Lactuca sativa
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A positive selection system using phosphomannose isomerase was employed for Agrobacterium tumefaciens mediated transformation of lettuce (Lactuca sativa L. var. 'Achát'). It was shown that the mannose-based selection system works very well with the lettuce genotype used, reaching up to 25% transformation efficiency on the medium with 20 g/L mannose and 20 g/L sucrose. The best transformation efficacy with the commonly-used kanamycin at 100 mg/L as a selection agent was 21%. Southern blot analyses of thirteen chosen mannose-resistant regenerants revealed that some of them have clonal origin, about one-half harbour a single T-DNA copy and one plant contains an incomplete T-DNA segment with only the left part of T-DNA with the pmi gene present in the genomic DNA. The following Northern analysis showed transcriptional activity of the introduced pmi gene in all plants analysed with very high differences in the level of pmi specific mRNA. The results demonstrate that both mannose and kanamycin provide comparable transformation efficiencies in our lettuce genotype. An alternative selection method with mannose as a selection agent is now available for lettuce transgenosis.
- MeSH
- Agrobacterium tumefaciens genetika MeSH
- geneticky modifikované rostliny MeSH
- mannosa-6-fosfátisomerasa genetika MeSH
- salát (hlávkový) enzymologie genetika růst a vývoj MeSH
- selekce (genetika) MeSH
- transformace genetická MeSH
- výhonky rostlin enzymologie genetika růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The biologically active molecules karrikinolide (KAR1) and trimethylbutenolide (TMB) present in wildfire smoke play a key role in regulating seed germination of many plant species. To elucidate the physiological mechanism by which smoke-water (SW), KAR1, and TMB regulate seed germination in photosensitive 'Grand Rapids' lettuce (Lactuca sativa), we investigated levels of the dormancy-inducing hormone abscisic acid (ABA), three auxin catabolites, and cytokinins (26 isoprenoid and four aromatic) in response to these compounds. Activity of the hydrolytic enzymes α-amylase and lipase along with stored food reserves (lipids, carbohydrate, starch, and protein) were also assessed. The smoke compounds precisely regulated ABA and hydrolytic enzymes under all light conditions. ABA levels under red (R) light were not significantly different in seeds treated with TMB or water. However, TMB-treated seeds showed significantly inhibited germination (33%) compared with water controls (100%). KAR1 significantly enhanced total isoprenoid cytokinins under dark conditions in comparison with other treatments; however, there was no significant effect under R light. Enhanced levels of indole-3-aspartic acid (an indicator of high indole-3-acetic acid accumulation, which inhibits lettuce seed germination) and absence of trans-zeatin and trans-zeatin riboside (the most active cytokinins) in TMB-treated seeds might be responsible for reduced germination under R light. Our results demonstrate that SW and KAR1 significantly promote lettuce seed germination by reducing levels of ABA and enhancing the activity of hydrolytic enzymes, which aids in mobilizing stored reserves. However, TMB inhibits germination by enhancing ABA levels and reducing the activity of hydrolytic enzymes.
- MeSH
- furany farmakologie MeSH
- fytochrom metabolismus MeSH
- gama-butyrolakton analogy a deriváty farmakologie MeSH
- klíčení účinky léků MeSH
- kouř * MeSH
- lékové interakce MeSH
- pyrany farmakologie MeSH
- regulátory růstu rostlin metabolismus MeSH
- salát (hlávkový) účinky léků metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cellular homeostasis of S-nitrosoglutathione (GSNO), a major cache of nitric oxide bioactivity in plants, is controlled by the NADH-dependent S-nitrosoglutathione reductase (GSNOR) belonging to the family of class III alcohol dehydrogenases (EC 1.1.1.1). GSNOR is a key regulator of S-nitrosothiol metabolism and is involved in plant responses to abiotic and biotic stresses. This study was focused on GSNOR from two important crop plants, cauliflower (Brassica oleracea var. botrytis, BoGSNOR) and lettuce (Lactuca sativa, LsGSNOR). Both purified recombinant GSNORs were characterized in vitro and found to exists as dimers, exhibit high thermal stability and substrate preference towards GSNO, although both enzymes have dehydrogenase activity with a broad range of long-chain alcohols and ω-hydroxy fatty acids in presence of NAD+. Data on enzyme affinities to their cofactors NADH and NAD+ obtained by isothermal titration calorimetry suggest the high affinity to NADH might underline the GSNOR capacity to function in the intracellular environment. GSNOR activity and gene expression peak during early developmental stages of lettuce and cauliflower at 20 and 30 days after germination, respectively. GSNOR activity was also measured in four other Lactuca spp. genotypes with different degree of resistance to biotrophic pathogen Bremia lactucae. Higher GSNOR activities were found in non-infected plants of susceptible genotypes L. sativa UCDM2 and L. serriola as compared to resistant genotypes. GSNOR and GSNO were localized by confocal laser scanning microscopy in vascular bundles and in epidermal and parenchymal cells of leaf cross-sections. The presented results bring new insight in the role of GSNOR in the regulation of S-nitrosothiol levels in plant growth and development.
MAIN CONCLUSION: Resistant Lactuca spp. genotypes can efficiently modulate levels of S-nitrosothiols as reactive nitrogen species derived from nitric oxide in their defence mechanism against invading biotrophic pathogens including lettuce downy mildew. S-Nitrosylation belongs to principal signalling pathways of nitric oxide in plant development and stress responses. Protein S-nitrosylation is regulated by S-nitrosoglutathione reductase (GSNOR) as a key catabolic enzyme of S-nitrosoglutathione (GSNO), the major intracellular S-nitrosothiol. GSNOR expression, level and activity were studied in leaves of selected genotypes of lettuce (Lactuca sativa) and wild Lactuca spp. during interactions with biotrophic mildews, Bremia lactucae (lettuce downy mildew), Golovinomyces cichoracearum (lettuce powdery mildew) and non-pathogen Pseudoidium neolycopersici (tomato powdery mildew) during 168 h post inoculation (hpi). GSNOR expression was increased in all genotypes both in the early phase at 6 hpi and later phase at 72 hpi, with a high increase observed in L. sativa UCDM2 responses to all three pathogens. GSNOR protein also showed two-phase increase, with highest changes in L. virosa-B. lactucae and L. sativa cv. UCDM2-G. cichoracearum pathosystems, whereas P. neolycopersici induced GSNOR protein at 72 hpi in all genotypes. Similarly, a general pattern of modulated GSNOR activities in response to biotrophic mildews involves a two-phase increase at 6 and 72 hpi. Lettuce downy mildew infection caused GSNOR activity slightly increased only in resistant L. saligna and L. virosa genotypes; however, all genotypes showed increased GSNOR activity both at 6 and 72 hpi by lettuce powdery mildew. We observed GSNOR-mediated decrease of S-nitrosothiols as a general feature of Lactuca spp. response to mildew infection, which was also confirmed by immunohistochemical detection of GSNOR and GSNO in infected plant tissues. Our results demonstrate that GSNOR is differentially modulated in interactions of susceptible and resistant Lactuca spp. genotypes with fungal mildews and uncover the role of S-nitrosylation in molecular mechanisms of plant responses to biotrophic pathogens.
- MeSH
- aldehydoxidoreduktasy metabolismus MeSH
- konfokální mikroskopie MeSH
- nemoci rostlin mikrobiologie MeSH
- odolnost vůči nemocem fyziologie MeSH
- oomycety patogenita MeSH
- polymerázová řetězová reakce MeSH
- regulace genové exprese u rostlin MeSH
- S-nitrosothioly metabolismus MeSH
- salát (hlávkový) enzymologie fyziologie MeSH
- western blotting MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Lettuce (Lactuca sativa L.) is a crop that is very sensitive to herbicide contamination owing to its short growing season. The use of long-residual herbicides and non-woven fabric coverings could therefore influence pendimethalin concentrations in soil and lettuce. RESULTS: The pendimethalin half-life in soil ranged between 18 and 85 days and was mainly affected by season (i.e. weather), and especially by soil moisture. Pendimethalin degradation in soil was slowest under dry conditions. A longer pendimethalin half-life was observed under the non-woven fabric treatment, but the effect of varying application rate was not significant. Pendimethalin residue concentrations in lettuce heads were significantly influenced by pendimethalin application rate and by non-woven fabric cover, especially at the lettuce's early growth stages. The highest pendimethalin concentration at final harvest was determined in lettuce grown on uncovered plots treated with pendimethalin at an application rate of 1200 g ha(-1) (7-38 µg kg(-1) ). Depending on growing season duration and weather conditions, pendimethalin concentrations in lettuce grown under non-woven fabric ranged from 0 to 21 µg kg(-1) . CONCLUSION: Use of transparent non-woven fabric cover with lettuce can help to reduce application rates of soil herbicides and diminish the risk of herbicide contamination in the harvested vegetables. © 2016 Society of Chemical Industry.
- MeSH
- aniliny analýza MeSH
- půda chemie MeSH
- rezidua pesticidů analýza MeSH
- salát (hlávkový) chemie MeSH
- textilie * MeSH
- Publikační typ
- časopisecké články MeSH
Phytopathology is a highly complex scientific discipline. Initially, its focus was on the study of plant-pathogen interactions in agricultural and forestry production systems. Host-pathogen interactions in natural plant communities were generally overlooked until the 1970s when plant pathologists and evolutionary biologists started to take an interest in these interactions, and their dynamics in natural plant populations, communities, and ecosystems. This article introduces the general principles of plant pathosystems, provides a basic critical overview of current knowledge of host-pathogen interactions in natural plant pathosystems, and shows how this knowledge is important for future developments in plant pathology especially as it applies in cropping systems, ecology, and evolutionary biology. Plant pathosystems can be further divided according to the structure and origin of control, as autonomous (wild plant pathosystems, WPPs) or deterministic (crop plant pathosystems, CPPs). WPPs are characterized by the disease triangle and closed-loop (feedback) controls, and CPPs are characterized by the disease tetrahedron and open-loop (non-feedback) controls. Basic general, ecological, genetic, and population structural and functional differences between WPPs and CPPs are described. It is evident that we lack a focus on long-term observations and research of diseases and their dynamics in natural plant populations, metapopulations, communities, ecosystems, and biomes, as well as their direct or indirect relationships to CPPs. Differences and connections between WPPs and CPPs, and why, and how, these are important for agriculture varies. WPP and CPP may be linked by strong biological interactions, especially where the pathogen is in common. This is demonstrated through a case study of lettuce (Lactuca spp., L. serriola and L. sativa) and lettuce downy mildew (Bremia lactucae). In other cases where there is no such direct biological linkage, the study of WPPs can provide a deeper understanding of how ecology and genetics interacts to drive disease through time. These studies provide insights into ways in which farming practices may be changed to limit disease development. Research on interactions between pathosystems, the "cross-talk" of WPPs and CPPs, is still very limited and, as shown in interactions between wild and cultivated Lactuca spp.-B. lactucae associations, can be highly complex. The implications and applications of this knowledge in plant breeding, crop management, and disease control measures are considered. This review concludes with a discussion of theoretical, general and specific aspects, challenges and limits of future WPP research, and application of their results in agriculture.
The aim of this work was to determine the ability of rhodococci to transform 3,5-dichloro-4-hydroxybenzonitrile (chloroxynil), 3,5-dibromo-4-hydroxybenzonitrile (bromoxynil), 3,5-diiodo-4-hydroxybenzonitrile (ioxynil) and 2,6-dichlorobenzonitrile (dichlobenil); to identify the products and determine their acute toxicities. Rhodococcus erythropolis A4 and Rhodococcus rhodochrous PA-34 converted benzonitrile herbicides into amides, but only the former strain was able to hydrolyze 2,6-dichlorobenzamide into 2,6-dichlorobenzoic acid, and produced also more of the carboxylic acids from the other herbicides compared to strain PA-34. Transformation of nitriles into amides decreased acute toxicities for chloroxynil and dichlobenil, but increased them for bromoxynil and ioxynil. The amides inhibited root growth in Lactuca sativa less than the nitriles but more than the acids. The conversion of the nitrile group may be the first step in the mineralization of benzonitrile herbicides but cannot be itself considered to be a detoxification.
- MeSH
- amidohydrolasy metabolismus MeSH
- amidy metabolismus toxicita MeSH
- benzamidy metabolismus MeSH
- biotransformace MeSH
- dehydratasy metabolismus MeSH
- herbicidy chemie metabolismus MeSH
- hydrolýza MeSH
- jodbenzeny metabolismus MeSH
- kořeny rostlin účinky léků růst a vývoj metabolismus MeSH
- nitrily chemie metabolismus toxicita MeSH
- Rhodococcus metabolismus MeSH
- salát (hlávkový) účinky léků růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The dissipation, partitioning dynamics and biouptake was measured for selected hazardous current-used pesticides (conazole fungicides: epoxiconazole, flusilazole, tebuconazole; prochloraz, chlorpyrifos, pendimethalin) and for a transformation product (2-hydroxyatrazine) in agricultural soil and quartz sand as representatives of a real and a worst-case scenario. Dissipation, uptake to Lactuca sativa and the freely dissolved concentration along with the organic carbon-normalized sorption coefficients (Koc) were determined on days 12, 40, and 90 following the application of compounds at three fortification levels (0.1-1.0-10 mg/kg). Conazole fungicides showed similar dissipation patterns and were more persistent in soil than prochloraz, chlorpyrifos and pendimethalin. 2-Hydroxyatrazine showed a concentration-depended decrease in persistency in soil. Lettuce roots were shown to accumulate higher amounts than shoots where the extent of root uptake was driven by compound partitioning. This was evidenced by the ability of freely dissolved concentration (Cfree) to reliably (r2 = 0.94) predict root uptake. Concentration in leaves did not exceed the maximum residue levels (MRLs) for lettuce, which was likely given by the low root-to-shoot translocation factors (TFs) of the tested compounds varying between 0.007 and 0.14. Koc values were in the range of literature values. Sorption to soil was higher than to sand for all compounds, yet following the Koc dynamics compounds did not appear to be sequestered in soil with increasing residence time. From these results, it follows that the tested compounds may persist in soil but since they did not accumulate in lettuce above MRLs, contamination of the food web is unlikely.
- MeSH
- atrazin analogy a deriváty analýza farmakokinetika MeSH
- kořeny rostlin metabolismus MeSH
- látky znečišťující půdu analýza MeSH
- listy rostlin metabolismus MeSH
- pesticidy analýza farmakokinetika MeSH
- průmyslové fungicidy analýza farmakokinetika MeSH
- salát (hlávkový) metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Sludge coming from remediation of groundwater contaminated by industry is usually managed as hazardous waste despite it might be considered for further processing as a source of nutrients. The ecotoxicity of phosphorus rich sludge contaminated with arsenic was evaluated after mixing with soil and cultivation with Sinapis alba, and supplementation into composting and vermicomposting processes. The Enchytraeus crypticus and Folsomia candida reproduction tests and the Lactuca sativa root growth test were used. Invertebrate bioassays reacted sensitively to arsenic presence in soil-sludge mixtures. The root elongation of L. sativa was not sensitive and showed variable results. In general, the relationship between invertebrate tests results and arsenic mobile concentration was indicated in majority endpoints. Nevertheless, significant portion of the results still cannot be satisfactorily explained by As chemistry data. Composted and vermicomposted sludge mixtures showed surprisingly high toxicity on all three tested organisms despite the decrease in arsenic mobility, probably due to toxic metabolites of bacteria and earthworms produced during these processes. The results from the study indicated the inability of chemical methods to predict the effects of complex mixtures on living organisms with respect to ecotoxicity bioassays.
- MeSH
- arsen chemie toxicita MeSH
- biotest MeSH
- členovci účinky léků MeSH
- ekotoxikologie MeSH
- hořčice rodu Sinapis růst a vývoj MeSH
- kompostování * MeSH
- kroužkovci účinky léků MeSH
- látky znečišťující půdu chemie toxicita MeSH
- odpadní vody chemie MeSH
- půda chemie MeSH
- rozpustnost MeSH
- salát (hlávkový) účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Iceberg lettuce is one of the most consumed leafy vegetables, which is often treated by different pesticides against pests and diseases. The aim of this study was to describe the fate of 25 pesticides (16 fungicides, 7 insecticides and 2 herbicides) based on quantitative analysis of the parent compounds and targeted screening of their (bio)transformation products. Mathematical models describing a decrease in pesticide residue levels were proposed for 24 pesticides using a first-order kinetic equation. These models provide the data needed to predict consumer exposure associated with the consumption of conventionally grown iceberg lettuce. At harvest, concentrations of most pesticides were dropped under the established EU maximum residue levels, except for flonicamid, fluazifop and pyriproxyfen. A total of 113 pesticide metabolites and degradation products were detected and tentatively identified in extracts prepared by an optimized extraction procedure, i.e., the acidified QuEChERS method. Several products of reactions such as hydrolysis, dealkylation, dehalogenation and/or oxidation-reduction, originated either from various physicochemical processes, or within Phase I pesticide metabolism were detected. Additionally, numerous conjugates with hexose, malonic acid or acetic acid formed during PhaseII of pesticide metabolism were found. In this way, a deeper understanding of specific pesticide degradation mechanisms is facilitated. In addition, it is easier to track the history of pesticide treatment.
