Mode of action of gibberellic Acid and light on lettuce seed germination

The seeds of lettuce (Lactuca sativa L. cv. Grand Rapids) germinate in darkness at 25 C when treated by gibberellic acid (GA₃) for 1 hour following 2 hours of imbibition. The time of GA₃ application influences the rate and the final percentage of seeds that germinate. In contrast, red light illumination given at different times affects only the rate and not the final germination percentage. The early process(es) of germination initiated by GA₃ or light treatment can be arrested by subjecting the treated seeds to a nongerminative temperature of 35 C. The results suggest differences in the mode of action of light and GA₃ during germination. They indicate that different kinds of processes are involved in the biochemical control of germination.     

Differential effects of actinomycin d and cordycepin in lettuce seed germination and RNA synthesis

Intact lettuce seed germination was inhibited by cordycepin but not by actinomycin D; however, when seeds were clipped at the cotyledonary end, actinomycin D partially inhibited germination. Uptake studies with intact seeds using ³H-actinomycin D showed that it was unable to reach the embryo prior to radical protrusion. ³H-Cordycepin uptake studies using intact seeds showed that cordycepin was able to reach the embryo during the first 3 hours of incubation and at subsequent times. The pericarp and endosperm offered resistance to penetration of cordycepin into the embryo. In contrast to actinomycin D, cordycepin markedly inhibited ³H-uridine incorporation into RNA of intact seeds during the first 10 and 12 hours of incubation. About 60% of ³H-adenosine incorporation into poly A-RNA was inhibited by cordycepin during 12 hours of incubation, whereas actinomycin D had little effect. RNA synthesis appears to be essential for seed germination.     

RNA synthesis and the germination of light-sensitive lettuce seeds

Summary The germination of lettuce seeds is inhibited by the nucleotide base analogue 6-methylpurine. RNA synthesis has been measured during imbibition and germination as ³²P-phosphate incorporation into RNA species as fractionated by polyacrylamide gel electrophoresis. Seeds were surface sterilized and imbibed in the presence of various antibiotics. RNA preparations from lettuce seeds were coelectrophoresed with ³H-RNA prepared from bacteria to check for bacterial contamination of the seeds. There is a much higher rate of RNA synthesis in illuminated, germinating seeds as compared to dark, non-germinating seeds. This difference does not develop until after 12 hours of imbibition at 27°, which is the time of onset of germination and radicle growth.This investigation was supported by a contract from the United States Department of Agriculture (No. 616-15-3). Journal paper of the Purdue Agriculture Experiment Station.     

Stimulation of lettuce seed germination by ethylene

Ethylene increased the germination of freshly imbibed lettuce (Lactuca sativa L. var. Grand Rapids) seeds. Seeds receiving either red or far-red light or darkness all showed a positive response to the gas. However, ethylene was apparently without effect on dormant seeds, those which failed to germinate after an initial red or far-red treatment. Carbon dioxide, which often acts as a competitive inhibitor of ethylene, failed to clearly reverse ethylene-enhanced seed germination. While light doubled ethylene production from the lettuce seeds, its effect was not mediated by the phytochrome system since both red and far-red light had a similar effect.     

Mediation of phytochrome in the inductive action of low temperature on dark germination of lettuce seed at supra-optimal temperature

The induction of dark germination in light-requiring lettuce (Lactuca sativa) seed at supraoptimal temperatures by cold treatment (in darkness) was partly reversed by a brief far-red irradiation made at time of transfer, and even more so when the irradiation was made at the beginning of the cold pretreatment. When the inhibitory far-red irradiation was followed by additional cold treatment, the promotion was greatly restored. The promotive effects of brief irradiations with red light were further enhanced by a following cold period, before transfer to the supraoptimal temperature. These results are interpreted as indicating that the active (far-red absorbing) form of phytochrome is pre-existing in the dry seed, and interacts with a co-factor which is built-up during imbibition. The rate of build-up of this co-factor, as well as of the dark inactivation of active phytochrome increase with temperature. The products of the interaction pass through a photo-labile thermo-stable phase, before becoming photo-stable as well.     

Actions of gibberellic Acid and phytochrome on the germination of grand rapids lettuce seeds

Red light and gibberellic acid were about equally effective in promoting germination of Grand Rapids lettuce (Lactuca sativa L.) seeds. With initial far red light treatment more than 80% remained dormant in subsequent dark storage. After 2 days of dark storage, red light effectively promoted germination, while gibberellic acid action was weak. With between 2 and 10 days of dark storage, gibberellic acid had little effect, while promotion by red light decreased slowly and finally disappeared. After 10 days of dark storage, both gibberellic acid and red light were required for germination. The dark storage treatment interferes with phytochrome-independent germination processes and cannot be overcome by added gibberellic acid. However, storage may also decrease the effectiveness of endogenous gibberellins. Phytochrome-dependent germination seems to require only low levels of endogenous gibberellin activity or the addition of gibberellic acid. Gibberellins and red light appear to act on germination by regulation of sequential sites of a branched-looped pathway.     

The effect of growth retardants on phytochrome-induced lettuce seed germination

Experiments were carried out to explore the involvement of the plant hormone gibberellin (GA) in the light-induced germination of lettuce seeds. Three growth retardants known to be inhibitors of GA biosynthesis were tested for their effect on red-light-induced germination. Chlormequat chloride (CCC) and AMO-1618 had no effect, but ancymidol was strongly inhibitory. Moreover, the inhibition caused by ancymidol was completely overcome by GA₃. CCC and AMO-1618 inhibit the formation ofent-kaurene, while ancymidol blocks the oxidation ofent-kaurene toent-kaurenoic acid. Ancymidol also was found to inhibit GA-induced dark germination of lettuce seeds, and this inhibition was partially reversed by higher levels of GA. Therefore, the results suggest two possibilities for the relationship between phytochrome and GA in this system: first, the rate-limiting step in the germination of light-sensitive lettuce seeds, that which is regulated by phytochrome, is the oxidation ofent-kaurene toent-kaurenoic acid. Alternatively, red-light treatment may result in the release of active GAlike substances which, in turn, induce germination. In either case the results presented here support the view that phytochrome exerts its effect on lettuce seed germination by means of GA rather than via an independent pathway.     

Morphactin-kinetin interaction in lettuce seed germination and seedling growth

Summary Butylester of morphactin (n-butyl-9-hydroxy-fluorence-(9)-carboxylate) greatly inhibited seed germination and seedling growth of two strains of lettuce, Attraktion and Hohlblättriger Butter. The inhibitory effect of morphactin on seed germination was completely overcome by kinetin but the latter chemical was ineffective in reversing the morphactin induced inhibition of seedling growth. Thus it appears probable that the two substances effect seedling growth independently.     

Morphactin-gibberellin interaction in lettuce seed germination and seedling growth

Morphactin-butylester (a flourene-9-carboxylic acid derivative) inhibited seed germination of two strains of lettuce. Morphactin induced inhibition of germination could be partially or wholly reversed by simultaneous addition of gibberellic acid. However, gibberellic acid played very little part in reversing the inhibitory effect of morphactin on seedling growth. It is concluded that gibberellin can not reverse all the growth effects induced by morphactin.     

Promotive effects of organic solvents and kinetin on dark germination of lettuce seeds

Significant promotion in dark germination was observed when Grand Rapids lettuce (Lactuca sativa L.) seeds were soaked in acetone or dichloromethane, vacuum-dried, and imbibed at 25 C. Permeation of kinetin via these organic solvents further enhanced the dark germination. Those seeds that were affected by acetone and acetone-kinetin treatments and germinated in the dark escaped red-far red photocontrol of germination. Although abscisic acid was not detected in the organic solvent leachates, they did contain other inhibitory substances affecting lettuce seedling growth. In the light, acetone and acetone-kinetin treatments also enhanced the rate of germination and the increased germination by acetone-kinetin treatment was correlated with increased polyribosome formation. The possible mechanisms involved in promotion of lettuce seed germination by organic solvents and kinetin are discussed.     

Independent mode of action of cyanide and light on lettuce seed germination

Gaseous hydrogen cyanide stimulated subsequent lettuce seed ( Lactuca sativa L. cv. Grand Rapids) germination in darkness when applied for 1 or 22 h. Optimum concentrations were 5 × 10-5 M and 10-6 M, respectively. However, seeds did not germinate in the presence of HCN even at 10-8 M. The effects of unsaturating red light (R) and HCN (1 μM) showed a slight synergism. On the other hand, there was no difference between the effects of the sequences HCN – R and R – HCN. Stimulation of lettuce germination by an HCN pulse was practically not affected by far-red illumination, independently of the sequence of treatments. It was concluded that the primary stimulatory effect of HCN is of a regulatory character. Cyanide controls a regulatory point different from that affected by activated phytochrome.     

Inhibition of lettuce seed germination and seedling growth by antimetabolites of nucleic acids,and reversal by nucleic acid precursors and gibberellic acid

Summary Germination of White Paris lettuce seeds is inhibited by 2-thiouracil up to 24 hours. This inhibition is reversed by RNA precursors only. Seedling growth of lettuce is inhibited by 2-thiouracil and 5-fluorouracil; and white the effect of 2-thiouracil is counteracted by RNA precursors, inhibition due to 5-fluorouracil is not reversed significantly by any nucleic acid precursors. Possibly 2-thiouracil controls germination and seedling growth by interfering with RNA synthesis, while the effect of 5-fluorouracil is non-specific.In the presence of gibberellic acid, 5-fluorouracil and 2-thiouracil are relatively ineffective in causing inhibition of hypocotyl growth. Mechanism of gibberellic acid action remains obscure.     

Lettuce seed germination: modulation of pregermination protein synthesis by gibberellic Acid, abscisic Acid, and cytokinin

Protein synthesis in gibberellin-treated lettuce (Lactuca sativa) seeds has been studied during the lag phase between the beginning of imbibition and the first signs of radicle protrusion. When compared to the water-imbibed controls, both polyribosome populations and radioactive leucine incorporation into protein increase in the embryos of GA₃- induced seeds early in the imbibition period. Since these results are contradictory to previously published studies, the reasons for the differences are outlined and various alternative possibilities eliminated. The protocol for protein extraction, particularly the speed at which the supernatant from the seed homogenate is cleared, is important for demonstrating the GA₃-mediated changes. Embryos maintained in the dormant state by abscisic acid still conduct considerable amounts of protein synthesis, and this is enhanced by concentrations of 6-benzylaminopurine which also promote germination. Therefore, the actions of GA₃, abscisic acid, and cytokinin on lettuce seed germination are mediated, directly or indirectly, via protein synthesis.     

Interaction of triacontanol with gibberellin in control of lettuce seed germination

Triacontanol at concentrations from 2.3 × 10^-9 M to 2.3 × 10^-7 M did not affect the germination of lettuce ( Lactuca sativa L., cv. Grand Rapids) seeds in darkness, stimulated by light at 25°C or by benzyladenine at 31°C. Stimulation of seed germination by gibberellin A₃ (10^-5 M ) was significantly inhibited by triacontanol; the most effective concentration was 4.6 × 10^-8 M. Pulse experiments demonstrated that triacontanol was ineffective when applied later than gibberellin, whereas an inverse sequence of treatment caused an inhibition comparable to that resulting from continuous treatment of seeds with both factors. Possible interaction of triacontanol with gibberellin receptor is discussed.