Seeds of Kosteletzkya virginica (Malvaceae): their structure,germination, and salt tolerance. II. Germination and salt tolerance

Kosteletzkya virginica (L.) Presl. (Malvaceae) is a perennial that grows in saline or brackish water, and is salt-tolerant in its mature state, but less tolerant during germination. The seeds show a very low permeability to water that increases during storage. The permeability to water differs in seeds harvested in different years. Optimal temperature for germination is 28–30 C. The effect of salinity on imbibition is largely osmotic, but germination is inhibited, apparently, by the combined osmotic and “ionic” effects, especially at high NaCl concentrations. Inhibition of germination by high NaCl concentrations is relatively more severe in scarified than in intact seeds, indicating that the seed coat acts as a partial barrier to Na⁺ influx. External application of proline or betaine did not improve germination under saline conditions. Dry seeds contain a significant amount of betaine and low levels of proline, but during germination and in the presence of NaCl the betaine content decreased while the proline content increased. Thus, the likely compatible solute in the germinating seed seems to be proline.     

Effects of salinity levels and seed mass on germination in Australian species of Frankenia L. (Frankeniaceae)

Halophyte species demonstrate differing levels of salt tolerance. Understanding interspecific variation to salinity levels is of value from both the scientific perspective, which includes the identification of traits associated with salinity tolerance, as well as from an applied perspective, which includes identifying plant species for specific salinity restoration and remediation projects. This paper investigates the effects of salinity on germination of 12 Australian species of the plant genus Frankenia L. (Frankeniaceae). We use saline solutions that corresponded to the average soil–water salinity concentrations in the arid zones of inland Australia. These solutions consisted of 10 mM calcium chloride, 30 mM magnesium sulphate, and 450 mM sodium chloride. The aims of our study were: (1) to investigate the germination (germination rates, germination success) of Frankenia seeds to four salinity levels (0%, 10%, 20%, 30%), (2) to test for possible interaction effects between seed mass, germination, and salinity, and (3) to examine the effect of salinity levels on the inhibition of germination and/or seed damage. Species varied in their salt tolerance for germination rates and success. Species with larger seeds had higher germination rates and germination success for high salinity levels. Several species did not germinate well at any salinity level. Finally, no seeds were adversely affected by exposure to high salinity levels pre-germination. There is potential for including some Frankenia species in remediation and revegetation projects in areas affected by salinity, and also as garden plants in saline regions.     

Cytokinin profile during early phase of germination of gram seeds after application of benzylaminopurine

Qualitative and quantitative differences in cytokinin levels of gram seeds were observed during germination of control and 6-benzylaminopurine (BAP; 4.44 x 10^t-5 M) treated seeds. Zeatin riboside (ZR), zeatin (Z), dihydrozeatin (DHZ), isopentenyladenosine (iPA) and isopentenyladenine (iP) were determined using high pressure liquid chromatography. In the control seeds the ZR content increased markedly only after 24 h of germination. In BAP treated seeds, the increase in ZR content was stronger than in control seeds and it persisted during further germination. Z content increased during germination in both control and treated seeds. After 48 h of germination iP appeared in control and especially in treated seeds. Acknowledgements: The senior author is thankful to University Grants Commision, New Delhi for award of Research Associateship.     

Influence of Salinity and Temperature on the Germination of Haloxylon recurvum Bunge ex. Boiss.

The stem succulent perennial halophyteHaloxylon recurvum Bungeex. Boiss. grows and produces seed under highly saline conditionsand seeds are deposited in saline soils. Experiments were conductedto determine the effect of salinity and temperature on the germinationof seeds. Results indicate that seeds can germinate at veryhigh salt concentrations (500 mM). However, highest germinationpercentages were obtained in distilled water. Cooler thermoperiodspromoted germination, while high temperatures significantlyinhibited the germination of seeds at all NaCl concentrationstested. Rate of germination decreased with increases in salinity.At higher thermoperiods the rate of germination was significantlylower in comparison to lower thermoperiods. Seeds recover afterbeing transferred to distilled water and recovery was higherfrom higher salinity concentrations and lower thermoperiods.Final recovery germination percentages in high salt treatmentswere significantly lower than non-saline controls, indicatingthat exposure to high concentration of NaCl permanently inhibitedgermination. Germination; halophyte; Haloxylon recurvum recovery of germination; salinity; temperature     

Seed germination of GA-insensitive sleepy1 mutants does not require RGL2 protein disappearance in Arabidopsis

We explore the roles of gibberellin (GA) signaling genes SLEEPY1 (SLY1) and RGA-LIKE2 (RGL2) in regulation of seed germination in Arabidopsis thaliana, a plant in which the hormone GA is required for seed germination. Seed germination failure in the GA biosynthesis mutant ga1-3 is rescued by GA and by mutations in the DELLA gene RGL2, suggesting that RGL2 represses seed germination. RGL2 protein disappears before wild-type seed germination, consistent with the model that GA stimulates germination by causing the SCF(SLY1) E3 ubiquitin ligase complex to trigger ubiquitination and destruction of RGL2. Unlike ga1-3, the GA-insensitive sly1 mutants show variable seed dormancy. Seed lots with high seed dormancy after-ripened slowly, with stronger alleles requiring more time. We expected that if RGL2 negatively controls seed germination, sly1 mutant seeds that germinate well should accumulate lower RGL2 levels than those failing to germinate. Surprisingly, RGL2 accumulated at high levels even in after-ripened sly1 mutant seeds with 100% germination, suggesting that RGL2 disappearance is not a prerequisite for seed germination in the sly1 background. Without GA, several GA-induced genes show increased accumulation in sly1 seeds compared with ga1-3. It is possible that the RGL2 repressor of seed germination is inactivated by after-ripening of sly1 mutant seeds.     

Protein repair L-isoaspartyl methyltransferase in plants. Phylogenetic distribution and the accumulation of substrate proteins in aged barley seeds.

Protein L-isoaspartate (D-aspartate) O-methyltransferases (MTs; EC 2.1.1.77) can initiate the conversion of detrimental L-isoaspartyl residues in spontaneously damaged proteins to normal L-aspartyl residues. We detected this enzyme in 45 species from 23 families representing most of the divisions of the plant kingdom. MT activity is often localized in seeds, suggesting that it has a role in their maturation, quiescence, and germination. The relationship among MT activity, the accumulation of abnormal protein L-isoaspartyl residues, and seed viability was explored in barley (Hordeum vulgare cultivar Himalaya) seeds, which contain high levels of MT. Natural aging of barley seeds for 17 years resulted in a significant reduction in MT activity and in seed viability, coupled with increased levels of "unrepaired" L-isoaspartyl residues. In seeds heated to accelerate aging, we found no reduction of MT activity, but we did observe decreased seed viability and the accumulation of isoaspartyl residues. Among populations of accelerated aged seed, those possessing the highest levels of L-isoaspartyl-containing proteins had the lowest germination percentages. These results suggest that the MT present in seeds cannot efficiently repair all spontaneously damaged proteins containing altered aspartyl residues, and their accumulation during aging may contribute to the loss of seed viability.     

SEED GERMINATION STUDIES IN MUSA. II. ALTERNATING TEMPERATURE REQUIREMENT FOR THE GERMINATION OF MUSA BALBISIANA

Stotzky , G., and Elsie A. Cox . (Central Research Labs., United Fruit Co., Norwood, Mass.) Seed germination studies in Musa. II. Alternating temperature requirement for the germination of Musa balbisiana. Amer. Jour. Bot. 49(7): 763–770. Illus. 1962.—Alternating temperatures were found to be required for the germination of seeds of Musa balbisiana. The temperature differentials optimal for germination in soil are dependent upon both the high and low temperatures, and range from 8–23 C. Germination is maximal when the seeds are held 6–12 hr at the high (27–35 C) and 12–18 hr at the low (12–18 C) temperatures. Some germination can be induced by short exposures to alternating temperatures followed by constant high temperatures, but continuous exposure to alternating temperatures is necessary for maximum germination. Excised embryos develop better at constant than at alternating temperatures, showing that the mechanisms affected by alternating temperatures reside elsewhere in the seed. Alternating temperatures are also required for germination of mechanically scarified seeds, although the temperature differentials are less than those necessary for intact seeds, indicating that the action of alternating temperatures is not on the permeability of the integuments.     

IAA production during germination of Orobanche spp. seeds

Broomrapes (Orobanche spp.) are parasitic plants, whose growth and development fully depend on the nutritional connection established between the parasite and the roots of the respective host plant. Phytohormones are known to play a role in establishing the specific Orobanche-host plant interaction. The first step in the interaction is seed germination triggered by a germination stimulant secreted by the host-plant roots. We quantified indole-3-acetic acid (IAA) and abscisic acid (ABA) during the seed germination of tobacco broomrape (Orobanche ramosa) and sunflower broomrape (O. cumana). IAA was mainly released from Orobanche seeds in host-parasite interactions as compared to non-host-parasite interactions. Moreover, germinating seeds of O. ramosa released IAA as early as 24 h after the seeds were exposed to the germination stimulant, even before development of the germ tube. ABA levels remained unchanged during the germination of the parasites' seeds. The results presented here show that IAA production is probably part of a mechanism triggering germination upon the induction by the host factor, thus resulting in seed germination.     

The Lack of Correlation between ATP Accumulation in Seeds at the Early Stage of Germination and Seed Quality

Seeds of various species were treated with polyethyleneglycolin the absence or presence of AMP and phosphoenolpyruvate, redriedand examined for ATP accumulation at the early stage of germinationand for the rate of germination under suboptimal temperatures.The pretreatments resulted in seeds with various levels of accumulatedATP but in most comparative experiments no correlation was foundbetween the ATP accumulation and the rate of germination. Similarpretreatments with naturally aged seeds led to the same conclusion.The fact that ATP accumulation is a result of ATP synthesisas well as ATP utilization was demonstrated by a protein inhibitionexperiment which showed a possibility to produce seeds withrelatively high ATP accumulation levels at the early stage ofgermination, but with very low germination ability. Key words: AMP, PEP, Polyethyleneglycol, Pretreatment     

Role and regulation of cAMP in seed germination of Phacelia tanacetifolia

Although adenosine 3',5'-cyclic monophosphate (cAMP) is known as a key second messenger in many living organisms, regulating a wide range of cellular responses, its biological function in higher plants is not well understood. In this study, the role and the regulation mechanism of cAMP in seed germination of Phacelia tanacetifolia Benth. were examined. The cAMP level of the seeds incubated under optimal conditions for germination showed a transient elevation before germination. When the seeds were exposed to light or supraoptimal temperature during incubation, elevation of cAMP levels as well as germination of the seeds were inhibited. Addition of membrane-permeable cAMP to the medium restored the germination rates of these seeds, suggesting that cAMP functions during germination. Treatment of the seeds with gibberellin (GA) was also effective to restore the elevation of cAMP levels and germination of the seeds. Uniconazole, a potent inhibitor of GA biosynthesis, blocked elevation of cAMP level under optimal conditions for germination. These results suggest that cAMP plays a role in the regulation of germination and that the cAMP level is regulated by GA in P. tanacetifolia seeds.     

Amino acids,polyamines and proteins during seed germination of two species of Dipterocarpaceae

Summary Amino acid, polyamine and protein concentrations in seeds and their evolution during seed germination of two dipterocarp species, Hopea odorata and Dipterocarpus alatus, were determined with the help of a multianalytical system. Glutamic acid and glutamine were the major amino compounds present. Hopea seeds also contain high levels of aspartic acid/asparagine, serine, threonine, arginine and alanine, while those of Dipterocarpus contain high levels of alanine, arginine and threonine. These species were quite different in their germination behavior and thus in their protein and amine metabolism rates. In Hopea, polyamines increased during the first 3 days of germination and reached a maximum by the 3rd day, 1 day before maximum germination rate. In Dipterocarpus polyamines reached their maximum at the 6th day while maximum germination rate is observed by the 7th day. This suggests that polyamine compounds could play a role in the early part of the germination process in Hopea and Dipterocarpus seeds. The possibility that control of polyamine biosynthesis could be used for the establishment of biochemical methods to improve seed storage and to control germination of these recalcitrant seeds is discussed.     

The dynamics of indole-3-acetic acid in Acer platanoides seeds during stratification and germination

During stratification at 5°C indole-3-acetic acid (IAA) levels in embryos of Acer platanoides decreased during the early stages but subsequently increased again throughout the remainder of a 144 day period. The reduction in IAA levels in embryos of fruits stored at 17°C was even more pronounced, and in addition, no increase was observed after longer storage periods at this temperature, the levels of IAA remaining very low. Germination in seeds maintained at 5°C was not observed until after 120 days or longer, but germination potential increased at an earlier stage, as shown by the fact that seeds transferred to 20°C gave appreciable increases in germination after much shorter chilling periods. Endogenous IAA levels in embryos from seeds transferred to 20°C after a chilling period, long enough to break dormancy, increased within 24 h, i.e. before visible germination, to levels similar to those observed in embryos from seeds chilled continuously for 144 days. Embryos from seeds chilled for 120 days, i.e. when the samples already showed visible germination and when the endogenous IAA content was already high, showed no further increase in endogenous IAA during a three day incubation at 20°C. None of the treatments employed was effective in inducing germination of seeds or embryos from fruits stored at 17°C.     

High temperature-induced abscisic acid biosynthesis and its role in the inhibition of gibberellin action in Arabidopsis seeds

Suppression of seed germination at supraoptimal high temperature (thermoinhibiton) during summer is crucial for Arabidopsis (Arabidopsis thaliana) to establish vegetative and reproductive growth in appropriate seasons. Abscisic acid (ABA) and gibberellins (GAs) are well known to be involved in germination control, but it remains unknown how these hormone actions (metabolism and responsiveness) are altered at high temperature. Here, we show that ABA levels in imbibed seeds are elevated at high temperature and that this increase is correlated with up-regulation of the zeaxanthin epoxidase gene ABA1/ZEP and three 9-cis-epoxycarotenoid dioxygenase genes, NCED2, NCED5, and NCED9. Reverse-genetic studies show that NCED9 plays a major and NCED5 and NCED2 play relatively minor roles in high temperature-induced ABA synthesis and germination inhibition. We also show that bioactive GAs stay at low levels at high temperature, presumably through suppression of GA 20-oxidase genes, GA20ox1, GA20ox2, and GA20ox3, and GA 3-oxidase genes, GA3ox1 and GA3ox2. Thermoinhibition-tolerant germination of loss-of-function mutants of GA negative regulators, SPINDLY (SPY) and RGL2, suggests that repression of GA signaling is required for thermoinibition. Interestingly, ABA-deficient aba2-2 mutant seeds show significant expression of GA synthesis genes and repression of SPY expression even at high temperature. In addition, the thermoinhibition-resistant germination phenotype of aba2-1 seeds is suppressed by a GA biosynthesis inhibitor, paclobutrazol. We conclude that high temperature stimulates ABA synthesis and represses GA synthesis and signaling through the action of ABA in Arabidopsis seeds.     

Promotion by 5-Aminolevulinic Acid of Germination of Pakchoi (Brassica campestris ssp. chinensis var. communis Tsen et Lee) Seeds Under Salt Stress

The seed germination and seedling growth of pakchoi (Brassica campestris ssp. chinensis var.communis Tsen et Lee cv. Hanxiao) were not significantly inhibited until the concentration of NaCl was increased to150 mmol/L. Treatment of pakchoi seeds with exogenous 5-aminolevulinic acid (ALA), at concentrations ranging from 0.01 to 10.00 mg/L, promoted seed germination when seeds were stressed by salinity, whereas levulinic acid (LA), an inhibitor of ALA dehydrase, significantly inhibited seed germination and seedling growth, suggesting that metabolism of ALA into porphyrin compounds was necessary for seed germination and seedling growth. Determination of respiratory rate during seed germination showed that ALA increased seed respiration under both normal conditions and salt stress. Furthermore, salt stress decreased levels of endogenous ALA, as well as heme, in etiolated seedlings. More salt-tolerant cultivars of pakchoi contained higher relative levels of endogenous ALA and heme under conditions of salt stress.These results indicate that salt stress may inhibit the biosynthesis of endogenous ALA and then heme,which is necessary for seed germination, and treatment of seeds with exogenous ALA prior to germination may be associated with the biosynthesis of heme.     

Germination of Tagetes minuta L. II. Role of gibberellins

The effect of the application of gibberellins to Tagetes minuta L. achenes (seeds) was determined at both 25°C, the optimal germination temperature, and 35°C, at which temperature the achenes are thermoinhibited. Both GA₃ and GA₄₊₇ accelerated germination at 25°C. Seed germination at 25°C was inhibited by paclobutrazol, but on subsequent application of GA₄₊₇ rapid germination was induced. Following application of GA₃ or GA₄₊₇ to thermoinhibited seeds, a significantly higher final germination percentage was observed than in the distilled water control. However, endogenous gibberellin levels in germinating and thermoinhibited seeds did not differ significantly.     

Free radical generation in Pinus sylvestris and Larix decidua seeds primed with polyethylene glycol or potassium salt solutions.

Electron paramagnetic resonance (EPR) spectra of Pinus sylvestris and Larix decidua seeds show that priming with PEG+200 mg kg(-1) gibberelic acid (GA(3)) results in appreciably higher free radical contents than in unprimed control samples. Only relatively minor changes in the free radical levels were observed in seeds primed with K(+) salts. However, both priming treatments have been reported previously to result in faster germination rates compared to controls without changing the germination percentage. In measurements on individual seeds of L. decidua, there were no significant differences between the mean free radical levels in viable and non-viable seeds within each treatment group. Thus, the elevation in free radical levels in the PEG+GA(3) treatments appear to be a direct consequence of the priming treatment and do not correspond to the initiation of germination.     

Seed germination and seedling emergence of Euryodendron excelsum H. T. Chang: implications for species conservation and restoration

Euryodendron excelsum H. T. Chang is a critically endangered plant from the family Theaceae that is endemic to China. It is now present in only one remnant population in a very narrow range and with a highly isolated and fragmented distribution pattern. The species is distributed close to the local villages and faces extinction because of the high level of anthropogenic disturbance. Thus, conservation and restoration of this species is urgent. The species reproduces by seed, but its germination requirements and seedling emergence are rarely understood. In this study, the germination requirements, desiccation tolerance and seedling emergence of the species were studied under controlled laboratory conditions. The results indicated that seeds of E. excelsum were non‐dormant. Optimal temperatures for germination were 15°C to 25°C; the germination percentage decreased and the mean germination time increased at high temperature. Seed germination was inhibited in the dark, suggesting that the seeds were positive photoblastic. Water stress also significantly inhibited germination percentage; no seeds germinated at 15% polyethylene glycol 6000. The fresh seeds had moderate moisture content of 28.6% and showed strong tolerance of dehydration. Thus, the seeds of E. excelsum tended to be orthodox. Seeds on the soil surface had the highest emergence percentage, which declined with depth of burial. Soil types significantly affected seedling emergence; seeds had higher emergence percentage in sandy soil than in mixed soil and clayed soil. Our study demonstrated that ex situ conservation and reintroduction of E. excelsum using seedling propagation from seeds is feasible.     

GERMINATION ECOLOGY OF PHACELIA DUBIA VAR. DUBIA IN TENNESSEE GLADES

The germination characteristics of a population of the winter annual Phacelia dubia (L.) Trel. var. dubia from the middle Tennessee cedar glades were investigated in an attempt to define the factor(s) regulating germination in nature. Factors considered were changes in physiological response of the seeds (after-ripening), temperature, age, light and darkness, and soil moisture. At seed dispersal (late May to early June), approximately 50 % of the seeds were non-dormant but, would germinate only at low temperatures (10–15 C). As the seeds aged from June to September, there was an increase in rate and total percent of germination at 10, 15, and 20 C, and the maximum temperature for germination increased to 25 C. Little or no germination occurred at the June, July, and August temperatures in 0- to 2-month-old seeds, even in seeds on soil that was kept continuously moist during this 3-month period. At the September, October, and November temperatures 3- to 5-month-old seeds germinated to high percentages. In all experiments seeds germinated better at a 14-hr photoperiod than in constant darkness. Inability of 0- to 2-month-old seeds to germinate at high summer temperatures allows P. dubia dubia to pass the dry summer in the seed stage, while increase in optimum and maximum temperatures for germination during the summer permits seeds to germinate in late summer and early fall when conditions are favorable for seedling survival and eventual maturation.     

Delayed germination and dispersal in desert annuals: Escape in space and time

Summary A model is developed to consider the interplay between dispersibility and delayed germination in desert annuals. The model explores the effect of low levels of dispersal, considered realistic for annual plants, on optimal germination fraction. The model also demonstrates the effect of the amount and accuracy of predictive (responsive to the environment) dormancy on the optimal innate germination fraction (not responsive to environmental conditions).Optimal germination fraction is found to be very sensitive to changes in despersibility especially at the limited dispersibilities that are realistic for annual plants. As dispersibility increases, optimal germination fraction increases. If plants make two kinds of seeds with differing despersibility, reproduction is maximized if the low dispersal seeds have delayed germination and the high dispersal seeds have quick germination. If dormancy mechanisms permit seeds to germinate when environmental conditions allow successful maturation, and remain dormant when environmental conditions do not permit successful maturation, what fraction of seeds should remain dormant under predicted good conditions as a hedge against inaccurate prediction of the environment? If environmental cues that break dormancy are uncorrelated with environmental conditions that permit successful maturation, predictive dormancy has little or no effect on the optimal innate germination fraction. When predictive dormancy lowers the probability of germinating when environmental conditions preclude successful maturation, the optimal innate germination fraction increases with increasing germination control by predictive dormancy. With a moderate degree of germination control by predictive dormancy, the optimal innate dormancy is still sensitive to changes in dispersal in the low dispersal ranges characteristic of annual plants.Evidence is presented from plant species that have both dispersal and germination dimorphisms to support the predicted correlation of high germination fractions with high dispersal.     

Factors influencing the storability of Fagus sylvatica L. seeds after release from dormancy

Seeds of beech (Fagus sylvatica L.) that have been subjected to dormancy breaking consisting of 10 weeks of prechilling at 3 °C and 34 % water content (WC) and then desiccation to 10 % WC, are non-dormant (ND). ND seeds are characterised by greater sensitivity to storage conditions, than no prechilled, dormant (D) seeds. The aim of the present work was to investigate factors affecting the loss of seed viability during storage of D and ND beech seeds at different temperatures (4 and 20 °C) and humidity levels (45 and 75 % RH) for 3 weeks. In general, both D and ND seeds maintained a high germination capacity after storage at 4 °C. At 20 °C and 45 and 75 % RH the germination capacity of D seeds diminished to 80 and 28 %, respectively. Under the same conditions, ND seeds lost germination capacity to a greater degree, with only 62 and 7 % germinated seeds, respectively. At 20 °C, an increase in production of reactive oxygen species was observed, and the increase was significantly higher in ND seeds. The loss of germination capacity was coincident with an increase in electrolyte leakage and accumulation of free fatty acids, which suggests that membrane deterioration was the cause of the decline in germinability. ND seeds stored at 20 °C and 45 and 75 % RH showed a greater decrease than D seeds in contents of the primary phospholipids phosphatidylcholine (PC) and phosphatidylethanolamine (PE) as well as in polyunsaturated fatty acids (18:2 and 18:3). ND seeds possessed more unsaturated fatty acids, especially 18:3, than D seeds in the phospholipid fraction before storage. D seeds were characterised by a significantly higher level of α-tocopherol and UV-absorbing phenols. The level of ascorbate was similar in D and ND seeds. D seeds contained glutathione in both reduced (GSH) and oxidised (GSSG) forms, and GSSG dominated GSH. ND seeds contained more GSSG form than D seeds. We concluded that the membranes of ND seeds are exposed to greater oxidative stress during storage due to higher levels of unsaturation and lower levels of α-tocopherol, the main antioxidant that protects membranes against free radical attack.