Effects of ethylene and some other environmental factors on different stages of germination in red root pigweed (Amaranthus retroflexus L.) seeds*

Abstract. Ethylene was found to promote two distinct processes during germination of redroot pigweed (Amarantus retroflexus L.) seeds: embryo expansion that splits the seed coat (incomplete germination), and radicle penetration through the more elastic endosperm (complete germination). The two events can be separated in time by subjecting seeds to low water potential or low CO₂ levels, which arrest germination of some seeds at the incomplete stage. Ethylene applications to incompletely germinated seeds promote complete germination, with a response threshold near 0.02 cm³ m^?3 and saturation near 0.5 cm³ m^?3. Higher ethylene concentrations (0.5 to 50 cm³ m^?3) given during the first day of seed imbibition also increase the percentage of seeds which initiate embryo expansion and split the seed coat. Light and elevated CO₂ also promote radicle penetration of the endosperm in seeds incubated under water stress. The results support the view that the germination pause at the incomplete stage is an adaptation to environmental stresses that can be overcome with exogenous ethylene or certain other stimuli.     

Response of Amaranthus retroflexus L. seeds to gibberellic acid, ethylene and abscisic acid depending on duration of stratification and burial

Freshly harvested, dormant seeds of Amaranthus retroflexus were unable to germinate at 25 and 35 °C. To release their dormancy at the above temperatures, the seeds were stratified at a constant temperature (4 °C) under laboratory conditions or at fluctuating temperatures in soil or by outdoor burial in soil. Fully dormant, or seeds stratified or buried (2006/2007 and 2007/2008) for various periods were treated with exogenous gibberellic acid (GA₃), ethephon and abscisic acid (ABA). Likewise, the effects of these regulators, applied during stratification, on seed germination were determined. The results indicate that A. retroflexus seed dormancy can be released either by stratification or by autumn–winter burial. The effect of GA₃ and ethylene, liberated from ethephon, applied after various periods of stratification or during stratification, depends on dormancy level. GA₃ did not affect or only slightly stimulated the germination of non-stratified, fully dormant seeds at 25 and 35 °C respectively. Ethylene increased germination at both temperatures. Seed response to GA₃ and ethylene at 25 °C was increased when dormancy was partially removed by stratification at constant or fluctuating temperatures or autumn–winter burial. The response to GA₃ and ethylene increased with increasing time of stratification. The presence of GA₃ and ethephon during stratification may stimulate germination at 35 °C. Thus, both GA₃ and ethylene can partially substitute the requirement for stratification or autumn–winter burial. Both hormones may also stimulate germination of secondary dormant seeds, exhumed in September. The response to ABA decreased in parallel with an increasing time of stratification and burial up to May 2007 or March 2008. Endogenous GA_n, ethylene and ABA may be involved in the control of dormancy state and germination of A. retroflexus. It is possible that releasing dormancy by stratification or partial burial is associated with changes in ABA/GA and ethylene balance and/or sensitivity to these hormones.     

The levels of GA3 and GA20 may be associated with dormancy release in Onopordum nervosum seeds

Endogenous levels of two gibberellins, GA₃ andGA₂₀, were quantified in unimbibed Onopordumnervosum seeds collected from two different populations, whichshoweddifferences in their germination capacity. After purifying the seed extracts,gibberellin levels were evaluated by gas chromatography mass spectrometry byusing selected ion monitoring (GC-MS-SIM) adding deuterated gibberellins asinternal standards. The intraspecific differences in germination capacity wereassociated with differences in the endogenous levels of both gibberellins. Thecontents of GA₃ and GA₂₀ in seeds with high germinationrate were twice and five times higher, respectively, than those from seeds witha low germination rate, indicating a possible role of gibberellins in dormancyrelease in this plant species.     

Changes in sensitivity of Amaranthus retroflexus L seeds to ethylene during preincubation. II. Effects of alternating temperature and burial in soil

Abstract. The effects of diurnally alternating temperatures and of prolonged burial in the soil on germination response of redroot pigweed ( Amaranthus retroflexus L.) seeds to ethylene were investigated. Percentage germination in a 12 h/12 h, 23° C/35° C temperature regime roughly equalled that observed at constant 35° C, and greatly exceeded that observed at 30°C. Preincubation for 61 d in alternating temperatures, which were gradually increased to simulate soil warming in spring, caused little germination in the absence of ethylene, but considerably enhanced sensitivity to ethylene. Seeds kept in soil in the same temperature regime failed to show the response to ethylene, and the soil itself removed ethylene from the soil atmosphere.
After burial in a field plot either over winter or during the summer, seeds had a very low ethylene response threshold (0.01−0.05 cm³ m⁻³) and strong response to ethylene (70–95% germination at 51 cm³ m⁻³ compared to 1–20% without ethylene). Germinability of seeds buried overwinter declined between 10 May (85%) and 24 May (7%), and 90% of those recovered on or after 24 May had a visible rupture in the seed coat. Apparently, germination had begun during burial, but was arrested by unknown causes in an early phase and was followed by seed deterioration.
Although the role of ethylene in germination of buried seeds remains uncertain, the greatly enhanced sensitivity to ethylene observed in pigweed seeds after burial deserves further investigation.     

Responsiveness of Amaranthus retroflexus seeds to ethephon, 1-aminocyclopropane 1-carboxylic acid and gibberellic acid in relation to temperature and dormancy

Dormant Amaranthus retroflexus seeds do not germinate in the dark at temperatures below 35°C. Fully dormant seeds germinate only at 35–40°C whereas non-dormant ones germinate within a wider range of temperatures (15 to 40°C). Germination of non-dormant seeds requires at least 10% oxygen, but the sensitivity of seeds to oxygen deprivation increases with increasing depth of dormancy. 10^–6 to 10^–4 M ethephon, 10^–3 M 1-aminocyclopropane 1-carboxylic acid (ACC) and 10^–3 M gibberellic acid (GA₃) break this dormancy. In the presence of 10^–3 M GA₃ dormant seeds are able to germinate in the same range of temperatures as non-dormant seeds. The stimulatory effect of GA₃ is less dependent on temperature than that of ethephon, while ACC stimulates germination only at relatively high temperatures (25–30°C). The results obtained are discussed in relation to the possible involvement of endogenous ethylene in the regulation of germination of A. retroflexus seeds.Abbreviations ACC 1-aminocyclopropane 1-carboxylic acid - GA₃ gibberellic acid - SD standard deviation     

Some effects of nitrate-treated soil upon the sensitivity of buried redroot pigweed (Amaranthus retroflexus L.) seeds to ethylene, temperature, light and carbon dioxide

Abstract Fresh dormant redroot pigweed (Amaranthus retroflexus L.) seeds were buried 5 cm deep in the field at Stoneville, MS in November 1981. Potassium nitrate (200 kg ha ¹) or nothing was applied to the soil in the fall of 1981 and the late winter of 1982. Seeds were recovered at intervals under darkness during the following 2 years and tested for responses to ethylene, temperature, light and carbon dioxide. During the first overwintering, nitrate enhanced loss of primary dormancy and increases seed sensitivity to temperature, light and ethylene. The loss of dormancy reached a maximum at 25 to 30 weeks (early summer) after burial. Examination of the recovered seeds indicated that about 80% of the non-treated seeds and 98% of the nitrate-treated seeds germinated in situ during the period of maximum loss of dormancy. Thus, after one overwintering period, about 20% of the original buried seed population remained dormant in nontreted soil and 2% remained dormant in the nitratetreated soil. After the second overwintering, the percentages of dormant seeds remaining in nontreated or treated soil were both only 1–2%. Nitrate reduced dormancy and enhanced germination in early summer following the first overwintering. Regardless of treatment, the remaining 1 2% of seeds in soil after the second year were of low sensitivity to the germination stimuli (ethylene, temperature, light) and constituted the long-lived portion of the original seed population.     

The role of gibberellic acid (GA3) in the removal of dormancy inFraxinus excelsior L. seeds

The seeds ofFraxinus excelsior L. were stratified at 17-20 °C (warm stratification), at 4-6 °C (cold stratification) and at alternating temperature (warm — cold stratification). The seeds subjected to warm stratification only, remained dormant. The seeds stratified only at 4-6 °C germinated gradually during a long period of time. The seeds subjected to warm — cold stratification, however, germinated with great intensity within a relatively short period of time. GA₃ was shown to stimulate the growth of embryos markedly, and its effect on the germination of seeds depended on the temperature of stratification. GA₃ applied during the cold stratification accelerated the removal of dormancy by shortening the period of stratification and by influencing the germination of seeds. The results obtained indicate a similarity between the effect of temperature 17-20 °C during the warm stratification and that of gibberellic acid. Both those factors applied separately affect favourably after-ripening of the embryos and accelerate the germination of seeds.     

Changes in Phytochrome Expressed by Germination of Amaranthus retroflexus L. Seeds

Effects of red (600 to 680 nanometers) and far red (700 to 760 nanometers) irradiances on Amaranthus retroflexus L. seeds indicate that synthesis of phytochrome in the red-absorbing form takes place in water-imbibed nongerminating seeds at 35 C. After 96 hours in darkness, conversion of about 0.10% phytochrome to the far red-absorbing form induces 50% germination. Continuous far red radiation at 35 C with an irradiance of 0.4 × 10⁻¹⁰ Einsteins per square centimeter per second caused photoinactivation of phytochrome about equal to the rate of synthesis. Germination of seeds at 35 C, following far red irradiation adequate to establish the photostationary state, is enhanced by holding at 26 C for 16 minutes. Germination is unaffected relative to controls at constant temperature, if the period at 26 C precedes irradiation. The results indicate a quick response to action of phytochrome in a germination process.     

Comparison of carbon dioxide fixation and the fine structure in various assimilatory tissues of Amaranthus retroflexus L

The pattern for primary products of CO₂-fixation and the chloroplaststructure of Amaranthus retrqflexus L., a species which incorporatescarbon dioxide into C₄ dicarboxylic acids as the primary productof photosynthesis, were compared in various chlorophyll containingtissues,i.e., foliage leaves, stems, cotyledons and pale-greencallus induced from stem pith. Despite some morphological differencesin these assimilatory tissues, malate and aspartate were identifiedas the major compounds labelled during a 10 sec fixation of¹⁴CO₂ in all tissues. Whereas, aspartate was the major componentin C₄-dicarboxylic acids formed in foliage leaves, malate predominatedas the primary product in stems, cotyledons and the pale-greencallus. The percentage of ¹⁴C-radioactivity incorporated intoPGA and sugar-P esters increased and ¹⁴C-sucrose was detectedin the prolonged fixation of ¹⁴CO₂ in the light, not only infoliage leaves, but also in stems and cotyledons. ¹ This work was supported by a Grant for Scientific ResearchNo. 58813, from the Ministry of Education, Japan. ² Present address: Institute of Applied Microbiology, Universityof Tokyo, Tokyo, Japan. ³ Present address: Department of Biochemistry, University ofGeorgia, Athens 30601. Georgia, U. S. A. (Received July 10, 1971; )     

Impacts of fire cues on germination of Brassica napus L. seeds with high and low secondary dormancy

• Agricultural burning is used in farm management operations; however, information about the impact of fire cues on the release and/or induction of secondary dormancy in crop seeds is scarce.
• Seeds from two oilseed rape cultivars were induced for high (HD) or low (LD) secondary dormancy using polyethyleneglycol (PEG) pre‐treatment, and their germination after exposure to various fire cues was compared to control PEG pre‐treated and non‐dormant seeds.
• Non‐dormant seed germination was unaffected by various fire cues. Low doses of aerosol smoke released secondary dormancy in HD seeds, while higher doses increased dormancy of LD seeds. Dilute smoke water also released HD seed secondary dormancy, but concentrated smke water enhanced dormancy in both LD and HD seeds. The concentrated aqueous extracts from charred oilseed rape straw only promoted germination of HD seeds, while dilution inhibited LD seed germination. Heat shock (80 °C, 5 min) released secondary dormancy in HD seeds; however, higher temperatures and/or increased exposure time was associated with seed death. GC‐MS analyses of smoke water revealed two butenolides and an array of monoaromatic hydroxybenzene compounds with potential germination inhibitor or promoter activity.
• The extent of secondary dormancy induction in seeds affects their subsequent responses to fire cues. Both aerosol smoke and smoke water have both germination promoter and inhibitor activity. Lacking any butenolides, aqueous extracts of charred straw contain a potential germination stimulating steroid, i.e. ergosterol. The significance of fire‐derived cues on behaviour of oilseed rape seeds in the soil seed bank is discussed.

     

Physical forces in dormancy and germination of xanthium seeds

The germination of seeds of Xanthium pensylvanicum Wallr. occurs in 2 phases, an initial passive phase of water uptake followed by an active phase of growth. These 2 phases have been separated experimentally, and shown to occur similarly in isolated cotyledons and embryonic axes. Measurements of the physical thrust generated by the entire seed and its separate components of cotyledon and axis reveal that non-dormant Xanthium seeds develop more than twice the thrust of dormant seeds, and that this difference develops principally in the second phase of enlargement of the axis. Measurement of the forces required for piercing the testa of these seeds establishes that whereas the thrust developed by non-dormant seed is adequate to cause testa rupture, that developed by dormant seeds is not. It is concluded that the dormancy of Xanthium involves an inadequacy in the embryo for rupture of the testa.     

A kinetic analysis of the facultative photoperiodic response in Amaranthus retroflexus L.

The ontogenetic change taking place in the facultative photoperiodic response of A. retroflexus to inductive short-day (SD) conditions was studied by exposing plants to continuous induction after different initial exposures to long-days (LD), and comparing the kinetics of their developmental responses (cumulative number of plants with reproductive apices, flowering stage, and height of the apical dome). As the plants progressed from emergence to autonomous flowering (i.e., in non-inductive conditions), their response to continuous induction became progressively more rapid. Reproductive development was initiated following a progressively shorter lag-phase after the start of induction, but its subsequent rate remained unchanged. Until the onset of reproductive development, the undifferentiated upper part of the shoot apex (apical dome) elongated much more rapidly in SD than in LD. However, in both cases reproductive development was initiated when the apex had elongated to about the same extent, after which its elongation accelerated considerably, but to similar rates in both photoperiods. The data indicate that progress towards reproductive development takes place in inductive (SD), as well as in non-inductive (LD) photoperiods, but one cycle of the latter is as effective as 0.20–0.25 of a cycle of the former. —Plants induced at different stages in ontogeny started to change their subsequent branching pattern (ratio of leafy to leafless branches) as soon as induction was delayed beyond autonomous flowering.Abbreviations LD long-days - SD short-days - RGR Relative Growth Rate     

Sodium-Stimulated NO(3) Uptake in Amaranthus tricolor L. Plants

Nitrate uptake of Na⁺ -deficient Amaranthus tricolor L. cv Tricolor seedlings from complete culture solution was stimulated by about 210% within 5 hours by application of 0.5 millimolar NaCl. From a Na⁺ -preloading experiment, intracellular Na⁺ was shown to be responsible for the stimulation of NO₃⁻ uptake. The results suggest a possible role of Na⁺ in NO₃⁻ uptake in C₄ plants.     

Stimulatory effect of ethephon,ACC, gibberellin A3 and A4+7 on germination of methyl jasmonate inhibited Amaranthus caudatus L. seeds

Methyl jasmonate (JA-Me) inhibited or retarded germination of Amaranthus caudatus seeds in darkness at 24°C, Ethephon, ACC and gibberellins (GA₃ or GA₄₊₇) partially or completely reversed this inhibition depending on the concentration of JA-Me applied. Both ethephon and the gibberellins were more effective than ACC. Both GA₃ and GA₄₊₇ enhanced the stimulatory effect of ethephon or ACC on germination of seeds inhibited by JA-Me.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - JA jasmonic acid - JA-Me methyl jasmonate     

Free and conjugated gibberellins in dormancy and germination of apple seeds

Halińska, A. and Lewak, St. 1987. Free and conjugated gibberellins in dormancy and germination of apple seeds.
The presence of gibberellin A₄ (GA₄) was confirmed in partly stratified seeds of apple ( Malus domestica Borb., cv. Antonówka) by mass spectrometry of the methyl ester. Levels of free and conjugated gibberellins A₄₊₇ and A₉ changed during drying of mature seeds, during cold and warm stratification, as well as during germination of dormant and non-dormant embryos. The temporary rise in GA₄₊₇ during cold stratification and during the culture of dormant embryos as well as the lack of it under conditions of warm stratification, allowed us to postulate a role for GA₄₊₇ in the removal of dormancy. In addition, GA₉ was absent in dormant embryos and increased during cold stratification and during the culture of non-dormant embryos. This suggests the involvement of GA₉, in induction of normal development of the seedling. The equivalence between changes in free and conjugated GAs suggests that formation and hydrolysis of conjugates are involved in the control of the physiologically active levels of free GA₄₊₇ and GA₉.     

Seed dormancy in Acer: Endogenous germination inhibitors and dormancy in Acer pseudoplatanus L.

Summary Dormant seeds of Acer pseudoplatanus L. contain two zones of inhibition on paper chromatograms in 10:1:1 as detected by the lettuce and cress seed germination, and the wheat coleoptile bioassays. One zone at R_f 0.6–0.8 was partitioned into ethyl acetate at acid pH and was shown to contain ABA by its behaviour on GLC and isomerization under ultra-violet light. The other zone at R_f 0.9 was detected only in the germination bioassays and was partitioned into ethyl acetate over a range of pH indicating the presence of one or more neutral compounds.The inhibitors present in the embryo of dormant sycamore seeds inhibited the germination of non-dormant sycamore seeds at relatively low concentrations. A comparison with the effects of application of exogenous ABA indicated that endogenous ABA could not solely account for the inhibitory activity of seed extracts, which appeared to be due partly to the presence of ABA and partly to that of neutral compounds present in the embryo. Leaching treatments that removed dormancy led to a decrease in the level of inhibitors present mainly in the basic fraction. The exogenous application of kinetin to dormant sycamore seeds increased germination whereas gibberellic acid had no effect. Similar responses were obtained with lettuce seeds inhibited by the basic fraction of dormant sycamore seeds.It is suggested that an inhibitor-cytokinin interaction may be involved in the dormancy of sycamore seeds.     

Changes in sensitivity of Amaranthus retrof/exus L. seeds to ethylene during preincubation. I. Constant temperatures

Abstract. Germination responses of redroot pigweed ( Amaranthus retroflexus L.) seeds to ethylene were determined at 25, 30, 35, or 40° C after preincubation at various temperatures (15–35° C) for different periods (0.5–32 d). After 7 d preincubation, seeds showed a log-linear germination response to ethylene concentration in most of the temperature treatments. Sensitivity to ethylene increased with longer preincubation; response thresholds of 0.03−0.09 cm³ m⁻³ were observed after 32 d, compared to 0.18−1.6 cm³ m⁻³ after 7 d of preincubation. Preincubation at 15 or 20° C generally enhanced germinability, whereas 25 or 30° C produced secondary dormancy, which was readily broken with ethylene. Temperature during preincubation also significantly influenced the slope of the dose-response curve. The responses of preincubated redroot pigweed seeds to ethylene suggested that, in the field, seeds would probably not lose their sensitivity to this gas during prolonged burial in soil.     

AtPER1 enhances primary seed dormancy and reduces seed germination by suppressing the ABA catabolism and GA biosynthesis in Arabidopsis seeds

Seed is vital to the conservation of germplasm and plant biodiversity. Seed dormancy is an adaptive trait in numerous seed‐plant species, enabling plants to survive under stressful conditions. Seed dormancy is mainly controlled by abscisic acid (ABA) and gibberellin (GA) and can be classified as primary and secondary seed dormancy. The primary seed dormancy is induced by maternal ABA. Here we found that AtPER1, a seed‐specific peroxiredoxin, is involved in enhancing primary seed dormancy. Two loss‐of‐function atper1 mutants, atper1‐1 and atper1‐2, displayed suppressed primary seed dormancy accompanied with reduced ABA and increased GA contents in seeds. Furthermore, atper1 mutant seeds were insensitive to abiotic stresses during seed germination. The expression of several ABA catabolism genes (CYP707A1, CYP707A2, and CYP707A3) and GA biosynthesis genes (GA20ox1, GA20ox3, and KAO3) in atper1 mutant seeds was increased compared to wild‐type seeds. The suppressed primary seed dormancy of atper1‐1 was completely reduced by deletion of CYP707A genes. Furthermore, loss‐of‐function of AtPER1 cannot enhance the seed germination ratio of aba2‐1 or ga1‐t, suggesting that AtPER1‐enhanced primary seed dormancy is dependent on ABA and GA. Additionally, the level of reactive oxygen species (ROS) in atper1 mutant seeds was significantly higher than that in wild‐type seeds. Taken together, our results demonstrate that AtPER1 eliminates ROS to suppress ABA catabolism and GA biosynthesis, and thus improves the primary seed dormancy and make the seeds less sensitive to adverse environmental conditions.     

Biomass production and nitrate metabolism of Atriplex hortensis L. (C3 plant) and Amaranthus retroflexus L. (C4 plant) in cultures at different levels of nitrogen supply

Summary Pure and mixed cultures of the dicotyledons Atriplex hortensis L. (C₃ plant) and Amaranthus retroflexus L. (C₄ plant) were maintained under open air conditions in standard soil at low and high nitrogen supply levels.A comparison of shoot dry weight and shoot length in the various series shows that the growth of the aboveground parts of both species was severely reduced under low N conditions. In both pure and mixed cultures the differences resulting from low N vs. high N conditions was less pronounced with Atriplex (C₃ plant) than with Amaranthus (C₄ plant). The root dry weight of the two species was not reduced so much under low N conditions as was the shoot dry weight. The low N plants were found to contain a larger proportion of their biomass in the roots than did the high N plants. In general the root proportion of Atriplex was greater than that of Amaranthus. The contents of organic nitrogen and nitrate and the nitrate reductase activity (NRA) per g dry weight of both species decreased continually throughout the experiments. With the exception of young plants, the low N plants always had tower contents of organic nitrogen and nitrate and nitrate reductase activities than did the high N plants. The highest values of NRA were measured in the leaf laminae. The eaves also exhibited the highest concentrations of organic nitrogen. The highest nitrate concentrations, however, were observed in the shoot axis, and in most cases the lowest nitrate values were found in the laminae. At the end of ne growing season this pattern was found to have been reversed with Atriplex, but not with Amaranthus. Thus Atriplex was able to maintain a higher NRA in the laminae than Amaranthus under low N conditions.The transpiration per leaf area of the C₄ plant Amaranthus during the course of a day was substantially lower than that of the C₃ plant Atriplex. There were no significant differences in transpiration between the low N and high N series of Amaranthus. The low N plants of Atriplex, however, clearly showed in most cases higher transpiration rates than the corresponding high N plants. These different transpiration rates of the high N and the low N Atriplex plants were also reflected in a distinct ¹³C discrimination.The sum of these results points to the conclusion that the C₃ plant Atriplex hortensis can maintain a better internal inorganic nitrogen supply than the C₄ plant Amaranthus retroflexus under low N conditions and an ample water supply, due to the larger root proportion and the more pronounced and flexible transpiration of the C₃ plant.Dedicated to Prof. Dr. Karl Mägdefrau, Deisenhofen, on the ocasion of his 80th birthday