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Lactuca saligna, a non-host for lettuce downy mildew ( Bremia lactucae), harbors a new race-specific Dm gene and three QTLs for resistance.

Lactuca sativa (lettuce) is susceptible to Bremia lactucae (downy mildew). In cultivated and wild Lactuca species, Dm genes have been identified that confer race-specific resistance. However, these genes were soon rendered ineffective by adaptation of the pathogen. Lactuca saligna (wild lettuce) is resistant to all downy mildew races and can be considered as a non-host. Therefore, L. saligna might be an alternative source for a more-durable resistance to downy mildew in lettuce. In order to analyze this resistance, we have developed an F(2) population based on a resistant L. saligna x susceptible L. sativa cross. This F(2) population was fingerprinted with AFLP markers and tested for resistance to two Bremia races NL14 and NL16. The F(2) population showed a wide and continuous range of resistance levels from completely resistant to completely susceptible. By comparison of disease tests, we observed a quantitative resistance against both Bremia races as well as a race-specific resistance to Bremia race NL16 and not to NL14. QTL mapping revealed a qualitative gene ( R39) involved in the race-specific resistance and three QTLs ( RBQ1, RBQ2 and RBQ3) involved in the quantitative resistance. The qualitative gene R39 is a dominant gene that gives nearly complete resistance to race NL16 in L. saligna CGN 5271 and therefore it showed features similar to Dm genes. The three QTLs explained 51% of the quantitative resistance against NL14, which indicated that probably only the major QTLs have been detected in this F(2) population. The perspectives for breeding for durable resistance are discussed.

Journal Article↗

Evidence for abrasion and enhanced growth of Ulva lactuca L. in the presence of colliery waste particles.

Previous studies have highlighted a reduction in occurrence and biomass of Ulva lactuca L. (Chlorophyceae) on shores where inputs of colliery waste occur. It was postulated that this was owing to an abrasive effect of colliery waste on macroalgal fronds. To test this, individual U. lactuca plants were exposed to colliery waste (three different grain size categories: < 500 microm, 500-2000 microm, and 0-2000 microm) in both shaken (turbulent) and still conditions in the laboratory. Over an 8-day period, U. lactuca plants lost weight when colliery waste was present and gained weight when no colliery waste was present. Weight loss was most pronounced in U. lactuca exposed to colliery waste of 500-2000 microm grain sizes in shaken conditions (- 18.8% +/- 4.65 SE, n = 4). However, greatest weight gain (+ 26.8% +/- 6.26 SE, n = 4) also occurred in the presence of colliery waste (0-2000 microm), but in still conditions. Weight gain was also observed after 60 days in still conditions in the presence of colliery waste (grain sizes 0-2000 microm); U. lactuca showed significant growth (+ 69% +/- 67 SE of starting weight, n = 5) compared with controls (- 51% +/- 41 SE, n = 5). These results suggest that 'large' grains of colliery waste act as a physical abrading agent on macroalgae when in turbulent conditions, and may be responsible for lowering of species richness of macroalgae where colliery waste inputs occur. However, by contrast, colliery waste in still conditions promotes the growth of U. lactuca, suggesting that, for example, rock pool flora may benefit from its presence

Journal Article↗

Use of Ulva lactuca to identify ammonia toxicity in marine and estuarine sediments.

Toxicity identification evaluation (TIE) methods are being developed for use with whole sediments. Although a phase I TIE method has been developed to characterize ammonia toxicity in aqueous samples using the marine macroalga Ulva lactuca, the relationship between amphipod and mysid mortality and uptake by U. lactuca of bedded sediment ammonia had not been explored. Additionally, it was not known how interactions in whole sediments between metal and organic contaminants with U. lactuca and ammonia would affect TIE interpretation. The current study showed that ammonia toxicity to amphipods and mysids was reduced significantly in marine sediments in the presence of U. lactuca. The alga slightly affected metal concentrations but did not alter the concentration of organic contaminants in either overlying or interstitial waters. Conversely, ammonia uptake by U. lactuca was not affected by the presence of metal or organic contaminants in the sediments. When used with other TIE manipulations, U. lactuca can be utilized in a whole-sediment, phase I TIE to remove toxicity due to ammonia.

Ammonia↗

A Lactuca universal hybridizer, and its use in creation of fertile interspecific somatic hybrids.

A Lactuca sativa cv. Ardente line heterozygous for a gene encoding resistance to kanamycin, a positive and dominant trait, was crossed with cv. Girelle, which is heterozygous for a recessive albinism marker. The resulting seeds yielded 25% albino seedlings, of which 50% were also resistant to kanamycin. Such plantlets (KR, a) grown in vitro were used for preparation of universal hybridizer protoplasts, since green buds that can develop on kanamycin containing-medium should result from fusion with any wild-type protoplast. To test the practicability of this selection scheme, we fused L. sativa KR, a protoplasts with protoplasts derived from various wild Lactuca as well as various other related species. Protoplast-derived cell colonies were selected for resistance to kanamycin at the regeneration stage. Green buds were regenerated after fusion with protoplasts of L. tatarica and of L. perennis. So far, 9 interspecific hybrid plants have been characterized morphologically. In addition, random amplified polymorphic DNA (RAPD) analysis with selected primers confirmed that these plants are indeed interspecific hybrids. Some plants are female-fertile and production of backcross progenies with L. sativa is in progress. Since many desirable traits such as resistances to viruses, bacteria and fungi (Bremia lactucae) have been characterized in wild Lactuca species, the use of somatic hybridization in breeding programmes now appears a practical possibility.

Base Sequence↗

Metabolite profiling of sesquiterpene lactones from Lactuca species. Major latex components are novel oxalate and sulfate conjugates of lactucin and its derivatives.

Wounding leaves or stems of Lactuca species releases a milky latex onto the plant surface. We have examined the constituents of latex from Lactuca sativa (lettuce) cv. Diana. The major components were shown to be novel 15-oxalyl and 8-sulfate conjugates of the guaianolide sesquiterpene lactones, lactucin, deoxylactucin, and lactucopicrin. The oxalates were unstable, reverting to the parent sesquiterpene lactone on hydrolysis. Oxalyl derivatives have been reported rarely from natural sources. The sulfates were stable and are the first reported sesquiterpene sulfates from plants. Unusual tannins based on 4-hydroxyphenylacetyl conjugates of glucose were also identified. Significant qualitative and quantitative variation was found in sesquiterpene lactone profiles in different lettuce varieties and in other Lactuca spp. The proportions of each conjugate in latex also changed depending on the stage of plant development. A similar profile was found in chicory, in which oxalyl conjugates were identified, but the 8-sulfate conjugates were notably absent. The presence of the constitutive sesquiterpene lactones was not correlated with resistance to pathogens but may have a significant bearing on the molecular basis of the bitter taste of lettuce and related species. The induced sesquiterpene lactone phytoalexin, lettucenin A, was found in the Lactuca spp. but not in chicory.

Chromatography, High Pressure Liquid↗

[Lettuce, lactuca sp., as a medicinal plant in polish publications of the 19th century].

Mentions of lettuce Lactuca sp. that have appeared since antiquity contained similar information on its curative properties, but such properties were ascribed to different species or varieties. Apart from the wild and poisonous lettuce, also garden or common lettuce were identified as having curative action, and some publications lacked information enabling the precise identification of the lettuce in question. In the 19th century, attempts were made to put some order into the knowledge of lettuce as a medicinal plant. Information contained in Polish medical studies of the 19th century on lettuce points to the poisonous species, Lactuca virosa, and the common or garden lettuce, Lactuca sativa v. Lactuca hortensis, as being used as a medicinal plant. In that period, lettuce and especially the the desiccated lactescent juice obtained from it, lactucarium, were considered to be an intoxicant, and were used as a sedative and an analgesic. The action of the substance was weaker than that of opium but free of the side-effects, and medical practice showed that in some cases lactucarium produced better curative effects than opium. To corroborate those properties of lettuce and its lactescent juice, studies were undertaken to find the substance responsible for the curative effects of the juice. However, such studies failed to produce the expected results, and the component responsible for the curative properties of letuce was not identified. Medical practice thus had to restrict itself to the uses of the desiccated lactescent juice and extracts obtained from it. The possibility of obtaining lactucarium from plants cultivated in Poland caused Polish pharmacists and physicians to take an interest in the stuff and launch their own research of lettuce and the lactescent juice obtained from it. Results of research on lettuce were published in 19th-century journals by, among others, Jan Fryderyk Wolfgang, Florian Sawiczewski and Józef Orkisz.

History, 19th Century↗

Removal of ammonia toxicity in marine sediment TIEs: a comparison of Ulva lactuca, zeolite and aeration methods.

Toxicity Identification Evaluations (TIEs) can be used to determine the specific toxicant(s), including ammonia, causing toxicity observed in marine sediments. Two primary TIE manipulations are available for characterizing and identifying ammonia in marine sediments: Ulva lactuca addition and zeolite addition. In this study, we compared the efficacy of these methods to (1) remove NH(x) and NH(3) from overlying and interstitial waters and (2) reduce toxicity to the amphipod Ampelisca abdita and mysid Americamysis bahia using both spiked and environmentally contaminated sediments. The utility of aeration for removing NH(x) and NH(3) during a marine sediment TIE was also evaluated preliminarily. In general, the U. lactuca and zeolite addition methods performed similarly well at removing spiked NH(x) and NH(3) from overlying and interstitial waters compared to an unmanipulated sediment. Toxicity to the amphipod was reduced approximately the same by both methods. However, toxicity to the mysid was most effectively reduced by the U. lactuca addition indicating this method functions best with epibenthic species exposed to ammonia in the water column. Aeration removed NH(x) and NH(3) from seawater when the pH was adjusted to 10; however, very little ammonia was removed at ambient pHs ( approximately 8.0). This comparison demonstrates both U. lactuca and zeolite addition methods are effective TIE tools for reducing the concentrations and toxicity of ammonia in whole sediment toxicity tests.

Ammonia↗

Genetic analysis of the fungus, Bremia lactucae, using restriction fragment length polymorphisms.

Restriction fragment length polymorphisms (RFLPs) were developed as genetic markers for Bremia lactucae, the biotrophic Oomycete fungus which causes lettuce downy mildew. By using 55 genomic and cDNA probes, a total of 61 RFLP loci were identified among three heterothallic isolates of B. lactucae. Of these 61 RFLP loci, 53 were heterozygous in at least one of the three strains and thus were informative for linkage analysis in at least one of two F1 crosses that were performed. Analysis of the cosegregation of these 53 RFLPs, eight avirulence loci and the mating type locus allowed the construction of a preliminary genetic linkage map consisting of 13 small linkage groups. Based on the extent of linkage detected among probes, the genome of B. lactucae can be estimated to be approximately 2000 cM. Linkage was detected between a RFLP locus and an avirulence gene, providing a potential starting point for chromosome walking to clone an avirulence gene. The high frequency of DNA polymorphism in naturally occurring isolates and the proper Mendelian segregation of loci detected by low copy number probes indicates that it will be possible to construct a detailed genetic map of B. lactucae using RFLPs as markers. The method of analysis employed here should be applicable to many other outbreeding, heterozygous species for which defined inbred lines are not available.

Blotting, Southern↗

The disease resistance gene Dm3 is infrequent in natural populations of Lactuca serriola due to deletions and frequent gene conversions at the RGC2 locus.

Resistance genes can exhibit heterogeneous patterns of variation. However, there are few data on their frequency and variation in natural populations. We analysed the frequency and variation of the resistance gene Dm3, which confers resistance to Bremia lactucae (downy mildew) in 1033 accessions of Lactuca serriola (prickly lettuce) from 49 natural populations. Inoculations with an isolate of Bremia lactucae carrying avirulence gene Avr3 indicated that the frequency of Dm3 in natural populations of L. serriola was very low. Molecular analysis demonstrated that Dm3 was present in only one of the 1033 wild accessions analysed. The sequence of the 5' region of Dm3 was either highly conserved among accessions, or absent. In contrast, frequent chimeras were detected in the 3' leucine-rich repeat-encoding region. Therefore low frequency of the Dm3 specificity in natural populations was due to either the recent evolution of Dm3 specificity, or deletions of the whole gene as well as variation in 3' region caused by frequent gene conversions. This is the most extensive analysis of the prevalence of a known disease resistance gene to date, and indicates that the total number of resistance genes in a species may be very high. This has implications for the scales of germplasm conservation and exploitation of sources of resistance.

5' Untranslated Regions↗

Seaweeds along KwaZulu-Natal coast of South Africa-3: elemental uptake by Ulva lactuca (Sea lettuce).

The elemental uptake by Ulva lactuca (Sea lettuce), a marine macro-algae (chlorophyta, green alga) grown richly along KwaZulu-Natal coastline. The total elemental concentrations of seven important elements, namely manganese, iron, arsenic, boron, titanium, zinc and mercury, selected based on their abundance in U. lactuca were investigated for one year cycle (June 2002 to May 2003). The four selected sampling sites, Zinkwasi, Ballito, Treasure Beach and Park Ryrie are spread over 150 km wide along the KwaZulu-Natal coastline from North to South. The Ulva lectuca possess good manganese and arsenic accumulating ability and an excellent bio-indicator for most of the metals studied. A typical U. lectuca sample at Zinkwasi (in winter) recorded Mn (25.3 +/- 1.16 ppm), Fe (21.0 +/- 0.85 ppm), As (6.2 +/- 0.30 ppm), B (935 +/- 14 ppb), Ti (863 +/- 34 ppb), Zn (421 +/- 21 ppb), and Hg (61.3 +/- 1.2 ppb). The general trend found at all sites was high elemental concentrations in winter and a decrease in concentrations from winter to spring and summer. Iron uptake was lowest in summer and autumn at all sites. Ulva lactuca recorded highest mercury levels (>400 ppb) during the spring season at the Treasure Beach site near Durban.

Arsenic↗

Phosphorus-31 and Nitrogen- 14 NMR Studies of the Uptake of Phosphorus and Nitrogen Compounds in the Marine Macroalgae Ulva lactuca.

Cytoplasmic phosphomonoesters and inorganic phosphate, as well as vacuolar inorganic phosphate and polyphosphates, gave rise to the major peaks in (31)P nuclear magnetic resonance (NMR) spectra of the marine macroalgae Enteromorpha sp., Ceramium sp., and Ulva lactuca which were collected from the sea. In contrast, NMR-visible polyphosphates were lacking in Pylaiella sp. and intracellular vacuolar phosphate seemed to act as the main phosphorus store in this organism. In laboratory experiments, polyphosphates decreased in growing U. lactuca which was cultivated in continuous light under phosphate-deficient conditions. In contrast, the same organism cultivated in seawater with added phosphate and ammonium, accumulated phosphate mainly in the form of polyphosphates. When nitrate was provided as the only nitrogen source, accumulation of polyphosphates in the algae decreased with increasing external nitrate concentration. From the chemical shift of the cytoplasmic Pi peak, the cytoplasmic pH of superfused preparations of Ulva was estimated at 7.2. The vacuolar pH, determined from the chemical shifts of the vacuolar Pi and the terminal polyphosphate peaks, was between 5.5 and 6.0. The intracellular nitrate and ammonium levels in U. lactuca were determined by (14)N NMR. Both nitrogen sources were taken up and stored intracellularly; however, the uptake of ammonium was much faster than that of nitrate.

Journal Article↗

Utilization of Inorganic Carbon by Ulva lactuca.

Thalli discs of the marine macroalga Ulva lactuca were given inorganic carbon in the form of HCO(3) (-), and the progression of photosynthetic O(2) evolution was followed and compared with predicted O(2) evolution as based on calculated external formation of CO(2) (extracellular carbonic anhydrase was not present in this species) and its carboxylation (according to the K(m)(CO(2)) of ribulose-1,5-bisphosphate carboxylase/oxygenase), at two different pHs, assuming a photosynthetic quotient of 1. The K(m)(inorganic carbon) was some 2.5 times lower at pH 5.6 than at the natural seawater pH of 8.2, whereas V(max) was similar under the two conditions, indicating that the unnaturally low pH per se had no adverse effect on U. lactuca's photosynthetic performance. These results, therefore, could be evaluated with regard to differential CO(2) and HCO(3) (-) utilization. The photosynthetic performance observed at the lower pH largely followed that predicted, with a slight discrepancy probably reflecting a minor diffusion barrier to CO(2) uptake. At pH 8.2, however, dehydration rates were too slow to supply CO(2) for the measured photosynthetic response. Given the absence of external carbonic anhydrase activity, this finding supports the view that HCO(3) (-) transport provides higher than external concentrations of CO(2) at the ribulose-1,5-bisphosphate carboxylase/oxygenase site. Uptake of HCO(3) (-) by U. lactuca was further indicated by the effects of potential inhibitors at pH 8.2. The alleged band 3 membrane anion exchange protein inhibitor 4,4'-diisothiocyanostilbene-2,2'disulphonate reduced photosynthetic rates only when HCO(3) (-) (but not CO(2)) could be the extracellular inorganic carbon form taken up. A similar, but less drastic, HCO(3) (-)-competitive inhibition of photosynthesis was obtained with Kl and KNO(3). It is suggested that, under ambient conditions, HCO(3) (-) is transported into cells at defined sites either via facilitated diffusion or active uptake, and that such transport is the basis for elevated internal [CO(2)] at the site of ribulose-1,5-bisphosphate carboxylase/oxygenase carboxylation.

Journal Article↗

Occupational dermatitis from Lactuca sativa (lettuce) and Cichorium (endive). Simultaneous occurrence of immediate and delayed allergy as a cause of contact dermatitis.

Four patients with occupational contact dermatitis to Lactuca sativa had cross-sensitivity to Cichorium endivia. One of the patients also had contact urticaria to Lactuca and Cichorium, and another reacted positively to scratch tests with these plants as a sign of immediate allergy. In two cases such immediate allergy was considered the cause of a vesicular, intense itching eruption within a few minutes of contact with fresh leaves of Lactuca on previously eczematous skin. The severe chronic dermatitis of the hands of these patients is ascribed to combined delayed and immediate allergy.

Adult↗

Toxicity identification evaluation of five metals performed with two organisms (Daphnia magna and Lactuca sativa).

When trying to identify the main toxicants in effluents, natural waters, sediments, soil leachates, and leachates from products, the Toxicity Identification Evaluation (TIE) procedure has proven useful. To enhance the use of this procedure for soil, sewage, and sediment samples, we wanted to evaluate this TIE procedure, regarding metal toxicity, for the 96-h root elongation test performed with Lactuca sativa (lettuce) seeds. We also wanted to evaluate the effect of TIE treatment on the toxicity of Mn and Fe to Daphnia magna. Bioassays were performed with Daphnia magna (48-h immobility) and lettuce seeds (96-h root elongation) to determine the effect concentrations for both organisms of Ag, Cu, Fe, Mn, and Zn. The TIE was then performed at the determined Daphnia 48-h EC(84) and Lactuca 96-h EC(50) for each metal. Our results showed that the order of the metal toxicity was Ag>Cu>Zn>Fe>Mn, for Daphnia and Ag = Zn = Fe = Cu > Mn for lettuce seeds. We also found that toxicity of the metals for Daphnia magna was reduced according to the prevailing knowledge regarding Cu, Zn, and Ag. However, the toxicity of Ag and Cu for Daphnia was also reduced by filtration through a C18 resin. Toxicity of Mn and Fe was reduced by filtration through a CM resin and increase of pH. For lettuce seeds, toxicity of the metals was reduced by the same treatments as for Daphnia magna with the exception of EDTA addition, which did not affect Cu toxicity to lettuce seeds. No effects were found for filtration through a C18 resin. We suggest that the TIE procedure using lettuce seeds can be used in toxicity identification of metals. However, the effects of pH manipulations were often stronger with lettuce and should be interpreted with care.

Animals↗

Identification and comparison of natural rubber from two Lactuca species.

Renewed interest in the identification of alternative sources of natural rubber to Hevea brasiliensis has focused on the Compositae family. In our search for Compositae models for rubber synthesis, we extracted latex from stems of two lettuce species: Lactuca serriola, prickly lettuce, and Lactuca sativa cv. Salinas, crisphead lettuce. Both species contained cis-1,4-polyisoprene rubber in the dichloromethane-soluble portions of their latex, and sesquiterpene lactones in their acetone-soluble portions. The rubber from both species and their progeny had molecular weights in excess of 1,000,000g/mol, and polydispersity values of 1.1. Rubber transferase activity was detected across a range of farnesyl diphosphate initiator concentrations, with decreased activity as initiator concentrations exceeded putative saturation. These results add lettuce to the short list of plant species that produce high molecular weight rubber in their latex. Due to the genomic and agronomic resources available in lettuce species, they provide the opportunity for further dissection of natural rubber biosynthesis in plants.

Lactones↗

Cloning and expression of sesquiterpene synthase genes from lettuce (Lactuca sativa L.).

Sesquiterpenoid lactones (SLs) from lettuce (Lactuca sativa L.) include constitutive components of latex such as lactucin and the induced phytoalexin, lettucenin A. A redundant primer strategy was used to recover two full length cDNA clones (LTC1 and LTC2) encoding sesquiterpene synthases from a cDNA library derived from seedlings with the red spot disorder, which accumulate phytoalexins. Recombinant enzymes produced from LTC1 and LTC2 in Escherichia coli catalysed the cyclisation of farnesyl diphosphate to germacrene A, potentially an early step in the biosynthesis of SLs. RT-PCR analysis showed LTC1 and LTC2 were expressed constitutively in roots, hypocotyls and true leaves but not in cotyledons. Expression in cotyledons was induced by challenge with the downy mildew pathogen Bremia lactucae in the disease resistant cultivar Diana. Southern hybridisation experiments showed that LTC1 and LTC2 were not part of a multigene family. The germacrene A synthases provide targets for modified expression to generate beneficial modifications to the SL profile in lettuce.

Amino Acid Sequence↗

SNP-based codominant markers for a recessive gene conferring resistance to corky root rot (Rhizomonas suberifaciens) in lettuce (Lactuca sativa).

The analysis of F2 progeny and derived F3 families of Lactuca sativa segregating for resistance to corky root rot caused by Rhizomonas suberifaciens permitted the identification of restriction fragment length polymorphism (RFLP) and single nucleotide polymorphism (SNP) markers linked to the recessive resistance gene cor. PCR-based markers were identified by bulked segregant analysis (BSA). Allele-specific primers were generally designed with the 3 terminal base coinciding with an SNP, matching one of the alleles and mismatching the other, and with an additional subterminal 3 base mismatching both alleles. Codominant, robust, and inexpensive molecular markers were obtained that used standardized PCR conditions. Some of the markers could be analyzed in multiple Lactuca mapping populations that did not segregate for disease resistance allowing the cor locus to be located on several maps. The consistent low density of markers around cor in these maps suggests that cor may be in an area with an elevated rate of recombination. Evaluation of these markers in a large sample of cultivars and landraces identified pairs of flanking polymorphic markers that can be used for marker-assisted selection of corky root resistance.

Chromosome Mapping↗

Microsatellite retrieval in lettuce (Lactuca sativa L.).

By using enriched genomic libraries, microsatellite-containing sequences were isolated from lettuce (Lactuca sativa) with high efficiency. With this approach, a sizeable fraction (up to 55%) of the clones contained a microsatellite. In about half of these clones, primers could be designed for PCR amplification of the microsatellite. This yielded 28 primer sets amplifying unambiguously scorable products, of which 26 showed polymorphisms in a test set of six lettuce varieties. Practically all microsatellite-amplifying primer sets yielded products in lettuce's nearest relative, L. serriola, but only half of the primer sets yielded products in the more distant species L. saligna and L. virosa. An average polymorphism information content (PIC) value of 0.55 and an average number of 3.5 alleles per locus were in the normal range for a self-fertilizing species like lettuce. In addition, the incidental cloning of a microsatellite-containing repeat family, apparently specific for Lactuca, is reported and the implications for the efficient retrieval of single-locus microsatellite sequences are discussed. The microsatellite loci isolated will be useful for distinguishing lettuce cultivars and for screening diversity of genetic resources.

Alleles↗