Hormonal regulation of tomato seed germination at a supraoptimal temperature

Germination of tomato cv. New Yorker seed is inhibited at 35°C. This thermoinhibition was partially counteracted by application of GA₄₊₇ alone, the compound applied in combination with ACC or ethephon markedly enhancing the process. The latter compound alone was not able to induce germination at 35 °C. Thermoinhibition of seeds at 35 °C was also counteracted by fluridone, an inhibitor of ABA biosynthesis. At 25 °C, an optimal temperature, ABA inhibited germination of New Yorker seeds. Although another known growth inhibitor MeJA, when applied at an optimal temperature (25 °C), had also a slightly inhibitory effect on germination of those seeds and clearly delayed the process, inhibitors of its biosynthetic pathway (ibuprofen, indoprofen, antypiryne and salicylic acid) did not remove thermoinhibition at 35 °C. An increase in endo-β-mannanase activity after 24 hours of incubation at 35 °C was observed in the seeds incubated in the presence of gibberellins, ACC, ethephon, fluridone used alone and in combinations, but it was not clearly correlated with the effects of these compounds on alleviation of seed germination. However, fluridone present in the same incubation medium at 35 °C with ABA was able to counteract the inhibitory effect of ABA on endo-β-mannanase activity. The results of our study suggest that gibberellins, ethylene (produced from ACC or ethephon) and ABA, but not jasmonates, regulate tomato seed germination at supraoptimal temperatures. Alleviation of thermoinhibition of New Yorker seed germination by plant growth regulators and fluridone is partially associated with their controlling endo-β-mannanase activity.     

Abscisic acid in the thermoinhibition of lettuce seed germination and enhancement of its catabolism by gibberellin

Germination of lettuce (Lactuca sativa L. cv. 'Grand Rapids') seeds was inhibited at high temperatures (thermoinhibition). Thermoinhibition at 28 degrees C was prevented by the application of fluridone, an inhibitor of abscisic acid (ABA) biosynthesis. At 33 degrees C, the sensitivity of the seeds to ABA increased, and fluridone on its own was no longer effective. However, a combined application of fluridone and gibberellic acid (GA3) was able to restore the germination. Exogenous GA3 lowered endogenous ABA content in the seeds, enhancing catabolism of ABA and export of the catabolites from the intact seeds. The fluridone application also decreased the ABA content. Consequently, the combined application of fluridone and GA3 decreased the ABA content to a sufficiently low level to allow germination at 33 degrees C. There was no significant temperature-dependent change in endogenous GA1 contents. It is concluded that ABA is an important factor in the regulation of thermoinhibition of lettuce seed germination, and that GA affects the temperature responsiveness of the seeds through ABA metabolism.     

Control of seed dormancy in Nicotiana plumbaginifolia: post-imbibition abscisic acid synthesis imposes dormancy maintenance

The physiological characteristics of seed dormancy in Nicotiana plumbaginifolia Viv. are described. The level of seed dormancy is defined by the delay in seed germination (i.e the time required prior to germination) under favourable environmental conditions. A wild-type line shows a clear primary dormancy, which is suppressed by afterripening, whereas an abscisic acid (ABA)-deficient mutant shows a non-dormant phenotype. We have investigated the role of ABA and gibberellic acid (GA₃) in the control of dormancy maintenance or breakage during imbibition in suitable conditions. It was found that fluridone, a carotenoid biosynthesis inhibitor, is almost as efficient as GA₃ in breaking dormancy. Dry dormant seeds contained more ABA than dry afterripened seeds and, during early imbibition, there was an accumulation of ABA in dormant seeds, but not in afterripened seeds. In addition, fluridone and exogenous GA₃ inhibited the accumulation of ABA in imbibed dormant seeds. This reveals an important role for ABA synthesis in dormancy maintenance in imbibed seeds. Received: 31 December 1998 / Accepted: 9 July 1999     

Genetic variation for lettuce seed thermoinhibition is associated with temperature-sensitive expression of abscisic Acid, gibberellin, and ethylene biosynthesis, metabolism, and response genes

Lettuce (Lactuca sativa ‘Salinas’) seeds fail to germinate when imbibed at temperatures above 25°C to 30°C (termed thermoinhibition). However, seeds of an accession of Lactuca serriola (UC96US23) do not exhibit thermoinhibition up to 37°C in the light. Comparative genetics, physiology, and gene expression were analyzed in these genotypes to determine the mechanisms governing the regulation of seed germination by temperature. Germination of the two genotypes was differentially sensitive to abscisic acid (ABA) and gibberellin (GA) at elevated temperatures. Quantitative trait loci associated with these phenotypes colocated with a major quantitative trait locus (Htg6.1) from UC96US23 conferring germination thermotolerance. ABA contents were elevated in Salinas seeds that exhibited thermoinhibition, consistent with the ability of fluridone (an ABA biosynthesis inhibitor) to improve germination at high temperatures. Expression of many genes involved in ABA, GA, and ethylene biosynthesis, metabolism, and response was differentially affected by high temperature and light in the two genotypes. In general, ABA-related genes were more highly expressed when germination was inhibited, and GA- and ethylene-related genes were more highly expressed when germination was permitted. In particular, LsNCED4, a gene encoding an enzyme in the ABA biosynthetic pathway, was up-regulated by high temperature only in Salinas seeds and also colocated with Htg6.1. The temperature sensitivity of expression of LsNCED4 may determine the upper temperature limit for lettuce seed germination and may indirectly influence other regulatory pathways via interconnected effects of increased ABA biosynthesis.     

Triangular interplay between ROS,ABA and GA in dormancy alleviation of <Emphasis Type="Italic">Bunium persicum</Emphasis> seeds by cold stratification

Seeds of Bunium persicum (Boiss.) B. Fedtsch. have complex physiological dormancy that can be released by 15 weeks stratification. The present study revealed that cold stratification enhanced content of H₂O₂, O^{2 -·} and application of GA₃ and ROS donors (Fenton reagent, H₂O₂, methylviologen and menadione) did not affect or only slightly promoted the germination of non-stratified, fully dormant seeds. Dormancy was markedly decreased by ROS-generating reagents, GA₃ and fluridone (an inhibitor of ABA biosynthesis) and was enhanced by ROS-decreasing compounds (DMTU, Tiron, SB and DPI), diniconazole (Dinc, an inhibitor of ABA catabolism) and paclobutrazol (PAC, an inhibitor of GA biosynthesis) when dormancy was partially removed by cold stratification. The response to these compounds reduced with increasing time of stratification. ABA inhibited germination by repressing of NADPH oxidase activity and ROS accumulation and conversely, GA triggered germination by promoting an increase of NADPH oxidase activity and ROS levels. Data in this study, for the first time suggest releasing deep complex physiological dormancy by cold stratification is associated with interplay between ROS and ABA/GA.     

Germination response of black and yellow seed coated canola (<Emphasis Type="Italic">Brassica napus</Emphasis>) lines to chemical treatments under cold temperature conditions

Seed quality is a key critical component to produce well established and vigorous seedlings under cool soil (<10°C) conditions experienced in Western Canada. A simple, relatively quick germination assay is required to separate small differences in seed germination which can have a significant impact on seedling growth. It has long been established that phytohormones regulate seed germination: abscisic acid inhibits germination whereas gibberellins enhance germination. We investigated the effects of ABA, GA, ethylene and inhibitors of these phytohormones alone and in combination on the germination rate of a black and a yellow seed canola (Brassica napus) imbibed at 8°C. The effects of either saline solutions, osmotic solutions, fusicoccin or testa on the germination of canola seeds imbibed at 8°C were also investigated. This temperature is representative of the soil temperatures experienced in the early spring of Western Canada. The two canola seed lines, especially the yellow seed line, were very sensitive to increasing concentration of saline solutions at 8°C, but not at 23°C; however, iso-osmotic solutions that reduced water potential were more inhibitory. The seed coat (testa) including the endosperm was a major factor affecting the germination rate of the yellow seed line at 8°C, however, GA₄₊₇ overcame the inhibitory effect of the testa, whereas ABA exacerbated it. Fusicoccin was more stimulatory to germination than GA₄₊₇, however, unlike GA₄₊₇, it was unable to overcome the inhibitory effect of paclobutrazol, a GA biosynthesis inhibitor. Fluridone, an ABA biosynthesis inhibitor, was unable to overcome the inhibitory effects of a saline solution suggesting that the inhibitory effect was not due to elevated ABA levels. Ethylene, a stimulator of germination did not appear to be involved in the germination of these two lines. Controlled deterioration at 35°C, 85% RH could be either partially or completely overcome by exogenous GA₄₊₇. This study demonstrated the effect of hormones, salinity and testa on the germination of canola seeds under less than ideal environmental conditions.     

Abscisic acid regulates seed germination of Vellozia species in response to temperature

• The relationship between the phytohormones, gibberellin (GA) and abscisic acid (ABA) and light and temperature on seed germination is still not well understood. We aimed to investigate the role of the ABA and GA on seed germination of Vellozia caruncularis, V. intermedia and V. alutacea in response to light/dark conditions on different temperature.
• Seeds were incubated in GA (GA₃ or GA₄) or ABA and their respective biosynthesis inhibitors (paclobutrazol – PAC, and fluridone – FLU) solutions at two contrasting temperatures (25 and 40 °C). Furthermore, endogenous concentrations of active GAs and those of ABA were measured in seeds of V. intermedia and V. alutacea during imbibition/germination.
• Exogenous ABA inhibited the germination of Vellozia species under all conditions tested. GA, FLU and FLU + GA₃ stimulated germination in the dark at 25 °C (GA₄ being more effective than GA₃). PAC reduced seed germination in V. caruncularis and V. alutacea, but did not affect germination of V. intermedia at 40 °C either under light or dark conditions. During imbibition in the dark, levels of active GAs decreased in the seeds of V. intermedia, but were not altered in those of V. alutacea. Incubation at 40 °C decreased ABA levels during imbibition in both V. caruncularis and V. alutacea.
• We conclude that the seeds of Vellozia species studied here require light or high temperature to germinate and ABA has a major role in the regulation of Vellozia seed germination in response to light and temperature.

     

Changes in endogenous abscisic acid levels during dormancy release and maintenance of mature seeds: studies with the Cape Verde Islands ecotype,the dormant model of<Emphasis Type="Italic"> Arabidopsis thaliana</Emphasis>

Mature seeds of the Cape Verde Islands (Cvi) ecotype of Arabidopsis thaliana (L.) Heynh. show a very marked dormancy. Dormant (D) seeds completely fail to germinate in conditions that are favourable for germination whereas non-dormant (ND) seeds germinate easily. Cvi seed dormancy is alleviated by after-ripening, stratification, and also by nitrate or fluridone treatment. Addition of gibberellins to D seeds does not suppress dormancy efficiently, suggesting that gibberellins are not directly involved in the breaking of dormancy. Dormancy expression of Cvi seeds is strongly dependent on temperature: D seeds do not germinate at warm temperatures (20–27°C) but do so easily at a low temperature (13°C) or when a fluridone treatment is given to D seeds sown at high temperature. To investigate the role of abscisic acid (ABA) in dormancy release and maintenance, we measured the ABA content in both ND and D seeds imbibed using various dormancy-breaking conditions. It was found that dry D seeds contained higher amounts of ABA than dry ND after-ripened seeds. During early imbibition in standard conditions, there was a decrease in ABA content in both seeds, the rate of which was slower in D seeds. Three days after sowing, the ABA content in D seeds increased specifically and then remained at a high level. When imbibed with fluridone, nitrate or stratified, the ABA content of D seeds decreased and reached a level very near to that of ND seeds. In contrast, gibberellic acid (GA₃) treatment caused a transient increase in ABA content. When D seeds were sown at low optimal temperature their ABA content also decreased to the level observed in ND seeds. The present study indicates that Cvi D and ND seeds can be easily distinguished by their ability to synthesize ABA following imbibition. Treatments used here to break dormancy reduced the ABA level in imbibed D seeds to the level observed in ND seeds, with the exception of GA₃ treatment, which was active in promoting germination only when ABA synthesis was inhibited.Abbreviations ABA Abscisic acid - Cvi Cape Verde Islands - D Dormant - GA Gibberellin - GA₃ Gibberellic acid - ND Non dormant     

Cellular alteration and differential protein profile explain effects of GA3 and ABA and their inhibitor on Trichocline catharinensis (Asteraceae) seed germination

Seed physiology of wild species has not been studied as deeply as that of domesticated crop species. Trichocline catharinensis (Asteraceae) is an endemic wildflower species from the high-altitude fields of southern Brazil. This species is of interest as a source of genes to improve cultivated Asteraceae because of its ornamental features, disease resistance and ability to tolerate drought and poor soil conditions. We studied the effects of abscisic acid (ABA) and gibberellic acid (GA₃) and their inhibitors, fluridone (FLU) and paclobutrazol (PAC), on seed germination. We individually assessed ultrastructural changes and differential protein accumulation. The principal component analysis explained 69.66% of differential accumulation for 32 proteins at phase II of seed germination in response to hormone and inhibitor treatment. GA₃-imbibed seed germination (98.75%) resulted in increased protein accumulation to meet energy demand, redox regulation, and reserve metabolism activation. FLU-imbibed seeds showed a higher germination speed index as a consequence of metabolism activation. ABA-imbibed seeds (58.75%) showed osmotolerance and flattened cells in the hypocotyl-radicular axis, suggesting that ABA inhibits cell expansion. PAC-imbibed seeds remained at phase II for 300 h, and germination was suppressed (7.5%) because of the increased signaling proteins and halted reserve mobilization. Therefore, our findings provide insight into the behavior of Asteraceae non-dormant seed germination, which broadens our knowledge of seed germination in a wild and endemic plant species from a threatened ecosystem.     

Requirement for Ethylene Synthesis and Action during Relief of Thermoinhibition of Lettuce Seed Germination by Combinations of Gibberellic Acid, Kinetin, and Carbon Dioxide

Application of exogenous ethylene in combination with gibberellic acid (GA₃), kinetin (KIN), and/or CO₂ has been reported to induce germination of lettuce seeds at supraoptimal temperatures. However, it is not clear whether endogenous ethylene also plays a mediatory role when germination under these conditions is induced by treatment regimes that do not include ethylene. Therefore, possible involvement of endogenous ethylene during the relief of thermoinhibition of lettuce (Lactuca sativa L. cv Grand Rapids) seed germination at 32°C was investigated. Combinations of GA₃ (0.5 millimolar), KIN (0.05 millimolar), and CO₂ (10%) were used to induce germination. Little germination occurred in controls or upon treatment with ethylene, KIN, or CO₂. Neither KIN nor CO₂ affected the rate of ethylene production by seeds. Both germination and ethylene production were slightly promoted by GA₃. Treatments with GA₃+CO₂, GA₃+KIN, or GA₃+CO₂+KIN resulted in approximately 10-to 40-fold increases in ethylene production and 50 to 100% promotion of germination as compared to controls. Initial ethylene evolution from the treated seeds was greater than from the controls and a major surge in ethylene evolution occurred at the time of visible germination. Application of 1 millimolar 2-aminoethoxyvinyl glycine (AVG), an inhibitor of ethylene synthesis, in combination with any of above three treatments inhibited the ethylene production to below control levels. This was accompanied by a marked decline in germination percentage. Germination was also inhibited by 2,5-norbornadiene (0.25-2 milliliters per liter), a competitive inhibitor of ethylene action. Application of exogenous ethylene (1-100 microliters per liter) overcame the inhibitory effects of AVG and 2,5-norbornadiene on germination. The results demonstrate that endogenous ethylene synthesis and action are essential for the alleviation of thermoinhibition of lettuce seeds by combinations of GA₃, KIN, and CO₂. It also appears that these treatment combinations do not act exclusively via promotion of ethylene evolution as the application of exogenous ethylene alone did not promote germination.     

Profile of Plant Hormones and their Metabolites in Germinated and Ungerminated Canola (Brassica napus) Seeds Imbibed at 8°C in either GA4+7, ABA, or a Saline Solution

Abscisic acid (ABA) and gibberellins (GAs) are two major phytohormones that regulate seed germination in response to internal and external factors. In this study we used HPLC-ESI/MS/MS to investigate hormone profiles in canola (Brassica napus) seeds that were 25, 50, and 75% germinated and their ungerminated counterparts imbibed at 8°C in either water, 25 μM GA₄₊₇, a 80 mM saline solution, or 50 μM ABA, respectively. During germination, ABA levels declined while GA₄ levels increased. Higher ABA levels appeared in ungerminated seeds compared to germinated seeds. GA₄ levels were lower in seeds imbibed in the saline solution compared to seeds imbibed in water. Ungerminated seeds imbibed in ABA had lower GA₄ levels compared to ungerminated seeds imbibed in water; however, the levels of GA₄ were similar for germinated seeds imbibed in either water or ABA. The ABA metabolites PA and DPA increased in seeds imbibed in either water, the saline solution, or ABA, but decreased in GA₄₊₇-imbibed seeds. In addition, ABA inhibited GA₄ accumulation, whereas GA had no effect on ABA accumulation but altered the ABA catabolism pathway. Information from our studies strongly supports the concept that the balance of ABA and GA is a major factor controlling germination.     

Restoration of Seed Germination at Supraoptimal Temperatures by Fluridone, an Inhibitor of Abscisic Acid Biosynthesis

Fluridone, an inhibitor of ABA biosynthesis, restored the seedgermination of lettuce (Lactuca sativa L. cv. Grand Rapids)and many other plant species at supra-optimal temperatures.ABA content in lettuce seeds after imbibition quickly decreasedat 23°C, but not at 33°C (a supraoptimal temperature).Fluridone caused a decrease in ABA content at 33°C, whichsuggests that the maintenance of high ABA content could be responsiblefor high-temperature inhibition of germination of lettuce seeds.This probably results from an increase in the rate of ABA biosynthesisat the higher temperature. The present study indicates thatABA plays a decisive role in the regulation of seed germinationat supraoptimal temperatures. ¹ Corresponding author: fax 81-22-717-8834; e-mail yoshi@bios.tohoku.ac.jp     

ABA, GA3, and nitrate may control seed germination of Crithmum maritimum (Apiaceae) under saline conditions

Impaired germination is common among halophyte seeds exposed to salt stress, partly resulting from the salt-induced reduction of the growth regulator contents in seeds. Thus, the understanding of hormonal regulation during the germination process is a main key: (i) to overcome the mechanisms by which NaCl-salinity inhibit germination; and (ii) to improve the germination of these species when challenged with NaCl. In the present investigation, the effects of ABA, GA₃, NO⁻₃, and NH⁺₄ on the germination of the oilseed halophyte Crithmum maritimum (Apiaceae) were assessed under NaCl-salinity (up to 200 mM NaCl). Seeds were collected from Tabarka rocky coasts (N-W of Tunisia). The exogenous application of GA₃, nitrate (either as NaNO₃ or KNO₃), and NH₄Cl enhanced germination under NaCl salinity. The beneficial impact of KNO₃ on germination upon seed exposure to NaCl salinity was rather due to NO⁻₃ than to K⁺, since KCl failed to significantly stimulate germination. Under optimal conditions for germination (0 mM NaCl), ABA inhibited germination over time in a dose dependent manner, but KNO₃ completely restored the germination parameters. Under NaCl salinity, the application of fluridone (FLU) an inhibitor of ABA biosynthesis, stimulated substantially seed germination. Taken together, our results point out that NO⁻₃ and GA₃ mitigate the NaCl-induced reduction of seed germination, and that NO⁻₃ counteracts the inhibitory effect of ABA on germination of C. maritimum. To cite this article: A. Atia et al., C. R. Biologies 332 (2009).     

Regulation of Medicago sativa L. somatic embryos regeneration by gibberellin A3 and abscisic acid in relation to starch content and α-amylase activity

Somatic embryo quality is an important factor decisive for the efficiency of somatic embryogenesis. Addition gibberellic acid (GA₃) at a concentration of 1 μM to germination medium improved the regeneration of alfalfa somatic embryos. Inhibitory effect of ancymidol, an inhibitor of gibberellin biosynthesis, on germination and conversion may indicate that those processes require endogenous GAs. Since fluridone, an ABA biosynthetic inhibitor, at a concentration of 1 μM, had a slight stimulatory effect on germination of somatic embryos, it may be presumed that embryos contain a too high level of residual ABA after maturation phase (20 μM ABA is used at that phase). The observed improvement of regeneration of somatic embryos by GA₃ was correlated with acceleration of starch hydrolysis through α-amylase activity enhancement by GA₃. In contrast, the inhibitory effect of ABA on the above processes was probably related to inhibition of α-amylase activity and, in consequence, to delayed starch hydrolysis. It is suggested that α-amylase activity can be considered a good marker of the quality of Medicago sativa L. somatic embryos.     

The effects of gibberellic acid,fluridone, abscisic acid and paclobutrazol on anther culture of Brussels sprouts

Both gibberellic acid (GA₃) and fluridone, a non-specific inhibitor of ABA biosynthesis, promoted embryo production in anther cultures of Brussels sprouts cv. Hal, but not in cv. Gower. Abscisic acid (ABA) and the gibberellin-biosynthesis inhibitor paclobutrazol inhibited embryo production in both cultivars.     

Effects of gibberellic acid,ethephon, and 1-aminocyclopropane-1-carboxylic acid on germination ofAmaranthus caudatus seeds inhibited by paclobutrazol

The specific inhibitor of gibberellin biosynthesis, (2RS,3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)pentan-3-ol (paclobutrazol), inhibited germination ofAmaranthus caudatus L. seeds. Addition of gibberellic acid (GA₃), 2-chloroethylphosphonic acid (ethephon), or 1-aminocyclopropane-1-carboxylic acid (ACC) effectively antagonized inhibition. Ethephon was found to be the most efficient antagonist. The transfer of seeds after 1 day's incubation in paclobutrazol to solutions of GA₃ or ethephon reversed the inhibition, the effect increasing with increasing concentration of GA₃ or ethephon. Seeds incubated in paclobutrazol for 5 days decreased sensitivity to GA₃ and ethephon.     

Evidence for the accumulation of a germination inhibitor during progressive thermoinhibition of seeds of celery (Apium graveolens L.)

The germination of seeds of celery (Apium graveolens L.) becomes progressively thermoinhibited on incubation in the dark at high temperatures, the inhibitory temperature being dependent on the cultivar used. In two high-dormancy cultivars of celery, the production of germination inhibitors in seeds incubated in the dark at 26°C gradually increased over a 7-day period. Inhibitor production was measured by incubating seeds of the low-dormancy cultivar Florida 683 in homogenates of the thermoinhibited seeds of the high-dormancy cultivars and recording germination either in the light or with the gibberellins A₄ and A₇ (GA_4/7) in the dark. Most Florida 683 seeds which failed to germinate in the homogenates after 15 days were induced to germinate by addition of N⁶-benzyladenine (BA). The presence of BA in addition to GA_4/7 throughout incubation in the dark completely overcame the inhibitory effects of homogenates. This indicates that thermoinhibition of celery seeds is associated with the accumulation of a germination inhibitor which interacts with cytokinins. This does not appear to be abscisic acid (ABA) since ABA levels in thermoinhibited seeds were lower than in untreated seeds and did not increase with duration of high temperature treatment.Abbreviations ABA Abscisic acid - BA N⁶-benzyladenine - GA_4/7 a mixture of the gibberellins A₄ and A₇ - HTP high-temperature pretreatment     

Functional relationships of <Emphasis Type="Italic">phytoene synthase 1</Emphasis> alleles on chromosome 7A controlling flour colour variation in selected Australian wheat genotypes

Thermoinhibition, or failure of seeds to germinate when imbibed at warm temperatures, can be a significant problem in lettuce (Lactuca sativa L.) production. The reliability of stand establishment would be improved by increasing the ability of lettuce seeds to germinate at high temperatures. Genes encoding germination- or dormancy-related proteins were mapped in a recombinant inbred line population derived from a cross between L. sativa cv. Salinas and L. serriola accession UC96US23. This revealed several candidate genes that are located in the genomic regions containing quantitative trait loci (QTLs) associated with temperature and light requirements for germination. In particular, LsNCED4, a temperature-regulated gene in the biosynthetic pathway for abscisic acid (ABA), a germination inhibitor, mapped to the center of a previously detected QTL for high temperature germination (Htg6.1) from UC96US23. Three sets of sister BC₃S₂ near-isogenic lines (NILs) that were homozygous for the UC96US23 allele of LsNCED4 at Htg6.1 were developed by backcrossing to cv. Salinas and marker-assisted selection followed by selfing. The maximum temperature for germination of NIL seed lots with the UC96US23 allele at LsNCED4 was increased by 2–3°C when compared with sister NIL seed lots lacking the introgression. In addition, the expression of LsNCED4 was two- to threefold lower in the former NIL lines as compared to expression in the latter. Together, these data strongly implicate LsNCED4 as the candidate gene responsible for the Htg6.1 phenotype and indicate that decreased ABA biosynthesis at high imbibition temperatures is a major factor responsible for the increased germination thermotolerance of UC96US23 seeds.     

Isolation and characterization of high temperature-resistant germination mutants of Arabidopsis thaliana

Temperature is a primary environmental cue for seed germination of many weeds and vegetables. To investigate the mechanism of germination regulation by temperature, we selected five high temperature (thermoinhibition)-resistant germination mutants (TRW lines) from 20,000 T-DNA insertion lines of Arabidopsis. Segregation analyses indicated that each of the five lines had single locus recessive mutations. The seeds of TRW134-15 and TRW187 showed reduced sensitivity to ABA and also to the gibberrellin biosynthesis inhibitor, paclobutrazol. Genetic and nucleotide sequencing analyses indicated that TRW187 is a new allele of abi3 (abi3-14). TRW71-1 exhibited a maternal effect for both thermoinhibition-resistant and transparent testa phenotypes, and genetic analysis revealed that the mutation was allelic to tt7 (tt7-4 sib). Interestingly, the seeds of reduced dormancy mutants rdo1, rdo2, rdo3 and rdo4 were also thermoinhibition tolerant, and all the TRW seeds showed reduced dormancy. Like rdo3, TRW13-1 had shorter siliques and slightly shorter stems than the wild type. The mutation of TRW13-1 was mapped to the bottom arm of chromosome 1 where rdo3 has also been mapped, but the two mutants are not allelic. We designated TRW13-1 as thermoinhibition-resistant germination 1 (trg1). We also mapped the ABA-insensitive mutation of TRW134-15 to the bottom arm of chromosome 5 and named it trg2. These results show that both embryo/endosperm and maternal factors contribute to germination inhibition at supraoptimal temperatures in Arabidopsis. In addition, we confirm the role of ABA in thermoinhibition of seed germination and a link between seed physiological dormancy and response to high temperature.     

Hormonal and temperature regulation of seed dormancy and germination in <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Fluctuating temperature plays a critical role in determining the timing of seed germination in many plant species. However, the physiological and biochemical mechanisms underlying such a response have been paid little attention. The present study investigated the effect of plant growth regulators and cold stratification in regulating Leymus chinensis seed germination and dormancy response to temperature. Results showed that seed germination was less than 2 % at all constant temperatures while fluctuating temperature significantly increased germination percentage. The highest germination was 71 % at 20/30 °C. Removal of the embryo enclosing material of L. chinensis seed germinated to 74 %, and replaced the requirement for fluctuating temperature to germinate, by increasing embryo growth potential. Applications of GA₄₊₇ significantly increased seed germination at constant temperature. Also, inhibition of GA biosynthesis significantly decreased seed germination at fluctuating temperatures depending upon paclobutrazol concentration. This implied GA was necessary for non-dormant seed germination and played an important role in regulating seed germination response to temperature. Inhibition of ABA biosynthesis during imbibition completely released seed dormancy at 20/30 °C, but showed no effect on seed germination at constant temperature, suggesting ABA biosynthesis was important for seed dormancy maintenance but may not involve in seed germination response to temperature. Cold stratification with water or GA₃ induced seed into secondary dormancy, but this effect was reversed by exogenous FL, suggesting ABA biosynthesis during cold stratification was involved in secondary dormancy. Also, cold stratification with FL entirely replaced the requirement of fluctuating temperature for germination with seeds having 73 % germination at constant temperature. This appears to be attributed to inhibition of ABA biosynthesis and an increase of GA biosynthesis during cold stratification, leading to an increased embryo growth potential. We suggest that fluctuating temperature promotes seed germination by increasing embryo growth potential, mainly attributed to GA biosynthesis during imbibitions. ABA is important for seed dormancy maintenance and induction but showed less effect on non-dormant seed germination response to temperature.