Ontogenetic variation in response to temperature change in the control of seed dormancy of Rumex obtusifolius L. and Rumex crispus L.

Abstract After the onset of imbibition, the dormant seeds of Rumex obtusifolius and R. crispus are stimulated to germinate by a change from an initial low temperature to a warmer temperature for a relatively brief period: the warmer that temperature the shorter is the optimum period spent at it, and this optimum value is unaffected by the initial temperature. The optimum period is more critical in R. crispus than in R. obtusifolius (about 1 h and 2.5 to 4 h, respectively, for a warmer temperature of 35°C in the dark); in the light the length of the period at the warmer temperature is less critical in both species. The sensitivity of the seeds to the change to the warmer temperature increases with time from the start of imbibition at a rate which is positively related to the initial temperature. In R. obtusifolius maximum sensitivity was typically reached after 3 to 5 d when the initial temperature was 20°C and then remained constant, or declined only slightly, over the period investigated (10 d). At the same initial temperature, however, R. crispus showed a cyclical pattern of sensitivity with peaks occurring at 3–4 d intervals from the start of imbibition.     

Effects of low temperatures on the loss of innate dormancy and the development of induced dormancy in seeds of Rumex obtusifolius L. and Rumex crispus L.

Abstract It is possible to remove the innate dormancy of seeds of Rumex crispus L and Rumex obtusifolius L. by an initial period of low-temperature stratification, providing the seeds are then transferred to a higher temperature. The lower the initial temperature within the range 1.5°-15°C, the greater the germination; there is no stratification effect at 20°C. Although 10°C and 15°C were shown to be suitable for both stratification and for the process of germination itself, neither temperature results in any germination if given constantly: a change from a lower to a higher temperature is essential. The optimum period for stratification depends on two separate processes which occur during the treatment–a rapid loss of innate or primary dormancy and a slower development of induced or secondary dormancy. Within the range 1.5°-15°C the rate of loss of innate dormancy appears to be independent of light and is probably independent of temperature. In contrast, the rate of induction of secondary dormancy increases with increase in temperature, and is more rapid in the dark than the light. The rate of induction of secondary dormancy during stratification is greater in R. crispus than in R. obtusifolius. As a consequence, maximum germination was obtained in both species after stratification at 1.5°C in the light, the optimum period of treatment being about 4 weeks in R. Obtusifolius and 6 weeks in R. crispus, while the maximum germination obtained and the optimal period of stratification decrease in both species with increase in stratification temperature.     

Interaction of light and temperature on seed germination of Rumex obtusifolius L.

Germination of Rumex obtusifolius L. seeds (nutlets) is low in darkness at 25° C. Germination is stimulated by exposure to 10 min red light (R) and also by a 10-min elevation of temperature to 35° C. A 10-min exposure to far-red light (FR) can reverse the effect of both R (indicating phytochrome control) and 35° C treatment. Fluence-response curves for this reversal of the effect of R and 35° C treatments are quantitatively identical. Treatment for 10 min with light of wavelenght 680, 700, 710 and 730 nm, after R and 35° C treatment, demonstrates that germination induced by 35° C treatment results from increased sensitivity to a pre-existing, active, far-red-absorbing form of phytochrome (Pfr) in the seeds.Abbreviations FR far-red light - P phytochrome - Pr red-absorbing form of P - Pfr far-red-absorbing form of P - R red light     

Biochemical recognition in seeds: Germination of Rumex obtusifolius is promoted by leaves of facilitative adult conspecifics

Seeds express various germination behaviors in response to competitor plants. However, germination behaviors in response to facilitator plants are not yet well understood. Rumex obtusifolius seedlings usually appear on the ground near adult conspecific plants, and their survival rate under the canopy of adult conspecifics is higher than that outside the canopy, indicating that adult R. obtusifolius plants facilitate their seedling establishments. We hypothesized that emergence of R. obtusifolius seedlings is promoted by cues from adult conspecifics, but emergence of heterospecific seedlings is not. To test this, we investigated emergence responses of seedlings of R. obtusifolius and three other species that grow with R. obtusifolius in the presence of R. obtusifolius leaf phytochemicals. Emergence of R. obtusifolius seedlings was promoted by the presence of R. obtusifolius leaves. In contrast, emergence of other species seedlings was not promoted by R. obtusifolius leaves. We conclude that germination of R. obtusifolius seeds is facilitated in the presence of conspecifics, via water-soluble chemical exposure, and that recognizing these chemicals has adaptive value.     

Analysis of the Effect of Light and Temperature on the Fluence Response Curves for Germination of Rumex obtusifolius

Both red light (10 minutes) and 35°C treatment (60 minutes) stimulate the germination of seeds of Rumex obtusifolius otherwise maintained in darkness at 25°C. Fluence response curves were determined for the effect of red light to stimulate germination of seeds with and without 35°C treatment. The endogenous far-red absorbing form (Pfr) level in the seeds was determined using short saturating fluences of wavelengths of light which maintain different proportions of phytochrome as Pfr at equilibrium. In the seed batches investigated, the endogenous Pfr level was found to be 4% or less of the total phytochrome. High dark germination after 35°C treatment does not result from an increase in sensitivity of the whole population to Pfr. Calculated fluence response curves for germination which best fit the experimental data suggest that seeds germinate in darkness after 35°C treatment because of a nonphytochrome-related process (overriding factor).     

Water Content and Phytochrome-induced Potential Germination Responses in Lettuce Seeds

Grand Rapids lettuce seeds (Lactuca sativa L.) with 6 to 8% water content show no light-induced germination responses, whereas in seeds with 15% or more water content, germination is promoted or retarded by red and far red light respectively. By adjusting seed water content, persistent potentiated responses to light are induced in the seeds at seed water levels much below that required for germination itself. Alternate moistening and drying of seeds in conjunction with red and far red irradiations show that potentiated responses may be phototransformed only with sufficient seed water. However, the process of drying and remoistening has no effect on potentiated responses.     

Performance and Mortality of Rumex obtusifolius and R. crispus in Managed Grasslands are Affected by Nutrient Availability

The reasons for the greater incidence of Rumex obtusifolius in grasslands compared with R. crispus have never been investigated. In a small-plot field experiment in which seedlings of R. obtusifolius and R. crispus were transplanted into a sward dominated by Dactylis glomerata, the growth and survival of seedlings were monitored over three years under control and P, N, NP and NPK fertilizer treatments. The highest plants of R. obtusifolius and R. crispus were generally recorded in the N, NP and NPK treatments and the lowest in the control and P treatment. The maximum recorded heights of R. obtusifolius were 100?cm and of R. crispus 80?cm. The number of leaves per plant, weight of individual plants, cover and fertility were generally higher for R. obtusifolius than for R. crispus. 30% to 80% of transplanted R. obtusifolius plants flowered in the first (seeding) year and this contrasted with no flowering of transplanted R. crispus plants. After cutting, substantially better regrowth of R. obtusifolius was recorded compared to that of R. crispus and surviving R. crispus plants tended to show a gradual reduction in leaf number. The mortality of R. obtusifolius over three years ranged from 0% in the NPK treatment to 13% in the control, with an average of 8% over all treatments. The mortality of R. crispus ranged from 19% in the control to 94% in the NPK treatment with an average of 64% over all treatments. We concluded that R. obtusifolius is better adapted to growing in highly productive temperate grasslands than R. crispus because of its perennial character, larger plant size and higher fertility.     

The comparative germination ecology of nine Rumex species

The germination requirements, dormancy cycle and longevity of nine Rumexspecies were studied in field conditions and laboratory experiments to show theadaptations of the related species to their specific habitat. Within one genus,rather striking differences were observed in germination ecology. However, theclosely related species, R. acetosa and R.scutatus, are very similar: they fruit in early summer; theirseeds can germinate immediately after dispersal, and they are nondormant andshort-lived. R. acetosella also has fruits insummer, but the seeds do not germinate the first season after dispersal. Theyare long-lived, but buried seeds do not show a dormancy cycle; they mightgerminate in different seasons after exposure to light. Seeds of four species (R. conglomeratus,R. maritimus, R. sanguineus andR. crispus) are long-lived and undergo aseasonal dormancy cycle, with a low level of dormancy in winter and early springand a deep dormancy in summer as was already known for R.obtusifolius. These seeds are shed in the autumn, and they germinatemainly in the spring in consecutive years. R. maritimusalso germinates in summer and autumn on drying muddy soils. The seeds of R. hydrolapathum only germinate onwaterlogged soils, which explains its growth at the edge of streams and ponds.Its seeds are rather short-lived. The seeds of the species on very wetplaces require a higher temperature for germination.     

Rapid effects of red light on the isopentenyladenosine content in scots pine seeds

Red light (R) stimulates germination in Scots pine seed (Pinus sylvestris L.). The response is far red (FR) reversible. The dynamics of cytokinin changes following light treatment was investigated. Extracts were purified by immunoaffinity and high performance liquid chromatography. N⁶-(Δ²-Isopentenyl) adenosine (iPA) and trans-zeatin riboside (ZR) were quantified by both UV-absorbance of high performance liquid chromatography peaks and by enzyme-linked immunosorbent assay. Identification of iPA was accomplished by gas chromatography-mass spectrometry. Levels of cytokinins were low in seeds imbibed in the dark. Exposure of seeds imbibed in the dark for 5 hours to R for 15 minutes induced a strong, immediate but transitory increase in iPA content. This increase was not observed when the R treatment was followed by 10 minutes of FR or by storage in darkness before extraction. No ZR was detected during the first 8 hours of imbibition in any treatment. Addition of iPA via acetone enhanced seed germination in the dark. The results suggest that iPA may be involved in the R-mediated release of dormancy of Scots pine seed.     

Interaction of light and temperature on the germination of Rumex obtusifolius L.

Seeds (nutlets) of Rumex obtusifolius L. fail to germinate in darkness at 25° C, but are stimulated by short exposure to red light (R) the effectiveness of which can be negated by a subsequent short exposure to far red light (F) indicating phytochrome control. Short periods of elevated temperature treatment (e.g. 5 min at 35° C) can induce complete germination in darkness. Although short F cannot revert the effect of 35° C treatment, cycling the phytochrome pool by exposure to short R before short F results in reversion of at least 50% of the population. Prolonged or intermittent F can also revert the germination induced by 35° C treatment. The effect of elevated temperature treatment is interpreted on the basis of two possible models; (i) that it increases the sensitivity of the seeds to a low level of pre-existing active form of phytochrome (Pfr) (ii) that it induces the appearance of Pfr in the dark. In both cases it is envisaged that elevated temperature treatment and Pfr control germination at a common point in the series of reactions that lead to germination.Abbreviations D Dark - F far red light - P phytochrome - Pr red absorbing form of P - Pfr far red absorbing form of P - R red light     

Sheep grazing vs. cutting: regeneration and soil nutrient exploitation of the grassland weed Rumex obtusifolius

Broad-leaved dock (Rumex obtusifolius L., Polygonaceae) is an agronomically important perennial weed causing decreases in pasture yields and fodder quality. Non-chemical control measures for dock are often limited to frequent pulling and cutting, additionally it is usually avoided by grazing animals. Here, the regeneration of R. obtusifolius in a Rumex-infested grassland grazed by a sheep breed that explicitly feeds on dock (Ovis aries L. cv. East Prussian Skudden) was compared to cutting. Therefore, regeneration of 90 R. obtusifolius plants of three different size classes was monitored in three plots during three grazing and cutting cycles. Plant height and number of fruit-stands of regrown R. obtusifolius was significantly lower, number of leaves significantly higher after grazing than after cutting, while plant diameter was unaffected. Initially medium and large-sized plants (>40 cm diameter) were significantly more sensitive to grazing or cutting than initially smaller sized plants. Soil nitrate and ammonium concentrations in the vicinity of R. obtusifolius correlated with some regrowth parameters but were not affected by grazing or cutting. Sheep-grazed grassland communities comprised fewer legumes (p = 0.002), more grasses (p = 0.010) and fewer sward gaps (p = 0.025) than cut grassland. At the end of the experiment, abundance of R. obtusifolius in sheep grazed plots was lower than in cut plots (p = 0.089) suggesting that regrowth potential of this weed was depleted by continuous grazing and higher sward density. In conclusion, these data suggest that sheep could be considered in grassland management schemes to both directly and indirectly control Rumex infestations.     

Inhibition of Prechill-induced Dark Germination in Sorghum halepense (L.) Pers. Seeds by Phytochrome Transformations

A 10 C dark prechilling of johnsongrass [Sorghum halepense (L.) Pers.] seeds, when terminated by a 2-hr, 40 C temperature shift, potentiates about 40% germination at 20 C in darkness. Irradiation of the seeds before, during, and at the end of prechilling with far red light reduces the subsequent germination, although red irradiation after the far red can overcome some of the inhibition. However, either brief red or far red irradiation given immediately after the temperature shift inhibits subsequent germination by one-third to one-half. The results suggest that the far red-absorbing form of phytochrome is a factor in the prechill-induced dark germination and that phytochrome participates in the inhibition of germination by irradiations immediately after the temperature shift.     

An antagonist treatment in combination with tracer experiments revealed isocitrate pathway dominant to oxalate biosynthesis in Rumex obtusifolius L.

Oxalate accumulates in leaves of certain plants such as Rumex species (Polygonaceae). Oxalate plays important roles in defense to predator, detoxification of metallic ions, and in hydrogen peroxide formation upon wounding/senescence. However, biosynthetic pathways of soluble oxalate are largely unknown. In the present study we analysed Rumex obtusifolius L. treated with itaconate (an antagonist to isocitrate). Comprehensive metabolome analysis using capillary electrophoresis-mass spectrometry showed that oxalate content of “new leaves” was notably down-regulated by itaconate, as opposed to the accumulation of citrate. The ¹³CO₂ feeding experiment revealed that oxalate accumulation in new leaves was affected by citrate translocation from stems. The results suggested that excess oxalate in new leaves of R. obtusifolius was synthesized primarily via the isocitrate pathway utilizing citrate delivered from stems.     

Variation in germination and amino Acid leakage of seeds with temperature related to membrane phase change

Leakages of amino acids and/or fluorescent material as functions of temperature between 15 and 40 C are reported for imbibed seeds of Avena fatua L., Lactuca sativa L., Barbarea vulgaris R. Br., Amaranthus albus L., Abutilon theophrasti Medic., Lychnis alba Mill., Daucus carota L., Setaria faberi Herrm., Setaria viridis (L.) Beauv., and Datura stramonium L. The leakage indicates prominent increase in permeability of the plasmalemma in the 30 to 35 C range for 8 of the 10 kinds of seeds studied. Germination of the seeds at constant temperatures or with daily shifts in temperature is related to the membrane transition temperature for permeation by amino acids. Seeds of A. albus and A. theophrasti, which did not show membrane changes in the 25 to 40 C range, germinated best at 35 to 40 C; the other seeds germinated best below 30 C. Seeds of B. vulgaris showed rapid permeation of limiting membranes upon initial wetting with water, which was indicative of membrane disorder when dry. Leakage under anaerobiosis was observed for S. faberi seeds.     

Targeted metabolomics in an intrusive weed, <Emphasis Type="Italic">Rumex obtusifolius</Emphasis> L., grown under different environmental conditions reveals alterations of organ related metabolite pathway

This study was intended to analyze the metabolic pathway of Rumex obtusifolius L. (Broad-leaved dock), destructive weeds worldwide, in relation to major environmental factors (light and temperature). It was found that R. obtusifolius can be classified as plants in accumulating major organic acids such as oxalate in leaves and citrate in stems (Miyagi et al., Metabolomics 6:146–155 2010). The organ specific accumulation of certain metabolites was dissected by metabolomics approach in relation to metabolic pathway. Light or dark experiments showed that in the case of the oxalate accumulation, the major or the most dominated pathway was found to be the citrate-isocitrate-oxalate shunt. Furthermore, under the dark and/or low temperature (5°C) leaves showed sustainable growth with normal accumulation of TCA metabolites. Unlike leaves, there was a different pattern of metabolite accumulation in stems. Other metabolites such as amino acids also showed the organ specific alterations under the different ambient environments.     

Factors affecting the induction of secondary dormancy in lettuce

The relationship between the temperature at which germination of 50% of the seeds is inhibited in the light (GT₅₀ Light) and secondary dormancy was investigated in three cultivars of Lactuca sativa L. Seeds were incubated for varying periods under non-germinating conditions and subsequent germination in response to red light (R) was determined over a wide range of temperatures. Dark incubation at 32 C reduced the GT₅₀ Light of cv. New York but did not affect germination at temperatures below 24 C. Dark, 32 C incubation had no effect on the GT₅₀ Light of cv. Great Lakes. In cv. Grand Rapids, dark incubation at 15, 24, 32, or 35 C initially reduced the GT₅₀ Light. However, longer incubations induced a secondary dormancy, i.e., the seeds became unable to germinate at all temperatures in response to R given after the high temperature incubation. A single exposure to R at the beginning of a 32 C incubation slowed the induction of secondary dormancy. Repeated exposures to R prevented the induction of secondary dormancy, but did not prevent a decline in the GT₅₀ Light. GA₃ mimicked the effect of repeated R.     

Environmental factors controlling seed germination and seedling recruitment of Stipa bungeana on the Loess Plateau of northwestern China

Germination studies are important for collecting information on field seedling recruitment, plant conservation and restoration. This study investigated the role of light, temperature, nitrogen, water stress and burial depth in controlling germination of Stipa bungeana seeds. S. bungeana seeds are photo-inhibited; light significantly decreased seed germination regardless of temperature and water conditions. Seeds germinated at 10–30° C, and the highest germination was 72 % and 88 % at 20° C in light and dark, respectively. Thermal model analysis showed that presence of light significantly increased average thermal requirement [θ _T (50)] from 105°Cd to 186°Cd at sub-optimal temperature, implying that light delays seed germination. Hydrotime model analysis showed that presence of light caused a shift in the median base water potential [Ψ _b(50)] from ?0.68 to ?0.26 MPa, which partly explains why light decreased both percentage and speed of germination, even at optimal conditions. As burial depth increased, seedling emergence initially increased and then decreased; the highest seedling emergence recruitment was 43 %, for seeds buried at a depth of 1 cm. Field observations showed that seedling emergence occurred primarily from July to September, and scarcely occurred from April to June. These results suggest that the light inhibitory effect is an adaptive mechanism that prevents S. bungeana seeds from germinating on the soil surface. To attain highest seedling establishment, seeds of S. bungeana should be sown at a soil depth of 1 cm prior to the rainy season, using seeds stored for 1 year.     

Soil chemical properties affect the concentration of elements (N,P, K,Ca, Mg,As, Cd,Cr, Cu,Fe, Mn,Ni, Pb,and Zn) and their distribution between organs of Rumex obtusifolius

Background and aims

The ionome (elemental composition) of grassland species has rarely been studied at the level of individual organs and little is known about effects of soil chemical properties on the ionome. Using the model oxalate plant Rumex obtusifolius, we asked how its biomass production and the distribution of elements between its organs is affected by soil chemical properties.

Methods

We established a pot experiment with R. obtusifolius planted in acidic non-contaminated control and in slightly acidic and alkaline soils anthropogenically contaminated by the risk elements As, Cd, Pb, and Zn. Both contaminated soils were untreated and treated by lime and superphosphate. We determined biomass production and the concentrations of elements in its organs.

Results

Biomass production was negatively related to the mobility of micro- and risk elements. Restricted transport of micro- and risk elements from belowground organs into leaves was recorded in untreated contaminated soils. In both lime-treated soils and in superphosphate-treated alkaline soil, elevated transport of micro- and risk elements from belowground organs into leaves was recorded in comparison to untreated contaminated soils. The lowest concentrations of micro- and risk elements were recorded in stems and seeds, followed by belowground organs and leaves.

Conclusions

R. obtusifolius is an As-, Cd-, Pb-, and Zn-excluder and is sensitive to high availability of micro- and risk elements in the soil. Soil chemical properties affect the distribution of essential elements within the plant greatly.     

Release and Modification of nod-Gene-Inducing Flavonoids from Alfalfa Seeds

Traces of luteolin, an important rhizobial nod gene inducer in Rhizobium meliloti, are released by alfalfa (Medicago sativa L.) seeds, but most luteolin in the seed exudate is conjugated as luteolin-7-O-glucoside (L7G). Processes affecting the production of luteolin from L7G in seed exudate are poorly understood. Results from this study establish that (a) seed coats are the primary source of flavonoids, including L7G, in seed exudate; (b) these flavonoids exist in seeds before imbibition; and (c) both the host plant and the symbiotic R. meliloti probably can hydrolyze L7G to luteolin. Glycolytic cleavage of L7G is promoted by glucosidase activity released from sterile seeds during the first 4 hours of imbibition. Thus, L7G from imbibing alfalfa seeds may serve as a source of the nod-gene-inducing luteolin and thereby facilitate root nodulation by R. meliloti.