Evolução de etileno e atividade da enzima endo-beta-mananase durante a germinação de sementes de alface sob altas temperaturas

Altas temperaturas durante a embebição das sementes de alface podem atrasar ou inibir a germinação e o endosperma parece ser o responsável na restrição da protrusão da radícula. O envolvimento da enzima endo-beta-mananase durante a germinação de sementes de alface a 35°C e a influência do etileno na regulagem desta enzima foram estudados. Sementes das cultivares Dark Green Boston (DGB) e Everglades (EVE) foram germinadas em água ou em soluções de 10 mmol L-1 de 1-aminociclopropano-1-ácido carboxilico (ACC), 10 mmol L-1 de amino-etoxi-vinil-glicina (AVG), ou 20 mmol L-1 de tiossulfato de prata (STS). Sementes foram ainda osmoticamente condicionadas em soluções de polietilenoglicol (PEG), ou PEG + ACC, PEG + AVG, ou PEG + STS. Sementes não tratadas germinaram 100% a 20°C. A 35°C, EVE germinou 100%, enquanto DGB germinou somente 33%. A germinação a 35°C aumentou em sementes osmoticamente condicionadas ou sementes que receberam ACC durante a incubação. Maior evolução de etileno foi detectada em EVE do que em DGB durante a germinação a 35°C. AVG não inibiu a germinação de DGB a 35°C, enquanto que STS inibiu. Maior atividade de endo-beta-mananase nas sementes foi observada em EVE quando comparado com DGB. Fornecimento de ACC tanto durante o condicionamento osmótico como durante a germinação, aumentou a atividade de endo-beta-mananase, enquanto que AVG e STS proporcionaram um decréscimo ou ausência da atividade enzimática. O etileno pode minimizar o efeito inibitório de altas temperaturas na germinação de sementes de genótipos sensíveis de alface devido ao aumento da enzima endo-beta-mananase, possivelmente levando ao enfraquecimento do endosperma.High temperatures during lettuce seed imbibition can delay or completely inhibit germination and the endosperm layer appears to restrict the radicle protrusion. The role of endo-beta-mannanase during lettuce seed germination at 35°C and the influence of ethylene in endo-beta-mannanase regulation were investigated. Seeds of 'Dark Green Boston' (DGB) and 'Everglades' (EVE) were germinated in water, or 10 mmol L-1 of 1-aminocyclopropane-1-carboxylic acid (ACC), or 10 mmol L-1 of aminoethoxyvinylglycine (AVG), or 20 mmol L-1 of silver thiosulphate (STS). Seeds were also primed in polyethylene glycol (PEG), or PEG + ACC, PEG + AVG, or PEG + STS. Untreated seeds germinated 100% at 20°C. At 35°C, EVE seeds germinated 100%, whereas DGB seeds germinated only 33%. Seed priming or adding ACC during incubation increased germination at 35°C. Higher ethylene evolution was detected in EVE than in DGB during germination at 35°C. AVG did not inhibit seed germination of DGB at 35°C, but STS did. Higher endo-beta-mannanase activity was observed in EVE compared with DGB seeds. Providing ACC either during priming or during germination increased endo-beta-mannanase activity, whereas AVG and STS led to decreased or no activity. Ethylene may overcome the inhibitory effect of high temperature in thermosensitive lettuce seeds due to increased endo-beta-mannanase, possibly leading to weakening of the endosperm

Thermotolerance in lettuce seeds: association with ethylene and endo-B-mannanase.

Weakening of the endosperm tissue around the radicle tip before radicle protrusion and a potential role of endo-β-mannanase during germination of lettuce seeds (Lactuca sativa L.) at high temperature (35 °C) were investigated. Seeds from the thermotolerant genotypes ‘Everglades’ and PI 251245 had greater endo-β-mannanase activity before radicle protrusion at 35 °C than the thermosensitive genotypes ‘Dark Green Boston’, ‘Valmaine’ and ‘Floricos 83’. Thermotolerant genotypes also generated more ethylene at high temperature. At 35 °C, germination of ‘Dark Green Boston’ and ‘Everglades’ seeds produced at days/nights of 20/10 °C was 10% and 32%, respectively, whereas germination of seeds produced at days/nights of 30/20 °C was 67% and 83%, respectively. Higher endo-β-mannanase activity was observed before radicle protrusion in ‘Dark Green Boston’ seeds produced at 30/20 °C compared with those produced at 20/10 °C. A relationship between seed germination at high temperature, ethylene production, and an increase in endo-β-mannanase activity before radicle protrusion was confirmed

Bioassay for detection of glyphosate or kanamycin resistance in lettuce plants.

Trabalho apresentado no 41° Congresso Brasileiro de Olericultura, 2001

Iso-osmotic regulation of nitrate accumulation in lettuce (Lactuca sativa L.)

Concerns about possible health hazards arising from human consumption of lettuce and other edible vegetable crops with high concentrations of nitrate have generated demands for a greater understanding of processes involved in its uptake and accumulation in order to devise more sustainable strategies for its control. This paper evaluates a proposed iso-osmotic mechanism for the regulation of nitrate accumulation in lettuce (Lactuca sativa L.) heads. This mechanism assumes that changes in the concentrations of nitrate and all other endogenous osmotica (including anions, cations and neutral solutes) are continually adjusted in tandem to minimise differences in osmotic potential of the shoot sap during growth, with these changes occurring independently of any variations in external water potential. The hypothesis was tested using data from six new experiments, each with a single unique treatment comprising a separate combination of light intensity, N source (nitrate with or without ammonium) and nitrate concentration carried out hydroponically in a glasshouse using a butterhead lettuce variety. Repeat measurements of plant weights and estimates of all of the main soluble constituents (nitrate, potassium, calcium, magnesium, organic anions, chloride, phosphate, sulphate and soluble carbohydrates) in the shoot sap were made at intervals from about 2 weeks after transplanting until commercial maturity, and the data used to calculate changes in average osmotic potential in the shoot. Results showed that nitrate concentrations in the sap increased when average light levels were reduced by between 30 and 49 % and (to a lesser extent) when nitrate was supplied at a supra-optimal concentration, and declined with partial replacement of nitrate by ammonium in the external nutrient supply. The associated changes in the proportions of other endogenous osmotica, in combination with the adjustment of shoot water content, maintained the total solute concentrations in shoot sap approximately constant and minimised differences in osmotic potential between treatments at each sampling date. There was, however, a gradual increase in osmotic potential (ie a decline in total solute concentration) over time largely caused by increases in shoot water content associated with the physiological and morphological development of the plants. Regression analysis using normalised data (to correct for these time trends) showed that the results were consistent with a 1:1 exchange between the concentrations of nitrate and the sum of all other endogenous osmotica throughout growth, providing evidence that an iso-osmotic mechanism (incorporating both concentration and volume regulation) was involved in controlling nitrate concentrations in the shoot

A genetic locus and gene expression patterns associated with the priming effect on lettuce seed germination at elevated temperatures

Seeds of most cultivated varieties of lettuce (Lactuca sativa L.) fail to germinate at warm temperatures (i.e., above 25–30°C). Seed priming (controlled hydration followed by drying) alleviates this thermoinhibition by increasing the maximum germination temperature. We conducted a quantitative trait locus (QTL) analysis of seed germination responses to priming using a recombinant inbred line (RIL) population derived from a cross between L. sativa cv. Salinas and L. serriola accession UC96US23. Priming significantly increased the maximum germination temperature of the RIL population, and a single major QTL was responsible for 47% of the phenotypic variation due to priming. This QTL collocated with Htg6.1, a major QTL from UC96US23 associated with high temperature germination capacity. Seeds of three near-isogenic lines (NILs) carrying an Htg6.1 introgression from UC96US23 in a Salinas genetic background exhibited synergistic increases in maximum germination temperature in response to priming. LsNCED4, a gene encoding a key enzyme (9-cis-epoxycarotinoid dioxygenase) in the abscisic acid biosynthetic pathway, maps precisely with Htg6.1. Expression of LsNCED4 after imbibition for 24 h at high temperature was greater in non-primed seeds of Salinas, of a second cultivar (Titan) and of NILs containing Htg6.1 compared to primed seeds of the same genotypes. In contrast, expression of genes encoding regulated enzymes in the gibberellin and ethylene biosynthetic pathways (LsGA3ox1 and LsACS1, respectively) was enhanced by priming and suppressed by imbibition at elevated temperatures. Developmental and temperature regulation of hormonal biosynthetic pathways is associated with seed priming effects on germination temperature sensitivity

Quantitative trait loci associated with longevity of lettuce seeds under conventional and controlled deterioration storage conditions

Lettuce (Lactuca sativa L.) seeds have poor shelf life and exhibit thermoinhibition (fail to germinate) above ∼25°C. Seed priming (controlled hydration followed by drying) alleviates thermoinhibition by increasing the maximum germination temperature, but reduces lettuce seed longevity. Controlled deterioration (CD) or accelerated ageing storage conditions (i.e. elevated temperature and relative humidity) are used to study seed longevity and to predict potential seed lifetimes under conventional storage conditions. Seeds produced in 2002 and 2006 of a recombinant inbred line (RIL) population derived from a cross between L. sativa cv. Salinas×L. serriola accession UC96US23 were utilized to identify quantitative trait loci (QTLs) associated with seed longevity under CD and conventional storage conditions. Multiple longevity-associated QTLs were identified under both conventional and CD storage conditions for control (non-primed) and primed seeds. However, seed longevity was poorly correlated between the two storage conditions, suggesting that deterioration processes under CD conditions are not predictive of ageing in conventional storage conditions. Additionally, the same QTLs were not identified when RIL populations were grown in different years, indicating that lettuce seed longevity is strongly affected by production environment. Nonetheless, a major QTL on chromosome 4 [Seed longevity 4.1 (Slg4.1)] was responsible for almost 23% of the phenotypic variation in viability of the conventionally stored control seeds of the 2006 RIL population, with improved longevity conferred by the Salinas allele. QTL analyses may enable identification of mechanisms responsible for the sensitivity of primed seeds to CD conditions and breeding for improved seed longevity