Palmitoyl-protein thioesterase (PPT), involved in the fatty acid synthesis and the de-palmitoylation of protein, was induced under ecological cooling treatment in hydrated lettuce seeds. However, there was no significant difference in fatty acid levels between the control and the cooled samples. To further study the function of PPT, 2-bromopalmitic acid (2-Bp), an inhibitor of protein palmitoylation, was applied during the imbibition of hydrated lettuce seeds, which was followed by slow-cooling treatment (−3°C h−1). The application of 2-Bp (1 mM) significantly increased the survival rate of seeds from 6.70% (control imbibition) to 22.67% (2-Bp imbibition) after slow cooling to −20°C. Differential scanning calorimetry (DSC) analysis indicated that 2-Bp led to earlier onset of ice crystals in the endosperm than the control group. Two-dimensional electrophoresis (2D) confirmed that 2-Bp could promote the hydrolysis of seed globulins and the accumulation of globulin peptides with small molecular weights. High-efficiency hydrolysis of globulin induced by mercaptoethanol improved the freezing tolerance of hydrated lettuce seeds and led to the accumulation of small globulin peptides, which further proved the positive function of small globulin polypeptides in enhancing the freezing tolerance of hydrated lettuce seeds. DSC of small globulin peptides showed that the smaller the molecular weight, the earlier the appearance of ice crystals and the higher the enthalpy of heat release. For the smallest peptides, the 2-Bp-4 in 2-Bp group exhibited higher enthalpy in exothermic peak than the control group (c-4). In conclusion, the hydrolysis of seed globulins and accumulation of small-molecule globulin peptides could be the major reason for improving the freezing tolerance of hydrated seeds after de-palmitoylation treatment.

This study aimed to evaluate the acaricidal activity of lettuce (Lactuca sativa) extracts against carmine spider mites (Tetranychus cinnabarinus Boisd.) and isolate the acaricidal components. Acaricidal activities of lettuce extracts isolated from different parts (the leaf, root and seed) using various solvents (petroleum ether, acetone and methanol) were evaluated with slide-dip bioassay and relatively high median lethal concentration (LC50) values were detected. Acetone extracts of lettuce leaves harvested in July and September were fractionated and isolated with silica gel and thin-layer chromatography. Consequently, acetone extracts of lettuce leaves harvested in July exhibited higher acaricidal activity than those harvested in September, with an LC50 value of 0.268 mg ml−1 at 72 h post-treatment. A total of 27 fractions were obtained from the acetone extract of lettuce leaves harvested in July, and mite mortalities with the 11th and 12th fractions were higher than those with the other 25 fractions (LC50: 0.751 and 1.258 mg ml−1 at 48 h post-treatment, respectively). Subsequently, active acaricidal components of the 11th fraction were identified by infrared, nuclear magnetic resonance and liquid chromatography/mass spectrometry. Five components were isolated from the 11th fraction, with components 11-a and 11-b showing relatively high acaricidal activities (LC50: 0.288 and 0.114 mg ml−1 at 48 h post-treatment, respectively). Component 11-a was identified as β-sitosterol. In conclusion, acetone extracts of lettuce leaves harvested in July might be used as a novel phytogenic acaricide to control mites.

The relative sensitivity of various crops and weeds to celery root residue incorporated into soil was determined in greenhouse experiments. A range of concentrations was used to determine the percentage of celery residue needed to cause a 50% reduction in plant growth (I50). The most sensitive weed was spiny amaranth and the least sensitive was common purslane. The most sensitive crop was radish and the least sensitive was celery. Selectivity was calculated as crop I50/weed I50 and varied from a maximum of 13 for celery and spiny amaranth to a low of 0.4 for radish and common purslane. Selective weed suppression might be obtained when celery residue is present.

Under simulated no-till conditions in the greenhouse, rye (Secale cereale L. ‘Wheeler’) residues reduced emergence of lettuce (Lactuca sativa L. ‘Ithaca’) and proso millet (Panicum miliaceum L. # PANMI) by 58 and 35%, respectively, over that obtained with a wood shaving control mulch. Rye shoot tissue inhibited lettuce germination 52% more than root tissue. Petri dish bioassays of residue in soil confirmed phytotoxicity on cress (Lepidium sativum L. ‘Curly’), lettuce, barnyardgrass (Echinochloa crus-galli L. Beauv. # ECHCG), and proso millet. Radicle elongation was a more senstive measure of phytotoxicity than germination per se. Phytotoxicity increased as the distance from seeds to rye residues decreased. While rye shoot residues caused 40% greater inhibition of cress radicle elongation in sterile soil than nonsterile soil, barnyardgrass was equally inhibited in both soil regimes. Phytotoxicity decreased in nonsterile soil, indicating that the compounds were degraded by microorganisms. Phytotoxic compounds in shoots were water extractable. Cress and barnyardgrass responded similarly to both aqueous rye extracts and to residues added to soil.

Sublethal amounts of 2,4-D [(2,4-dichlorophenoxy) acetic acid] were applied to several vegetable crops in order to determine effect on yield and quality in a simulated drift experiment. Tomato (Lycopersicon esculentum Mill.) and root crops were most sensitive to 2,4-D; as little as 2.1 g/ha of acid distorted tomato fruit shape and elongated radish (Raphanus sativus L.) roots. All root crops were rendered unmarketable by 10.4 g/ha and gross yields were reduced by exposure to 104 g/ha. Yield of peppers (Capsicum frutescens L. var. grossum) was increased by exposure to 2.1 g/ha and severely depressed by 104 g/ha. Cucumber (Cucumis sativus L.) fruit shape was distorted at 11 g/ha and yield was reduced slightly. Lettuce (Lactuca sativa L.), onion (Allium cepa L.), and cabbage (Brassica oleracea L. var. capitata) were least sensitive to 2,4-D. Exposure to 20.8 g/ha did not reduce yields of lettuce or cabbage. Onion yields were reduced by application of 104 g/ha. Bush bean (Phaseolus vulgaris L.) yield was decreased by exposure to 22 g/ha, but potato (Solarium tuberosum L.) yield increased at 16 g/ha. Herbicide residues in the crop foliage were proportional to the degree of exposure. Residue analysis combined with foliar symptoms may be useful in predicting crop damage following 2,4-D drift.

Dark germination of light-requiring lettuce seeds (Lactuca sativa L. ‘Grand Rapids’) was stimulated by SAN 9789 [4-chloro-5-(methylamino-2-(α,α,α-trifluoro-m-tolyl)-3(2H)-pyridazinone] and to a minor degree by BASF 13761 [4-chloro-5-methoxy-2-phenyl-3(2H)-pyridazinone] and BASF 44521 [4-chloro-5-methoxy-2-(α,α,α-trifluoro-m-tolyl)-3(2H)-pyridazinone], but not by’ pyrazon [5-amino-4-chloro-2-phenyl-3(2H)-pyridazinone], SAN 9785 [4-chloro-5-(dimethylamino)-2-phenyl-3 (2H)-pyridazinone], SAN 9774 [5-amino-4-chloro-2-(α,α,α-trifluoro-m-tolyl)-3(2H)-pyridazinone], or SAN 6706 [4-chloro-5-(dimethylamino)-2-(α,α,α-trifluoro-m-tolyl)-3(2H)-pyridazinone]. SAN 9789 stimulation was inhibited by cis-4-cyclohexene-1,2-dicarboximide (CHDC), and abscisic acid (ABA) at 1 × 10-4 M. Red light nullified the inhibitory effect of CHDC (1 × 10-4 M) but not the inhibitory effect of ABA (1 × 10-4 M) on SAN 9789 stimulated germination. Gibberellic acid (GA3) and kinetin (6-furfurylaminopurine) increased the germination stimulatory effect of SAN 9789 in darkness. Temperatures above 25 C decreased the effect of SAN 9789, with a temperature of 35 C completely inhibiting germination. The inhibitory effect of CHDC was strongly decreased at temperatures below 20 C. SAN 9789-induced germination in darkness was always the same (25 to 26% units increase in germination) even though the red light-stimulated germination differed with the seed batch.

Hydrophobic exudate from roots of sorghum [Sorghum bicolor (L.) Moench. #3 SORVU ‘IS 8768’] contain four p-benzoquinones which in the dihydroquinone form are active as germination stimulants of witchweed [Striga asiatica (L.) Kuntz. # STRLU]. The three minor p-benzoquinones were partially characterized and found to be structurally similar to sorgoleone, the major p-benzoquinone of this exudate. Herbicidal activity of the hydrophobic exudate was due to concentration- and pH-dependent inhibition of root elongation in some but not all weeds tested. Witchweed has apparently adapted these “defense” compounds of sorghum as host-specific germination stimulants.

Under greenhouse conditions, vaporization of oxyfluorfen [2-chloro-1-(3-ethoxy-4-nitrophenoxy)-4-(trifluoromethyl)benzene] formulations (2 EC, 25 WP, 1G) was studied on cabbage (Brassica oleracea L. var. capitata L. F. alba DC. ‘Market Prize’), tomato (Lycopersicon esculentum Mill. ‘Campbell 37’), cucumber (Cucumis sativus L. ‘Straight 8’), and lettuce (Lactuca sativa L. ‘Salad Bowl’). The emulsifiable concentrate and granular formulations caused vapor injury to plants exposed under glass bell jars. Vapor injury to all species increased when exposure times were increased from 8 to 72 h. Lettuce was the most susceptible test species. Exposure temperatures of 18 and 29 C did not alter the amount of vapor injury apparent on tomato foliage. Oxyfluorfen vapor injury did not reduce dry weight of any test species.

Lettuce (Lactuca sativa L.) and onion (Allium cepa L.) seeds have relatively short longevity during storage and their germination is sensitive to environmental stress. Seed priming (controlled hydration followed by drying) can improve seed germination under stressful conditions, inducing faster and more uniform germination over broader temperature ranges, but it can also reduce seed longevity in storage. Controlled deterioration (CD) tests are often employed to study longevity by ageing seeds rapidly at elevated temperature and moisture content, and primed seeds are particularly sensitive to CD conditions. As reactive oxygen (O2) species are thought to be involved in seed deterioration, we tested whether storage under reduced O2 atmospheres (0 and 2% O2) would extend the longevity of primed and non-primed seeds under low relative humidity (RH) (33% RH+37°C) and CD (75% RH+50°C) storage conditions. The longevity of both non-primed and primed lettuce seeds in low RH storage was extended by anaerobic environments, but the effect of O2 was much less under CD conditions. In onion, only primed seeds exhibited a beneficial effect of low O2 atmospheres under both types of ageing conditions. In both species, storage under anaerobic conditions was beneficial for extending the longevity of primed seeds, but was not able to ameliorate fully the negative effect of priming on storage life.

To reduce the level of redundancy in a collection of cultivated lettuce, data from 160 amplified fragment length polymorphism (AFLP) fragments and 10 polymorphic microsatellites were used in combination with passport data and morphological data, the latter obtained from an experimental field trial performed for verification purposes. Based on the observed distribution of the number of marker differences between and within accessions, a minimum of three AFLP differences and two microsatellite differences were regarded as levels warranting distinction between accessions in the redundancy analysis. The strategy followed in the redundancy analysis was mainly based on the confirmation of duplication by each of two independently generated data sources. The molecular data were used for the validation as well as the identification of potential duplicates, revealing a total number of 198 redundancies, corresponding to 12.9% of the total collection. Trueness to type, number of characterization and evaluation data, and collection management considerations, such as available seed quantities and germination percentages, were used as primary, secondary and tertiary criteria to decide which accession from duplication groups to maintain in the collection. Removal of accessions showed negligible effects on total collection diversity, as quantified for AFLPs and microsatellites, characterization and evaluation traits and resistance profiles against downy mildew pathotypes, indicating that the applied strategy was effective.

Seed germination of Grand Rapids lettuce (Lactuca sativa L.) was inhibited at 38°C (thermoinhibition). Thermoinhibition was totally relieved by a combination of 100% oxygen and 10 mg I−1 kinetin, to a lesser extent by a combination of oxygen and ethylene but not at all by oxygen plus GA3. The combination of oxygen with kinetin or ethylene was more effective in preventing thermoinhibition than a combination of all three hormones and CO2 in air. The beneficial effect of oxygen plus kinetin was not related to either increased production of CO2 or ethylene by the seeds. In air, germination was partially inhibited by 2-aminoethoxyvinyl glycine (AVG) or 2,5-norbornadiene (NBD) and totally inhibited by AVG plus NBD. Oxygen plus kinetin almost completely alleviated their inhibitory effects. It was concluded that oxygen plus kinetin caused seeds to bypass an ethylene requirement for germination or increased the sensitivity of seeds to ethylene. In air at 38°C, seeds exhibited a high level of ethanolic fermentation, while in oxygen plus kinetin they respired aerobically. At 38°C in air, contents of ATP and total adenylates were reduced, while in seeds treated with oxygen plus kinetin these contents were similar or higher than in 25°C controls. A similar response was also observed for adenylate energy charge. It was concluded that part of the mechanism of thermoinhibition could be the failure of ATP content to reach a sufficient level to satisfy the requirement for germination at 38°C. It appeared possible that the prevention of thermoinhibition by oxygen plus kinetin could be related to increased ATP content in seeds caused by this treatment.

A recent hypothesis states that the accumulation of amino-carbonyl reaction products of reducing sugars with proteins (Maillard products) is related to the loss of seed vigour and viability during ageing. Since Maillard products are biochemical end products, their accumulation should provide an index of seed deterioration. A simple fluorescence assay has been used to estimate the quantity of Maillard products in aqueous seed extracts. We tested whether an accumulation of fluorescent compounds correlates with the loss of seed viability during ageing. Fluorescence of extracts from lettuce (Lactuca sativa) embryonic axes increased after controlled deterioration, but was not correlated with ageing period. Fluorescence of extracts from whole broccoli (Brassica oleracea var. italica) seeds was either unchanged or declined during controlled deterioration. Extracts from naturally aged whole seeds of carrot (Daucus carota), tomato (Lycopersicon esculentum), and cauliflower (B. oleracea var. botrytis) had higher fluorescence than controls, but there was no consistent increase in fluorescence in proportion to the loss of viability. Extracts of naturally aged onion (Allium cepa) seeds exhibited less fluorescence than extracts from seeds with higher viability. We conclude that the fluorescence assay does not have general utility as an index of seed deterioration.

Prolonged imbibition and incubation of lettuce seeds at a supraoptimal temperature induces secondary dormancy. Such seeds no longer germinate when returned to conditions optimal for germination of non-dormant seeds. The possibility that inhibitors are involved in the induction of thermodormancy was investigated.

Washing of thermodormant seeds restored germination to a small extent. However, continuous leaching of seeds during high-temperature treatment with distilled water, largely prevented the induction of thermodormancy. Such seeds were qualitatively similar to nondormant seeds, i.e. they required only light to germinate at a lower temperature and germinated in the dark if given GA3. The germination water from thermodormant seeds inhibited the germination of non-dormant lettuce seeds. The inhibition was not an osmotic effect. Absorbance in the ultraviolet region was higher in germination water from thermodormant seeds than that of non-dormant seeds. It is concluded that inhibitors are involved in the induction of thermodormancy of lettuce seeds.

On the premise that seed ageing may be largely a result of free-radical lipid autoxidation, a study was made of the relationship between lipid stability and longevity in seeds of soybean (Glycine max), lentil (Lens culinaris), mungbean (Vigna radiata), chickpea (Cicer arietinum), broadbean (Vicia faba), pea (Pisum sativum), lettuce (Lactuca sativa), and tomato (Lycopersicon esculentum). Seed lipids were examined for α-, γ- and δ-tocopherols, and levels of unsaturated and saturated fatty acids. Using this information, analysis was attempted using linear, multiplicative or exponential models to correlate aspects of lipid stability with seed longevity values from the published literature. No statistically significant correlations could be found between longevity and total lipid unsaturation, tocopherol levels or two protection formulae obtained from the oil chemistry literature. When values for tomato were excluded, a good correlation (r = 0.89, P = 0.007) was obtained using a multiplicative regression model for levels of linolenic acid per unit of total tocopherols in relation to longevity. Possible factors contributing to a lessening of the relationship between lipid stability and seed longevity are discussed.

At the time of protrusion, radicles of lettuce seeds became susceptible to desiccation damage. Concomitant with radicle elongation, both the reducing sugar content and the amount of lipid-aldehydes increased. The role of the hydroxy- and lipid-aldehydes in browning reactions that occurred during desiccation and re-imbibition was analysed. It was concluded that both types of aldehydes reacted with amino-groups during desiccation and that the increase in lipid-aldehydes during germination could be due to the augmented reducing sugar content. Therefore, the high reducing sugar content in radicles of germinated lettuce seeds may be the primary source of hydroxyl radicals, and thereby, a principal cause of desiccation damage.

Hybridization and introgression between crops and wild relatives may have important evolutionary and ecological consequences such as gene swamping or increased invasiveness. In the present study, we investigated hybridization under field conditions between crop lettuce (Lactuca sativa) and its wild relative prickly lettuce (L. serriola), two cross-compatible, predominantly autogamous and insect pollinated species. In 2003 and 2004, we estimated the rates of hybridization between L. sativa and L. serriola in close-to-reality field experiments carried out in two locations of Northern Switzerland. Seeds set by the experimental wild plants were collected and sown (44 352 in 2003 and 252 345 in 2004). Progeny was screened morphologically for detecting natural hybrids. Prior to the experiment, specific RAPD markers were used to confirm that morphological characters were reliable for hybrid identification. Hybridization occurred up to the maximal distance tested (40 m), and hybridization rates varied between 0 to 26%, decreasing with distance. More than 80% of the wild plants produced at least one hybrid (incidence of hybridization, IH) at 0 m and 1 m. It equaled 4 to 5% at 40 m. In sympatric crop-wild populations, cross-pollination between cultivated lettuce and its wild relative has to be seen as the rule rather than the exception for short distances.

The effects of seed priming on germination and endo-β-mannanase activity at inhibitory and non-inhibitory temperatures in thermosensitive ‘Dark Green Boston’ (DGB) and in thermotolerant ‘Everglades’ (EVE) lettuce were investigated. A single-seed endo-β-mannanase assay was used to follow the enzyme activity during priming. Seeds were primed at 15°C in aerated solutions of polyethylene glycol (PEG) with constant light, then redried. Primed and non-primed seeds germinated 100% at 20°C. At 35°C, non-primed and primed EVE seeds germinated 100%, whereas non-primed seeds of DGB germinated only 4%. During priming, endo-β-mannanase activity increased between 24 and 48 h in EVE and between 24 and 72 h in DGB after the beginning of osmotic imbibition. Endo-β-mannanase activity persisted in primed seeds following seed drying, was detected before radicle protrusion and was present in the micropylar region in front of the radicle tip. Higher enzyme activity was observed in primed seeds and EVE compared with non-primed and DGB seeds. The results suggest that priming may overcome the inhibitory effect of high temperature in thermosensitive lettuce seeds due to increased endo-β-mannanase activity, possibly leading to a weakening of endosperm, thus overcoming thermodormancy.

The impact of rehydration on a multicellular organism was studied in lettuce (Lactuca sativa L.) embryos, using cryo-scanning electron microscopy (cryo-SEM). Naked embryos were sensitive to imbibitional stress, whereas embryos with an intact, thick-walled endosperm were not. Imbibitional injury to naked embryos was mainly confined to the outer 2–3 cell layers of the axis. These cells failed to swell and appeared poorly hydrated. Deeper layers were not affected even after extended periods of cold rehydration. The proportion of damaged cells (6–7% of total) roughly corresponded with the additional K+ that gradually leached from the embryos. Damaged embryos were able to survive the loss of their surface layers and form adventitious roots. The swelling of inner tissues caused the dead surface layers to rupture into patches. Plasma membranes in dried embryos showed normal bilayer structure with a homogeneous distribution of intra-membrane particles (IMPs), also after non-injurious rehydration. Imbibitionally damaged plasma membranes showed many irregularities, such as globular insertions, that probably resulted from malfusions in the ruptured membrane, but the IMPs were still randomly distributed.

Molecular mobility is hypothesised to be a key factor influencing storage stability of seeds because it may control the rate of deteriorative reactions responsible for reduced shelf life. The relationship between the longevity of lettuce seeds and the molecular mobility was investigated in the cytoplasm of lettuce radicles. Longevity of lettuce seeds was predicted using the viability equation of Ellis and Roberts, and the molecular mobility was determined by saturation transfer electron paramagnetic resonance spectroscopy measurements of rotational motion of a polar spin probe inserted into the cytoplasm. Increasing the temperature resulted in faster rotational motion and shorter longevity. There was a linear relationship between the logarithms of rotational motion and estimated seed longevity for temperatures above 5°C. Below 5°C, there was a deviation from linearity. Based on the hypothesis that the molecular mobility in the cytoplasm determines the rate of detrimental reactions, seed longevity at sub-zero temperatures was predicted by extrapolation of the rotational motion using the linear relationship. Predictions of longevity at sub-zero temperatures based on rotational motion indicated longer survival times than those based on the viability equation. A kinetic approach to ageing using molecular mobility measurements is expected to improve our understanding of seed storage stability and might eventually lead to realistic predictions of longevity at low temperatures.