Proteomics Analysis Indicates Greater Abundance of Proteins Involved in Major Metabolic Pathways in Lactuca sativa cv. Salinas than Lactuca serriola Accession US96UC23.

Lettuce (Lactuca sativa), cultivated mainly for its edible leaves and stems, is an important vegetable crop worldwide. Genomes of cultivated lettuce (L. sativa cv. Salinas) and its wild relative L. serriola accession US96UC23 are sequenced, but a clear understanding of the genetic basis for divergence in phenotypes of the two species is lacking. Tandem mass tag (TMT) based mass spectrometry is used to quantitatively compare protein levels between these two species. Four-day old seedlings is transplanted into 500 mL pots filled with soil. Plants are grown for 8 weeks under 250 µmol m-2 sec-1 continuous light, 20 °C and relative humidity between 50-70%. Leaf discs (1 cm diameter) from three individuals per biological replicate are analyzed. A total of 3000 proteins are identified, of which the levels of 650 are significantly different between 'Salinas' and US96UC23. Pathway analysis indicated a higher flux of carbon in 'Salinas' than US96UC23. Many essential metabolic pathways such as tetrapyrrole metabolism and fatty acid biosynthesis are upregulated in 'Salinas' compared with US96UC23. This study provides a reference proteome for researchers interested in understanding lettuce biology and improving traits for cultivation.

Preponderance of Oxygenated Sesquiterpenes and Diterpenes in the Volatile Oil Constituents of Lactuca serriola L. Revealed Antioxidant and Allelopathic Activity.

Using synthetic chemicals in industry and agriculture has led to several environmental problems. Thus, plant products derived from volatile oils (VOs) could be a potential green source for bioherbicides. Little is known about the VOs of Lactuca serriola. Hence, the present study aimed to characterize the VOs chemical composition from the aerial parts of L. serriola, assessment of antioxidant activity, and evaluate allelopathic potential against the noxious weed Bidens pilosa. The VOs were extracted by hydrodistillation and analyzed by GC/MS. The VOs from the Egyptian ecospecies of L. serriola were found to comprise 34 compounds mainly oxygenated sesquiterpenes and diterpenes. The major compounds were isoshyobunone (64.22 %), isocembrol (17.35 %), and alloaromadendrene oxide-1 (7.32 %). So, L. serriola can be considered as a good source for isoshyobunone, considering that it has a much higher concentration than any other plants. Also, this plant has a high content of the oxygenated diterpene compound, isocembrol, which is rarely found in the VOs of most plants. The VOs expressed strong antioxidant activity. Also, for the first time, our results showed a strong allelopathic activity of VOs from L. serriola on germination and seedling growth of the noxious weed, B. pilosa. We suppose that the activity of the VOs from L. serriola could be attributed to these previously mentioned major compounds, as they represent about 89 % of the total identified oil constituents. Nevertheless, to evaluate these compounds as new allelochemicals, further study is needed to test the allelopathic activity of authenticated standard of these compounds either singular or in combination on several weeds as well as evaluate the safety, and improve the efficacy and stability at the field scale.

Does co-inoculation of Lactuca serriola with endophytic and arbuscular mycorrhizal fungi improve plant growth in a polluted environment?

Phytoremediation of polluted sites can be improved by co-inoculation with mycorrhizal and endophytic fungi. In this study, the effects of single- and co-inoculation of Lactuca serriola with an arbuscular mycorrhizal (AM) fungus, Rhizoglomus intraradices, and endophytic fungi, Mucor sp. or Trichoderma asperellum, on plant growth, vitality, toxic metal accumulation, sesquiterpene lactone production and flavonoid concentration in the presence of toxic metals were evaluated. Inoculation with the AM fungus increased biomass yield of the plants grown on non-polluted and polluted substrate. Co-inoculation with the AM fungus and Mucor sp. resulted in increased biomass yield of plants cultivated on the polluted substrate, whereas co-inoculation with T. asperellum and the AM fungus increased plant biomass on the non-polluted substrate. In the presence of Mucor sp., mycorrhizal colonization and arbuscule richness were increased in the non-polluted substrate. Co-inoculation with the AM fungus and Mucor sp. increased Zn concentration in leaves and roots. The concentration of sesquiterpene lactones in plant leaves was decreased by AM fungus inoculation in both substrates. Despite enhanced host plant costs caused by maintaining symbiosis with numerous microorganisms, interaction of wild lettuce with both mycorrhizal and endophytic fungi was more beneficial than that with a single fungus. The study shows the potential of double inoculation in unfavourable environments, including agricultural areas and toxic metal-polluted areas.

Molecular Biogeography of Prickly Lettuce (Lactuca serriola L.) Shows Traces of Recent Range Expansion.

Prickly lettuce (Lactuca serriola L., Asteraceae), a wild relative of cultivated lettuce, is an autogamous species which greatly expanded throughout Western and Northern Europe during the last 2 centuries. Here, we present a large-scale biogeographic genetic analysis performed on a dataset represented by 2622 individuals from 110 wild European populations. Thirty-two maternally inherited chloroplast RFLP-markers and 10 nuclear microsatellite loci were used. Microsatellites revealed low genetic variation and high inbreeding coefficients within populations, as well as strong genetic differentiation between populations, which was in accordance with the autogamous breeding system. Analysis of molecular variance based clustering indicated the presence of 3 population clusters, which showed strong geographical patterns. One cluster occupied United Kingdom and part of Northern Europe, and characterized populations with a single predominant genotype. The second mostly combined populations from Northern Europe, while the third cluster grouped populations particularly from Southern Europe. Kriging of gene diversity for L. serriola corroborated northwards and westwards spread from Central (Eastern) Europe. Significant lower genetic diversity characterized the newly colonized parts of the range compared to the historical ones, confirming the importance of founder effects. Stronger pattern of isolation by distance was assessed in the newly colonized areas than in the historical areas (Mantel's r = 0.20). In the newly colonized areas, populations at short geographic distances were genetically more similar than those in the historical areas. Our results corroborate the species' recent and rapid northward and westward colonization from Eastern Europe, as well as a decrease of genetic diversity in recently established populations.

Evaluation of the Anxiolytic and Anti-Epileptogenic Potential of Lactuca Serriola Seed Using Pentylenetetrazol-Induced Kindling in Mice and Metabolic Profiling of Its Bioactive Extract.

This study aimed to assess the potential of Lactuca serriola (Asteraceae) seed n-hexane, chloroform, methanol, and aqueous extracts as anticonvulsant, sedative, anticonvulsant and antiepileptic agents in Swiss albino mice. Different doses of each extract were evaluated for the anxiolytic potential using the hole-board, the elevated plus maze and the light/dark test. A phenobarbitone-induced sleep test was employed for the evaluation of sedative potential. Acute anticonvulsant activity was evaluated by picrotoxin and strychnine-induced convulsion models. All extracts significantly reduced the number of head dips where n-hexane extract (400 mg/kg) showed 96.34% reduction in the tendency of head dipping when compared with the control. Mice treated with extracts preferred elevated plus maze open arms and were shown to lack open arms evasion, especially n-hexane extract (400 mg/kg)-which showed 456.14%-increased the duration of open arm stay with the respective control group. By reducing sleep latency and greatly lengthening sleep duration, L. serriola enhanced the effects of barbiturate-induced sleep. A significant increase in convulsion latency and decrease in convulsions induced by picrotoxin and strychnine duration was observed in all extract-treated groups. All the extracts exhibited anti-epileptogenic potential as the seizure score in pentylenetetrazol (PTZ)-induced kindling in mice was reduced significantly. Maximum protection was afforded by chloroform extract that reduced the seizure score by 79.93% compared with the PTZ group. Chloroform executed antioxidant effect by elevating super oxide dismutase (SOD) by 126%, catalase (CAT) by 83.53%, total glutathione (tGSH) by 149%, and reducing malondialdhyde (MDA) levels by 36.49% in the brain tissues that is further consolidated by histopathological examination. Metabolic profiling of the most active chloroform extract using Gas chromatography coupled with mass showed the presence of 16 compounds. This anti-epileptic activity was further confirmed via in silico molecular modelling studies in the active site Gamma-aminobutyric acid aminotransferase (GABA-AT) where all of the tested metabolites illustrated a potent inhibitory potential towards GABA-AT with hexadecanoic acid, 15-methyl-, methyl ester followed by octadecanoic acid, methyl ester showed the best fitting. The results indicated the possible anxiolytic and anti-epileptogenic potential of the plant and further consolidated the ethnopharmacological use of L. serriola seeds.

Sequence of the Mitochondrial Genome of Lactuca virosa Suggests an Unexpected Role in Lactuca sativa's Evolution.

The involvement of the different Lactuca species in the domestication and diversification of cultivated lettuce is not totally understood. Lactuca serriola is considered as the direct ancestor and the closest relative to Lactuca sativa, while the other wild species that can be crossed with L. sativa, Lactuca virosa, and Lactuca saligna, would have just contributed to the latter diversification of cultivated typologies. To contribute to the study of Lactuca evolution, we assembled the mtDNA genomes of nine Lactuca spp. accessions, among them three from L. virosa, whose mtDNA had not been studied so far. Our results unveiled little to no intraspecies variation among Lactuca species, with the exception of L. serriola where the accessions we sequenced diverge significantly from the mtDNA of a L. serriola accession already reported. Furthermore, we found a remarkable phylogenetic closeness between the mtDNA of L. sativa and the mtDNA of L. virosa, contrasting to the L. serriola origin of the nuclear and plastidial genomes. These results suggest that a cross between L. virosa and the ancestor of cultivated lettuce is at the origin of the actual mitochondrial genome of L. sativa.

The use of strip-seeding for management of two late-season invasive plants.

The spread and persistence of weedy plants in rangelands highlight the need for refinement of existing management techniques and development of novel strategies to address invasions. Strip-seeding - the strategic seeding of a portion of an invaded area to reduce costs and enhance success - is an underutilized management approach that holds promise for reducing weed dominance in grassland habitats. A strip-seeding experiment was established in 2011 in a California grassland where portions (between 0-100%) of invaded plots were seeded with native grasses. In 2016, we assessed the height, above-ground biomass and flower production of two late-season invasive plants: field bindweed and prickly lettuce. We found significant reductions in plant height and flower production (for both target invasives), and biomass (for field bindweed) in many of the seeded strips compared to the unseeded strips. Smaller seed applications demonstrated similar or better utility for weed control compared to greater seed applications, suggesting that this approach can be effective while reducing labor and materials cost of typical restoration management approaches. We did not find evidence that seeded strips provided invasion resistance to unseeded strips. This is possibly due to the lag in native species dispersal and establishment into contiguous unseeded strips, and suggests that strip-seeding might not provide invasion resistance to unseeded strips on timescales that are relevant to managers. However, this work does suggest that strip-seeding native species that overlap in phenology with target invasives can reduce late-season weed dominance on rangelands.

Genetic and biochemical evaluation of natural rubber from Eastern Washington prickly lettuce (Lactuca serriola L.).

Alternative sources of natural rubber are of importance due to economic, biological, and political threats that could diminish supplies of this resource. Prickly lettuce (Lactuca serriola L.) synthesizes long-chain natural rubber and was studied to determine underlying genetic and phenotypic characteristics of rubber biosynthesis. Genotypic and phenotypic analysis of an F2 segregating population using EST-SSR markers led to the discovery of genetic regions linked to natural rubber production. Interval mapping (IM) and multiple QTL mapping (MQM) identified several QTL in the mapping population that had significance based on LOD score thresholds. The discovered QTL and the corresponding local markers are genetic resources for understanding rubber biosynthesis in prickly lettuce and could be used in marker-assisted selection (MAS) breeding. Prickly lettuce is an excellent candidate for elucidating the rubber synthesis mechanism and has potential as a crop plant for rubber production.