Investigating and modeling the combined effects of pH and osmotic pressure on seed germination for use in phytoactivity and allelopathic research

Phytoactivity and allelopathic studies are heavily dependent on germination bioassays of water solutions of allelochemical(s), which necessarily imply that pH and osmotic pressure vary among treatments and between treatments and controls and are therefore a confounding factor in the assessment of seed germination responses to allelochemical(s). When the contribution of pH and osmotic pressure to seed germination responses is considered in experimental designs their effects are almost without exceptions examined separately being assumed, without any evidences, that pH and osmotic pressure act independently on seed germination responses. The objectives of this work were to examine experimentally such assumption using wheat, lettuce, and subterranean clover cultivars to evaluate and model the combined effects on germination of pH and osmotic pressure in the range between 3.0–6.0 and 0–100 mOsmol kg^?1, respectively. Empirical equations are fitted, discussed, and the need to consider the simultaneous effects of pH and osmotic pressure firmly established. Finally, the use of the equations fitted and its impact on conclusions is exemplified in a dose-response bioassay of water extracts of Cistus ladanifer on seed germination using subterranean clover as target species where hormesis was found before allelochemical effects were corrected for pH and osmotic pressure values of control and extracts.     

FACTORS AFFECTING THE TOXICITY OF SEVERAL LICHEN ACIDS: EFFECT OF PH AND LICHEN ACID CONCENTRATION

The effect of pH 5–8 and lichen acid concentration gradients (2.7 times 10⁻² - 2.7 times 10⁻⁶ m ) on the toxicity of the following lichen acids: usnic, lecanoric, evernic, vulpinic, stictic, fumarpro-tocetraric, psoromic, and atranorin, on spores of Funaria hygrometrica was tested. Percent germination and sporeling growth were used as indicators of toxicity. None of the lichen acids were significantly toxic, for either percent germination or sporeling growth at concentrations equal to or below 2.7 times 10⁻⁵ m at pH 7.0, but many of the lichen acids which increased in toxicity at values different from pH 7 may have been toxic at lower concentrations if a different pH was used for the assay. Lichen acid toxicity showed a good correlation with pH for the parameter of spore germination, or sporeling growth, or both. Some lichen acids did not inhibit germination but were effective in retarding sporeling growth, or vice versa. This observation is discussed in relation to changing fatty acids and other lipid composition as germination occurs. Two of the three O-methylated lichen acids (evernic and psoromic) were among the most effective in inhibiting growth over all, but at lower pH values these were less effective than non-O-methylated lichen acids. Stictic, which is also an O-methylated lichen acid, was the least effective inhibitor over all the pH values for both parameters, while vulpinic was the most toxic over all the pH values. The order of relative toxicity for the lichen acids is different, depending on the pH and concentration at which they are tested and depending on the parameter measured. Thus, in an ecological sense, it is difficult to evaluate the adaptive significance of a particular compound or group of compounds without knowing what factors influence the toxicity of those compounds and how these factors vary in the organism's habitat.     

Regulatory Changes in the Intracellular pH and Cl– Efflux at Early Stages of Pollen Grain Germination in vitro

The regulatory role of intracellular pH changes and of transmembrane Cl^– transport in the activation of Nicotiana tabacum L. pollen grains at a stage preceding in vitro germination was studied. The acidification of the cytosol with propionic acid hindered the germination of pollen grains, whereas its alkalization by fusicoccin-stimulated H⁺-ATPase activity of plasma membranes sharply increased the germination frequency with respect to control values. The activation of pollen grains was accompanied by the Cl^– efflux. The blockage of Cl^– efflux with 1 mM ethacrynic acid significantly decreased the intracellular pH and fully inhibited germination. The results allow assumption that the intracellular pH rise and Cl^– efflux are prerequisites for pollen grain activation.     

Germination response to fire-related factors of seeds from non-serotinous and serotinous cones

Pinus halepensis, a Mediterranean pine tree, is a partially serotinous species: individual trees of this species carry both non-serotinous and serotinous cones. Serotinous cones open mainly after fire, whereas non-serotinous cones open in absence of fire. In this study we addressed the question, whether or not this cone response is linked with the germination response of seeds to fires. Two main factors associated with fire are heating of seeds and soil pH. A combination of high heat and high pH simulates a scenario with fire, whereas low heat and low pH simulates a scenario without fire. We assessed the separate and combined effects of heat and pH on the germination rate and the percentage of germination of seeds from non-serotinous cones and two age classes of serotinous cones of P. halepensis. Heat had no effect on the percentage of germination of seeds from any of the cone types, but did positively affect the germination rates of seeds from both age-classes of serotinous cones. High pH negatively affected the germination rate of seeds from all cone types as well as the percentage of germination of seeds from non-serotinous cones. The combinations of heat and pH had different effects on the three cone types: percentage of germination and rate of germination of seeds from non-serotinous cones was higher in the combination high heat-high pH than in the combination low heat-low pH. In the combination high heat-high pH, seeds from serotinous cones germinated better than seeds from non-serotinous cones. The different germination responses of seeds from non-serotinous and serotinous cones could not be attributed to differences in cone age. Our results indicate that the cone response is linked to the germination response of the seeds in P. halepensis, with seeds from serotinous cones being more tolerant to fire related factors. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of simulated acid rain on the pollen germination and pollen tube growth of apple (Malus sylvestris Miller cv. Golden)

The pollens of apple flowers have been treated with simulated acid rain solutions in range of pHs 2.9 to 5.0 in order to determine the threshold proportion values that lead the observed symptoms of detriments of acid rain. Compared to controls (pH 6.5), pollen germination decreased by 41.75% at pH 3.3 and pollen tube elongation decreased by 24.3% at pH 3.4. Acid rain threshold proportion value was around pH 3.3 and 3.4 for apple pollen germination and pollen tube elongation, respectively. Furthermore, pollen tube elongation was determined to be more sensitive to acid rain than pollen germination. The pH values below 3.1 resulted in complete destruction of pollen tubes. Pollen germination entirely stopped at around pH 3.0. Finally, it has been shown that the acid rain has a blocking effect on pollen germination and pollen tube elongation in apple. The conclusion is that not only pH value but also the quantity of acid rain is important factor in germination. The results were found statistically significant through the LSD test at levels of p < 0.05 and p < 0.01.     

Synergistic and Antagonistic Effects of Salinity and pH on Germination in Switchgrass (Panicum virgatum L.)

The effects of salt-alkaline mixed stress on switchgrass were investigated by evaluating seed germination and the proline, malondialdehyde (MDA) and soluble sugar contents in three switchgrass (Panicum virgatum L.) cultivars in order to identify which can be successfully produced on marginal lands affected by salt-alkaline mixed stress. The experimental conditions consisted of four levels of salinity (10, 60, 110 and 160 mM) and four pH levels (7.1, 8.3, 9.5 and 10.7). The effects of salt-alkaline mixed stress with equivalent coupling of the salinity and pH level on the switchgrass were explored via model analyses. Switchgrass was capable of germinating and surviving well in all treatments under low-alkaline pH (pH≤8.3), regardless of the salinity. However, seed germination and seedling growth were sharply reduced at higher pH values in conjunction with salinity. The salinity and pH had synergetic effects on the germination percentage, germination index, plumular length and the soluble sugar and proline contents in switchgrass. However, these two factors exhibited antagonistic effects on the radicular length of switchgrass. The combined effects of salinity and pH and the interactions between them should be considered when evaluating the strength of salt-alkaline mixed stress.     

Emergence and germination response of Sonchus oleraceus and Rapistrum rugosum to different temperatures and moisture stress regimes

Sonchus oleraceus and Rapistrum rugosum are two rapidly emerging weeds of the northern grain region of Australia. To understand the ability of these weeds regarding their germination response to temperature and different soil moisture regimes, experiments were undertaken on the germination of these weeds at varying osmotic potential and temperature regimes. The experiment was conducted as a split-plot design with alternating day/night temperature regimes (15/5, 20/10, 25/15 and 30/20°C) as a main plot and osmotic potential regimes (0.0, −0.1, −0.2, −0.4, −0.6, −0.8 and −1 MPa) as a subplot. At different temperature regimes, there was 65–91% germination of S. oleraceus in water (0 MPa). There was 0–4% germination at −0.8 MPa and no germination at −1.0 MPa. Osmotic potential values that can cause 50% reduction in germination of S. oleraceus based on a sigmoid regression model ranged from −0.38 to −0.48 MPa. There was 33–81% germination of R. rugosum in distilled water (0 MPa), 1–3% germination at −0.8 MPa and no germination at −1.0 MPa. Osmotic potential values that can cause 50% reduction in germination of R. rugosum based on a sigmoid model ranged from −0.26 to −0.54 MPa. Results of the study were related to the emergence pattern of weeds during field survey and soil moisture profiles estimated by the Australian Landscape Water Balance Model and explain the emergence of these weeds outside the normal seasonal window of prevalence as a response to changes in weather.     

不同pH值土壤及其浸提液对羊草种子萌发和幼苗生长的影响

研究了不同pH值土壤以及重度盐碱土和非盐碱土浸提液对羊草(Leymus chinensis)种子萌发及幼苗生长的影响。不同pH值土壤由配土和调酸两种方式取得,前者由重度盐碱土(pH=10．24)与非盐碱土(pH=7．49)按不同比例配制,后者是由重度苏打盐碱土经硫酸调酸处理得到。配土发芽的实验结果表明,当pH值在7．49-9．14时,种子的发芽率均在50％以上,且幼苗能够正常生长;pH=9．53时,羊草种子的发芽率低于50％,仅部分幼苗个体能够成活;当pH>9．86时,幼苗在萌发后50天左右全部枯死。说明羊草种子个体萌发期最大耐受pH值在9．14-9．53之间。重度苏打盐碱土调酸发芽实验结果表明,当调酸后的土壤pH值降至7．0-10．0时,均能显著促进羊草种子的萌发,且幼苗生长正常。确定了羊草种子萌发的最适pH值为8．0-8．5。土／水比为1:1的非盐碱土浸提液能够显著提高羊草种子发芽率,而重度苏打盐碱土浸提液对羊草种子萌发没有产生显著抑制。     

不同pH 值土壤及其浸提液对羊草种子萌发和幼苗生长的影响

研究了不同pH值土壤以及重度盐碱土和非盐碱土浸提液对羊草(Leymus chinensis)种子萌发及幼苗生长的影响。不同pH值土壤由配土和调酸两种方式取得, 前者由重度盐碱土(pH = 10.24)与非盐碱土(pH = 7.49)按不同比例配制, 后者是由重度苏打盐碱土经硫酸调酸处理得到。配土发芽的实验结果表明, 当pH值在7.49-9.14时, 种子的发芽率均在50%以上, 且幼苗能够正常生长; pH = 9.53时, 羊草种子的发芽率低于50%, 仅部分幼苗个体能够成活; 当pH > 9.86时, 幼苗在萌发后50天左右全部枯死。说明羊草种子个体萌发期最大耐受pH值在9.14-9.53之间。重度苏打盐碱土调酸发芽实验结果表明, 当调酸后 的土壤pH值降至7.0-10.0时, 均能显著促进羊草种子的萌发, 且幼苗生长正常。确定了羊草种子萌发的最适pH值为8.0-8.5。土/水比为1: 1的非盐碱土浸提液能够显著提高羊草种子发芽率, 而重度苏打盐碱土浸提液对羊草种子萌发没有产生显著抑制。     

模拟酸雨对茄科3种蔬菜种子萌发的影响

采用pH 2.0、3.0、4.0、5.0的模拟酸雨和pH 6.5的中性溶液(ck)处理番茄、茄子、辣椒3种蔬菜种子,研究酸雨胁迫强度对3种蔬菜种子萌发的影响。结果表明：从发芽率、发芽势、发芽指数3个萌发指标来看,番茄种子在pH 2.0～3.0的强酸雨胁迫下,萌发受到轻微抑制;茄子种子在pH 2.0～5.0的酸雨胁迫下,萌发一直受到抑制,尤其是在pH 2.0的强酸雨胁迫时表现出萌发几乎完全受到抑制,而辣椒种子在酸雨胁迫下,萌发几乎没有受到抑制。3个萌发指标均随着酸雨pH值的减小而减少。pH 2.0～5.0酸雨胁迫下,辣椒吸水值的变幅（13.75）﹤番茄（19.91）﹤茄子（20.58）,同等酸雨胁迫强度下,根长抑制指数、蛋白质含量降幅、丙二醛增幅辣椒﹤番茄﹤茄子。这表明抗酸雨胁迫能力：辣椒>番茄>茄子。     

Effects of Ruta graveolens leaves on soil characteristics and on seed germination and early seedling growth of four crop species

Effects of rue (Ruta graveolens) leaves on soil characteristics and on germination and seedling growth of four crop species (radish, zucchini, cauliflower and tomato) were investigated. Soil not previously associated with rue was mulched and amended with its fresh leaves. The objectives of the present study were to determine: (i) the changes in soil chemical characteristics after addition of rue leaves, (ii) the growth responses in soils treated with R. graveolens leaves, and (iii) whether the soil chemistry changes affect plant growth. Results indicate that soil incorporated with rue leaves had higher values of water‐soluble phenolics, electrical conductivity and nitrate, and lower pH values compared to untreated soil. Seed germination of radish and tomato was suppressed in covered and amended soils compared to control soil. Seedling development for all the species was suppressed in incorporated and amended soil, compared to the rue‐free soil. Correlations between soil chemical characteristics, germination and seedling growth of plants are also discussed.     

Effects of acidity on the insoluble polysaccharide content of germinating pollen of Pinus pinea L. and Pinus pinaster Aiton

The PAS reaction was used to stain total insoluble polysaccharides in pollen of two species of Pinus collected in two different years and germinated in vitro at different pH values. PAS-detectable polysaccharides are localized in the intine, cytoplasm and amyloplasts; PAS-undetectable (callose) in the middle of the intine, but only in the distal part of the grain. The total PAS stained polysaccharide content was quantified with a microdensitometer. Values are correlated with germination percentages and pollen tube length. The total polysaccharide content, as well as pollen tube length, varies in each species according to the year, probably due to environmental conditions. Also the various pHs deeply influence pollen germination and tube length, the lowest pH being the most unfavourable condition. At this pH (3.5) the total polysaccharide content is higher than in the control (pH 5.5): this is probably due to a lack of callose synthesis for tube wall or to a depolymerization of this polysaccharide, and a subsequent storage in a PAS-detectable form.     

Prediction of cotton seedling germination against pre-emergence damping off on the basis of environmental factors and seed applied fungicides

Cotton is an important fibre crop chiefly attacked by a number of diseases. Pre-emergence damping off seedling is an important disease of cotton. Efficacy of three fungicides (Dynesty, Topsin-M and Antracol) was assessed on early mid and late sowings. Topsin-M and Antracol were evaluated at 1X and 2X concentrations, Dynasty was only used at standard dose. Significant increase in seed germination was seen, maximum germination was observed in late sowing. Antracol (2X) significantly improved seed germination rate in all three sowings as compared to other treatments. Impact of environmental parameters on germination rate was assessed relating to seed dressing fungicides at 1X and 2X concentrations. Significant correlation was observed between soil temperature and seedling germination rate by all treatments, 30–31 °C favoured germination rate. Maximum seedling germination rate was observed at 24–26 and 22–23 °C and 45–50% by average air temperature, minimum air temperature and relative humidity respectively. Regression model was contrived to predict seedling germinations impacted by seed dressing fungicides relating to environmental factors. Closed resemblance between observed and predicted values states that model can be used to predict seed germination by treating with fungicides regarding to environmental factors.     

Phytotoxicity of iron in relation to its solubility conditions and the effect of ionic strength

The effect of iron on root elongation and seed germination of Lactuca sativa was investigated within the limits of the plant's sensitivity to an acidic environment and iron solubility under experimental conditions. Because ionic strength was found to have phytotoxic effects, the same pH and ionic strength conditions were kept constant over the entire iron concentration range, so as to evaluate solely the effect of iron. 0.14 mM Fe³⁺ was found to be the lowest effective concentration inhibiting root elongation. The low solubility of iron at a pH of 3 (the lowest value which could be applied for root elongation) excluded the testing of higher iron concentrations. Due to the lower sensitivity of seed germination to acidity, a pH of 2.6 could be applied in this case. The lowest effective iron concentration (the lowest applied concentration causing a significant decrease in germination as compared to the control, P < 0.05), EC50 (the concentration at which germination was 50% of that of the control specimens) and the completely effective iron concentration (the concentration that completely inhibited seed germination) were 0.6, 0.92 and 2.0 mM, respectively.     

Responses of maternal siblings ofPinus densiflora to simulated add rain

Because most research on acid rain does not consider the genetic composition of seeds and the inherent variations in seed mass and germination date, the potentially diverse patterns of plant response are neglected within a population. Therefore, we examined the effects of both family identity and simulated acid rain onPinus densiflora seedlings over a 34-week period, beginning at germination. We were able to evaluate the importance of family and pH treatment independent of seed mass and germination date by considering the latter two variables as covariates. Seeds were collected from six randomly chosen trees that were assumed to represent different genotypes. After germination in a controlled environment, the seedlings were moved outdoors. Each was then subjected twice a week to 40 mL of simulated acid rain, at a pH of either 3.0 or 5.6. We examined variations in germination date, seedling height, needle numbers and lengths, and root and shoot masses. The maternal family effect was significant for all characters over the entire test period. Strong effects of pH treatment were detected only at the later growth stages. Artificial rain with a pH of 3.0 enhanced growth rates, particularly aboveground, compared with seedlings treated with pH 5.6 rain. However, regardless of pH, certain groups of maternal siblings consistently performed better in almost all parameters. The exception was for mean germination date, which differed among families only at a pH of 3.0. Therefore, although we demonstrated that acid rain affected both the date of germination as well as seedling growth rates inP. densiflora, the responses differed among maternal families, and were also associated with seed mass and germination date.     

Effect of temperature,hydrogen ion concentration and osmotic potential on oospore germination of fivePythium spp. isolated from pond water

Oospore germination occurred over a temperature ranging of 15–35°C forPythium coloratum, 10–35°C forP. diclinum, 15–30°C forP. dissotocum, 7–30°C forP. monospermum, and 10–30°C forP. pleroticum. Optimum temperature was 25°C for all species tested. In case of pH, oospore germination occurred over a range of 4.76–8.55 with an optimum of 6.40–7.40. The least germination occurred at pH 4.76 forP. coloratum, P. diclinum, P. monospermum andP. pleroticum, whileP. dissotocum germinated from pH 5.02. Oospores of the all tested pythia were able to germinate at –0.13 to –1.65 MPa and could not germinate at –3.40 MPa, with the highest germination rate at –0.27 to –0.47 MPa. The effect of temperature, pH and osmotic potential on oospore germination was discussed in relation to pollution of pond water.     

Effect of certain environmental factors on zygospore germination ofSpirogyra hyalina

The present study focusses on the effects of pH, temperature, intensity of white, red far-red light on zygospore germination in the filamentous green algaSpirogyra hyalina. Maximum germination of zygospores occurred at pH 8.0 and 25°C. Germination of zygospores was favored by white light at an intensity of 3–4 klx. Red light resulted in higher germination when applied at the beginning or in the middle of the dark period, while far-red light decreased zygospore germination. Red light in the middle of the dark period was found to be most effective for germination. These observations suggest a possible presence of the phytochrome system in the test alga contributing to its morphogenetic response.     

Factors affecting spore germination in Aspergillus niger

Temperature markedly affected germination and germ tube length of A. niger. More than 90% of the spores were germinated in the range 30°–34 °C and formed maximum length of germ tubes. At temperatures from 38° to 43 °C, the proportion of the spores that germinated as well as the germ tube length were both gradually decreased. However, at 47°C germ tube formation was completely inhibited up to 15 hrs. after inoculation.High relative humidity was found necessary for the spore germination of A. niger. Germination failed to occur at 76% relative humidity. At 78 and 81% relative humidity germination was detected 15 hrs. after inoculation while at the higher humidities germination was started after 6 hrs. only.Conidiospores of A. niger were very sensitive to changes in the hydrogen ion concentration, pH. Complete inhibition of germination was found at pH less than 3.5. The germination and the length of the formed germ tubes increased with pH to reach their maximum rates at pH 4.5.     

Effect of environmental factors on seed germination and emergence of Lepidium vesicarium

Lepidium vesicarium is a weed species with a wide distribution in the rangelands and dry‐land farming in East Azarbaijan, Iran. The experiments were undertaken to assay the effects of light, temperature, pH, osmotic potential, NaCl concentration and burial depth on seed germination and emergence of L. vesicarium. Germination was maintained at high levels (> 80%) over a wide day/night temperature range (10/5 to 30/20°C), but a severe reduction in the germination rate of L. vesicarium was found below 20/10°C. Germination of L. vesicarium was influenced by different light/dark regimes, as the germination rate was highest at 16 h light for the all treatments (0, 8, 12, 16 and 24 h light). Germination was 92–95% over a wide range of pH (2‐10). Germination was >50% at a water potential of ?0.7 MPa and salinity of 21 dS/m, indicating that drought and salt conditions have a minimal impact on seed germination. With increasing burial depth from 0 to 2 cm, the number of days required for 50% emergence increased and no germination was observed at burial depths deeper than 3 cm. This suggests that L. vesicarium would become troublesome in the rangelands and for growers in reduced‐tillage cropping systems. The ability to emerge from shallow depths, coupled with tolerance of a wide pH range, drought and salinity at germination, should be taken into account when managing this weed species.