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.     

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.     

Effects of temperature, water potential, and light on germination responses of redroot pigweed seeds to ethylene

The responses of redroot pigweed (Amaranthus retroflexus L.) seeds to nine ethylene concentrations between 0.5 and 50 microliters per liter were assessed at different temperatures and water potentials and in either continuous white light or darkness. Under all experimental treatments, the probit-transformed percentages increased linearly with the log of the ethylene concentration. In dormant seeds, the slope of the response line was unaffected by either light or water potential but increased with decreasing temperature. Conversely, the slope increased with increasing temperature in a partially afterripened seed lot.     

Germinative response of redroot pigweed (Amaranthus retroflexus L.) to environmental conditions: Is there a seasonal pattern?

Experiments on redroot pigweed (Amaranthus retroflexus L.) were conducted to investigate whether the germinative response to environmental conditions is affected by the time of seed set. Seeds were collected in the same field (Sicily, Southern Italy) in May, July and October; each lot was dry-stored from 15 to 400 days after harvest (DAH) and submitted to germination assays from 15 to 40°C, both in continuous darkness (D) and in alternate light/darkness regime (L/D). For the three lots, over 15 DAH, the response to temperature and light regime was strongly affected by harvesting time. Seeds set in May, negatively affected by L/D, showed a high germination capability (>80%) at 95 DAH from 25 to 40°C. Seeds set in July were favoured by L/D and required at least 170 DAH to reach 80% germination capability. Seeds set in October were also favoured by L/D and gave a good germination capability only at 300 and 400 DAH. These results prove that seed germination behaviour in redroot pigweed is not independent of the time of the year in which seeds are produced and is due to both the environmental conditions experienced by the mother plant during seed maturation and those experienced by seeds after seed set.     

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.     

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.     

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.     

Interference between Redroot Pigweed (Amaranthus retroflexus L.) and Cotton (Gossypium hirsutum L.): Growth Analysis

Redroot pigweed is one of the injurious agricultural weeds on a worldwide basis. Understanding of its interference impact in crop field will provide useful information for weed control programs. The effects of redroot pigweed on cotton at densities of 0, 0.125, 0.25, 0.5, 1, 2, 4, and 8 plants m^-1 of row were evaluated in field experiments conducted in 2013 and 2014 at Institute of Cotton Research, CAAS in China. Redroot pigweed remained taller and thicker than cotton and heavily shaded cotton throughout the growing season. Both cotton height and stem diameter reduced with increasing redroot pigweed density. Moreover, the interference of redroot pigweed resulted in a delay in cotton maturity especially at the densities of 1 to 8 weed plants m^-1 of row, and cotton boll weight and seed numbers per boll were reduced. The relationship between redroot pigweed density and seed cotton yield was described by the hyperbolic decay regression model, which estimated that a density of 0.20–0.33 weed plant m^-1 of row would result in a 50% seed cotton yield loss from the maximum yield. Redroot pigweed seed production per plant or per square meter was indicated by logarithmic response. At a density of 1 plant m^-1 of cotton row, redroot pigweed produced about 626,000 seeds m^-2. Intraspecific competition resulted in density-dependent effects on weed biomass per plant, a range of 430–2,250 g dry weight by harvest. Redroot pigweed biomass ha^-1 tended to increase with increasing weed density as indicated by a logarithmic response. Fiber quality was not significantly influenced by weed density when analyzed over two years; however, the fiber length uniformity and micronaire were adversely affected at density of 1 weed plant m^-1 of row in 2014. The adverse impact of redroot pigweed on cotton growth and development identified in this study has indicated the need of effective redroot pigweed management.     

Allelopathic effects of switchgrass on redroot pigweed and crabgrass growth

Non-native invasive plant species influence plant community composition and competitively eradicate native species. However, there is doubt regarding how global invasive species increase and explosively interfere with native plants. Invasive plants always have strong allelopathic potential. In this study, allelopathic effects of switchgrass on redroot pigweed and crabgrass growth were investigated by field and laboratory experiments. Within a 0.4-m distance of switchgrass, density and shoot biomass of native species were significantly suppressed in the field, with 95.1% and 93.0% inhibition on density of redroot pigweed and crabgrass and with 99.0% and 97.7% inhibition on shoot biomass, respectively, during the third growing season. Significant inhibitory effects on shoot and root biomass were observed at the 5:5 (switchgrass–native species) proportion in glass bottles, by 41.57% and 51.21% for shoot and root biomass of redroot pigweed and by 33.42% and 56.95% for shoot and root biomass of crabgrass, respectively. Results of a glass bottle experiment showed that shoot and root biomass of redroot pigweed and crabgrass could be significantly inhibited by contact with switchgrass root. Results of a Petri dish experiment showed that aqueous extracts of switchgrass significantly inhibited germination process of both species at high concentrations, with 90.74% and 18.62% inhibition on germination rate and plumule length of redroot pigweed and with 63.59%, 16.38%, and 19.92% inhibition on germination rate, plumule, and radicle lengths of crabgrass, respectively, at the concentration of 0.1 g·mL^?1. This report demonstrated that switchgrass had allelopathic effects on redroot pigweed and crabgrass growth.

     

Germination and Dormancy of Reed Canary-Grass Seeds (Phalaris arundinacea)

Effects of various chemical and physical factors on the germination of several seed lots of reed canary-grass (Phalaris arundinacea L.) have been studied. Germination at the optimum constant temperatures of 24 to 27°C was significantly stimulated by the following treatments: moist chilling in light, red light given during the first 3 days of imbibition, three 2-h periods at 12°C given during the second day of imbibition, ethylene, increased oxygen tension and soaking in aerated water for 4 days. Dry storage at 20–30°C had no effect on the germination ability of the seeds. No significant quantities of germination inhibitors were found either in water or methanol extracts of seed dispersal units. By comparing three cultivars with various degrees of seed dormancy, respiration measurements showed that there was a significant positive correlation between oxygen uptake prior to visible germination and germination capacity. Similarly, germination-stimulating treatment significantly enhanced oxygen uptake prior to visible germination.     

Inhibitory effects of redroot pigweed and crabgrass on switchgrass germination and growth—from lab to field

Interactions between weeds and crops often occur by resource competition or allelopathy. However, it is still unknown how local weed species influence artificially introduced switchgrass. In this study, four experiments were conducted to evaluate the inhibitory effects of redroot pigweed (Amaranthus retroflexus) and crabgrass (Digitaria sanguinalis) on germination and growth of the lowland tetraploid switchgrass cultivar ‘Alamo’ (Panicum virgatum cv. Alamo). Switchgrass germination was significantly inhibited in Petri dishes, with 48.1% and 33.9% inhibitions on germination rate by redroot pigweed and crabgrass root aqueous extracts, respectively, at 0.1 g mL^?1 concentration. Significant inhibitory effects on switchgrass seedling biomass were observed at 5:5 ratio with redroot pigweed and crabgrass in glass jars, with 61.6% and 53.4% inhibitions on plant biomass, respectively. Under the same root segregation, redroot pigweed had a stronger inhibitory effect on switchgrass seedling growth than crabgrass. Growth of transplanted switchgrass seedlings was significantly inhibited by local weeds in the field, with 46.2% and 11.7% inhibitions on shoot biomass during the first and second growing seasons, respectively. However, no significant growth reduction in switchgrass was detected in the third growing season. These findings further our understanding of weed–crop interactions and could help develop weeds management strategies with ecological security.

     

The impact of elevated CO2 on yield loss from a C3 and C4 weed in field-grown soybean

Soybean (Glycine max) was grown at ambient and enhanced carbon dioxide (CO₂, + 250 μL L^?1 above ambient) with and without the presence of a C3 weed (lambsquarters, Chenopodium album L.) and a C4 weed (redroot pigweed, Amaranthus retroflexus L.), in order to evaluate the impact of rising atmospheric carbon dioxide concentration [CO₂] on crop production losses due to weeds. Weeds of a given species were sown at a density of two per metre of row. A significant reduction in soybean seed yield was observed with either weed species relative to the weed‐free control at either [CO₂]. However, for lambsquarters the reduction in soybean seed yield relative to the weed‐free condition increased from 28 to 39% as CO₂ increased, with a 65% increase in the average dry weight of lambsquarters at enhanced [CO₂]. Conversely, for pigweed, soybean seed yield losses diminished with increasing [CO₂] from 45 to 30%, with no change in the average dry weight of pigweed. In a weed‐free environment, elevated [CO₂] resulted in a significant increase in vegetative dry weight and seed yield at maturity for soybean (33 and 24%, respectively) compared to the ambient CO₂ condition. Interestingly, the presence of either weed negated the ability of soybean to respond either vegetatively or reproductively to enhanced [CO₂]. Results from this experiment suggest: (i) that rising [CO₂] could alter current yield losses associated with competition from weeds; and (ii) that weed control will be crucial in realizing any potential increase in economic yield of agronomic crops such as soybean as atmospheric [CO₂] increases.     

Salicylhydroxamic Acid Potentiates Germination of `Waldmann''s Green'' Lettuce Seed

A combination of salicylhydroxamic acid (SHAM) + cyanide (CN) is known to stimulate dark germination of Lactuca sativa L. seeds. Further studies were done to characterize SHAM and CN action in stimulating dark germination of lettuce seed. Germination was stimulated slightly by either SHAM or CN, whereas when SHAM and CN were combined germination was greatly enhanced. Treatment of seeds with SHAM + CN only during the first 8 hours of hydration stimulated germination as much as did treatment for 72 hours. During the first 8 hours of incubation in SHAM + CN, potentiation (i.e. dormancy-breaking) of germination occurs. SHAM alone stimulated potentiation nearly to the level of SHAM + CN but inhibited subsequent radicle elongation, thereby decreasing germination when present for 72 hours. Oxygen must be present for SHAM or SHAM + CN to potentiate dark germination. The ability of SHAM and SHAM + CN to potentiate germination is influenced by O₂ concentration and the timing of chemical treatment.     

Physiology of Oil Seeds: IV. Role of Endogenous Ethylene and Inhibitory Regulators during Natural and Induced Afterripening of Dormant Virginia-type Peanut Seeds

To further elucidate the regulation of dormancy release, we followed the natural afterripening of Virginia-type peanut (Arachis hypogaea L.) seeds from about the 5th to 40th week after harvest. Seeds were kept at low temperature (3 ± 2 C) until just prior to testing for germination, ethylene production, and internal ethylene concentration. Germination tended to fluctuate but did not increase significantly during the first 30 weeks; internal ethylene concentrations and ethylene production remained comparatively low during this time. When the seeds were placed at room temperature during the 30th to 40th weeks after harvest, there was a large increase in germination, 49% and 47% for apical and basal seeds, respectively. The data confirm our previous suggestion that production rates of 2.0 to 3.0 nanoliters per gram fresh weight per hour are necessary to provide internal ethylene concentrations at activation levels which cause a substantial increase of germination. Activation levels internally must be more than 0.4 microliter per liter and 0.9 microliter per liter for some apical and basal seeds, respectively, since dormant-imbibed seeds containing these concentrations did not germinate. Abscisic acid inhibited germination and ethylene production of afterripened seeds. Kinetin reversed the effects of ABA and this was correlated with its ability to stimulate ethylene production by the seeds. Ethylene also reversed the effects of abscisic acid. Carbon dioxide did not compete with ethylene action in this system. The data indicate that ethylene and an inhibitor, possibly abscisic acid, interact to control dormant peanut seed germination. The inability of CO₂ to inhibit competitively the action of ethylene on dormancy release, as it does other ethylene effects, suggests that the primary site of action of ethylene in peanut seeds is different from the site for other plant responses to ethylene.     

Investigating seed dormancy in cotton (Gossypium hirsutum L.): understanding the physiological changes in embryo during after‐ripening and germination

• The dormancy of seeds of upland cotton can be broken during dry after‐ripening, but the mechanism of its dormancy release remains unclear.
• Freshly harvested cotton seeds were subjected to after‐ripening for 180 days. Cotton seeds from different days of after‐ripening (DAR) were sampled for dynamic physiological determination and germination tests. The intact seeds and isolated embryos were germinated to assess effects of the seed coat on embryo germination. Content of H₂O₂ and phytohormones and activities of antioxidant enzymes and glucose‐6‐phosphate dehydrogenase were measured during after‐ripening and germination.
• Germination of intact seeds increased from 7% upon harvest to 96% at 30 DAR, while embryo germination improved from an initial rate of 82% to 100% after 14 DAR. Based on T₅₀ (time when 50% of seeds germinate) and germination index, the intact seed and isolated embryo needed 30 and 21 DAR, respectively, to acquire relatively stable germination. The content of H₂O₂ increased during after‐ripening and continued to increase within the first few hours of imbibition, along with a decrease in abscisic acid (ABA) content. A noticeable increase was observed in gibberellic acid content during germination when ABA content decreased to a lower level. Coat removal treatment accelerated embryo absorption of water, which further improved the accumulation of H₂O₂ and changed peroxidase content during germination.
• For cotton seed, the alleviation of coat‐imposed dormancy required 30 days of after‐ripening, accompanied by rapid dormancy release (within 21 DAR) in naked embryos. H₂O₂ acted as a core link between the response to environmental changes and induction of other physiological changes for breaking seed dormancy.

     

Changes in the activity of phosphoenolpyruvate carboxylating enzymes in germinating yellow lupin seeds

The rate of phosphoenolpyruvate carboxylation by extracts from germinating lupin seeds was measured through the H¹⁴CO₃ fixation. PEP carboxylation in seed axes increased during their imbibition, mainly as a result of the increase in the activity of PEP carboxylase [EC 4.1.1.31]. However, the activity of PEP carboxykinase [EC 4.1.1.38], present during the first 3 hours of imbibition, as well as the activity of PEP-carboxykinase [EC 4.1.1.49], after 24 hours of imbibition, have also been shown. Possible physiological role of the changes in the activity of PEP carboxylases during lupin seeds germination is discussed.     

Seed Development in Phaseolus vulgaris L. cv Seminole: II. Precocious Germination in Late Maturation

Seeds of Phaseolus vulgaris L. cv Seminole in late maturation phase germinated precociously in vitro. Germination occurred in the absence of free water after 5 days but within 24 to 48 hours in contact with water. Excised axes germinated within 12 hours and embryos by 48 hours only if supplied with water. Ethylene accelerated the germination of seeds and embryos irrespective of water availability. There was no effect of ethylene on the rate of axis germination. Ethylene was equally effective within the range 0.5 to 1000 parts per million and 1 hour exposure was fully effective. Induction of precocious germination in vivo was observed by manipulating water content inside pods or by ethylene injection, whether pods were attached to the parent plant or not. These results demonstrate the importance of endogenous regulation of water supply in suppressing precocious germination. Ethylene is identified as a powerful antagonist to the natural control.     

Enhancement of germination rate of aged seeds by ethylene

Naturally and artificially aged seeds of rape, Brassica napus L., produced less ethylene than freshly harvested seed during the early stage of germination. With freshly harvested seeds one peak of ethylene production was observed during germination, which coincided with the emergence and elongation of root and cotyledon, accompanied by splitting of the seed coat. Application of exogenous ethylene was effective in accelerating germination in aged seeds but did not significantly improve the percentage of germination. Ethylene as a hormone was considered to serve as a stimulator of germination and growth. One of the factors causing seed aging might be the degeneration of an ethylene-producing system in the seed. Exogenous ethylene may be effective only for the seeds in which the ethylene-producing system is weakened but the following responding systems are still functional.     

Control of Seed Germination by Abscisic Acid: I. Time Course of Action in Sinapis alba L

The germination process of mustard seeds (Sinapis alba L.) has been characterized by the time courses of water uptake, rupturing of the seed coat (12 hours after sowing), onset of axis growth (18 hours after sowing), and the point of no return, where the seeds lose the ability to survive redesiccation (12 to 24 hours after sowing, depending on embryo part). Abscisic acid (ABA) reversibly arrests embryo development at the brink of radicle growth initiation, inhibiting the water uptake which accompanies embryo growth. Seeds which have been kept dormant by ABA for several days will, after removal of the hormone, rapidly take up water and continue the germination process. Seeds which have been preincubated in water lose the sensitivity to be arrested by ABA after about 12 hours after sowing. This escape from ABA-mediated dormancy is not due to an inactivation of the hormone but to a loss of competence to respond to ABA during the course of germination. The sensitivity to ABA can be restored in these seeds by redrying. It is concluded that a primary action of ABA in inhibiting seed germination is the control of water uptake of the embryo tissues rather than the control of DNA, RNA, or protein syntheses.     

Elevated CO2 decreases seed germination in Arabidopsis thaliana

The impact of elevated [CO₂] on seed germination was studied in different genotypes of Arabidopsis thaliana from natural populations. Two generations of seeds were studied: the maternal generation was produced in the greenhouse (present-day conditions), the offspring generation was produced in two chambers where the CO₂ concentration was either the present atmospheric concentration (about 350 ppm) or elevated (700 ppm). The seeds were tested for proportion of germinated seeds and mean germination time in both chambers to study the impact of elevated [CO₂] during seed production and germination. Elevated [CO₂] during maturation of seeds on the mother-plants decreased the proportion of germinated seeds, while elevated [CO₂] during germination had no effect on the proportion of germinated seeds. However, when seeds were both produced and germinated under elevated [CO₂] (situation expected by the end of next century), germination was slow and low. Moreover, the effect of the [CO₂] treatment differs among genotypes of Arabidopsis: there is a strong treatment × genotype interaction. This means that there is ample genetic variance for a selective response modiying the effects of high levels of [CO₂] in natural populations of Arabidopsis thaliana. The outcome at the community level will depend on what seeds are available, when they germinate and the resulting competition following germination.