Lettuce Roots Research Articles

Plantibacter flavus, Curtobacterium herbarum, Paenibacillus taichungensis, and Rhizobium selenitireducens Endophytes Provide Host-Specific Growth Promotion of Arabidopsis thaliana, Basil, Lettuce, and Bok Choy Plants.

A collection of bacterial endophytes isolated from stem tissues of plants growing in soils highly contaminated with petroleum hydrocarbons were screened for plant growth-promoting capabilities. Twenty-seven endophytic isolates significantly improved the growth of Arabidopsis thaliana plants in comparison to that of uninoculated control plants. The five most beneficial isolates, one strain each of Curtobacterium herbarum, Paenibacillus taichungensis, and Rhizobium selenitireducens and two strains of Plantibacter flavus were further examined for growth promotion in Arabidopsis, lettuce, basil, and bok choy plants. Host-specific plant growth promotion was observed when plants were inoculated with the five bacterial strains. P. flavus strain M251 increased the total biomass and total root length of Arabidopsis plants by 4.7 and 5.8 times, respectively, over that of control plants and improved lettuce and basil root growth, while P. flavus strain M259 promoted Arabidopsis shoot and root growth, lettuce and basil root growth, and bok choy shoot growth. A genome comparison between P. flavus strains M251 and M259 showed that both genomes contain up to 70 actinobacterial putative plant-associated genes and genes involved in known plant-beneficial pathways, such as those for auxin and cytokinin biosynthesis and 1-aminocyclopropane-1-carboxylate deaminase production. This study provides evidence of direct plant growth promotion by Plantibacter flavusIMPORTANCE The discovery of new plant growth-promoting bacteria is necessary for the continued development of biofertilizers, which are environmentally friendly and cost-efficient alternatives to conventional chemical fertilizers. Biofertilizer effects on plant growth can be inconsistent due to the complexity of plant-microbe interactions, as the same bacteria can be beneficial to the growth of some plant species and neutral or detrimental to others. We examined a set of bacterial endophytes isolated from plants growing in a unique petroleum-contaminated environment to discover plant growth-promoting bacteria. We show that strains of Plantibacter flavus exhibit strain-specific plant growth-promoting effects on four different plant species.

Uptake, translocation and ligand of silver in Lactuca sativa exposed to silver nanoparticles of different size, coatings and concentration

The broad use of silver nanoparticles (AgNPs) in daily life products enhances their possibilities to reach the environment. Therefore, it is important to understand the uptake, translocation and biotransformation in plants and the toxicological impacts derived from these biological processes. In this work, Lactuca sativa (lettuce) was exposed during 9 days to different coated (citrate, polyvinylpyrrolidone, polyethylene glycol) and sized (60, 75, 100 nm) AgNPs at different concentrations (1, 3, 5, 7, 10, 15 mg L−1). Total silver measurements in lettuce roots indicated that accumulation of AgNPs is influenced by size and concentration, but not by nanoparticle coating. On the other hand, nanosilver translocation to shoots was more pronounced for neutral charged and large sized NPs at higher NP concentrations. Single particle inductively coupled plasma mass spectrometry analysis, after an enzymatic digestion of lettuce tissues indicated the dissolution of some NPs. Ag K-edge X-ray absorption spectroscopy analysis corroborated the AgNPs dissolution due to the presence of less Ag-Ag bonds and appearance of Ag-O and/or Ag-S bonds in lettuce roots. Toxicological effects on lettuces were observed after exposure to nanosilver, especially for transpiration and stomatal conductance. These findings indicated that AgNPs can enter to edible plants, exerting toxicological effects on them.

Mechanistic study on uptake and transport of pharmaceuticals in lettuce from water

The dissemination of pharmaceuticals in agroecosystems originating from land application of animal manure/sewage sludge and irrigation with treated wastewater in agricultural production has raised concern about the accumulation of pharmaceuticals in food products. The pathways of pharmaceutical entries via plant roots, transport to upper fractions, and the factors influencing these processes have yet been systematically elucidated, thus impeding the development of effective measures to mitigate pharmaceutical contamination in food crops. In this study, lettuce uptake of thirteen commonly used pharmaceuticals was investigated using a hydroponic experimental setting. Pharmaceutical sorption by lettuce roots was measured in order to evaluate the influence on pharmaceutical transport from roots to shoots. Small-sized pharmaceuticals e.g., caffeine and carbamazepine with molecular weight (MW) <300 g mol−1 and a low affinity to lettuce roots (sorption coefficient Kp < 0.05 L g−1) manifested substantial transport to shoots. Small-sized molecules lamotrigine and trimethoprim had a relatively strong affinity to lettuce roots (Kp > 12.0 L g−1) and demonstrated a reduced transport to shoots. Large-sized pharmaceuticals (e.g. MW >400 g mol−1) including lincomycin, monensin sodium, and tylosin could be excluded from cell membranes, resulting in the predominant accumulation in lettuce roots. Large-sized oxytetracycline existed as zwitterionic species that could slowly enter lettuce roots; however, the relatively strong interaction with lettuce roots limits its transport to shoots. The mass balance analysis revealed that acetaminophen, β-estradiol, carbadox, estrone and triclosan were readily metabolized in lettuce with >90% loss during 144-h exposure period. A scheme was proposed to describe pharmaceutical uptake and transport in plant, which could reasonably elucidate many literature-reported results. Molecular size, reactivity and ionic speciation of pharmaceuticals, as well as plant physiology, collectively determine their uptake, transport and accumulation in plants.

Cadmium Content and Accumulation in Lettuce Grown on Different Types of Soil

This study was conducted on a pseudogley (pH 5.0) and a vertisol (pH 6.9) to analyze their total and available Cd levels and Cd accumulation by lettuce for risk assessment of potential human health impacts. Total Cd levels in both soil types were below maximum allowable concentrations, and showed a high dependence on soil pH (R=-0.974). Monoammonium phosphate (MAP) fertilizers containing various amounts of cadmium led to an increase in cadmium uptake, with higher concentrations accumulated in lettuce roots than in leaves. The transfer factor showed that Cd uptake from the pseudogley was much higher than from the vertisol, and its values indicated that lettuce is a cadmium hyperaccumulator. Lettuce head weight decreased with increasing cadmium rates and was highly negatively correlated with Cd concentration in the leaves (R=-0.818).

Phytochemical analysis and effect of the essential oil of Psidium L. species on the initial development and mitotic activity of plants.

The use of allelopathic compounds is an alternative for weeds control, since they present low toxicity when compared with the synthetic herbicides, that may cause several damages, as the contamination of the environment. Our objective was to determine the chemical composition and allelopathic properties of the essential oils of Psidium cattleianum, P. myrtoides, P. friedrichsthalianum, and P. gaudichaudianum on the germination and root growth of Lactuca sativa and Sorghum bicolor, and to evaluate their action on the cell cycle of root meristematic cells of L. sativa. The main compound found in all the studied species was (E)-caryophyllene (P. cattleianum-23.4 %; P. myrtoides-19.3%; P. friedrichsthalianum-24.6% and P. gaudichaudianum-17.0%). The different essential oils were tested at different concentrations on L. sativa and S. bicolor, reducing germination, germination speed index, and root and shoot growth of lettuce and sorghum seedlings. The cytotoxicity and aneugenic potential of these oils were evidenced by the reduction of the mitotic index and increase of the frequency of chromosomal alterations in L. sativa. The essential oils of the species of Psidium studied have potential to be used in weeds control.

Effect of tungsten-resistant bacteria on uptake of tungsten by lettuce and tungsten speciation in plants.

Tungsten is an emerging contaminant because of its potential toxicity to humans. However, tungsten-plant-microbe interactions remains unknown. The objective of the study was to evaluate the effect of tungsten-resistant bacteria on tungsten species in plants and microbial community structure in soil. Although bacterial inoculation did not affect lettuce (Lactuca sativa L.) growth or tungsten uptake via root, tungsten-resistant bacteria increased translocation of tungsten from root to shoot. Bacterial inoculation slightly oxidized tungsten in lettuce based on tungsten L3 x-ray absorption near-edge structure (XANES). Tungsten in lettuce roots and shoots grown in tungsten(VI)-spiked soil existed as a mixture of tungsten(IV) and tungsten(VI). Tungsten accumulated as polytungstate in the root and monotungstate in the shoot. Inoculation with tungsten-resistant bacteria and plant growth increased microbial diversity in tungsten-contaminated soil. In tungsten-spiked soils without plants, metal-resistant or reducing bacteria were found while bacteria growing in rhizosphere were detected in soils supporting plant growth. These results indicate a role of the bacteria and plants in phytoremediation of tungsten-contaminated soil.

Strontium Uptake and Effect in Lettuce and Radish Cultivated Under Hydroponic Conditions.

The accumulation of strontium (Sr) in lettuce and radish under 0 (control), 0.5, 1, 2.5, 5, and 10mM Sr treatments in hydroponic solution at 16, 23 and 30days and the effects of Sr stress on six nutrient elements in plants were investigated. The results showed that Sr concentrations in plant aerial and underground parts increased in low-Sr treatments (0.5, 1 and 2.5mM) and fluctuated in high-Sr treatments (5 and 10mM) throughout the three sampling periods. Sr concentrations were higher in roots than in leaves, reaching 108.8 ± 14.7 and 134.1 ± 1.2mg/g in lettuce and radish roots, respectively, after 10mM Sr treatment. Translocation factor (TF) values (ratio of the Sr concentrations in aerial parts to that in roots) were inversely related to the Sr content in the hydroponic solution, and reached 1.45 ± 0.17 to 0.15 ± 0.03 and 1.06 ± 0.20 to 0.12 ± 0.004 for lettuce and radish. The variation in chlorophyll content was consistent with that in plant biomass.

Acid soil amended in contaminated conditions. Effect on cultivated lettuces (Lactuca sativa var. longifolia)

The waste amendment is a common practice to improve crop production on acid soils. This work aims to study the effectiveness of water potabilization plant sludge and lamb manure in acid soils remediation and the possible absorption of metals in cultivated lettuces, as well as the elements´distribution in roots and leaves after extra-contaminated irrigation. Al2(SO4)3-stabilized sludge, and lamb manure was added to an acid soil collected from Huasca de Ocampo, Hidalgo State, Mexico. The sludge (L), manure (M), control soil (C), unamended (S) and amended soils (SML) were characterized, and the metals´ concentrations determined. The SML was prepared by adding 2.62 % M and 5.25% L to fill up 600 L tanks. Lettuces (Lactuca sativa var. longifolia) seeds were sown in S and SML, and irrigated once by metal salts solution (Na, Mg, Ca, Fe, Ni, Cu, Zn, and Cd as sulfates, 200 mg kg-1 each). Lettuce germinated and grew satisfactorily in the SML, but they grew slowly in the unamended acid S, and the roots were affected. Leaves and roots were regularly analyzed along three months, and the metal concentrations showed higher values in S than the ones in SML. Pollutants were available from S, as they were present in roots, and desorbed after 40 days. The elements were present in higher concentrations in lettuce roots than in leaves, as expected. Conclusions. a) Acid soils can be amended with potabilization plants wastes in mining zones, but pH must be controlled to prevent the availability of metals. b) The soil texture was the principal cause of infertility due to root damage, and wastes eliminate the impairment. c) The element concentrations present in the leaves were acceptable for human nutrition.

Salmonella Establishment in Agricultural Soil and Colonization of Crop Plants Depend on Soil Type and Plant Species.

Human pathogenic bacteria, such as Salmonella enterica, are able to colonize crop plants. So far, not much is known about biotic and abiotic factors influencing this colonization in field soil. This understanding, however, is imperative for the provision of safe fresh produce to the consumer. In this study, we investigated the effects of soil type, organic fertilization, plant species and the way of Salmonella entry into the plant production system, on the survival of S. enterica in soil as well as the colonization of plants. The selected S. enterica serovar Typhimurium strain 14028s, S. Typhimurium strain LT2 and S. Senftenberg were able to persist in soil for several weeks. Salmonella’s persistence in soil was prolonged in loamy, if compared to sandy soil, and when applied together with organic fertilizer. The leaves of lettuce and corn salad were colonized by S. enterica providing evidence for internalization from the soil via the root. Colonization rates were affected by soil type, plant species and S. enterica strain. Overall, S. enterica was detected in leaves of 0.5–0.9% of the plants, while lettuce was more frequently colonized than corn salad. Plants grown in sandy soil were more often colonized than plants grown in loamy soil. After spray inoculation, S. enterica could be detected on and in leaves for several weeks by cultivation-depending methods, confirmed by confocal microscopy using GFP-labeled S. Typhimurium 14028s. On the one hand, transcriptome data from S. Typhimurium 14028s assessed in response to lettuce medium or lettuce root exudates showed an upregulation of genes associated with biofilm formation and virulence. On the other hand, lettuce inoculated with S. Typhimurium 14028s showed a strong upregulation of genes associated with plant immune response and genes related to stress response. In summary, these results showed that organic fertilizers can increase the persistence of Salmonella in soil and that soil type and plant species play a crucial role in the interactions between human pathogens and crop plants. This understanding is therefore a starting point for new strategies to provide safe food for the consumer.

Humus-Rich Compost Increases Lettuce Growth, Nutrient Uptake, Mycorrhizal Colonisation, and Soil Fertility

Sandy soils, typical of Australia's west, either have little or no habitat protection for microbes including arbuscular mycorrhizal (AM) fungi, which are essential for nutrient cycling. To minimize this problem, the application of organic matter, such as humus-rich composts, is necessary during vegetable crop production. This study aimed at determining the effects of humus-rich composts on either indigenous or inoculated AM fungal colonisation in roots, lettuce (Lactuca sativa L. var. Quechua) growth, and soil fertility improvement. Four different humus-rich composts with varying humus contents were applied at the same standard rate to lettuce grown under glasshouse conditions for 10 weeks after sowing and compared with two low-humus composts and non-amended soil (control). Humus-rich composts significantly increased lettuce shoot growth, root growth, and AM fungal colonisation in roots. Humus contents in the composts were also correlated with lettuce shoot and root growth. Soil dissolved organic carbon, microbial biomass carbon, and fertility were increased with the application of humus-rich composts. These humus-rich composts, especially the compost of higher humic acid with and without AM inoculation, might have a significant role in sustainable vegetable production, for example lettuce growth. Overall, the results indicate that supplementation with humus-rich compost is highly beneficial to enhance soil fertility and potentially maintain the sustainability of vegetable production.

Diffusive gradient in thin films with open and restricted gels for predicting mercury uptake by plants

The presence of mercury (Hg) in the environment is a major concern because mercury is toxic for living organisms. Estimating the Hg toxicity of environmental samples is difficult because contaminants occur in pools of various bioavailabilities, and thus, measuring the total concentration of a contaminant rarely reflects the real toxicity and risk. There is therefore a need to design advanced methods to measure the bioavailable fraction of the contaminants. Here, we used the diffusive gradient in thin films (DGT) method to determine the bioavailability of Hg in agricultural soils containing 0.02 and 0.64 mg kg−1 of Hg. Specifically, we used homemade manufactured open and restricted diffusive layers to distinguish between inorganic and organic bioavailable Hg species in soil. The findings demonstrate for the first time the successful use of restricted gel for the determination of inorganic Hg species in soil. Moreover, a model program has been used to calculate the effective concentration for predicting Hg concentration uptake by plants. For this purpose, lettuces were planted in soils to assess Hg soil-to-plant transfer. Concentrations of Hg obtained by DGT measurements in soils matched the Hg concentration found in lettuce roots, thus confirming the effectiveness of this technique in predicting Hg uptake by plants.

Isolation of isolactarane sesquiterpenes from a Phlebia tremellosa culture filtrate and their growth promotion effects on lettuce roots.

The ethyl acetate extract of the culture filtrate of Phlebia tremellosa promoted elongation of the lateral roots of lettuce seedlings at 250 µg/mL. We purified two compounds that promote root elongation by using activity-guided chromatographic fractionation. On the basis of spectroscopic analyses, these compounds were identified to be isolactarane sesquiterpenes derived from the dehydrogenation of merulactone, which was previously isolated from the same species. We named the purified compounds phlelactones A and B. Phlelactones A and B promoted primary root elongation at 100-300 and 10-30 µg/mL and the elongation and formation of lateral roots at 300-1000 and 30-100 µg/mL, respectively.

Antioxidative enzymes activity and thiol metabolism in three leafy vegetables under Cd stress

The enrichment of Cadmium in vegetables is threatening human health. The study aimed to screen Cd low-enriched leafy vegetables and explore whether antioxidative enzymes and heavy metal chelators are synergistic defensive mechanisms. In this paper, the Cd accumulation and translocation of garland chrysanthemum (Chrysanthemum coronarium L.), spinach (Spinacia oleracea L.), and lettuce (Lactuca sativa L.) were examined by soil pot culture and hydroponic experiments. The responses of oxidative stress markers, antioxidative enzymes activity, and thiol pool (cysteine, γ-glutamylcysteine, glutathione, and phytochelatins) content to Cd stress were assayed. The results showed that Garland chrysanthemum was Cd low-uptake species. The soil Cd safety thresholds for spinach, lettuce, and garland chrysanthemum were 0.41, 0.49, and 9.10 mg kg–1, respectively. The order of root phytochelatins (PCs) concentration was consistent with that of plant tolerance index (TI): garland chrysanthemum > spinach > lettuce. While the order of the ratio of shoot Cd to root Cd (SR ratio) was exactly the opposite of that of TI. In lettuce root, activity of superoxide dismutase, peroxidase, and catalase decreased significantly under Cd stress. Nevertheless those parameters in the roots of spinach and lettuce maintained steady, or even enhanced. In conclusion, the Cd translocation and partition in plant, antioxidative defense, and PCs homeostasis played an important role in the Cd tolerance of vegetables.

Demonstration of allelopathy of horseradish root extract on lettuce seed.

Allelopathy plays crucial roles in invasive plant viability and agricultural production systems. However, there is no well-established hands-on learning activity to teach the concept of allelopathy. Nor is there an activity which allows students to gain knowledge about glucosinolates and their corresponding enzyme, myrosinase, which are present in almost all Brassica crops. Lettuce germination was counted by the students from three different treatments including water treated with Parafilm sealing, horseradish treated with Parafilm sealing, and horseradish treated without Parafilm sealing. Additionally, lettuce root length was measured by students using ImageJ software from each treatment using pictures captured by students' smartphones. Students took an identical quiz as a pre-laboratory and a post-laboratory assignment. Their average scores on the pre-laboratory and post-laboratory quizzes were 3.14 and 6.56 out of 10, respectively, indicating the lab activity significantly improved students' understanding of allelopathy and glucosinolate-myrosinase system. In addition, students (n = 76) completed a survey post-laboratory to assess their self-efficacy. This simple and cost-effective laboratory activity improved students' knowledge and skill development as it made learning more inviting, meaningful, and fun. © 2019 International Union of Biochemistry and Molecular Biology, 47(3):333-340, 2019.

Application of MALDI-TOF mass spectrometry and specific PCR for tracking of E. coli O157:H\u2212 strain 431/97 in Batavia lettuce

BackgroundIn this study, lettuce roots and leaves were contaminated with enterohemorrhagic Escherichia coli O157:H− strain 431/97 under greenhouse conditions. Furthermore, the internalization of strain 431/97 in lettuce roots and leaves was examined. To track the inoculated bacteria during the experiments and to differentiate them from the autochthonous microbiota, a combined protocol including matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) and PCR was established.ResultsAt different time points after inoculation of Batavia lettuce with 5.5 × 106 cfu/mL (high inoculation level) or 3.0 × 101 cfu/mL (low inoculation level) E. coli O157:H− strain 431/97 as well as sterile 0.9% (w/v) sodium chloride solution (negative control), samples from the root and the leaf were taken and surface disinfected with gentamicin. After homogenization, microorganisms were isolated from the samples and analyzed by MALDI-TOF MS. Analysis of the root samples resulted in bacterial counts of 1.0 × 102–1.0 × 106 cfu/0.25 g depending on the inoculated viable counts and the incubation period. In the leaf samples, strain 431/97 was not detected. The investigation of the viable cell counts of E. coli O157:H− 431/97 following irrigation of the leaves resulted in bacterial counts of 102 cfu/0.25 g for the disinfected leaf samples.ConclusionsThus, the established protocol is suitable for detecting the investigated strain under greenhouse conditions in plant infection experiments. This strain may indeed survive in the soil, but did not enter the plant via the root in detectable numbers. Contrarily, viable counts exceeding the generally accepted infective dose of less than 100 cells for enterohemorrhagic E. coli were determined internalized after irrigation of the leaves. As this may pose a risk for the consumer, the present study provides a valuable set of tools for further research.

Uptake and metabolism of clarithromycin and sulfadiazine in lettuce

Antibiotics are introduced into agricultural fields by the application of manure or biosolids, or via irrigation using reclaimed wastewater. Antibiotics can enter the terrestrial food chains through plant uptake, which forms an alternative pathway for human exposure to antibiotics. However, previous studies mainly focused on detecting residues of the parent antibiotics, while ignoring the identification of antibiotics transformation products in plants. Here, we evaluated the uptake and metabolism of clarithromycin (CLA) and sulfadiazine (SDZ) in lettuce under controlled hydroponic conditions. The antibiotics and their metabolites were identified by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS/MS) and ultra-performance liquid chromatograph Micromass triple quadrupole mass spectrometry (UPLC−QqQ−MS/MS). The structure of CLA, SDZ and N-acetylated SDZ were confirmed with synthesized standards, verifying the reliability of the identification method. Eight metabolites of CLA and two metabolites of SDZ were detected in both the leaves and roots of lettuce. The metabolites of CLA included phases I and II transformation products, while only phase II metabolites of SDZ were observed in lettuce. The proportion of CLA metabolites was estimated to be greater than 70%, indicating that most of the CLA was metabolized in plant tissues. The proportion of SDZ metabolites was lower than 12% in the leaves and 10% in the roots. Some metabolites might have the ability to increase or acquire antibacterial activity. Therefore, in addition to the parent compounds, metabolites of antibiotics in edible vegetables are also worthy of study for risk assessment and to determine the consequences of long-term exposure.

Uptake and accumulation of microplastics in an edible plant

Microplastic (MP, 100 nm−5 mm) may present an attributable risk to ecosystem and human health, and its pollution has become a global environmental concern. Despite a wealth of information on the accumulation of MPs in aquatic species, there is no information on the uptake and accumulation of MPs by higher plants. Terrestrial edible plants are directly exposed to MPs when agricultural soil was applied with organic manure, sewage sludge as fertilizer or plastic mulching. In this paper, the uptake of two sizes of polystyrene (PS) microbeads (0.2 and 1.0 μm) and then their distribution and migration in an edible plant lettuce were firstly investigated based on laboratory experiments. We used fluorescent markers to track PS microbeads in plant tissues and found fluorescence to be a sensitive and reliable detection method. Sections from untreated control lettuce showed no autofluorescence. When roots were treated with fluorescently labeled PS microbeads, the microbeads could be identified by its fluorescence. Our main study investigated the uptake of 0.2 μm beads, as few luminescence signals were observed in lettuce roots for 1.0 μm beads in our experiment. We observed that 0.2 μm fluorescent microbeads were extracellularly trapped in the root cap mucilage (which is a highly hydrated polysaccharide) and a “dark green tip” (which was typical of lettuce roots exposed to label PS beads) was usually visible to the naked eye. Confocal images revealed that the PS luminescence signals were mainly located in the vascular system and on the cell walls of the cortex tissue of the roots, indicated that the beads passed through the intercellular space via the apoplastic transport system. Once inside the central cylinder, the 0.2 μm PS beads were transferred from the roots to the stems and leaves via the vascular system following the transpiration stream. We also observed that the PS beads adhered to one another and self-assembled systematically into “grape-like” and “(chain) string-like” clusters in the intercellular space of the root and stem vascular tissue of lettuce plant. In contrast to the root and stem, PS beads were dispersed in the leaf tissue. Here, for the first time we provide evidence of the adherence, uptake, accumulation, and translocation of submicrometer MPs within an edible plant. Our findings highlight the previously underappreciated human exposure pathway to MPs through the consumption of contaminated crops and emphasize the need for new management strategies to control the release of MPs waste products into the terrestrial environment. Ultimately, the potential impacts of low range sized MPs on food safety of crop plants and human health need to be urgently considered.

The response and tolerance mechanisms of lettuce (Lactuca sativa L.) exposed to nickel in a spiked soil system

Nickel contamination may lead to the destruction of food, ecological safety and its toxicity to plants remains to be studied in depth. In our present study, the translocation factors (TFsoil to root and TFroot to shoot) revealed a significant logarithmic decline with the increase of Ni exposure. In lettuce roots, NiHAC played an important protective role against high Ni stress and the ratio of Ni with high activity (NiE and NiW) in root decreased with the addition of Ni. The activities of antioxidant enzymes (CAT, POD and SOD) in the lettuce roots were increased and might be the way for lettuce to adapt Ni stress. CAT and POD can be great indicators of Ni pollution exhibiting better dose-effect relationships with Ni. Under high Ni stress, lettuce roots contained higher levels of MDA suffering greater pressure than shoots. Expression levels of gene GST 23-like indicated a remarkable (P < 0.05) down-regulation and then this trend would be alleviated after high Ni exposure, and it was positively correlated with GST concentrations (R2 = 0.704). We believe that our research would open up the new avenues for effective understanding ecological risks of Ni.

Detection and Quantification of Pythium tracheiphilum in Soil by Multiplex Real-Time qPCR

In Canada, head lettuce (Lactuca sativa capitata) is extensively produced in the muck soils of southwestern Québec. However, yields are increasingly affected by various soilborne pathogens, including Pythium spp., which cause wilt and damping off. In a survey conducted in Québec muck soils in 2010 and 2011, Pythium tracheiphilum Matta was identified as the predominant Pythium sp. in the root of head lettuce showing Pythium stunt symptoms. Therefore, to improve risk assessment and help further understanding of disease epidemiology, a specific and sensitive real-time quantitative polymerase chain reaction (qPCR) assay based on TaqMan-minor groove binder (MGB) technology was developed for P. tracheiphilum. The PCR primers along with a TaqMan-MGB probe were designed from the ribosomal internal transcribed spacer 2 region. A 100-bp product was amplified by PCR from all P. tracheiphilum isolates tested while no PCR product was obtained from 38 other Pythium spp. or from a selection of additional lettuce pathogens tested. In addition to P. tracheiphilum, the assay was multiplexed with an internal control allowing for the individual validation of each PCR. In artificially infested soils, the sensitivity of the qPCR assay was established as 10 oospores/g of dry soil. P. tracheiphilum was not detected in soils in which lettuce has never been grown; however, inoculum ranged from 0 to more than 200,000 oospores/g of dry soil in commercial lettuce fields. Also, disease incidence was positively correlated with inoculum concentration (r = 0.764). The results suggest that inoculum concentration should be considered when making Pythium stunt management decisions. The developed qPCR assay will facilitate reliable detection and quantification of P. tracheiphilum from field soil.

Phytotoxic activity of Chinese violet (Asystasia gangetica (L.) T. Anderson) and two phytotoxic substances

Chinese violet (Asystasia gangetica (L.) T. Anderson) is a perennial invasive weed belonging to Acanthaceae. Leaves of this weed have been suggested to possess phytotoxic activity. However, phytotoxic substances in this weed have not yet been reported. Therefore, the present study investigated phytotoxic activity of Chinese violet extracts and phytotoxic substances. The extracts of Chinese violet leaves inhibited the root and shoot growth of cress, lettuce, alfalfa, barnyard grass, ryegrass, and timothy, where the level of inhibition increased with increasing extract concentrations. Bioassay‐guided separations of the extracts led to isolation of two phytotoxic substances, indole‐3‐carboxaldehyde and (6R,9S)‐3‐oxo‐α‐ionol. Indole‐3‐carboxaldehyde significantly inhibited the root and shoot growth of cress at concentrations ≥100 and 30 μmol L−1, respectively, and concentrations of the substance required for 50% growth inhibition were 210 and 127 μmol L−1 for cress roots and shoots, respectively. The other substance, (6R,9S)‐3‐oxo‐α‐ionol, was reported to have strongly inhibited cress roots and shoots. The present results suggest that Chinese violet contains two phytotoxic substances indole‐3‐carboxaldehyde and (6R,9S)‐3‐oxo‐α‐ionol, and those substances may play an important role in the phytotoxic activity of Chinese violet.