Lettuce Root Elongation Research Articles

Superfície de resposta para quantificação dos efeitos de NaCl e pH em alface

Abstract The objective of this work was to evaluate the response surface methodology (RSM), with a large sample size, to investigate the effects of NaCl and pH on the root elongation of lettuce (Lactuca sativa) in vitro. For this, 20 lettuce seeds were placed in sterile Petri dishes with two filter papers at the bottom and 3.0 mL of the sample being tested. A rotatable 2x2 factorial arrangement was used, with two factors and a two-level matrix, incorporating two center points and ±1.41 α axial points. Each treatment group consisted of 40 replicates, resulting in 400 experimental observations. Increasing NaCl concentration from 0.0065 to 2,700 g L-1 caused a 19.3% decrease in lettuce root elongation. When pH decreased from 7.0 to 1.5, there was a 68.8% decrease in root elongation, but, when it increased from 7.0 to 12.5, there was a 44.9% decrease. The analysis of combined effects showed that the increase in NaCl concentration reduced the ability of the plant to resist changes in pH. The RSM was effective in evaluating the statistical significance of the NaCl and pH factors, including their second-order effects and interactions, as well as in quantifying their effects on the root elongation of lettuce under the experimental conditions. Therefore, the RSM is a valuable tool for interpreting the results and evaluating the sensitivity of multi-factor research, even when dealing with a large sample size.

The effect of metal oxide nanoparticle overexposure on lettuce root elongation and chlorophyll production

The effect of metal oxide nanoparticle overexposure on lettuce root elongation and chlorophyll production

Identification of a new gibberellin receptor agonist, diphegaractin, by a cell-free chemical screening system

Gibberellin (GA) is a phytohormone that regulates various developmental processes during the plant life cycle. In this study, we identify a new GA agonist, diphegaractin, using a wheat cell-free based drug screening system with grape GA receptor. A GA-dependent interaction assay system using GA receptors and DELLA proteins from Vitis vinifera was constructed using AlphaScreen technology and cell-free produced proteins. From the chemical compound library, diphegaractin was found to enhance the interactions between GA receptors and DELLA proteins from grape in vitro. In grapes, we found that diphegaractin induces elongation of the bunch and increases the sugar concentration of grape berries. Furthermore, diphegaractin shows GA-like activity, including promotion of root elongation in lettuce and Arabidopsis, as well as reducing peel pigmentation and suppressing peel puffing in citrus fruit. To the best of our knowledge, this study is the first to successfully identify a GA receptor agonist showing GA-like activity in agricultural plants using an in vitro molecular-targeted drug screening system.

Ecotoxicological risk assessment in soils contaminated by Pb and As 20 years after a mining spill

This study evaluates the potential toxicity of the soils of the Guadiamar Green Corridor (GGC) affected by the Aznalcóllar mine spill (Andalusia, Spain), one of the most important mining accidents in Europe in recent decades. Twenty years after the accident, although the area is considered to be recovered, residual contamination in soils persists, and the bioavailability of some contaminants, such as As, is showing trends of increasing. Therefore, the potential residual toxicity in 84 soil samples was evaluated by bioassays with lettuce (Latuca sativa L.), earthworms (Eisenia andrei) and determining the microbial activity by basal respiration and metabolic quotient. The selected soils sampled along the GGC were divided into 4 types according to their physicochemical properties. In the closest part of the mine two soil types appear (SS1 and SS2), originally decarbonated and loamy, with a reduction in lettuce root elongation of 57% and 34% compared to the control, as well as a the highest metabolic quotient (23.9 and 18.1 ng CcO2 μg Cmicrob−1 h−1, respectively) with the highest risk of Pb and As toxicity. While, located in the middle and final part of the affected area of the spill (SS3 and SS4), soils presented alkaline pH, finer textures and the lowest metabolic quotient (<9.5 ng CcO2 μg Cmicrob−1 h−1). In addition, due to Pb and As exceeded the Guideline values established in the studied area, the human toxicity risk was determined according to US-EPA methodology. Although the total contents were higher than the Guidelines established, the obtained hazard quotients for both contaminants were less than one, so the risk for human health was discarded. However, monitoring over time of the toxicity risks of the GGC soils would be advisable, especially due to the existence of areas where residual contamination persist, and soil hazard quotient obtained for As in children was higher and close to unity.

Antioxidant, antimicrobial and phytotoxic activities of Rhaponticum acaule DC. essential oil

The in-vitro antioxidant activity of Rhaponticum acaule essential oil (RaEO) was evaluated using β-carotene/linoleic acid bleaching, chelating activity, and lipid peroxidation inhibition (TBARS) assays. The antimicrobial activity of RaEO was assessed by disc diffusion and microdilution methods against 8 bacteria and 4 yeast. Finally, the allelopathic activity of RaEO on the seed germination and the shoot and root elongation of lettuce (Lactuca sativa L.) seedlings were investigated. According to our results, the RaEO exhibited significant antioxidant activity, similar to those of standards (BHT and ascorbic acid) with IC50 values of 0.042 and 0.045 mg/mL obtained by β-carotene bleaching and TBARS assays, respectively. On the other hand, despite its interesting ferrous chelating activity, RaEO possesses moderate IC50 value (0.35 mg/mL) as compared with that of EDTA (0.015 mg/mL). RaEO exhibited a strong antimicrobial activity against all the tested microorganisms, with IZ, MIC and MBC values being in the range of 7.67 ± 0.58 to 13.33 ± 0.58 mm, 1.25 to 5.00 and 5.00 to 10.00 mg/mL, respectively. The results reveled also that RaEO inhibited the shoot and root growth of Lactuca sativa L. seedlings. Our data suggested that the RaEO had pharmaceutical benefits and could be used as a potential natural herbicide resource.

Sandwich method applied to the screening of allelopathic action in Byrsonima spp. (Malpighiaceae)

In order to understand the allelochemical action of Byrsonima intermedia A. Juss. and B. verbascifolia Rich. ex A. Juss., the effects on germination, initial growth and cell cycle of Lactuca sativa L., exposed to leaves and fruits of this species, were evaluated using the sandwich method. Data were subjected to analysis of variance and means were compared by the Scott-Knott test (p < 0.05). For the parameters germination percentage and germination rate index, no difference between Byrsonima spp. was observed, and B. intermedia fruits were responsible for the highest inhibition of lettuce germination, compared to the leaves. Root elongation in lettuce exposed to B. verbascifolia organs showed stimulatory effects and B. intermedia organs showed inhibitory effects on root growth. The mitotic index showed the allelopathic effect of Byrsonima spp. by the reduction in cell division in lettuce root meristem, and the stimulus in root elongation was caused by B. verbascifolia, understood as a consequence of cell expansion, not mitotic activity. An aneugenic effect of Byrsonima spp. was observed, and the highest frequency of C-metaphase and stickness was found in B. verbascifolia fruits and B. intermedia leaves.

Phytotoxicity of veterinary antibiotics to seed germination and root elongation of crops

Large quantities of veterinary antibiotics (VAs) are being used worldwide in agricultural fields through wastewater irrigation and manure application. They cause damages to the ecosystem when discharged into the environment, but there is a lack of information on their toxicity to plants and animals. This study evaluated the phytotoxic effects of five major VAs, namely tetracycline (TC), sulfamethazine (SMZ), norfloxacin (NOR), erythromycin (ERY) and chloramphenicol (CAP), on seed germination and root elongation in lettuce, tomato, carrot and cucumber, and investigated the relationship between their physicochemical properties and phytotoxicities. Results show that these compounds significantly inhibited root elongation (p<0.05), the most sensitive endpoint for the phytotoxicity test. TC was associated with the highest level of toxicity, followed by NOR, ERY, SMZ and CAP. Regarding crop species, lettuce was found to be sensitive to most of the VAs. The median effect concentration (EC50) of TC, SMZ, NOR, ERY and CAP to lettuce was 14.4, 157, 49.4, 68.8 and 204mg/L, respectively. A quantitative structure-activity relationship (QSAR) model has been established based on the measured data. It is evident that hydrophobicity was the most important factor governing the phytotoxicity of these compounds to seeds, which could be explained by the polar narcosis mechanism. Lettuce is considered a good biomarker for VAs in the environment. According to the derived equation, phytotoxicities of selected VA compounds on different crops can be calculated, which could be applicable to other VAs. Environmental risks of VAs were summarized based on the phytotoxicity results and other persistent factors.

The sensitivity of an hydroponic lettuce root elongation bioassay to metals, phenol and wastewaters

The root elongation bioassay is one of the most straightforward test methods used for environmental monitoring in terms of simplicity, rapidity and economy since it merely requires filter paper, distilled water and Petri dishes. However, filter paper as a support material is known to be problematic as it can reduce the sensitivity of the test.The newly developed hydroponic method reported here differs from the conventional root elongation method (US EPA filter paper method) in that no support material is used and the exposure time is shorter (48h in this test versus 120h in the US EPA test). For metals, the hydroponic test method was 3.3 (for Hg) to 57 (for Cu) times more sensitive than the US EPA method with the rank orders of sensitivity, estimated from EC50 values, being Cu≥Cd>Ni≥Zn≥Hg for the former and Hg≥Cu≥Ni≥Cd≥Zn for the latter methods. For phenol, the results did not differ significantly; EC50 values were 124mgL−1 and 108–180mgL−1 for the hydroponic and filter paper methods, respectively. Lettuce was less sensitive than daphnids to wastewaters, but the root elongation response appears to be wastewater-specific and is especially sensitive for detecting the presence of fluorine.The new hydroponic test thus provides many practical advantages, especially in terms of cost and time-effectiveness requiring only a well plate, a small volume of distilled water and short exposure period; furthermore, no specialist expertise is required. The method is simpler than the conventional EPA technique in not using filter paper which can influence the sensitivity of the test. Additionally, plant seeds have a long shelf-life and require little or no maintenance.

Long-term toxicity assessment of soils in a recovered area affected by a mining spill

Residual pollution in the Guadiamar Green Corridor still remains after Aználcollar mine spill in 1998. The polluted areas are identified by the absence of vegetation, soil acidic pH and high concentrations of As, Pb, Zn and Cu. Soil toxicity was assessed by lettuce root elongation and induced soil respiration bioassays. In bare soils, total As and Pb concentrations and water-extractable levels for As, Zn and Cu exceeded the toxicity guidelines. Pollutants responsible for toxicity were different depending on the tested organism, with arsenic being most toxic for lettuce and the metal mixture to soil respiration. Soil properties, such as pH or organic carbon content, are key factors to control metal availability and toxicity in the area. According to our results, there is a risk of pollution to living organisms and the soil quality criteria established in the area should be revised to reduce the risk of toxicity.

Phytotoxicity and genotoxicity assessment of imazethapyr herbicide using a battery of bioassays.

The imazethapyr herbicide (formulation Verosil(®)) was evaluated for phytotoxicity and genotoxicity using a battery of bioassays: (1) the growth inhibition of the green alga Pseudokirchneriella subcapitata, (2) the root growth and germination of the higher plant Lactuca sativa, (3) the genetic damage using the Salmonella/microsome test, and (4) the aneugenic and clastogenic effects on Allium cepa. The Verosil(®) formulation was highly toxic to the non-target green alga (median effective concentration (EC50) = 1.05 ± 0.05 mg active ingredient (a.i.) L(-1)), and concentrations above 10 mg a.i. L(-1) inhibited root elongation in lettuce: relative growth index (RGI) between 0.28 ± 0.01 and 0.66 ± 0.10. No genotoxic effect was observed in S almonella typhimurium at 100 mg a.i. L(-1), either with or without the microsomal fraction. However, significant differences in the frequency of chromosomal aberrations in anaphases and telophases (bridges, chromosome fragments, and vagrants) were observed in A. cepa at concentrations between 0.01 and 1 mg a.i. L(-1) with respect to the control. The frequencies of micronuclei showed significant differences with respect to the control at concentrations between 0.001 and 0.1 mg a.i. L(-1). A very high mitotic index (MI = 93.8 ± 5.8) was observed associated with a high number of cells in the prophase stage at 100 mg a.i. L(-1), indicating cytotoxicity. These results showed that imazethapyr is toxic to the non-target populations in both aquatic and terrestrial ecosystems. This herbicide might also exert clastogenic and aneugenic mitotic damage in higher plants. Therefore, the imazethapyr formulation may constitute an environmental risk to plants.

Statistically significant deviations from additivity: What do they mean in assessing toxicity of mixtures?

There is increasing attention from scientists and policy makers to the joint effects of multiple metals on organisms when present in a mixture. Using root elongation of lettuce (Lactuca sativa L.) as a toxicity endpoint, the combined effects of binary mixtures of Cu, Cd, and Ni were studied. The statistical MixTox model was used to search deviations from the reference models i.e. concentration addition (CA) and independent action (IA). The deviations were subsequently interpreted as ‘interactions’. A comprehensive experiment was designed to test the reproducibility of the ‘interactions’. The results showed that the toxicity of binary metal mixtures was equally well predicted by both reference models. We found statistically significant ‘interactions’ in four of the five total datasets. However, the patterns of ‘interactions’ were found to be inconsistent or even contradictory across the different independent experiments. It is recommended that a statistically significant ‘interaction’, must be treated with care and is not necessarily biologically relevant. Searching a statistically significant interaction can be the starting point for further measurements and modeling to advance the understanding of underlying mechanisms and non-additive interactions occurring inside the organisms.

Evaluation of phytotoxicity of municipal landfill leachate before and after biological treatment.

In the present study, leachate toxicity of a municipal solid waste landfill located in the Sinos River Valley region (southern Brazil) was evaluated using plant bioassays. Leachate toxicity was assessed by analysis of seed germination and root elongation of lettuce (Lactuca sativa L.) and rocket plant Eruca sativa Mill.) and root elongation of onions Allium cepa L.). Bioassays were performed by exposing the seeds of L. sativa and E. sativa and the roots of A. cepa to raw leachate, treated leachate (biological treatment) and negative control (tap water). The levels of metals detected in both samples of leachate were low, and raw leachate showed high values for ammoniacal nitrogen and total Kjeldahl nitrogen. There is a reduction in the values of several physicochemical parameters, which demonstrates the efficiency of the treatment. Both L. sativa and A. cepa showed a phytotoxic response to landfill leachate, showing reduced root elongation. However, the responses of these two plant species were different. Root elongation was significantly lower in A. cepa exposed to treated leachate, when compared to negative control, but did not show any difference when compared to raw leachate. In L. sativa, seeds exposed to the raw leachate showed significant reduction in root elongation, when compared to treated leachate and negative control. Seed germination showed no difference across the treatments. The results of the study show that plant species respond differently and that municipal solid waste landfill leachate show phytotoxicity, even after biological treatment.

Chemical Stabilization of Metal-Contaminated Mine Soil: Early Short-Term Soil-Amendment Interactions and Their Effects on Biological and Chemical Parameters

Chemical stabilization is a cost-effective, environmentally friendly, in situ remediation technology based on the application of organic and/or inorganic amendments to reduce soil metal bioavailability. Our objective was to assess the early short-term effects of organic amendments (sheep manure—SHEEP, poultry litter—POULTRY, cow slurry—COW, paper mill sludge mixed with poultry litter—PAPER), in sterilized and non-sterilized form, on the microbial and chemical properties, as well as on the phytotoxicity, of a Cd, Pb and Zn contaminated soil. Our results provide useful information regarding (1) the effectiveness of amendments for chemical stabilization of mine soil and (2) the impact of microbial populations present in the amendments on soil native microbial communities. Microbial populations present in the amendments did not substantially modify soil microbial functional diversity, as reflected by Biolog EcoPlates™ data, except for PAPER-amended soils. We observed a good correlation between lettuce root elongation (phytotoxicity bioassay) and Cd, Pb, and Zn CaCl2-extractable concentrations in soil. SHEEP and PAPER amendments were particularly effective at increasing soil pH and reducing metal bioavailability and phytotoxicity, while POULTRY and COW led to higher values of soil microbial properties (respiration and functional diversity). Beneficial effects observed under POULTRY at the beginning of the experiment, due to the presence of easily degradable organic matter, were partially lost over time. Our results emphasize the importance of the early monitoring of soil properties (microbial and chemical) and phytotoxicity to properly identify bottlenecks during amendment selection for chemical stabilization, in terms of reduction in metal bioavailability and improvement in soil health.

Phytotoxic activity of Cachrys pungens Jan, a mediterranean species: separation, identification and quantification of potential allelochemicals

In continuous research for bioactive compounds obtained from plants to use for weed control in sustainable agriculture, the aerial parts of Cachrys pungens Jan (Umbelliferae) were extracted with methanol and then fractionated using hexane, chloroform (CHCl3) and ethyl acetate (AcOEt). The potential phytotoxicity of total methanolic extract and each fraction was assayed in vitro on seed germination and root elongation of lettuce (Lactuca sativa L.) and the most active fractions were assayed on three of the most common weeds (Lolium perenne, Amaranthus retroflexus, Echinochloa crus-galli). Non linear regression that allowed to obtain the ED50 index for both physiological processes was applied. The fraction bioassays indicated the following hierarchy of phytotoxicity for both processes: CHCl3 ≥ AcOEt > hexane. Moreover, in the present work was chemically characterized for the first time (through HPTLC) the polar fraction of this species pointing out the high presence of flavonoids and phenolic acids. In particular six of them have been chemically characterized and quantified (naringin, quercetin, catechin, caffeic acid, ferulic acid and gallic acid). These results make C. pungens Jan a potential source of natural compounds employable for an eco-friendly agriculture.

Main Alkaloids of Peganum harmala L. and Their Different Effects on Dicot and Monocot Crops

Alkaloids with allelopathic activity are not as well-known as other allelochemicals. Our study revealed that total alkaloids from seeds of the medicinal plant Peganum harmala L. possessed significant growth inhibitory effect on four treated plants, with dicot plants (lettuce and amaranth) being more sensitive than the tested monocot plants (wheat and ryegrass). Further investigation led to the isolation of harmaline and harmine as the main active ingredients in the total alkaloids of P. harmala seeds. Harmaline exerted potent inhibitory effects on seedling growth of treated plants, especially dicots, inhibiting root elongation of lettuce and amaranth by 31% and 47% at a very low concentration (5 µg/mL), whereas harmine exhibited much weaker non-selective inhibitory effect on the plants. Considering the high yield and poor utilization of P. harmala in China, we anticipate that this plant could be exploited as an alternative weed management tool in the future.

Rehabilitation of degraded soils containing aged PAHs based on phytoremediation with alfalfa ( Medicago sativa L.)

It has been hypothesized that the sorption of high-molecular-weight PAHs to soil and organic matter matrices reduces PAHs availability to biodegradation and phytoextraction. One of the main objectives of this pot-scale phytoremediation study was to enhance the disappearance of weathered PAHs, mainly recalcitrant benzo[ a]pyrene (B[ a]P), seeing that a previous 15-month bioremediation strategy simulating landfarming (unvegetated) could not achieve a complete removal of spiked hydrocarbons. In addition, landfarming coupled with sewage sludge compost (SSC) addition led also to salt accumulation and soil pH reduction, which are warning signs for degraded soils. In this subsequent six month phytoremediation study, alfalfa plants ( Medicago sativa L.) had a remarkable effect on ameliorating the physicochemical properties of SSC-degraded soils by reestablishing initial pH values (>7.0) and significantly reducing salinity (EC < 500 μS cm −1). These changes were reflected markedly by a steep decrease of soil toxicity toward lettuce root elongation and ostracod survival. Soil rehabilitation coupled with rhizosphere effect also impacted positively on microbial growth and soil enzyme activities as compared to the situation prior to alfalfa planting. The dissipation of residual B[ a]P was also enhanced by soil cropping, but only up to 64% for treatments initially spiked at the highest level of 100 mg B[ a]P kg −1 soil. Consequently, remaining B[ a]P in soil (2.5–4.3 mg kg −1) is not likely to be available for further degradation using non-destructive remediation strategies as applied throughout this long-term study.

Toxicity of Polyfluorinated and Perfluorinated Compounds to Lettuce (Lactuca sativa) and Green Algae (Pseudokirchneriella subcapitata)

Recently, polyfluorinated and perfluorinated compounds (PFCs) have been detected in most surface waters around the world. Because some PFCs are persistent and tend to accumulate in surface waters, their potential adverse effects to aquatic organisms have received increasing attention. Nevertheless, currently available toxicity information is limited. The aim of this study was to evaluate the toxicity effects of seven PFCs on root elongation of lettuce (Lactuca sativa) and photosynthesis of green algae (Pseudokirchneriella subcapitata). It was found that the toxicity profiles of both species tested were similar and had good relations with the fluorinated carbon-chain length of the PFCs investigated. One of the compounds tested, perfluorobutanoic acid, was found to be more toxic than expected in the algae test, which may be related with acidification of the test solution. It was concluded that because short-chained PFCs are becoming the predominant PFC pollutants in surface waters, their long-term toxicity and mixture toxicity with other PFCs should be studied in greater detail.

Using response surface methodology to assess the effects of iron and spent mushroom substrate on arsenic phytotoxicity in lettuce ( Lactuca sativa L.)

The effects of iron (Fe) and spent mushroom substrate (SMS) arsenic (As) phytotoxicity towards lettuce in artificial soils were investigated to separate the adverse soil parameters relating to As toxicity using a response surface methodology. SMS induced the root elongation of lettuce in both control and As-treated soils. However, in phytotoxicity test using a median effective concentration (EC 50) of As, Fe and the interaction between both parameters (Fe*SMS) significantly affected EC 50, which explained 71% and 23% of the response, respectively. The refined model was as follows: EC 50 of As (mg kg −1) = 10.99 + 60.03 × Fe − 10.50 × Fe*SMS. The results confirmed that the soil parameters relating to the As mobility in soils were important factors affecting its toxicity. In conclusion, Fe significantly reduced the As phytotoxicity. However, although SMS enhanced the root elongation, SMS in As-treated soils decreased EC 50 of As on the root growth via its interaction with Fe. Despite the limitations of the artificial soils and range of parameters studied, the application of this statistical tool can be considered a powerful and efficient technique for interpretation and prediction of the complicated results caused by the interactions between many factors within the soil environments.

Water-extractable organic matter of different composts: A comparative study of properties and allelochemical effects on horticultural plants

The water-extractable organic matter (WEOM) was obtained from a green compost (GC WEOM), a mixed compost (MC WEOM) and a coffee compost (CC WEOM) and then characterized by means of chemical and physico-chemical methods, such as pH, electrical conductivity, total organic carbon (TOC), E 4/E 6 ratio, fluorescence and Fourier Transform infrared (FT IR) spectroscopies and high performance liquid chromatography (HPLC). The comparative evaluation of the three WEOM samples highlighted significant differences among them. In particular, the TOC content, the E 4/E 6 ratio, the ε 280 value and the humification index followed the same order: CC WEOM > MC WEOM > GC WEOM. The fluorescence analysis of the three WEOM samples showed the presence of a common fluorophore unit possibly associated to simple aromatic units such as phenolic-like, hydroxy-substituted benzoic and cinnamic acid derivatives. The FT IR spectra of all WEOM samples indicated the presence of aromatic phenolic structures, while the HPLC analysis showed the presence of benzoic acid derivatives such as phthalic and salicylic acids. The allelochemical potential of each WEOM sample at two concentrations was tested on tomato and lettuce germination and early growth. In general, for both species, each WEOM sample at the two doses caused a significant increase of shoot length and plant fresh weight. The germination percentage of both plants and root elongation of lettuce resulted unaffected, whereas tomato roots resulted generally shorter in the presence of any WEOM sample, particularly at the higher dose.

Ecotoxicological risk assessment of undisturbed metal contaminated soil at two remote lighthouse sites

Ecotoxicological risk assessments of contaminated soil are commonly completed using guideline values based on total concentrations. However, only certain fractions of contaminants are bioavailable and pose a hazard to the environment. This paper investigates the relationship between measured metal concentrations in soil and soil leachate, and the effects in organisms exposed to intact, undisturbed soil cores (wheat, Tricum aestivum) and soil leachate (lettuce, Lactuca sativa, and water flea, Daphnia magna). Despite the samples containing metal concentrations significantly above guideline values, metals of concern (e.g. Pb and Zn) did not have a significant toxic effect on wheat or D. magna. During weeks with low leachate pH, an effect on lettuce root elongation was observed in the most contaminated samples. This study has shown that bioassays with intact soil cores can indicate metal bioavailability and provide a better estimate of ecological risk than total metal concentrations in the soil.