Laboratory-and Field-Phenotyping for Drought Stress Tolerance and Diversity Study in Lentil (<i>Lens culinaris</i> Medik.)

Drought susceptibility and low genetic variability are the major constraints of lentil (Lens culinaris Medik.) production worldwide. Development of an efficient pre-field drought phenotyping technique and identification of diversified drought tolerant lentil genotype(s) are therefore vital and necessary. Two separate experiments were conducted using thirty diverse lentil genotypes to isolate drought tolerant genotype(s) as well as to assess their diversity. In both of the experiments, significant (p ≤ 0.01) variation in genotype (G), treatment (T) and G X T was observed for most of the studied traits. In experiment I, genotypes were examined for drought tolerance at the seedlings stage under hydroponic conditions by assessing root and shoot traits. Among the 30 genotypes studied, BM-1247, BM-1227 and BM-502 were selected as highly tolerant to drought stress as they showed maximum seedling survivability and minimum reduction in growth parameters under drought stress. In experiment II, the genotypes were assayed for diversity and drought stress tolerance based on morphological traits grown under field condition. Drought stress caused a substantial reduction in yield attributing traits, however, the genotypes BM-1247, BM-981, BM-1227 and BM-502 were categorized as drought tolerant genotypes with less than 20% yield reduction. The field screening result of drought stress tolerance was coincided well with the results of laboratory screening. Genetic divergence study reflected the presence of considerable diversity among the genotypes. Considering laboratory and field screening results, the genotypes, BM-1247, BM-1227, BM-981 and BM- 502 were selected as the best drought tolerant genotypes. This information can be exploited for further breeding in developing drought tolerance in lentil.     

Screening and Assessment of Selected Chilli (<i>Capsicum annuum</i> L.) Genotypes for Drought Tolerance at Seedling Stage

This study was undertaken to investigate oxidative stress tolerant mechanisms in chilli (Capsicum annuum L.) under drought genotypes through evaluating morphological, physiological, biochemical and stomatal parameters. Twenty genotypes were evaluated for their genetic potential to drought stress tolerant at seedling stage. Thirty days old seedlings were exposed to drought stress induced by stop watering for the following 10 days and rewatering for the following one week as recovery. Based on their survival performance, two tolerant genotypes viz. BD-10906 and BD-109012 and two susceptible genotypes viz. BD-10902 and RT-20 were selected for studying the oxidative stress tolerance mechanism. Drought reduced root and shoot length, dry weight, ratio, petiole weight and leaf area in both tolerant and susceptible genotypes, and a higher reduction was observed in susceptible genotypes. Lower reduction of leaf area and photosynthetic pigments were also found in tolerant genotypes. Moreover, tolerant genotypes showed higher recovery than susceptible genotypes after the removal of stress. A higher reduction of relative water content (RWC) may cause an imbalance between absorbed and transpirated water in susceptible genotypes. Higher accumulation of proline in tolerant genotypes might be helpful to for better osmotic maintenance than that in susceptible genotypes. Tolerant genotypes showed higher antioxidant activity as they showed DPPH radical scavenging percentage than the susceptible genotypes. Moreover, closer stomata in tolerant genotypes than susceptible ones helped to avoid dehydration in tolerant genotypes. Thus, the above morphological, physiological, biochemical and stomatal parameters helped to show better tolerance in chilli under drought stress.     

Integrated Fertilization Regimes Boost Heavy Metals Accumulation and Biomass of <i>Sedum alfredii</i> Hance

The hyperaccumulator Sedum alfredii Hance (S. alfredii) may be employed for zinc (Zn) and cadmium (Cd)-polluted soil remediation. However, the low phytoremediation efficiency, related to the low biomass production, limits its use with that purpose. In this experiment, nitrogen (N), phosphorus (P), and potassium (K) fertilizers, and organic manure were applied to investigate the phytoremediation ability of S. alfredii. Hydroponic and pot experiments were conducted using Zn-Cd polluted soil. The hydroponic experiment indicated that appropriate fertilizer application could increase (p < 0.05) the amount of accumulated Zn and Cd in S. alfredii. When N supply ranged from 0.5 to 2.5 mmol L⁻¹, it could improve growth and accumulation of Zn and Cd in whole plants of S. alfredii. The 1 mmol L^-1 N was an optimal N dosage for shoot biomass production and Cd accumulation in shoots, while the 2.5 mmol L^-1 was an optimal N dosage for Zn accumulation in shoots. Both low (<0.05 mmol L^-1) and high (>0.8 mmol L^-1) P supply decreased growth, and Zn/Cd accumulation in whole plants of the studied species. The 0.1 mmol L^-1 P was an optimal dosage for S. alfredii biomass production and Zn/Cd accumulation in shoots. The supply levels within the range from 0.3 to 1 mmol L^-1 K could significantly improve the biomass production of S. alfredii and its capability to accumulate Zn and Cd in the biomass. The 0.5 mmol L^-1 K was an optimal dosage for the whole biomass production and Zn accumulation in shoots, while the 1 mmol L^-1 was an optimal K dosage for Zn accumulation in shoots, which was 17.2% higher than the control. Moreover, the soil pot experiment showed that the combination of organic (fermented manure) and inorganic fertilizers made significant effects on the Zn and Cd-polluted soil remediation by S. alfredii. These effects varied, however, with the application of different proportions of N, P, K and organic matter. The Zn accumulation by S. alfredii reached the highest efficiency ability under the highest fertilizer mixing rate (N: 50 mg kg^-1, P: 40 mg kg^-1, K: 100 mg kg^-1, organic matter: 1%). Even more, S. alfredii showed the strongest ability to accumulate Cd with a lower fertilizer mixing rate (N: 25mg kg^-1, P: 20mg kg^-1, K: 50 mg kg^-1, organic matter: 0.5%).

     

Changes in Growth,Photosynthetic Pigments,Cell Viability,Lipid Peroxidation and Antioxidant Defense System in Two Varieties of Chickpea (<i>Cicer arietinum</i> L.) Subjected to Salinity Stress

Salinity is one of the most severe abiotic stresses for crop production. The present study investigates the salinity-induced modulation in growth indicators, morphology and movement of stomata, photosynthetic pigments, activity of carbonic anhydrase as well as nitrate reductase, and antioxidant systems in two varieties of chickpea (Pusa-BG5023, and Pusa-BGD72). On 20^th day of sowing, plants were treated with varying levels of NaCl (0, 50, 100, 150 and 200 mM) followed by sampling on 45 days of sowing. Recorded observations on both the varieties reveal that salt stress leads to a significant decline in growth, dry biomass, leaf area, photosynthetic pigments, protein content, stomatal behavior, cell viability, activity of nitrate reductase and carbonic anhydrase with the rise in the concentration of salt. However, quantitatively these changes were less in Pusa-BG5023 as compared to Pusa-BGD72. Furthermore, salinity-induced oxidative stress enhanced malondialdehyde content, superoxide radicals, foliar proline content, and the enzymatic activities of superoxide dismutase, catalase, and peroxidase. The variety Pusa-BGD72 was found more sensitive than Pusa-BG5023 to salt stress. Out of different graded concentrations (50, 100, 150 and 200 mM) of sodium chloride, 50 mM was least toxic, and 200 mM was most damaging. The differential behavior of these two varieties measured in terms of stomatal behavior, cell viability, photosynthetic pigments, and antioxidant defense system can be used as prospective indicators for selection of chickpea plants for salt tolerance and sensitivity.     

Differential cadmium stress tolerance in five indian mustard (Brassica juncea L.) cultivars: An evaluation of the role of antioxidant machinery

The presence of Cadmium (Cd) in the agricultural soils affects horticultural cultivars and constrains the crop productivity. A pot experiment was performed using five cultivars of mustard (Brassica juncea L.) to evaluate the difference in their response to Cd toxicity under greenhouse conditions. The pots containing reconstituted soil were supplied with different concentration of CdCl₂ (0, 25, 50, 100 or 150 mg Cd kg⁻¹ soil). Increasing concentration of Cd in the soil resulted in decreased growth, photosynthesis and yield. Maximum significant reduction in growth, photosynthesis and yield were observed with 150 mg Cd kg⁻¹ soil in all the cultivars. Our results indicate that the cultivar Alankar is found to be more tolerant to Cd stress, recording higher plant dry mass, net photosynthesis rate, associated with high antioxidant activity and low Cd content in the plant leaves and thus less oxidative damage. Cultivar RH30 experienced maximum damage in terms of reduction in growth, photosynthesis, yield characteristics and oxidative damage and emerged as sensitive cultivar. The data of tolerance index of Alankar were found to be higher among all tested mustard cultivars which indicate its higher tolerance to Cd. Better coordination of antioxidants protected Alankar from Cd toxicity, whereas lesser antioxidant activity in RH30 resulted in maximum damage. Cultivars of mustard were ranked with respect to their tolerance to Cd: Alankar > Varuna > Pusa Bold > Sakha > RH30, respectively.Key words: antioxidants, cadmium, growth, mustard cultivars, photosynthesis, stress, yield     

Alleviation of Cadmium Toxicity to <i>Medicago Truncatula</i> by AMF Involves the Changes of Cd Speciation in Rhizosphere Soil and Subcellular Distribution

Arbuscular mycorrhizal fungi (AMF) can improve plant tolerance to several abiotic stresses, including heavy metals, drought or salinity exposure. However, the role of AMF in alleviation of soil cadmium (Cd)-induced toxicity to plants is still largely unknown. In this study, Cd speciation in soil and subcellular distribution of Cd were used to characterize the roles of application AM fungi in the alleviation of Cd toxicity in alfalfa plants. Our results showed that the addition of Glomus mosseae in Cd contaminated soil (10 mg/Kg) significantly increased soil pH, cation exchange capacity (CEC) and organic matter in rhizosphere soil with Medicago truncatula L., and then account for significantly decreased contents of exchangeable and carbonate-bounded Cd speciation in rhizosphere soil, indicating alleviation of plant toxicity by reduction of bioavailable fractions of Cd. Although there is no significant difference found in Cd accumulation by roots and shoots respectively between Cd and AM-Cd treatments, more portion of Cd was recorded compartmentalization in cell wall fraction of both root and shoot in treatment of Cd with AM application, indicating alleviation of Cd toxicity to plant cell. Herein, application of AM fungi in Cd treatments performed to inhibit the appearance of Cd toxicity symptoms, including the improvement of leaf electrolyte leakage, root elongation, seedling growth and biomass. This information provides a clearer understanding of detoxification strategy of AM fungi on Cd behavior with development and stabilization of soil structure and subcellular distribution of plant.     

Cadmium Removal Potential from Contaminated Soil by Tagetes erecta L. and Panicum maximum: Influence of Soil pH

The potential for cadmium (Cd) removal from contaminated soil by two species—marigold (Tagetes erecta L.) and Guinea grass (Panicum maximum)—was investigated in pot culture experiments in a greenhouse in triplicate. The concentration of Cd was varied from 50 to 200 mg kg^?1 and the pH was varied from 5.0 to 7.5 to investigate the effect of pH on Cd uptake. The results showed that total biomass of Guinea grass was around nine and seven times higher than that of marigold for Cd treatments of 50 and 100 mg kg^?1 at pH 5.0, respectively. Total cadmium uptake at Cd treatments of 50 and 100 mg kg^?1 at pH 5.0 by Guinea grass was 19.28 ± 3.14 and 36.06 ± 4.28 mg kg^?1, respectively, and for marigold was 15.66 ± 4.17 and 20.38 ± 3.24 mg kg^?1, respectively. The total Cd uptake by Guinea grass was 1.23 and 1.77 higher than that of marigold at Cd treatments of 50 and 100 mg kg^?1, respectively, at pH 5.0 due to higher biomass. The maximum Cd uptake by marigold and Guinea grass occurred at pH 5.0 at Cd treatment of 100 mg kg^?1. The results clearly show that the two species behave very differently for Cd uptake. Guinea grass is easy to grow, drought tolerant and, due to its higher biomass, it can be used for remediation of Cd-contaminated soil.     

Sulphur protects mustard (<Emphasis Type="Italic">Brassica campestris</Emphasis> L.) from cadmium toxicity by improving leaf ascorbate and glutathione

In a pot-soil culture ameliorative effect of sulphur (S) (0 or 40 mg S kg⁻¹ soil) on cadmium (Cd) (0, 25, 50 and 100 mg Cd kg⁻¹ soil)-induced growth inhibition and oxidative stress in mustard (Brassica campestris L.) cultivar Pusa Gold was studied. Cadmium at 100 mg kg⁻¹ soil caused maximum increase in the contents of Cd and thiobarbituric acid reactive substances (TBARS) in leaves. Maximum reductions in growth (plant dry mass, leaf area), chlorophyll content, net photosynthetic rate (P_N) and the contents of ascorbate (AsA), glutathione (GSH) were observed with 100 mg Cd kg⁻¹ soil compared to control. The application of S helped in reducing Cd toxicity, which was greater for 25 and 50 mg Cd kg⁻¹ soil) compared to 100 mg Cd kg⁻¹ soil. Addition of S to Cd-treated plants showed decrease in Cd and TBARS content in leaves and restoration of growth and photosynthesis through increase in the contents of AsA and GSH. Net photosynthetic rate and plant dry mass were strongly and positively correlated with the contents of AsA and GSH. It is suggested that S may ameliorate Cd toxicity and protects growth and photosynthesis of mustard involving AsA and GSH.     

Genotype-Dependent Effect of Exogenous Nitric Oxide on Cd-induced Changes in Antioxidative Metabolism, Ultrastructure, and Photosynthetic Performance in Barley Seedlings (Hordeum vulgare)

A greenhouse hydroponic experiment was performed using Cd-sensitive (cv. Dong 17) and Cd-tolerant (Weisuobuzhi) barley seedlings to evaluate how different genotypes responded to cadmium (Cd) toxicity in the presence of sodium nitroprusside (SNP), a nitric oxide (NO) donor. Results showed that 5 μM Cd increased the accumulation of O₂^•−, H₂O₂, and malondialdehyde (MDA) but reduced plant height, chlorophyll content, net photosynthetic rate (P _n), and biomass, with a much more severe response in the Cd-sensitive genotype. Antioxidant enzyme activities increased significantly under Cd stress in the roots of the tolerant genotype, whereas in leaves of the sensitive genotype, superoxide dismutase (SOD) and ascorbate peroxide (APX), especially cytosol ascorbate peroxidase (cAPX), decreased after 5–15 days Cd exposure. Moreover, Cd induces NO synthesis by stimulating nitrate reductase and nitric oxide synthetase-like enzymes in roots/leaves. A Cd-induced NO transient increase in roots of the Cd-tolerant genotype might partly contribute to its Cd tolerance. Exogenous NO dramatically alleviated Cd toxicity, markedly diminished Cd-induced reactive oxygen species (ROS) and MDA accumulation, ameliorated Cd-induced damage to leaf/root ultrastructure, and increased chlorophyll content and P _n. External NO counteracted the pattern of alterations in certain antioxidant enzymes induced by Cd; for example, it significantly elevated the depressed SOD, APX, and catalase (CAT) activities in the Cd-sensitive genotype after 10- and 15-day treatments. Furthermore, NO significantly increased stromal APX and Mn-SOD activities in both genotypes and upregulated Cd-induced decrease in cAPX activity and gene expression of root/leaf cAPX and leaf CAT1 in the Cd-sensitive genotype. These data suggest that under Cd stress, NO, as a potent antioxidant, protects barley seedlings against oxidative damage by directly and indirectly scavenging ROS and helps to maintain stability and integrity of the subcellular structure.     

Role of Organic Amendments to Mitigate Cd Toxicity and Its Assimilation in <i>Triticum aestivum</i> L.

In soil biota, higher and enduring concentration of heavy metals like cadmium (Cd) is hazardous and associated with great loss in growth, yield, and quality parameters of most of the crop plants. Recently, in-situ applications of eco-friendly stabilizing agents in the form of organic modifications have been utilized to mitigate the adverse effects of Cd-toxicity. This controlled experiment was laid down to appraise the imprints of various applied organic amendments namely poultry manure (PM), farmyard manure (FYM), and sugarcane press mud (PS) to immobilize Cd in polluted soil. Moreover, phytoavailability of Cd in wheat was also accessed under an alkaline environment. Results revealed that the addition of FYM (5–10 ton ha^-1 ) in Cd-contaminated soil significantly increased germination rate, leaf chlorophyll content, plant height, spike length, biological and grain yield amongst all applied organic amendments. Moreover, the addition of FYM (5–10 ton ha^-1 ) also reduced the phytoavailability of Cd by 73–85% in the roots, 57–83% in the shoots, and 81–90% in grains of wheat crop. Thus, it is affirmed that incorporation of FYM (5–10 ton ha^-1 ) performed better to enhance wheat growth and yield by remediating Cd. Thus, the application of FYM (5–10 ton ha^-1 ) reduced the toxicity induced by Cd to plants by declining its uptake and translocation as compared to all other applied organic amendments to immobilize Cd under sandy alkaline polluted soil.     

Application of sulfur fertilizer reduces cadmium accumulation and toxicity in tobacco seedlings (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis>)

Greenhouse pot experiments were carried out to illuminate the mitigation effects of sulfur (S) on cadmium (Cd) uptake and toxicity in tobacco using two levels of exogenous sulfur of S1 and S2 containing 47 and 38% total S. Results showed that Cd1 and Cd2 treatments of 1 and 5 mg Cd kg^?1 soil increased leaf Cd concentration and accumulation but reduced plant height, net photosynthetic rate (Pn) and biomass, with a much severe response in Cd2 treatment. Application of S2 fertilizer alleviates Cd toxicity, markedly decreased Cd concentration and improved photosynthesis, compared with Cd1 and Cd2 alone treatment; but S1 only alleviated Cd toxicity when tobacco was subjected to Cd1 stress. Our results suggest that S2 fertilizer was more effective in reducing Cd accumulation and toxicity in Tobacco than S1, and highlights a promising approach of S fertilizer application to lower leaf Cd accumulation in order to ensure product safety of tobacco grown in Cd polluted soils.     

<Emphasis Type="Italic">Cannabis sativa</Emphasis> L. growing on heavy metal contaminated soil: growth,cadmium uptake and photosynthesis

The effects of different cadmium concentrations [17 mg(Cd) kg⁻¹(soil) and 72 mg(Cd) kg^{− 1}(soil)] on Cannabis sativa L. growth and photosynthesis were examined. Hemp roots showed a high tolerance to Cd, i.e. more than 800 mg(Cd) kg⁻¹(d.m.) in roots had no major effect on hemp growth, whereas in leaves and stems concentrations of 50 – 100 mg(Cd) kg⁻¹(d.m.) had a strong effect on plant viability and vitality. For control of heavy metal uptake and xylem loading in hemp roots, the soil pH plays a central role. Photosynthetic performance and regulation of light energy consumption were analysed using chlorophyll fluorescence analysis. Seasonal changes in photosynthetic performance were visible in control plants and plants growing on soil with 17 mg(Cd) kg⁻¹(soil). Energy distribution in photosystem 2 is regulated in low and high energy phases that allow optimal use of light and protect photosystem 2 from overexcitation, respectively. Photosynthesis and energy dissipation were negatively influenced by 72 mg(Cd) kg⁻¹(soil). Cd had detrimental effects on chlorophyll synthesis, water splitting apparatus, reaction centre, antenna and energy distribution of PS 2. Under moderate cadmium concentrations, i.e. 17 mg(Cd) kg⁻¹(soil), hemp could preserve growth as well as the photosynthesis apparatus, and long-term acclimation to chronically Cd stress occurred.     

Aluminium localization and toxicity symptoms related to root growth inhibition in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) seedlings

We correlated root growth inhibition with aluminium (Al³⁺) localization and toxicity symptoms in rice roots using seedlings of two genotypes (tolerant and sensitive) that were exposed to different AlCl₃ concentrations. Al³⁺ localization was evaluated by hematoxylin in primary roots and by morin in cross-sections of the root tips. Neutral invertase enzyme activity and callose (1→3, β-d-glucan) accumulation were observed and compared with Al³⁺ accumulation sites. Root growth was inhibited by Al³⁺ in a concentration-specific manner and proportional to the increase of hematoxylin staining, being more pronounced in the sensitive genotype. Morin staining showed the presence of Al³⁺ deep within the roots of the sensitive genotype, indicating that the metal was able to penetrate beyond the first few cell layers. In the tolerant genotype, Al³⁺ penetration was restricted to the first two cell layers. Ruptures in exodermis and epidermis layers by lateral root protrusions in both genotypes allowed Al³⁺ to enter into the roots. More intense activity of invertase in roots of the tolerant genotype was also observed, which could be related to greater root growth of this cultivar when submitted to Al³⁺ stress. Moreover, Al³⁺-induced callose accumulation was a late response occurring in the same areas where Al³⁺ was present.     

Physiological and Biochemical Mechanisms Preventing Cd Toxicity in the New Hyperaccumulator <Emphasis Type="Italic">Abelmoschus manihot</Emphasis>

Abelmoschus manihot, an ornamental plant, was examined for phytoremediation purposes in accordance with the ability to accumulate cadmium and physiological mechanisms of cadmium tolerance. A net photosynthetic rate (A _N) glasshouse experiment for 60 days was conducted to investigate the influence of different cadmium amounts (0–100 mg kg^?1) on the growth, biomass, photosynthetic performance, reactive oxygen species (ROS) production, antioxidative enzyme activities, Cd uptake and accumulation of A. manihot. Exposure to cadmium enhanced plant growth even at 100 mg kg^?1, without showing symptoms of visible damage. The cadmium concentration of shoots (stems or leaves) and roots was more than the critical value of 100 mg kg^?1 and reached 126.17, 185.26 and 210.24 mg kg^?1, respectively. BCF values of A. manihot plants exceeded the reference value 1.0 for all the Cd treatments, and TF values were greater than 1 at 15–60 mg kg^?1 Cd treatment. The results also showed that cadmium concentrations of 60 mg kg^?1 or less induced a significant enhancement in plant net photosynthetic rate (A _N), stomatal conductance (G _s), transpiration rate (T _r), photosynthetic pigments and F _v/F _m. These parameters were slightly decreased at the higher concentration (100 mg kg^?1). The ROS production (O₂ ^?, H₂O₂) and antioxidative response including SOD, CAT and POD were significantly enhanced by increasing cadmium. These results suggest that A. manihot can be considered as a Cd-hyperaccumulator and the hormetic effects may be taken into consideration in remediation of Cd contamination soil.     

Salicylic acid-mediated alleviation of cadmium toxicity in hemp plants in relation to cadmium uptake,photosynthesis, and antioxidant enzymes

To assess the role of salicylic acid (SA) in alleviating cadmium (Cd) toxicity in hemp (Cannabis sativa L.) plants, the growth parameters, Cd accumulation, photosynthetic performance and activities of major antioxidant enzymes were investigated in hemp seedlings treated with 500 μM SA, under 0, 25, 50, and 100 mg Cd kg⁻¹ sands (DW) conditions, respectively. Cd exposure resulted in a small reduction in biomass (12.0–26.9% for root, and 8.7–29.4% for shoot, respectively), indicating hemp plants have innate capacity to tolerant Cd stress. This was illustrated by little inhibition in photosynthetic performance, unchanged malondialdehyde content, and enhancement of superoxide dismutase (SOD) and peroxidases (POD) activities in hemp plants. Cd content in root is 25.0–29.5 times’ higher than that in shoot, suggesting the plant can be classified as a Cd excluder. It is concluded that SA pretreatment counteracted the Cd-induced inhibition in plant growth. The beneficial effects of SA in alleviating Cd toxicity can be attributed to the SA-induced reduction of Cd uptake, improvement of photosynthetic capacity, and enhancement of SOD and POD activities.     

Differential Effects of Ammonium and Nitrate on Growth Performance of <i>Glechoma longituba</i> under Heterogeneous Cd Stress

Water, minerals, nutrients, etc., can be shared by physiological integration among inter-connected ramets of clonal plants. Nitrogen plays an important role in alleviating cadmium (Cd) stress for clonal plants. But how different forms of nitrogen affect growth performance of clonal plants subjected to heterogeneous Cd stress still remains poorly understood. A pot experiment was conducted to investigate the differential effects of ammonium and nitrate on growth performance of Glechoma longituba under heterogeneous Cd stress. In the experiment, parent ramets of Glechoma longituba clonal fragments were respectively supplied with modified Hoagland solution containing 7.5 mM ammonium, 7.5 mM nitrate or the same volume of nutrient solution without nitrogen. Cd solution with different concentrations (0, 0.1 or 2.0 mM) was applied to offspring ramets of the clonal fragments. Compared with control (N-free), nitrogen addition to parent ramets, especially ammonium, significantly improved antioxidant capacity [glutathione (GSH), proline (Pro), peroxidase (POD,) superoxide dismutase (SOD) and catalase (CAT)], PSII activity [maximum quantum yield of PSII (F_v/F_m) and effective quantum yield of PSII (ΦPSII)], chlorophyll content and biomass accumulation of the offspring ramets suffering from Cd stress. In addition, negative effects of nitrate on growth performance of whole clonal fragments were observed under Cd stress with high concentration (2.0 mM). Transportation or sharing of nitrogen, especially ammonium, can improve growth performance of clonal plants under heterogeneous Cd stress. The experiment provides insight into transmission mechanism of nitrogen among ramets of clonal plants suffering from heterogeneous nutrient supply. Physiological integration might be an important ecological strategy for clonal plants adapting to heterogeneous environment stress conditions.     

Selection <Emphasis Type="Italic">in vitro</Emphasis> and accumulation of phytochelatins in cadmium tolerant cell line of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

Cucumber (Cucumis sativus L.) cells from suspension culture were selected for their ability to grow and divide rapidly in toxic concentration of cadmium. As a result of selection a cell suspension tolerant to 100 M cadmium chloride (CdCl₂) was initiated. The selected tolerant line exhibited stable and repeatable increase in fresh and dry weight of cells in the presence of cadmium. The accumulated level of phytochelatins in cadmium sensitive (unselected) and tolerant cell line was measured by high performance liquid chromatography (HPLC) after 3, 24 h and 5 days of cadmium treatment. It was shown that in both cell lines Cd induced accumulation of phytochelatins and simultaneous glutathione depletion occurred. No distinct changes were found after 3 and 24 h of cadmium treatment whereas after 5 days of exposure to the metal, the level of phytochelatins was two times higher in the sensitive cell line as compared to the tolerant one. The accumulation of phytochelatins was correlated with cadmium concentration that increased in both cell lines during the course of cell exposure to metal. However, the level of cadmium was always lower in the tolerant cell line. The results showed no direct correlation between the tolerance of cucumber cells to Cd and the accumulated level of phytochelatins. Other mechanisms responsible for the increased tolerance of cucumber cells exposed to Cd are discussed.     

Effect of cadmium on the growth and antioxidant enzymes in two varieties of Brassica juncea

Increasing contamination and higher enrichment ratio of non-essential heavy metal cadmium (Cd) induce various toxic responses in plants when accumulated above the threshold level. These effects and growth responses are genotype and Cd level dependent. An experiment was conducted to analyze the effect of Cd toxicity in Brassica juncea [L] Czern and Coss by selecting its two varieties Varuna and RH-30. Cadmium (0, 25, 50 or 100 mg CdCl₂ kg⁻¹ of soil) fed to soil decreased the values of growth characteristics, activity of nitrate reductase and leaf water potential, whereas activities of antioxidant enzymes and proline content increased with the increasing concentration of Cd, observed at 30 and 60 day stages of growth, in both the varieties. Moreover, Cd uptake by the roots was higher in RH-30 than Varuna. Also the activity of antioxidant enzymes and proline accumulation were higher in Varuna with increasing soil level of Cd. Out of the two varieties, Varuna was more tolerant than RH-30 to Cd stress.     

Cadmium Tolerance and Accumulation of Elsholtzia argyi Origining from a Zinc/Lead Mining Site - A Hydroponics Experiment

In this study, a hydroponics experiment was conducted to investigate the characteristics of Cd tolerance and accumulation of Elsholtzia argyi natively growing on the soil with high levels of heavy metals in a Zn/Pb mining site. Seedlings of E. argyi grown for 4 weeks and then were treated with 0(CK), 5, 10, 15, 20, 25, 30, 40, 50, 100 μM Cd for 21days. Each treatment had three replications. No visual toxic symptoms on shoots of E. argyi were observed at Cd level ≤50 μM. The results indicated that the dry biomass of each tissue and the whole plants of the treatments with ≤40 μM cadmium were similar to that of the control, implying that E. argyi was a cadmium tolerant plant. The results also showed that the shoot Cd concentration significantly (P < 0.05) increased with the increase in the Cd level in nutrient solution. The shoot Cd concentration of the treatment with 40 μM Cd was as high as 237.9 mg kg^–1, which was higher than 100 mg kg^–1, normally used as the threshold concentration for identifying the Cd hyperaccumulating plant. It could be concluded that E. argyi was a Cd tolerant and accumulating plant species.