Cell-Wall Autohydrolysis in Isolated Endosperms of Lettuce (Lactuca sativa L.)

Cell walls prepared from the endosperm tissue of hydrated lettuce (Lactuca sativa L.) seeds undergo autohydrolysis. Release of carbohydrates is most rapid (0.4-0.6 [mu]g per endosperm) within the 1st h of incubation in buffer, but substantial autolysis is sustained for at least 10 h. Autolysis is temperature sensitive, and the optimum rate occurs at pH 5. The rate of autolysis increases markedly in the period just prior to radicle emergence. The cell-wall polysaccharide composition in micropylar and lateral endosperm regions differs significantly; the micropylar walls are rich in arabinose and glucose with substantially lower amounts of mannose. Although walls prepared from both micropylar and lateral regions undergo autolysis, micropylar walls release carbohydrates at a higher rate than lateral walls. Autolysis products elute as large polymers when subjected to size-exclusion chromatography, suggesting that endo-enzyme activity is responsible for release of fragments containing arabinose, galactose, mannose, and uronic acids. Arabinose, galactose, mannose, and glucose are also released as monomers. As a function of time, the ratio of polymers to monomers decreases, indicating that exo-enzyme activity is also present. Thermoinhibition or treatment with abscisic acid suppresses germination and reduces the rates of autolysis of walls isolated from the endosperm by about 25%. Treatments that alleviate thermoinhibition (kinetin and gibberellic acid) increase the rates of autolysis by 20 to 30% when compared to thermoinhibited controls.     

Glaucocalyxin A and B Regulate Growth and Induce Oxidative Stress in Lettuce (Lactuca sativa L.) Roots

Glaucocalyxin (Gla) A–C are major ent-kaurane diterpenoids isolated from Isodon japonicus var. glaucocalyx (Maxim.) H. W. Li. This study investigated the possible interference of these diterpenoids with root growth and its mechanism of action in lettuce (Lactuca sativa L.) seedlings. Results indicated the dual stimulatory and inhibitory effects of Gla A and B on root growth and their phytotoxic effects on root hair development. The promotion of root growth by lower levels of Gla A and B (20–40 μM) resulted from enhanced cell length and increased mitotic activity. However, higher concentrations (80–200 μM) of Gla A and B had inhibitory effects. In addition, Gla A and B inhibited root hair development of lettuce seedlings in a dose-dependent manner at concentrations between 20 and 200 μM. Exposure of lettuce roots to Gla A and B at 200 μM increased levels of malondialdehyde and the generation of O ₂ ^·? , indicating lipid peroxidation and induction of oxidative stress. Activities of the antioxidant enzymes superoxide dismutase, catalase, and peroxidase were significantly elevated. Reactive oxygen species (ROS) scavengers dihydroxybenzene disulfonic acid (Tiron) and dimethylthiourea at 100 μM could efficiently alleviate the phytotoxicity induced by Gla A and B at 200 μM. These results demonstrated that the deleterious effect of Gla A and B at higher concentrations (80–200 μM) on roots may occur through the imposition of oxidative stress on cell growth and cell division. Due to the lack of an α,β-unsaturated ketone in α-methylenecyclopentanone moiety, Gla C could not induce ROS generation and exhibited no effect on the roots, even at the highest concentration (200 μM). Therefore, the α-methylenecyclopentanone moiety in the ent-kaurene diterpenoids was presented as an essential possible active center for the phytotoxicity.     

Release of Esterase Following Germination of Lettuce Seed (Lactuca sativa L.)

Light-insensitive lettuce seeds, Lactuca sativa L. cv. Great Lakes, release esterases for a period following radicle protrusion. Very little or no enzymes are released prior to 24 hours or after 48 hours of germination. As compared to intact seeds, half-seeds readily release esterases and the release is not affected by far red irradiation. Bulk of the released esterases are derived from the endosperm tissue and presumably exists in the intact seed as a component of the extraembryonic fluid.     

On the Structure of Lettuce (Lactuca sativa L.) Endosperm during Germination

Endosperm cells of lettuce (Lactuca sativa L.) are characterizedby thick cell walls and dense cytoplasm which contains numerousprotein bodies. Other organelles such as nucleus, mitochondria,plastids and dictyosomes are typical of plant cells. Light andelectron microscopy reveal that before radicle emergence micropylarcells of endosperm tissue undergo drastic protoplast alterations.These alterations seem to be the only structural modificationsbefore rupturing of the tissue since the walls of the endospermcells seem to degrade only after radicle emergence. The differentialbehaviour of the micropylar area of the endosperm before radicleemergence and the observation that the micropylar cells remainmetabolically active long after radicle emergence while therest of the tissue is almost completely disintegrated, suggeststhat the endosperm cells of the micropylar area may have a roleother than being a main reserve site like the rest of the endosperm. Lactuca sativa L., endosperm structure, seed germination, lettuce     

Photorespiration Process and Nitrogen Metabolism in Lettuce Plants (Lactuca sativa L.): Induced Changes in Response to Iodine Biofortification

Iodine is vital to human health, and iodine biofortification programs help improve human intake through plant consumption. There is no research on whether iodine biofortification influences basic plant physiological processes. Because nitrogen (N) uptake, utilization, and accumulation are determining factors in crop yield, the aim of this work was to establish the effect of the application of different doses (20, 40, and 80 μM) and forms of iodine (iodate [IO₃ ⁻] vs. Iodide [I⁻]) on N metabolism and photorespiration. For this study we analyzed shoot biomass and the activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), aspartate aminotransferase (AAT), glutamate dehydrogenase (GDH), glycolate oxidase (GO), glutamate:glyoxylate aminotransferase (GGAT), serine:glyoxylate aminotransferase (SGAT), hydroxypyruvate reductase (HR) and catalase (CAT), nitrate (NO₃ ⁻), ammonium (NH₄ ⁺), organic and total N, amino acids, proteins, serine (ser), malate, and α-ketoglutaric acid in edible lettuce leaves. Application of I⁻ at doses of at least 40 μM reduced the foliar concentration of NO₃ ⁻ with no decrease in biomass production, which may improve the nutritional quality of lettuce plants. In contrast, the application of 80 μM of I⁻ is phytotoxic for lettuce plants, reducing the biomass, foliar concentration of organic N and NO₃ ⁻, and NR and GDH activities. HR activity is significantly inhibited with all doses of I⁻; the least inhibition was at 80 μM. This may involve a decrease in the incorporation of carbonated skeletons from photorespiration into the Calvin cycle, which may be partially associated with the biomass decrease. Finally, the application of IO₃ ⁻ increases biomass production, stimulates NO₃ ⁻ reduction and NH₄ ⁺ incorporation (GS/GOGAT), and optimizes the photorespiratory process. Hence, this appears to be the most appropriate form of iodine from an agronomic standpoint.     

Somatic Embryogenesis and Analysis of Peroxidase in Cultured Lettuce (Lactuca sativa L.) Cotyledons

Somatic embryos were induced in lettuce cotyledons culturedon Murashige and Skoog's (MS) medium containing either 2 mgl^–1 6-benzylaminopurine (BA) and 0.2 mg l^–1 naphthaleneaceticacid (NAA) or 0.2 mg l^–1 BA and 2 mg l^–1 NAA. Bothcombinations induced a frequency of over 70%. The explants culturedonly in the presence of 2,4-dichlorphenoxyacetic acid (2,4-D)did not produce somatic embryos. The development of the embryoidswas studied histologically and by scanning electron microscopy.Peroxidase activity was assayed and the isoenzyme pattern ofcalluses was determined by polyacrylamide gel electrophoresis.Callus from an embryogenic line showed a much higher peroxidaseactivity than that from a non-embryogenic line, one extra peroxidaseisozyme band being present and typical of the embryogenic callus.No qualitative differences were detectable between the embryogeniccalluses. Lactuca sativa L, lettuce, somatic embryogenesis, peroxidases, isoenzymes     

莴苣大孢子发生过程中钙的分布动态

莴苣胚囊发育为蓼型,减数分裂形成的4个大孢子中只有合点端的一个大孢子可继续发育,其余3个大孢子从珠孔端依次退化.大孢子母细胞中的钙沉淀颗粒很少,减数分裂后的四分体中的钙沉淀颗粒稍有增加.以后,4个大孢子中的钙沉淀颗粒在数量上有明显差异:即将退化的大孢子中钙明显减少,而未退化大孢子细胞质中则保持有较多的细小钙沉淀颗粒.大孢子的退化是一种细胞程序死亡现象,细胞中的钙浓度降低时可能启动了大孢子细胞的程序性死亡过程,而细胞中的钙浓度高时则保持大孢子细胞的继续发育.文章首次揭示了大孢子发生过程中钙的分布特征.     

The Role of Nitrate in the Osmoregulation of Lettuce (Lactuca sativa L.) Grown at Different Light Intensities

Blom-Zandstra, M. and Lampe, J. E. M., 1985. The role of nitratein the osmoregulation of lettuce (Lactuca sativa L.) grown atdifferent light intensities.—J. exp. Bot. 36: 1043–1052. The effect of different light intensities on the nitrate accumulationvis-à-vis the concentration of other solutes in plantsap expressed from lettuce leaves was studied. After growinglettuce plants under constant environmental conditions for 52d, they were transferred to different light intensities andharvested periodically. A quantitative analysis of componentsin solution in the expressed plant sap showed a decrease innitrate concentration and an increase in the organic acids (mainlymalate) and sugars (mainly glucose) with increasing light intensity.The light intensity only slightly increased the osmolarity ofthe expressed plant sap. The measured osmolarity correspondedvery well with the value estimated from the quantitative analysesimplying that all osmotically active compounds had been accountedfor. The decrease in nitrate concentration in the expressedplant sap was fully compensated for by an increase in the dissociatedorganic acids that partly dissociate twofold to sustain electroneutralityand by an increase in both organic acids and sugars to maintainthe osmolarity. The suggestion is supported that nitrate mayserve as osmoticum at low light conditions to compensate forthe shortage of carbohydrates resulting from suboptimal photosynthesis. Key words: Nitrate accumulation, osmoregulation, Lactuca saliva L.     

Survival and Vigour of Lettuce (Lactuca sativa L.) and Sunflower (Helianthus annuus L.) Seeds Stored at Low and Very-low Moisture Contents

Seeds of lettuce (Lactuca sativa L.) and sunflower (Helianthusannuus L.) were stored hermetically at 35 °C with 11 differentmoisture contents between 1·3 and 6·9%, and between1·3 and 7·1% of fresh mass, respectively. Germinationand vigour (mean germination time, root length, seedling dryweight) were determined after storage for 0, 8, or 16 weeks(sunflower) or 0, 8, 16, or 48 weeks (lettuce) in these environmentsfollowed by various humidification treatments (to avoid imbibitioninjury). The range of seed storage moisture contents over whichdeterioration was minimized depended upon the criterion of deteriorationused, and varied somewhat between species. Comparison of theseranges for seeds stored for the longest durations showed thatfor some criteria seed performance was poorer (P < 0·05)at both the lowest and highest moisture contents investigatedthan at certain of the intermediate storage moisture contents(e.g, most rapid germination occurred in sunflower followingstorage at 2·2-4·7% moisture content), whereasfor other criteria all the drier storage moisture contents weresuperior to the more moist (e,g. greatest seedling growth occurredin sunflower following storage at 1·3-5·1% moisturecontent). But none of these results suggested that lettuce andsunflower seeds stored hermetically at 2·5-3·0%or 2·2-2·5% moisture content, respectively, wereless vigorous than at any other moisture content tested. Inboth species, these storage moisture contents are in equilibriumwith about 8-10% relative humidity (r.h.) at 20 °C, whichis similar to and indeed marginally less than the 10-13% r.h.recommended following earlier studies on the longevity of seedsin hermetic storage at much warmer temperatures. Thus, theseresults show no evidence that the optimum seed moisture contentfor storage increases with decrease in temperature, at leastover the range 35-65 °C, as has been suggested elsewhere.We conclude that the international recommendation for the long-termseed storage for genetic conservation at 5 ± 1% moisturecontent should not be revised upwardly, and that in situationswhere refrigeration cannot be provided storage at even lowermoisture contents is worthy of further investigation for thoseseeds in which desiccation at 20 °C to equilibrium at 10%r.h. results in moisture contents well below 5%.Copyright 1995,1999 Academic Press Helianthus annuus L., sunflower, Lactuca sativa L., lettuce, desiccation, seed storage, seed vigour     

Characterization of the Germination Responses to Temperature of Lettuce (Lactuca sativa L.) Achenes

Germination tests were done on 23 cultivars of lettuce (Lactucasativa L.) comprising a variety of different morphological formsselected for cultivation at various seasons. Significant differences at the upper limit of temperature tolerancewere found between different cultivars: maximum temperaturesfor 50 per cent germination ranged from 23 to 32 °C, andsusceptibility to the induction of secondary dormancy by hightemperatures varied widely from one cultivar to another. Nocorrelations were established between germination responsesand heading type, achene colour or growing season The germination responses of all cultivars at temperatures fromjust above 0 to 20 °C were closely similar and results froma large number of experiments were used to produce a standardgermination reference curve foti culvars of L. sativa. Departfuresrom this curve were found to arise predominantly from variationsin seed quality or test conditions rather than from the genotypeof the cultivar. Detailed comparisons of germination at all possible combinationsof alternate and constant day/night temperatures are presentedfor two cultivars. One with relatively high tolerance to high-temperatureinducedinhibition, the other with low tolerance. The results are discussed in relation to the original distributionof the species as a wild plant and its history of cultivationby man. Lactuca sativa L., Lettuce, achene germination, temperature response     

Freeze Desiccation: a Second Mechanism for the Survival of Hydrated Lettuce (Lactuca sativa L.) Seed at Sub-zero Temperatures

Differential Thermal Analysis of hydrated lettuce cv. GreatLakes achenes using a rapid cooling rate (20 °C h^–1)produced two exotherms per achene. Both exotherms representedthe freezing of supercooled water. The high temperature exothermoccurred at –93 °C and was produced by freezing ofwater inside the pericarp but exterior to the endosperm. Thetemperature at which it occurred could be altered by the additionof nucleating agents. The low temperature exotherm produced by freezing of the embryooccurred at –162 °C and marked the death of the seed.Its temperature was not changed by the addition of nucleatingagents but its occurrence required the structural integrityof the endosperm. At low cooling rates (1 and 2 °C h^–1)low temperature exotherms were not recorded and samples removedat –25 °C had high viability. Slow cooling causeda redistribution of water within the seed whereby ice formingoutside the endosperm caused desiccation of the embryo and preventedits freezing. A mechanism is proposed, in terms of established supercoolingand nucleation theory, to explain the observed results and thevalue of freeze tolerance to the species in its natural habitatis discussed. Cooling rate, differential thermal analysis, freezing avoidance, Lactuca sativa L., lettuce, seed, supercooling, water migration     

Cadmium Uptake by Lettuce (Lactuca sativa L.) as Basis for Derivation of Risk Limits in Soils

The availability and uptake of Cd by lettuce (Lactuca sativa L.) in two common tropical soils (before and after liming) were studied in order to derive human health-based risk soil concentration. Cadmium concentrations ranging from 1 to 12 mg kg^?1 were added to samples from a clayey Oxisol and a sandy-loam Ultisol under glasshouse conditions. After incubation, a soil sample was taken from each pot, the concentration of Cd in the soil was determined, lettuce was grown during 36 d, and the edible parts were harvested and analyzed for Cd. A positive linear correlation was observed between total soil Cd and the Cd concentration in lettuce. The amount of Cd absorbed by lettuce grown in the Ultisol was about twice the amount absorbed in the Oxisol. Liming increased the soil pH and slightly reduced Cd availability and uptake. CaCl₂ extraction was better than DTPA to reflect differences in binding strength of Cd between limed and unlimed soils. Risk Cd concentrations in the Ultisol were lower than in the Oxisol, reflecting the greater degree of uptake from the Ultisol. The derived risk Cd values were dependent on soil type and the exposure scenario.     

Influence of After-ripening on Phytochrome Control of Seed Germination in Two Varieties of Lettuce (Lactuca sativa L.)

The reversible photoreaction on seed germination of two varieties of lettuce differs remarkably not only with the variety but also with the germination temperature and the physiological conditions of seeds caused by after-ripening.     

Effects of Germination-promoting Substances Given in Conjunction with Red Light on the Phytochrome-mediated Germination of Dormant Lettuce Seeds (Lactuca sativa L.)

Ethylene or thiourea can substitute for gibberellic acid but not for red light in breaking the secondary dormancy induced by extended dark storage of fully hydrated lettuce seeds (Lactuca sativa cv. Grand Rapids). After 10 days of storage, ethylene, thiourea, or gibberellic acid applied either separately or in any combination in conjunction with red light induced near maximal germination. When applied separately without red light, none of the substances promoted germination of seeds stored 10 days. Combinations of any two or all three of the substances in the absence of red light induced some germination but no combination was as effective as any single substance given with red light.     

Microsatellite fingerprinting in lettuce (Lactuca sativa L.) and wild relatives

Southern hybridisation with a single microsatellite probe, (TCT)₁₀, sufficed to discriminate between a representative set of cultivars/accessions of lettuce, Lactuca sativa L., and its wild relatives L. serriola, L. saligna and L. virosa. Variability within cultivars was tested in a relatively modern cultivar (Hector), where no variation was found, and in an older and morphologically more variable cultivar (Madrilene), where heterogeneity was observed in the TCT fingerprint. (TCT)₁₀ fingerprinting should be useful for variety identification and homogeneity testing in lettuce. Received: 25 July 1997 / Revision received: 5 August 1997 / Accepted: 30 August 1997     

The Effect of Arsenate and Other Inhibitors on Early Events during the Germination of Lettuce Seeds (Lactuca sativa L.)

The effect of arsenate, arsenite, 2,4-dinitrophenol, and anaerobiosis on early events in seed germination was investigated using both intact and punched seeds of lettuce (Lactuca sativa L.). It was found that punching the seed removes penetration barriers to the entrance of inhibitors without an undue loss of germination or light responses. The kinetics of the action of germination inhibitors were established by 2-hour pulse experiments. Arsenate and 2, 4-dinitrophenol have very different kinetics. The inhibition of germination in punched seeds by arsenate given in conjunction with phosphate compared with the lack of inhibition of arsenate plus phosphate on the growing seedling, suggest a distinct metabolic change in the germinating embryo at some time between the onset of germination and subsequent seedling growth.     

Regulation of Root Growth in Lactuca sativa L. Seedlings by the Ent-Kaurane Diterpenoid Epinodosin

Epinodosin, an ent-kaurane diterpenoid isolated from Isodon japonica var. galaucocalyx, had a biphasic, dose-dependent effect on root growth and a strong inhibitory effect on root hair development in Lactuca sativa L. seedlings. Lower levels of epinodosin (25–100 μM) significantly promoted root growth, but higher concentrations (150–200 μM), by contrast, had inhibitory effects. In addition, all of the tested concentrations (20–80 μM) inhibited root hair development of lettuce seedlings in a dose-dependent manner. Further investigations on the underlying mechanism revealed that the promotion effect of epinodosin (25–100 μM) resulted from increasing the cell length in the mature region and enhancing the mitotic activity of meristematic cells in lettuce seedling root tips. On the other hand, epinodosin at higher concentrations inhibited root growth by strongly affecting both the cell length in the mature region and the division of meristematic cells. Comet assay analysis demonstrated that the decrease of mitotic activity of root meristematic cells was due to DNA damage induced by higher levels of epinodosin.     

Occurrence of Amoebae on Oak Leaf Lettuce (Lactuca sativa var. crispa) and Boston Lettuce (L. sativa var. capitata)1

The colonization, distribution, population density, and species diversity of amoebae on leaves of Oak Leaf lettuce, Lactuca sativa var. crispa, and Boston lettuce, L. sativa var. capitata, were investigated. The role of soil in the colonization of Oak Leaf lettuce was determined by comparing numbers of amoebae present on basal leaves (those that pass through soil) with numbers on wrapped leaves (those that do not pass through soil). Amoebae were present in ten samples of basal leaves and ranged from 154–1510/g of leaf tissue. Wrapped leaves failed to yield amoebae in seven of ten trials and contained <4 amoebae/g of tissue. Mean values for the population density of amoebae on Oak Leaf basal leaves and Boston lettuce leaves were 484 ± 133 and 453 ± 93, respectively. The distribution of amoebae on green and white parts of leaves from both kinds of lettuce was studied. The occurrence of amoebae on rinsed, unrinsed, visibly clean, and visibly dirty samples of Boston lettuce leaves was established.     

Effects of co-cropping Bidens pilosa (L.) and Tagetes minuta (L.) on bioaccumulation of Pb in Lactuca sativa (L.) growing in polluted agricultural soils

Polluted agricultural soils are a serious problem for food safety, with phytoremediation being the most favorable alternative from the environmental perspective. However, this methodology is generally time-consuming and requires the cessation of agriculture. Therefore, the purpose of this study was to evaluate two potential phytoextractor plants (the native species Bidens pilosa and Tagetes minuta) co-cropped with lettuce growing on agricultural lead-polluted soils. The concentrations of Pb, as well as of other metals, were investigated in the phytoextractors, crop species, and in soils, with the potential risk to the health of consumers being estimated. The soil parameters pH, EC, organic matter percentage and bioavailable lead showed a direct relationship with the accumulation of Pb in roots. In addition, the concentration of Pb in roots of native species was closely related to Fe (B. pilosa, r = 0.81; T. minuta r = 0.75), Cu (T. minuta, r = 0.93), Mn (B. pilosa, r = 0.89) and Zn (B. pilosa, r = 0.91; T. minuta, r = 0.91). Our results indicate that the interaction between rhizospheres increased the phytoextraction of lead, which was accompanied by an increase in the biomass of the phytoextractor species. However, the consumption of lettuce still revealed a toxicological risk from Pb in all treatments.