The GerW Protein Is Not Involved in the Germination of Spores of Bacillus Species

Germination of dormant spores of Bacillus species is initiated when nutrient germinants bind to germinant receptors in spores’ inner membrane and this interaction triggers the release of dipicolinic acid and cations from the spore core and their replacement by water. Bacillus subtilis spores contain three functional germinant receptors encoded by the gerA, gerB, and gerK operons. The GerA germinant receptor alone triggers germination with L-valine or L-alanine, and the GerB and GerK germinant receptors together trigger germination with a mixture of L-asparagine, D-glucose, D-fructose and KCl (AGFK). Recently, it was reported that the B. subtilis gerW gene is expressed only during sporulation in developing spores, and that GerW is essential for L-alanine germination of B. subtilis spores but not for germination with AGFK. However, we now find that loss of the B. subtilis gerW gene had no significant effects on: i) rates of spore germination with L-alanine; ii) spores’ levels of germination proteins including GerA germinant receptor subunits; iii) AGFK germination; iv) spore germination by germinant receptor-independent pathways; and v) outgrowth of germinated spores. Studies in Bacillus megaterium did find that gerW was expressed in the developing spore during sporulation, and in a temperature-dependent manner. However, disruption of gerW again had no effect on the germination of B. megaterium spores, whether germination was triggered via germinant receptor-dependent or germinant receptor-independent pathways.     

Effect of fungi and light on seed germination of three Opuntia species from semiarid lands of central Mexico

Fungal attack under light reduces mechanical resistance of the testa of Opuntia seeds, making it easier for the embryo to emerge. However, the effect of fungi on Opuntia seed germination in darkness is unknown. We evaluated the combined effects of light and inoculation with Phoma medicaginis, Trichoderma harzianum, Trichoderma koningii, and Penicillium chrysogenum on germination of O. streptacantha, O. leucotricha, and O. robusta seeds, from central Mexico. We also evaluated the combined effects of seed age (2-, 3-, and 12-year-old seeds) and presence of fungi on the testa on O. streptacantha germination. All fungal species eroded the funicular envelope and promoted seed germination for O. leucotricha and O. streptacantha, but did more so in light than in darkness. For the latter species, younger seeds inoculated with fungi had lower germination than older ones. For O. robusta, we found that seeds inoculated with P. medicaginis and T. harzianum had similar germination in light and in darkness. Our results strongly indicate that deterioration of the testa by fungi is higher in light than in darkness.     

Effects of a warmer climate on seed germination in the subarctic

Background and Aims

In a future warmer subarctic climate, the soil temperatures experienced by dispersed seeds are likely to increase during summer but may decrease during winter due to expected changes in snow depth, duration and quality. Because little is known about the dormancy-breaking and germination requirements of subarctic species, how warming may influence the timing and level of germination in these species was examined.

Methods

Under controlled conditions, how colder winter and warmer summer soil temperatures influenced germination was tested in 23 subarctic species. The cold stratification and warm incubation temperatures were derived from real soil temperature measurements in subarctic tundra and the temperatures were gradually changed over time to simulate different months of the year.

Key Results

Moderate summer warming (+2·5 °C) substantially accelerated germination in all but four species but did not affect germination percentages. Optimum germination temperatures (20/10°C) further decreased germination time and increased germination percentages in three species. Colder winter soil temperatures delayed the germination in ten species and decreased the germination percentage in four species, whereas the opposite was found in Silene acaulis. In most species, the combined effect of a reduced snow cover and summer warming resulted in earlier germination and thus a longer first growing season, which improves the chance of seedling survival. In particular the recruitment of (dwarf) shrubs (Vaccinium myrtillus, V. vitis-idaea, Betula nana), trees (Alnus incana, Betula pubescens) and grasses (Calamagrostis lapponica, C. purpurea) is likely to benefit from a warmer subarctic climate.

Conclusions

Seedling establishment is expected to improve in a future warmer subarctic climate, mainly by considerably earlier germination. The magnitudes of the responses are species-specific, which should be taken into account when modelling population growth and migration of subarctic species.Key words: Climate change, cold stratification, dwarf shrubs, germination percentage, incubation temperature, mean germination time, seedling establishment, seeds, Silene, subarctic species, Vaccinium, warming     

Structural requirements of strigolactones for germination induction and inhibition of Striga gesnerioides seeds

Key message

Structure–activity relationship studies of strigolactones and Striga gesnerioides seed germination revealed strict structural requirements for germination induction and a new function of the plant hormones as germination inhibitors.

Abstract

Stereoisomers of the naturally occurring strigolactones, strigol, sorgolactone, orobanchol, sorgomol and 5-deoxystrigol, 36 in total, were prepared and screened for the ability to induce and/or inhibit the germination of Striga hermonthica and Striga gesnerioides seeds collected from mature plants that parasitized on sorghum and cowpea, respectively. All of the compounds induced S. hermonthica seed germination, albeit displayed differential activities. On the other hand, only a limited number of the compounds induced significant germination in S. gesnerioides, thus indicating strict structural requirements. Strigolactones inducing high germination in S. gesnerioides induced low germination in S. hermonthica. Strigolactones with the same configuration at C3a, C8b and C2′ as that in 5-deoxystrigol (9a) induced high germination of S. hermonthica seeds, but most of them inhibited the germination of S. gesnerioides. The differential response of S. gesnerioides to strigolactones may play an important role in the survival of the species. However, the compounds could be used as means of control if mixed cropping of cowpea and sorghum is adopted.     

Maternal tissue is involved in stimulant reception by seeds of the parasitic plant Orobanche

Background and Aims

A fundamental element in the evolution of obligate root-parasitic angiosperms is their ability to germinate only in response to chemical stimulation by roots, to ensure contact with a nearby nourishing host. The aim of this study was to explore inheritance of the unique germination control in this group of plants.

Methods

Analysis was made of the segregation of spontaneous (non-induced) germination that appeared in hybrid progenies derived from crosses between Orobanche cernua and O. cumana, which, like all other Orobanche species, are totally dependent on chemical stimulation for the onset of germination, and show negligible spontaneous germination in their natural seed populations.

Key Results and Conclusions

F₁ and F₂ seeds did not germinate in the absence of chemical stimulation, but significant spontaneous germination was found in some F₃ seed families. This indicates that the prevention of non-induced germination in Orobanche seeds, i.e. dependence on an external chemical stimulation for seed germination, is genetically controlled, that this genetic control is expressed in a seed tissue with maternal origin (presumably the perisperm that originates from the nucellus) and that genetic variation for this trait exists in Orobanche species. Similar segregation results were obtained in reciprocal crosses, suggesting that stimulated germination is controlled by nuclear genes.     

Temperature and water requirements for germination and effects of discontinuous hydration on germinated seed survival in Tillandsia recurvata L.

Tillandsia recurvata is an epiphytic bromeliad with a wide distribution in the Americas; however, little is known about the development of its post-seminal adaptations for survival in epiphytic environments. The purpose of this study was to define the temperature and water requirements for the germination of T. recurvata seeds. The absence of radicle emergence in T. recurvata seeds resulted in 2?stages of germination: swollen with broken seed coat (stage-1) and chlorophyllic embryos (stage-2). The effects of partial or discontinuous hydration on germinated seed survival were also assessed. The seeds were collected in a semi-arid shrubland of Mexico City. We explored: (1) whether water vapour can provide a sufficient water source for germination; (2) the temperature required for germination stage-1 and the optimal and critical osmotic potentials for germination in both germination stages; (3) the effect of seed incubation at different osmotic potentials that undergo subsequent dehydration on their survival in stage-2; and (4) the loss of dehydration tolerance during early post-seminal development. In addition, an image of T. recurvata seed anatomy was obtained to illustrate its structures. Germination stage-1 of T. recurvata seeds is rather similar across the tested temperature range. The seeds required to be in contact with liquid water to germinate. The interval of osmotic potential facilitating both germination stages was from 0 to ?0.6?MPa. Although germinated seeds displayed dehydration tolerance, this tolerance decreased in germination stage-2. The osmotic potential during germination affected the tolerance of the chlorophyllic embryos (stage-2) to subsequent dehydration.     

Phosphorus-mediated changes in life history traits of the invasive New Zealand mudsnail (Potamopyrgus antipodarum)

The longer soil has to stay moist to allow germination the more likely that seedlings experience favourable moisture conditions. Since theory predicts that fitness variance-reducing traits will be negatively correlated, we tested the hypothesis that time to germination is negatively correlated with the ability of radicle growth to keep up with the drying front. We measured time to germination and root elongation rate (RER) in 14 Kalahari annuals. We controlled for habitat (canopy association and sand content), germinability, median base water potential for germination (ψ ₅₀), seed mass and seed shape as a persistence surrogate. For species and phylogenetically independent contrasts (PICs), we did not find a relationship between time to germination and RER. However, we found a negative relationship of time to germination with RER for PICs when controlling for sand content and ψ ₅₀. Seed shape increased with time to germination which can be explained by reduced opportunities for germination in slow-germinating species that select for persistence. We found a positive relationship between time to germination and ψ ₅₀, suggesting a continuum of risky to cautious germination. ψ ₅₀ was not correlated with RER suggesting that variation in ψ ₅₀ reflects different drought-adapted traits. Probably the relationship of time to germination with RER is not mediated by seed mass, which was not correlated with time to germination as found by others, though RER was positively correlated with seed mass. Instead of the seed size-seed number trade-off, a trade-off with resource capture may explain variation in RER: more root hairs or rootlets may increase resource capturing surface while reducing RER. For habitat, we found a (positive) relationship with time to germination only for canopy association. This may be explained by maximization of resource capture at the expense of RER being favoured by the higher nutrient and water availability under canopies. Future studies should clarify which trade-offs govern variation in time to germination, focussing on a possible resource capture-RER trade-off.     

Pregnancy coloration in macaques may act as a warning signal to reduce antagonism by conspecifics

Co-varying traits in acorns such as seed size and germination schedule are important to influence the behavioural decisions of hoarding rodents. Using acorn pairs from cork oak (Quercus variabilis) (large size and short germination schedules) serrate oak (Q. serrata) (small size and short germination schedule) and qinggang (Cyclobalanopsis glauca) (small size and long germination schedule) with contrasting seed size and germination schedule, we conducted a series of experiments to investigate hoarding preferences in response to seed size and germination schedule by Edward's long-tailed rats (Leopoldamys edwardsi) and South China field mice (Apodemus draco) in semi-natural enclosures. We found that the seed size hypothesis was consistently supported: both rodent species ate more small acorns but hoarded more large ones regardless of germination schedules. However, the germination schedule hypothesis was also supported when similar sized acorns were simultaneously provided, e.g. Q. serrata versus C. glauca or germinating versus non-germinating Q. variabilis. Our results, contrary to the studies from North America, indicate that seed size is more important than germination schedules in determining whether the tested animals eat or hoard a given seed.     

Role of Ethylene in the Germination of the Hemiparasite Striga hermonthica

Seed germination of the hemiparasitic angiosperm Striga hermonthica (Del.) Benth is elicited by compounds present in the root exudates of the host plant. Although a variety of compounds can substitute for the host-derived signal, the mechanism through which these act is unknown. In the present study, an inhibitor of ethylene biosynthesis, aminoethoxyvinyl glycine, was found to inhibit germination. Addition of an intermediate in ethylene biosynthesis, 1-aminocyclopropane-1-carboxylic acid, was found to override this inhibition and to act as a substitute for the host-derived signal. 2,5-Norbornadiene, an inhibitor of ethylene action, was also found to inhibit germination. Ethylene is rapidly produced by Striga seeds after treatment with host root exudates. These results are consistent with a model for Striga seed germination in which host-derived signals and other compounds act by eliciting the synthesis of ethylene and in which ethylene itself initiates the biochemical changes leading to germination.     

Effects of seed storage on germination of two succulent desert halophytes with little dormancy and transient seed bank

Seeds of both Salsola imbricata and Haloxylon salicornicum have high germination level and germination speed, and form a transient seed bank in nature. The impacts of storage period and condition on germination level and speed were assessed in the two species. Storage for three months significantly increased both germination level and speed of seeds stored under the different conditions, compared to that of fresh seeds. In both species, nine months storage did not affect germination percentage in cold storage seeds, but completely inhibited it in field seeds. Storage for longer time in room and warm temperatures resulted in significant reduction or complete inhibition in the germination of the two species, so this was more pronounced in H. salicornicum. Storage significantly increased germinate rate index of seeds stored in all conditions till 17 months in S. imbricata and till 12 months in H. salicornicum. In both species, fridge storage had little effects on final germination and germination speed of seeds incubated at the different temperatures, compared to fresh seeds. However, room temperature and warm storages significantly reduced final germination and germination speed at the different temperatures, so the reduction was more pronounced at 35 °C, especially in H. salicornicum.     

Photocontrol of the germination of onoclea spores: I. Action spectrum

Light stimulates the germination of spores of the fern Onoclea sensibilis L. At high dosages, broad band red, far red, and blue light promote maximal germination. Maximal sensitivity to these spectral regions is attained from 6 to 48 hours of dark presoaking, and all induced rapid germination after a lag of 30 to 36 hours. Maximal germination is attained approximately 70 hours after irradiation. Dose response curves suggest log linearity. The action spectrum to cause 50% germination shows that spores are most sensitive to irradiation in the red region (620-680 nm) with an incident energy less than 1000 ergs cm⁻²; sensitivity decreases towards both shorter and longer wavelengths. Although the action spectrum is suggestive of phytochrome involvement, photoreversibility of germination between red and far red light has not been demonstrated with Onoclea spores. An absorption spectrum of the intact spores reveals the presence of chlorophylls and carotenoids. Since the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea does not inhibit germination, it is concluded that photosynthesis does not play a role in the germination process.     

Atmospheric condition controlling the seed germination of an achlorophyllous orchid,Galeola septentrionalis

The aseptic seed germination ofGaleola septentrionalis was observed only in an airtight vessel at 30 C, germination being enhanced with increasing air pressure in the vessel up to the optimum of 1.8 atm. The presence of O₂ and CO₂ was essential for germination, and their optimum concentrations were 5 and 8%, respectively. The optimum pressure for germination (1.8 atm) was not influence by the composition of the air. Ethylene added at concentrations ranging from 2 to 8μl/l enhanced germination and its removal reduced the germination. The atmospheric condition required for the seed germination of this orchidin vitro was discussed with regard to that in its natural habitat.     

The effect of growth retardants on phytochrome-induced lettuce seed germination

Experiments were carried out to explore the involvement of the plant hormone gibberellin (GA) in the light-induced germination of lettuce seeds. Three growth retardants known to be inhibitors of GA biosynthesis were tested for their effect on red-light-induced germination. Chlormequat chloride (CCC) and AMO-1618 had no effect, but ancymidol was strongly inhibitory. Moreover, the inhibition caused by ancymidol was completely overcome by GA₃. CCC and AMO-1618 inhibit the formation ofent-kaurene, while ancymidol blocks the oxidation ofent-kaurene toent-kaurenoic acid. Ancymidol also was found to inhibit GA-induced dark germination of lettuce seeds, and this inhibition was partially reversed by higher levels of GA. Therefore, the results suggest two possibilities for the relationship between phytochrome and GA in this system: first, the rate-limiting step in the germination of light-sensitive lettuce seeds, that which is regulated by phytochrome, is the oxidation ofent-kaurene toent-kaurenoic acid. Alternatively, red-light treatment may result in the release of active GAlike substances which, in turn, induce germination. In either case the results presented here support the view that phytochrome exerts its effect on lettuce seed germination by means of GA rather than via an independent pathway.     

Abscisic Acid and the developmental regulation of embryo storage proteins in maize

The relationship of abscisic acid (ABA) inhibition of precocious germination and ABA-induced storage protein accumulation was examined over the course of embryogenesis in wild-type and viviparous mutants of maize (Zea mays L.). We show that a high level of embryo ABA and the product of the Viviparous-1 gene are both required in early maturation phase for germination suppression and the accumulation of storage globulins encoded by the gene Glb1. Suppressing precocious germination with a high osmoticum is not sufficient to initiate Glb1 protein synthesis, although continued accumulation is contingent upon this inhibition; germination of immature or mature embryos leads to a decline in synthesis and the degradation of stored globulins. Late in embryogenesis, fragments of Glb1 protein accumulate, coinciding with the loss of ABA sensitivity. These results suggest that ABA influences storage globulin accumulation by initiating synthesis, suppressing degradation, and inhibiting precocious germination.     

Seed flotation and germination of salt marsh plants: The effects of stratification, salinity, and/or inundation regime

We examined the effects of cold stratification and salinity on seed flotation of eight salt marsh species. Four of the eight species were tested for germination success under different stratification, salinity, and flooding conditions. Species were separated into two groups, four species received wet stratification and four dry stratification and fresh seeds of all species were tested for flotation and germination. Fresh seeds of seven out of eight species had flotation times independent of salinity, six of which had average flotation times of at least 50 d. Seeds of Spartina alterniflora and Spartina patens had the shortest flotation times, averaging 24 and 26 d, respectively. Following wet stratification, the flotation time of S. alterniflora seeds in higher salinity water (15 and 36 ppt) was reduced by over 75% and germination declined by more than 90%. Wet stratification reduced the flotation time of Distichlis spicata seeds in fresh water but increased seed germination from 2 to 16% in a fluctuating inundation regime. Fresh seeds of Iva frutescens and S. alternflora were capable of germination and therefore are non-dormant during dispersal. Fresh seeds of I. frutescens had similar germination to dry stratified seeds ranging 25-30%. Salinity reduced seed germination for all species except for S. alterniflora. A fluctuating inundation regime was important for seed germination of the low marsh species and for germination following cold stratification. The conditions that resulted in seeds sinking faster were similar to the conditions that resulted in higher germination for two of four species.     

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.     

Nitrite-induced Germination of Putrefactive Anaerobe 3679h Spores

Sodium nitrite alone has been shown to stimulate germination of PA 3679h spores. The process was accelerated by using increased concentrations of sodium nitrite, a low pH, and a high temperature of incubation. At low concentrations of nitrite (0.01 to 0.2%), the delay of 36 to 48 hr occurred before germination commenced at 37 C. However, with 3.45% nitrite at 45 C and pH 6.0, most of the spores germinated within 1 hr. At pH 7.0, the germination rate decreased markedly, and at pH 8.0 it was nil. The greatest acceleration in germination rate occurred near 60 C. Hydroxylamine was completely inhibitory to nitrite-induced germination. Sodium nitrite, in turn, inhibited germination by l-alanine, the degree of inhibition being influenced by nitrite concentration and pH.     

Effects of cyclic nucleotides upon the germination ofMucor racemosus sporangiospores

This study describes the effects of exogenous cyclic nucleotides upon the germination of sporangiospores ofMucor racemosus in a defined medium containing a variety of carbon and nitrogen sources. Adenosine 3′5′-cyclic monophosphate (cAMP) enhanced germination, with maximal stimulation at concentrations greater than 5 mm.N⁶-Monobutyryl cAMP, 8-bromo-cAMP, and adenosine also stimulated germination but 5′-adenosine monophosphate (AMP) did not.N⁶,O^2′-Dibutyryl cAMP blocked spore germination in cellobiose, maltose, galactose, 2-ketoglutarate, leucine, and lysine, a result that was also obtained when sporangiospores were incubated in a glucose medium without an exogenous nitrogen source. In glucose, dibutyryl cAMP retarded but did not arrest germination. Dibutyryl cAMP effected rapid and synchronous germination in alanine, aspartate, glutamate, and glutamine media. We conclude that cAMP nonspecifically stimulated the rate of sporangiospore germination while dibutyryl cAMP has a specific stimulatory effect on the initiation of germination ofMucor sporangiospores.