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Antioxidative components from the aerial parts of Lactuca scariola L.

The antioxidant activity of Lactuca scariola (Compositae) was investigated by measuring the radical scavenging effect on DPPH (1,1-diphenyl-2-picrylhydrazyl) radical. The methanolic extract of the aerial parts of Lactuca scariola showed strong radical scavenging activity. The EtOAc soluble fraction exhibited a stronger activity than the others, and was purified by silica gel and Sephadex LH-20 column chromatography. Quercetin-3-O-beta-D-glucopyranoside, luteolin-7-O-beta-D-glucopyranoside, luteolin, quercetin and kaempferol, together with 1beta,13-dihydrolactucin were isolated from the EtOAc soluble fraction as active ingredients.

Antioxidants↗

Structure and expression of a gene encoding heat-shock protein Hsp70 from the Oomycete fungus Bremia lactucae.

A gene encoding a protein homologous to a 70-kDa heat-shock protein (Hsp70) was isolated from Bremia lactucae and its structure and pattern of expression were determined. This is the first report on the structure of a protein-coding gene from an Oomycete fungus. The cloned gene is a member of a small multigene family. The level of hsp70 mRNA in germlings increases from a low constitutive level in response to heat or cold treatment. A high level of the mRNA is also detected in spores. The hsp70 gene is expressed as a primary transcript of 2241 nucleotides (nt) and contains a continuous open reading frame of 2025 nt. Near the C terminus of the coding sequence is an unusual region that contains repeated enhancer-like sequences. This insert has not been described in other hsp70 genes and is not an intron. Upstream from the 5' terminus of the mRNA are multiple CCAAT motifs, a sequence similar to a consensus heat-shock regulatory element, and an A + T-rich putative 'TATA' box. A canonical polyadenylation recognition sequence is present downstream from the coding sequence. The deduced amino acid sequence is equally similar to yeast and maize Hsp70, providing further evidence of the dissimilarity between Oomycetes and true fungi. The cloning of this gene is part of our strategy to develop a transformation system for B. lactucae.

Amino Acid Sequence↗

Biosorption of mercury from aqueous solution by Ulva lactuca biomass.

The mercury biosorption onto non-living protonated biomass of Ulva lactuca, as an alternative method for mercury removal from aqueous solutions, was investigated. Batch equilibrium tests showed that at pH 3.5, 5.5 and 7 the maxima of mercury uptake values, according to Langmuir adsorption isotherm, were 27.24, 84.74 and 149.25 mg/g, respectively. The ability of Ulva lactuca biomass to adsorb mercury in fixed-bed column, was investigated as well. The influence of column bed height, flow rate and effluent initial concentration of metal was studied. The adsorbed metal ions were easily desorbed from the algal biomass with 0.3 N H(2)SO(4) solution. After acid desorption and regeneration with distilled water, the biomass could be reused for other biosorption assays with similar performances.

Adsorption↗

Photoinhibition of photosynthesis in Macrocystis pyrifera (Phaeophyceae), Chondrus crispus (Rhodophyceae) and Ulva lactuca (Chlorophyceae) in outdoor culture systems.

The effect of solar radiation on photosynthesis and chlorophyll fluorescence associated to photosystem II (PS II) was determined in the Phaeophyta Macrocystis pyrifera, the Rhodophyta Chondrus crispus and the Chlorophyta Ulva lactuca by oxygen evolution and pulse-amplitude-modulated (PAM) fluorescence. The algae were maintained in 1.2 m3 outdoor tanks with constant aeration and at 8, 26 and 100% incident irradiance (E(o)). All three species showed a decrease in deltaF/F'm values during solar noon compared to values in the morning and afternoon, suggesting a photoinhibition of photosynthesis. In general, photoinhibition was negatively correlated to increasing daily irradiance in all three species. Photoinhibition in C. crispus occurred in tissue incubated at 8, 26 and 100% E(o), while in M. pyrifera and U. lactuca a decrease in deltaF/F'm values was only observed in tissue incubated at 100% E(o). This suggests that species that naturally grow at greater depths might be more susceptible to excessive light when cultured in shallow waters compared to species that naturally inhabit shallower depths. In M. pyrifera, deltaF/F'm values were lower in the afternoon than those in the morning, suggesting slower repair mechanisms of the photosystem II compared to the other species. The results suggest that photoinhibition could be reduced by reducing incident irradiance to culture systems or increasing of biomass to promote self-shading. Gross oxygenic photosynthesis increased linearly at low electron transport rates after which it saturated in all three species. This suggests that chlorophyll fluorescence could be used as an indicator of the physiological status of macroalgae maintained in dense aquaculture systems.

Chlorophyll↗

Fractionation of platinum group elements in aqueous systems: comparative kinetics of palladium and platinum removal from seawater by Ulva lactuca L.

A marine macroalga, Ulva lactuca L., was used as a substrate to compare the kinetics of palladium (Pd) and platinum (Pt) removal from seawater. This work indicates that, while the equilibrium behaviors of Pd and Pt are in many respects similar, their kinetic behaviors are quite distinct. The removal of both Pt(II) and Pt(IV) from seawater by U. lactuca is slower than the removal of Pd(II) by approximately an order of magnitude. Relative Pd and Pt removal rates are strongly influenced by system hydrodynamics. Under quiescent conditions, lambda(Pd)/lambda(Pt), the ratio of Pd and Pt removal rates, is 7 +/- 2, whereas under turbulent conditions lambda(Pd)/lambda(Pt) can be as large as 27. These observations suggest that the disparate kinetic behaviors of Pd and Pt may produce considerable differences in the environmental dispersion of these elements.

Biodegradation, Environmental↗

Highly abundant and stage-specific mRNAs in the obligate pathogen Bremia lactucae.

Germinating spores of the obligate pathogen Bremia lactucae (lettuce downy mildew) contain several unusually abundant species of mRNA. Thirty-nine cDNA clones corresponding to prevalent transcripts were isolated from a library synthesized using poly(A)+ RNA from germinating spores; these clones represented only five distinct classes. Each corresponding mRNA accounted for from 0.4 to 9 percent by mass of poly(A)+ RNA from germinating spores and together represented greater than 20 percent of the mRNA. The expression of the corresponding genes, and a gene encoding Hsp70, was analyzed in spores during germination and during growth in planta. The Hsp70 mRNA and mRNA from one abundant cDNA clone (ham34) were expressed constitutively. Two clones (ham9 and ham12) hybridized only to mRNA from spores and germinating spores. Two clones (ham37 and ham27) showed hybridization specific to germinating spores. Quantification of the number of genes homologous to each cDNA clone indicated that four clones corresponded to one or two copies per haploid genome, and one hybridized to an approximately 11-member family of genes. A sequence of the gene corresponding to ham34 was obtained to investigate its function and to identify sequences conferring high levels of gene expression for use in constructing vectors for the transformation of B. lactucae.

Amino Acid Sequence↗

Mycoplasma lactucae sp. nov., a sterol-requiring mollicute from a plant surface.

Strain 831-C4T (T = type strain), isolated from the surface of lettuce plants (Lactuca sativa) obtained from a retail food market, was shown to be a sterol-requiring mollicute. Morphological examination of this organism by electron and dark-field microscopic techniques showed that it consists of small, nonhelical, nonmotile, pleomorphic coccoid cells, with individual cells surrounded by a single cytoplasmic membrane. No evidence of a cell wall was observed. The organism grew rapidly in all conventional culture medium formulations for mollicutes in either aerobic or anaerobic environments. The optimum temperature for growth was 30 degrees C, but multiplication occurred at 18 to 37 degrees C. Strain 831-C4T catabolized glucose, but hydrolysis of arginine or urea could not be demonstrated. The genome size of strain 831-C4T was determined to be about 569 megadaltons, while the base composition (guanine-plus-cytosine content) of the DNA was 30.0 mol%. Recent studies in which we compared the 16S rRNA sequences of strain 831-C4T with those of more than 40 other mollicutes indicated that this organism is phylogenetically related to the Spiroplasma-Mycoplasma mycoides clade. Strain 831-C4T was serologically unrelated to the type strains of previously described Mycoplasma species and to 18 other unclassified sterol-requiring isolates cultivated from various animal, plant, or insect sources. Strain 831-C4T (= ATCC 49193) is the type strain of Mycoplasma lactucae sp. nov.

Mycoplasma↗

Introduction of xylem differentiation in lactuca by ethylene.

Evidence was obtained to support the hypothesis that ethylene is involved in xylem differentiation in primary pith explants of Lactuca sativa L. cv Romaine cultured in vitro. Xylem elements differentiated when explants were supplied indole-3-acetic acid (IAA) in combination with either the ethylene biosynthetic precursor 1-aminocyclopropane-1-carboxylic acid (ACC), the ethylene-releasing agent 2-chloroethylphosphonic acid (CEPA), or kinetin. In contrast, no xylem elements differentiated in the presence of IAA, kinetin, ACC, or CEPA alone, or when kinetin was supplied together with ACC or CEPA. These results show that ethylene will substitute qualitatively for cytokinin during auxin-induced xylogenesis, and suggest that both ethylene and auxin are required for xylem differentiation in Lactuca.

Journal Article↗

Phosphorus-31 NMR Studies of Cell Wall-Associated Calcium-Phosphates in Ulva lactuca.

Phosphate concentrations in the range 0.1 to 2.0 millimolar induced the formation of extracellular amorphous calcium-phosphates in the cell wall of the marine macro algae Ulva lactuca when they were cultivated in light in seawater at 20 degrees C. A broad resonance representing these compounds as well as resonances for extracellular orthophosphate and polyphosphates could be followed by (31)P-nuclear magnetic resonance spectroscopy. The presence of the calcium-phosphate made the cells brittle and it inhibited the growth of the macro algae and caused mortality within 1 week. The formation of the calcium-phosphates was influenced by the external phosphate concentration, the extracellular pH and the nature and concentration of the external nitrogen source. Furthermore, no formation of these compounds was observed when Ulva lactuca was cultivated in the dark, at low temperatures (5 degrees C) or in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. The complex could be removed through washes with ethylenediaminetetraacetate; this treatment did not alter the intracellular pH or the orthophosphate and polyphosphate pools and it restored growth.

Journal Article↗

Necrotizing fasciitis associated with Acinetobacter lactucae and Bacillus paramobilis infections in 2 dogs.

Two immunocompetent dogs from separate households were presented to a tertiary referral hospital with soft tissue wounds attributable to severe bacterial infection. A 5-y-old, castrated male Golden Retriever dog (case 1) was presented with extensive hemorrhagic cellulitis of the forelimbs and neck that developed over 48-h and was attributable to Acinetobacter lactucae infection. Additionally, an 8-y-old, spayed female Labrador Retriever-mix dog (case 2) sustained a deep penetrating wound over the left hip that progressed over 24 h to necrotizing fasciitis, from which Bacillus paramobilis was isolated. Histopathologic findings in both cases included severe acute, skeletal muscle necrosis and necrosuppurative myofasciitis, dermatitis, and panniculitis with intralesional rod-shaped bacteria. Whole-genome sequencing and phylogenetic analysis of the bacterial isolates revealed numerous cytolytic toxins and other virulence genes carried by the isolates, expanding the profile of these 2 bacteria. To our knowledge, canine cases of A. lactucae and B. paramobilis associated with necrotizing fasciitis have not been reported previously. In both cases, progression from an inciting incident to septic disease was extremely rapid, occurring within 48 h in case 1 and 24 h in case 2, underscoring the fulminant nature of necrotizing fasciitis.

Animals↗

Flagellar roots, mating structure and gametic fusion in the green alga Ulva lactuca (Ulvales).

The slightly anisogamous gametes of Ulva lactuca exhibit a cruciate flagellar root system consisting of 4 microtubular roots (4-2-4-2 system) and an elaborate system of fibrous roots associated with the 2-stranded microtubular roots. Two fibres (32-nm striation periodicity; system I fibres) closely underlie each of the 2-stranded roots, while different fibres (150-nm striation periodicity; system II fibres) run parallel to the root microtubules, and are 150-200 nm more internally located. Female gametes have 4 system II fibres, 3 of which are combined into a compound fibre associated with one microtubular root, while the fourth fibre is associated with the opposite root. In male gametes only 2 system II fibres are present, each underlying one of the two 2-stranded roots. A special region of the plasmalemma of both gamete types about 0.5 mum away from the basal bodies and located between 2 adjacent microtubular roots is structurally specialized and acts as a mating structure in gametic fusion. The region is oval-shaped and up to I.I mum long with a maximum diameter of 0.7 mum. A continuous electron-dense boundary layer underlies the plasmalemma at the edges of the mating structure. In both gamete types the mating structure consists of a fuzzy layer of material underlying the plasmalemma and special granules (60 nm diameter) are associated with this layer on its cytoplasmic side. In addition diffuse material overlies the mating structure, especially in male gametes. The mating structure is connected to 3 different kinds of flagellar roots: the boundary layer is linked to a 2-stranded microtubular root and its associated system I fibre; the fuzzy layer of the mating structure is connected with a system II fibre; and in female gametes this is the compound system II fibre. The ultrastructural changes which occur after mixing the 2 gamete types have been followed. Mating structure activation involves contraction of system II fibres (change of striation periodicity to 100 nm), detachment of special granules from the fuzzy layer of the mating structure and their replacement by electron-transparent vesicles at the prospective cell fusion site. Furthermore, release of electron-dense contents from Golgi-derived vesicles in the anterior part of both gamete types precedes cell fusion. Cell fusion is exclusively initiated in a region delimited by the 2 mating structures. After partial dissolution the 2 plasma membranes unite within the mating structure regions. The ultrastructure of gametic fusion in Ulva lactuca is compared to that of other green algae and the significance of flagellar roots in the mating process of green algae is discussed.

Chlorophyta↗

From aerial drone to quantitative trait locus: leveraging next-generation phenotyping to reveal the genetics of color and height in field-grown Lactuca sativa.

In recent years, accurate and low-cost variant calling has enabled the genotyping of large diversity panels for genome-wide association studies. As a result, phenotyping rather than genotyping is now the rate-limiting step, especially in field experiments. This has created a strong need for high-throughput, accurate, and low-cost in-field phenotyping. Here, we present a genome-wide association study (GWAS) study on 194 field-grown accessions of lettuce (Lactuca sativa). These accessions were non-destructively phenotyped at two time points 15 days apart using a drone equipped with an RGB and multispectral (MSP) camera. Our high-throughput phenotyping approach integrates an RGB- and MSP camera to measure the color and height of lettuce in this large-scale field experiment. We used the mean and other summary statistics, such as median, quantiles, skewness, kurtosis, minimum, and maximum to quantify different aspects of color and height variation in lettuce from the drone images. Using these summary statistics as traits for GWAS, we confirm several previously described genetic associations, now under field conditions, and identify additional novel associations for color and height traits in lettuce.

Lactuca↗

Velvetbean (Mucuna pruriens) extracts: impact on Meloidogyne incognita survival and on Lycopersicon esculentum and Lactuca sativa germination and growth.

Velvetbean (Mucuna spp.) is a summer annual that has been used as a cover crop to reduce erosion, fix nitrogen and suppress weeds and plant-parasitic nematodes. Crude aqueous extracts (1:15 dry weight plant/volume water) were made from velvetbean plant parts, and various concentrations of the extracts were evaluated in vitro for toxicities to different stages of Meloidogyne incognita (Kofoid and White) Chitwood and for suppression of hypocotyl and root growth and inhibition of germination of tomato (Lycopersicon esculentum L.) and lettuce (Lactuca sativa L.). Germination was only affected by the full-strength extract from leaf blades. Lettuce root growth was the most sensitive indicator of allelopathic activity of the plant part extracts. Lettuce and tomato root growth was more sensitive to the extract from main roots than to extracts of other plant parts, with lethal concentration (LC50) values of 1.2 and 1.1% respectively. Meloidogyne incognita egg hatch was less sensitive to extracts from velvetbean than the juvenile (J2) stage. There was no difference among LC50 values of the extracts from different plant parts against the egg stage. Based on LC50 values, the extract from fine roots was the least toxic to J2 (LC50 39.9%), and the extract from vines the most toxic (LC50 7.8%). The effects of the extracts were nematicidal because LC50 values did not change when the extracts were removed and replaced with water.

Animals↗

Molecular analysis of irradiation-induced and spontaneous deletion mutants at a disease resistance locus in Lactuca sativa.

The major cluster of disease resistance genes in lettuce (Lactuca sativa) contains at least nine downy mildew resistance genes (Dm) spanning a genetic distance of 20cM and a physical distance of at least 6 Mb. Nine molecular markers that were genetically tightly linked to Dm3 were used to analyze nine independent deletion mutants and construct a map of the region surrounding Dm3. This analysis identified a linear order of deletion breakpoints and markers along the chromosome. There was no evidence for chromosomal rearrangements associated with the deletions. The region is not highly recombinagenic and the deletion breakpoints provided greater genetic resolution than meiotic recombinants. The region contains a mixture of high-and low-copy number sequences; no single-copy sequences were detected. Three markers hybridized to low-copy-number families of sequences that are duplicated predominantly close to Dm3. This was not true for sequences related to the triose-phosphate isomerase gene; these had been shown previously to be linked to Dm3, as well as to two independent clusters of Dm genes, and elsewhere in the genome. Two spontaneous mutants of Dm3 were identified; several markers flanking Dm3 are absent in one of these two mutants. The stability of the Dm3 region was also studied by analyzing the genotypes of diverse related cultivars. The 1.5 Mb region surrounding Dm3 has remained stable through many generations of breeding with and without selection for Dm3 activity.

Chromosome Mapping↗

Phytol, SSADH inhibitory diterpenoid of Lactuca sativa.

The succinic semialdehyde dehydrogenase (SSADH) inhibitory component was isolated from the EtOAc fraction of Lactuca sativa through repeated column chromatography; then, it was identified as phytol, a diterpenoid, based on the interpretation of several spectral data. Incubation of SSADH with the phytol results in a time-dependent loss of enzymatic activity, suggesting that enzyme modification is irreversible. The inactivation followed pseudo-first-order kinetics with the second-rate order constant of 6.15 x 10(-2) mM(-1) min(-1). Complete protection from inactivation was afforded by the coenzyme NAD+, whereas substrate succinic semialdehyde failed to prevent the inactivation of the enzyme; therefore, it seems likely that phytol covalently binds at or near the active site of the enzyme. It is postulated that the phytol is able to elevate the neurotransmitter GABA levels in central nervous system through its inhibitory action on one of the GABA degradative enzymes, SSADH.

Aldehyde Oxidoreductases↗

The development of lettuce backcross inbred lines (BILs) for exploitation of the Lactuca saligna (wild lettuce) germplasm.

Backcross inbred lines (BILs) were developed in which chromosome segments of Lactuca saligna (wild lettuce) were introgressed into L. sativa (lettuce). These lines were developed by four to five backcrosses and one generation of selfing. The first three generations of backcrossing were random. Marker-assisted selection began in the BC(4) generation and continued until the final set of BILs was reached. A set of 28 lines was selected that together contained 96% of the L. saligna genome. Of these lines, 20 had a single homozygous introgression (BILs), four had two homozygous introgressions (doubleBILs) and four lines had a heterozygous single introgression (preBILs). Segregation ratios in backcross generations were compared to distorted segregation ratios in an F(2) population, and the results indicated that most of the distorted segregations can be explained by genetic effects on pollen- or egg-cell fitness. By means of BIL association mapping we were able to map 12 morphological traits and hundreds of additional amplified fragment length polymorphic (AFLP) markers. The total AFLP map now comprises 757 markers. This set of BILs is very useful for future genetic studies.

Chromosome Mapping↗

Quantitative trait loci associated with seed and seedling traits in Lactuca.

Seed and seedling traits related to germination and stand establishment are important in the production of cultivated lettuce (Lactuca sativa L.). Six seed and seedling traits segregating in a L. sativa cv. Salinas x L. serriola recombinant inbred line population consisting of 103 F8 families revealed a total of 17 significant quantitative trait loci (QTL) resulting from three seed production environments. Significant QTL were identified for germination in darkness, germination at 25 and 35 degrees C, median maximum temperature of germination, hypocotyl length at 72 h post-imbibition, and plant (seedling) quality. Some QTL for germination and early seedling growth characteristics were co-located, suggestive of pleiotropic loci regulating these traits. A single QTL (Htg6.1) described 25 and 23% of the total phenotypic variation for high temperature germination in California- and Netherlands-grown populations, respectively, and was significant between 33 and 37 degrees C. Additionally, Htg6.1 showed significant epistatic interactions with other Htg QTL and a consistent effect across all the three seed production environments. L. serriola alleles increased germination at these QTL. The estimate of narrow-sense heritability (h2) of Htg6.1 was 0.84, indicating potential for L. serriola as a source of germination thermotolerance for lettuce introgression programs.

Breeding↗

A note on the measurement of genetic diversity within genebank accessions of lettuce (Lactuca sativa L.) using AFLP markers.

This paper discusses a statistical approach for measuring genetic diversity within genebank accessions of a self-fertilising species. This approach is applied to lettuce (Lactuca sativa L.), using AFLP marker data on a set of 1,390 accessions, representing six different lettuce types. Knowledge of the within-accession genetic diversity is important for decisions about the way accessions have to be maintained by genebanks. It is argued that if the within-accession diversity is small, as can be expected for a self-fertilising species like L. sativa, the best approach is to sample as many accessions as possible with only two plants per accession and estimate the within-accession diversity by the proportion of accessions of which the individuals are different.

DNA, Plant↗