nodules when inoculated to seeds of Vicia faba planted in a soil containing indigenous R. leguminosarum. The percentage of recovery of the inoculum strains in the nodules of field-grown fababeans was positively correlated with the competitiveness of the strains as estimated by the greenhouse test. This simple way of evaluating the nodulating competitiveness of strains of rhizobia being indicative of their competitive behaviour with indigenous rhizobia in the field could therefore be useful for screening a large number of strains for competitiveness.     

Effects of arbuscular mycorrhizal inoculation on growth,yield, nitrogen,and phosphorus nutrition of nodulating bean varieties in two soil substrates of contrasting fertility

A pot experiment was conducted to study the response of nodulated bean (Phaseolus vulgaris L.), vars. Ceca and Montalbán, to inoculation with the arbuscular‐mycorrhizae (AM) Glomus spurcum strain IES‐3 and G. mosseae strain IES‐8 in two non‐sterilized non‐amended soil substrates (S‐l and S‐2) of contrasting fertility. S‐l was an acid soil (pH 5.0) with 77% exchangeable aluminum (Al), low available phosphorus (P) (3.5 ug g^‐1 dry soil) and 1.8% organic matter. S‐2 was a soil with pH 7.0, 7.9 μg P g^‐1 dry soil, 9.6% organic matter and no exchangeable Al. Plants growing in S‐1 (S‐1 plants) and S‐2 (S‐2 plants) soils were sampled at the beginning of the flowering stage. S‐1 and S‐2 plants were nodulated by the rhizobial populations native of each soil. S‐1 plants of both varieties did not respond to AM inoculation in term of shoot and root mass, P content, relative abundance of ureides (RAU) and seed yields. Only the total chlorophyll, chlorophyll‐a, chlrophyll‐a/b ratio and nodule mass were significantly enhanced in AM‐inoculated plants. Increased total reducing sugars concentration was detected in roots of S‐1 plants inoculated with G. mosseae. S‐2 plants displayed significantly higher shoot and nodule mass as well as increased total chlorophyll, chlorophyll‐a, chlorophyll‐a/b ratio and P content than S‐1 plants, regardless of the variety and AM inoculation. S‐2 plants of the var Ceca showed a two fold increase in seed yields but similar RAU values (>60%) when compared to S‐1 plants of the same variety. In contrast, the significant increase in RAU detected in S‐2 plants of the var Montalban was not translated into higher seed yields. In S‐2, the productivity of plants of the var Ceca doubled that of the var Montalban. For both bean varieties the highest significant P content and seed yield were observed exclusively in S‐2 plants inoculated with G. mosseae. This Glomelean strain enhanced the sink‐source ratio of the S‐2 plants as evidenced by the higher total reducing sugar concentration in the root mass. Arbuscular‐mycorrhizae inoculation significantly decreased the acid phosphatase activity in the rhizosphere of S‐1 and S‐2 plants, respectively, pointing toward a negative effect of foreign AM on the native microbial biomass. The effectiveness of the rhizobial populations native of each soil type and the weak response elicited by G. mosseae in S‐2 plants do not justify, at present, the inclusion of foreign inocula in the bean crops carried out at S‐1 and S‐2 soils of the Sucre State of Venezuela. Results also indicated the higher adaptability of var Ceca to conditions prevailing in S‐1 and S‐2.     

COPPER EFFECTS ON PRUNUS PERSICA IN TWO DIFFERENT GRAFTING COMBINATIONS (P. PERSICA × P. AMYGDALUS AND P. CERASIFERA)

Growth, chlorophyll content, biomass dry weight (DW), nutrient uptake and copper (Cu) accumulation were evaluated in BigTop® peach (Prunus persica L. Batsch) grafted onto two different rootstocks [GF677 (P. persica × P. amygdalus) and Mr.S2/5 (P. cerasifera)] when grown in the presence of 0.1, 10 or 100 μM copper sulfate (CuSO₄). Although scion shoot length varied significantly in response to Cu concentration only with Mr.S2/5 rootstock (40%reduction at 100 μM Cu compared with control), Cu concentration in the scion grafted on GF677 was higher (18.2 mg kg^?1) compared to that grafted on Mr.S2/5 (10.5 mg kg^?1). In both graft combinations, the Cu concentrations in leaves were not statistically different although values ranged between 4.7 and 8.1 mg kg^?1. The DW of leaves and stems was not statistically different from control plants in both graft combinations. On the contrary, root DW of Mr.S2/5 increased, about 13% at 10 μM Cu and 43% at 100 μM Cu. In contrast, DW of GF677 roots at higher Cu treatment was significant lower than control. Chlorophyll reduction Cu induced was not found, suggesting that the degradation of these pigments is maintained at low levels. The uptake of sodium (Na), zinc (Zn), and iron (Fe) changed during Cu stress. The different behavior observed between GF677 and Mr.S2/5 regarding Cu accumulation proves that these two rootstocks have a different ability in translocation and accumulation of Cu in presence of this heavy metal. The use of GF677 rootstocks, in substrates particularly rich of Cu, insures a better development of scion growth when compared to Mr.S2/5, but determines a higher Cu concentration in the stem, while Mr.S2/5 seems to be able to stabilize the Cu concentrations in the scion.     

Improved growth and Cu tolerance of Cu excess-stressed white clover after inoculation with arbuscular mycorrhizal fungi

The effects of arbuscular mycorrhizal (AM) fungus, Glomus intraradices, on growth and copper (Cu) tolerance of white clover (Trifolium repens) were investigated in soils with different Cu amounts. The AM inoculation increased plant biomass and the total or bound Cu concentrations in shoots and roots but decreased the total Cu in soils and the exchangeable Cu in shoots, roots and soils at all Cu levels. Mycorrhizal plants had higher levels of root phosphorus and shoot zinc (Zn) at lower Cu levels and more nitrogen and Zn in roots and potassium, calcium and magnesium in shoots and roots at all Cu addition levels. Additionally, AM inoculation enhanced urease, acid phosphatase and catalase activities in rhizosphere soils and mycorrhizal roots showed higher levels of peroxidase, catalase, proline and soluble sugar at all Cu addition levels. These results indicate that mycorrhizal white clover is potentially suitable for Cu phytoremediation based on greenhouse studies.     

Inoculation with arbuscular mycorrhizal fungus Acaulospora mellea decreases Cu phytoextraction by maize from Cu-contaminated soil

A pot culture experiment was carried out to study the growth of and Cu uptake by maize (Zea mays) inoculated with or without arbuscular mycorrhizal (AM) fungus Acaulospora mellea in sterilized soil with different Cu amounts added (0, 100, 200, 400, 800 mg kg⁻¹). Root colonization rates were significantly lower with the addition of 400 and 800 mg kg⁻¹ Cu. AM inoculation increased shoot dry weights at 200 and 400 mg kg⁻¹ Cu added but showed no effects at other levels, while increased root dry weights at all Cu addition levels except 800 mg kg⁻¹. Compared with the nonmycorrhizal plants, shoot Cu concentrations in mycorrhizal plants were higher when no Cu was added but lower at other levels, while root Cu concentrations were lower at 400 and 800 mg kg⁻¹ Cu added but not affected at other levels. Thus, shoot Cu uptake in mycorrhizal plants increased with no Cu added but decreased at other levels, while mycorrhizal effects on root Cu uptake varied. Compared with nonmycorrhizal controls, Cu uptake efficiency and phytoextraction efficiency in mycorrhizal plants were higher when no Cu was added but lower at other levels, and Cu translocation efficiency was lower at all Cu addition levels. AM inoculation improved shoot and root P nutrition at all Cu addition levels. Soil pH was higher in mycorrhizal treatment than in the control when 200 mg kg⁻¹ or more Cu was added. These results indicate that A. mellea ZZ may be not suitable for Cu phytoextraction by maize, but shows a potential role in phytostabilization of soil moderately polluted by Cu.     

Impact of plants on the microbial activity in soils with high and low levels of copper

Elevated copper (Cu) concentrations have been shown to decrease the microbial activity in soils. Plants can have beneficial effects on the biological activity of soils mainly through their root exudates. In this study we investigated the impact of various plant species with different Cu tolerance levels on the microbial activity in two soils with low (10 mg/kg) and high (180 mg/kg) copper concentrations. The soil was a Kahangi Sandy Loam. Three different plants, Agrostis capillaris ‘Parys’ tolerant for Cu, Agrostis capillaris ‘Highland’ non-tolerant and Helianthus annuus tolerant and a hyper-accumulator for Cu were used. To increase the Cu availability to plants, EDTA was added to some of the pots 20 days after sowing. The effect of Cu contamination on the biological activity of soil in the presence and absence of plant growth was evaluated by measuring the dehydrogenase activity, the microbial biomass, the basal respiration, and the potential nitrification.Results show that plants increased the microbial activity in the low Cu soil. In the high Cu soil the microbial activity seemed to be related to the plant health. With the Cu-tolerant Agrostis capillaris ‘Parys’, the microbial activity increased faster than with the other plant species. Up to 50 days after sowing, the tolerant grass Agrostis capillaris ‘Parys’ had a higher plant biomass and was much healthier than the non-tolerant grass. Later on the growth of the non-tolerant Agrostis capillaris ‘Highland’ recovered, and the microbial activity of the soil reached close to those recorded for the soil treatments with the Cu-tolerant plant species. The addition of EDTA delayed the increase in microbial activity even further. The proportion of microbial biomass carbon in the organic fraction was higher in the low Cu soil than in the high Cu soil, with ratios ranging from 1.3 to 3.3 and from 0.5 to 1.7 respectively. The basal respiration rate in the original soil was significantly lower in the high Cu soil than in the low Cu soil, and was generally increased by the presence of plants.