Vegetative survival, akinete formation and germination in three blue-green algae and one green alga in relation to light intensity, temperature, heat shock and UV exposure

The mere vegetative survival was not sufficient but suitable growth conditions were required for akinete formation to occur in the blue-green algaeAnabœna iyengarii, Westiellopsis prolifica, Nostochopsis lobatus and in the green algaPithophora oedogonia. In all algae, akinetes were neither formed nor germinated in darkness, and while dim light of 300 lx was sufficient for most of akinetes to germinate and also to maintain vegetative survival, it was not adequate for optinum akinete formation. Although akinetes of all algae could germinate at 35°C, both the vegetative survival and akinete formation were markedly suppressed at this temperature. Heat or UV shock of any level, whether ineffective or effecting vegetative survival, did not promote akinete formation or germination in any alga tested. Akinetes of all algae under study were relatively tolerant to heat and also to some extent to UV. Both wet and dried akinetes of all algae were equally UV tolerant. In all algae, the viability of both wet and dried akinetes decreased more or less equally with storage time, but the decrease was more drastic when storage temperature was progressively lowered from 20 to 0°C. Hence the akinetes can tolerate dryness but not frost.     

Energetic and metabolic requirements for the germination of akinetes of the cyanobacteriumNostoc PCC 7524

Although akinetes ofNostoc PCC 7524 lost little of their main photosynthetic pigments, phycocyanin and chlorophyll, with increasing age after the onset of sporulation, they lost at least 90% of their photosynthetic and respiratory capacities. Germination needed the supply of light throughout the process, though previous dark metabolism accelerated the following light process. In standard conditions, both respiratory and photosynthetic capacities increased markedly during the first 9–10 h, a time sufficient for the first doublets to appear, but when pigment contents had not yet changed. However, while respiratory capacity could be reacquired without de nove metabolism, resumption of photosynthetic capacity needed RNA and protein synthesis. The energetic requirement for germination was not efficiently fulfilled by cyclic photosynthesis on PSI alone or respiration alone. In the presence of both PSI and respiratory activities only 21% of the akinetes germinated, their endogenous carbon reserves thus being inadequate to support the process to completion. The addition of sucrose to such cultures permitted all of the akinetes to germinate, but at a very slow rate. Rapid and complete germination was only observed when both photosystem operated.Abbreviations Chl Chlorophyll - Phy phycocyanin - DCMU 3-(3,4-chlorophenyl)-1,1-dimethylurea - DPC diphenylcarbazide - DPIP 2,6-dichlorophenylindophenol     

Influence of certain environmental factors on spore germination and spore differentiation inPithophora oedogonia

Of the light- and dark-stored spores ofPithophora oedogonia for over a period of one and half years, the latter exhibited complete failure of germination under controlled culture conditions. pH 8, blue light and 1 klx intensity of white light proved to be ideal conditions as they resulted in optimum germination of akinetes. pH 5, green light and 0.25 klx light intensity delayed the initiation of sporulation and also reduced percentage sporulation. All filaments died before sporulation in pH 4.     

ENVIRONMENTAL CONTROL OF PITHOPHORA OEDOGONIA (CHLOROPHYCEAE) AKINETE GERMINATION1

Abundance of Pithophora oedogonia akinetes displayed seasonal changes, being greatest in winter and lowest in summer. Akinete abundance showed significant (P < 0.001) negative correlations with photoperiod(r = -0.53) and water temperature (r= -0.75) during the period February 1978 through June 1979. Experiments in which akinete germination was studied in response to manipulations of nutrients (NO₃-N and PO₄-P), photoperiod and temperature indicated that temperature was the primary factor regulating the timing of the vernal flush of akinete germination observed in Surrey Lake.     

Vegetative survival,akinete and zoosporangium formation and germination in some selected algae as affected by nutrients,pH, metals,and pesticides

Lack of nitrogen, phosphorus, or all nutrients, extremes of pH (<4, >11), presence of ‘heavy’ metals (Co, Cu, Zn, Hg, Pb; 0.5–10 ppm) or pesticides (carbofuran, 2,4-D, dithane, phorate, or bavistin; 1–50 ppm) decreased to various extent or completely inhibited the survival of vegetative cells in all studied algae. The formation of akinetes, thier viability and germination inAnabaena iyengarii, Westiellopsis prolifica, Nostochopsis lobatus andPithophora oedogonia and the formation of zoosporangia, their viability, and the germination of zoospores inCladophora glomerata andRhizoclonium hieroglyphicum was affected. The formation of viable akinetes or zoosporangia was shown to be directly linked with vegetative cell survival and growth; it could not be induced by any chemical stress imposed.     

PHOTOSYNTHETIC CHARACTERIZATION OF DEVELOPING AND MATURE AKINETES OF APHANIZOMENON OVALISPORUM (CYANOPROKARYOTA)1

Akinetes, differentiated resting cells produced by many species of filamentous, heterocystous cyanobacteria, enable the organism to survive adverse conditions, such as cold winters and dry seasons, and to maintain germination capabilities until the onset of suitable conditions for vegetative growth. Mature akinetes maintain a limited level of metabolic activities, including photosynthesis. In the present study, we have characterized changes in the photosynthetic apparatus of vegetative cells and akinetes of the cyanobacterium Aphanizomenon ovalisporum Forti (Nostocales) during their development and maturation. Photosynthetic variable fluorescence was measured by microscope‐PAM (pulse‐amplitude‐modulated) fluorometry, and the fundamental composition of the photosynthetic apparatus was evaluated by fluorescence and immunological techniques. Vegetative cells and akinetes from samples of Aphanizomenon trichomes from akinete‐induced cultures at various ages demonstrated a gradual reduction, with age, in the maximal photosynthetic quantum yield in both cell types. However, the maximal quantum yield of akinetes declined slightly faster than that of their adjacent vegetative cells. Mature akinetes isolated from 6‐ to 8‐week‐old akinete‐induced cultures maintained only residual photosynthetic activity, as indicated by very low values of maximal photosynthetic quantum yields. Based on 77 K fluorescence emission data and immunodetection of PSI and PSII polypeptides, we concluded that the ratio of PSI to PSII reaction centers in mature akinetes is slightly higher than the ratio estimated for exponentially grown vegetative cells. Furthermore, the cellular abundance of these protein complexes substantially increased in akinetes relative to exponentially grown vegetative cells, presumably due to considerable increase in the biovolume of akinetes.     

Environmental influences on akinete germination and development in Nodularia spumigena (Cyanobacteriaceae), isolated from the Gippsland Lakes, Victoria, Australia

This study was carried out to investigate the genesis of N. spumigena blooms by specifically studying the effects of environmental variables (salinity, nitrogen, phosphorus and light) on the germination of N. spumigena akinetes. Optimal conditions for maximum germination and germling growth were determined by exposing akinetes to a range of salinities and nutrient (nitrogen and phosphorus) concentrations under two different irradiances. At pre-determined time periods, treatments were sampled and the percent germination and length of germlings assessed. The results indicated that akinete germination and germling growth were optimal at salinities from 5 to 25 and significantly reduced outside this range. A positive correlation in germination was observed with increasing nutrient (phosphorus and nitrate) concentration. Similarly, germling growth increased with increasing concentrations of both nutrients. Irradiance significantly influenced both germination and growth during salinity experiments, whereas in nutrient addition experiments, irradiance had no effect on germination; however, growth was significantly influenced during phosphorus addition experiments. Consequently, salinity and light appeared to be most critical in the germination process for N. spumigena akinetes, with phosphorus most important for germling growth. The study showed that N. spumigena may be able to germinate under environmental conditions outside its optimal range, but the growth of the germling is significantly reduced, which in turn suggests that its ability to form a bloom outside its optimal environmental conditions would also be greatly reduced.     

Environmental influences on akinete germination of Anabaena circinalis and implications for management of cyanobacterial blooms

Certain cyanobacteria, including the noxious bloom-forming species Anabaena circinalis Rabenhorst, produce thick-walled reproductive structures (akinetes) which may serve as a resting stage and ensure survival during adverse growth conditions. The effect of certain environmental variables (temperature, salinity and desiccation) on akinete germination of A. circinalis was investigated under laboratory conditions, to determine the conditions under which germination was inhibited. The overall aims were to provide a broader understanding of the life history and ecology of this species and to assess suppression of akinete germination as a potential management strategy for control of cyanobacterial blooms in the lower Murray River, Australia. The results indicated a marked threshold of temperature and salinity tolerance for germination of A. circinalis, but the latter was not within a range that could be successfully manipulated in a natural ecosystem. However, it was found that desiccation of akinetes for moderately short periods can significantly impair their capacity to germinate. It is, therefore, speculated that allowing periodic drying of shallow wetlands adjacent to the Murray River and in other areas may reduce the size of the inoculum for population growth by reducing viability of akinetes in surface sediments.     

Soil seed bank,fire season,and temporal patterns of germination in a seeder-dominated Mediterranean shrubland

Soil seed banks play a major role in the post-fire regeneration of Mediterranean shrublands. They vary throughout the year in species composition, abundance, and readiness to germinate. After fire, germination occurs mainly during the following fall to spring. Time of germination can determine recruitment success. It is unclear what factors control post-fire germination and its timing. We tested the effects of season and fire on the readily germinable soil seed bank of a seeder-dominated shrubland. Plots were burned early and late in the summer season (ES, LS). Soil samples were collected before and after fire, and germinated in a chamber simulating successively autumn, winter, and spring conditions. Samples were kept moistened at all times. Fire intensity was similar between ES and LS. Several species of Cistus and herbs, mostly annuals, were dominant. Most germination occurred during the simulated-autumn period, with little subsequent germination during the following two periods. Germination speed (T ₅₀) during simulated-autumn was similar for shrubs and herbs, and independent of season or fire. Germination was lower for two shrubs (Rosmarinus officinalis, Cistus salvifolius) and higher for herbaceous dicots in LS than in ES soils. Fire reduced monocots and enhanced Cistus. Germination period significantly interacted with fire and season in some groups or species, altering the simulated-autumn germination peak. We demonstrate that the seed bank can germinate swiftly under simulated-autumn conditions. Hence, water availability is the main controlling factor of germination. Fire season differentially affected some species or groups, and could affect the post-fire regeneration.     

AUTOINHIBITION OF SPORE GERMINATION IN NOSTOC SPONGIAEFORME (CYANOPHYCEAE)1

Medium in which cultures of Nostoc spongiaeforme Ag. have sporulated contains one or more substances which inhbit the germination of spores (akinetes) of this organism. Germination of spores occurs rapidly in the absence of these substances, but is virtually completely suppressed in their presence.     

In vitro pollen germination and pollen tube growth differences among Quercus robur L. clones in response to meteorological conditions

The impact of meteorological conditions on in vitro pollen germination and pollen tube growth during the initial phases of the development of male flowers in the Pedunculate Oak, Quercus robur, is studied. Phenological observations of male flowers and pollen sampling were performed on the field trial established with grafted Pedunculate Oak clones. During the investigation, weather conditions (absolute minimum and maximum daily air temperature, minimum absolute relative humidity of air and amount of precipitation) were recorded by an automatic meteorological station installed at the field trial. Influence of meteorological conditions on pollen germination and pollen tube growth was studied in the following stages of male flower: (I) during the last ten days of flower bud dormancy, (II) during swelling of the buds, (III) during bud burst and beginning of male catkins elongation, (IV) during the final stage of male flower catkins elongation. High temperatures and low relative air humidity during the bud burst and beginning of the male catkins elongation reduced pollen germination and pollen tube growth. Weather conditions did not significantly affect pollen germination and pollen tube growth during the swelling of flower buds, or in the final stage of male catkins elongation.     

Influence of Environmental Stress on the Germination of Anabaena vaginicola Akinetes

Germination of akinetes of Anabeana vaginicola v. fertilissimaPrasad in response to environmental stress was studied. Additionof nitrate to the medium induced early and maximum germination(96 per cent), whereas less than half of the akinetes germinatedwhen either nitrate or phosphate was omitted from the medium.The pH range over which germination occurred was 7.0–9.0.The desiccated akinetes after rehydration germinated after acertain lag period, depending upon the dehydration state. Thetemperature optimum for germination and vegetative growth wasthe same (25 °C) and germination did not occur at 5 °Cor above 35 °C. The limit of heat shock tolerated was 55°C for 4 min. In addition to white light, only the red partof the visible spectrum induced germination. Ultraviolet radiationreduced germination rate presumably by inducing thymine dimersin DNA. The photoreactivating system (s) in akinetes is certainlynon-photosynthetic. LD₅₀ photon flux densities were 300 Jm^–2for akinetes and 240 Jm^–2 for vegetative cells. Anabaena vaginicola, blue-green alga, akinete, germination, environmental stress     

The Germination of Vavraia culicis Spores1

Buffered solutions of KCl and NaCl were tested for their stimulatory effect on the germination of variously-aged spores of Vavraia culicis. Germination was optimal in 0.2 M KCl, pH 6.5 for one isolate, and, for another isolate, peaks of germination occurred at pH 7.0 and 9.5. Spores incubated for several hours in suboptimal solutions became unable to germinate under optimal conditions. After being returned to water, they regained their ability to germinate. Calcium chloride, magnesium chloride, and ammonium chloride inhibited germination. After ingestion by mosquito larvae, spores germinated near the posterior end of the midgut.     

DORMANCY AND GERMINATION IN SEEDS OF COMMON RAGWEED WITH REFERENCE TO BEAL'S BURIED SEED EXPERIMENT

Common ragweed (Ambrosia artemisiifolia L.) was one of 19 herbaceous weedy species used by Beal in his buried viable seed experiment started in 1879. No seeds germinated during the first 35 years of the experiment when germination tests were performed in late spring, summer or early autumn. Germination did occur in seeds buried for 40 years when seeds were exhumed and tested for germination in early spring. Data obtained in more recent research provide the probable explanation for these results. Seeds of common ragweed that do not germinate in spring enter secondary dormancy by mid to late spring and will not germinate until dormancy is broken the following late autumn and winter. Thus, during the first 35 years of the experiment seeds were dormant when tested for germination, whereas seeds buried for 40 years were nondormant. Seeds buried 50 years or longer did not germinate when tested in spring, probably because they had lost viability and/or seeds germinated during burial and seedlings died.     

Effects of UV-light andγ-rays on the survival,akinete formation and akinete germination inStigeoclonium pascheri

The damage produced by UV light to any of the three different stages of the life cycle of the parent generation of the green algaS. pascheri, i.e. akinetes, germinating akinetes and vegetative cells remained up to the stage of germination of akinetes of the first generation and no deleterious effect was reported thereafter. Lower doses of γ-rays (25–75 Gy) increased the percentage germination of akinetes and germinating akinetes of parent generation. The percentage germination of akinetes, germinating akinetes, survival of colonies originated from vegetative cells and sporulation of cells of the parent generation decreased with increasing doses from 100 to 300 Gy. The γ-induced effect to any of the three different stages was not transferred to the subsequent stage of algal generation.     

Germination of desiccated aged akinetes of alkaliphilic cyanobacteria

Morphological and biochemical changes associated with synchronous germination of mature, aged and desiccated akinetes of two alkaliphilic cyanobacteria, Cyanospira rippkae and Cyanospira capsulata, are described. Akinetes of both strains proved to be highly resistant to desiccation, being able to germinate, in the presence of either N₂ or nitrate as nitrogen source, with a germination frequency of more than 90% after seven years of storage in a dried state. The first cell division occurred after 8–10 h of incubation, thereafter the germlings of the two strains followed a different pattern of cell differentiation. Heterocysts were first noted, in a terminal position, at 16–18 h in three-celled germlings of C. capsulata and at 21–24 h in C. rippkae, when germlings were at least seven cells in length. Akinetes of both species possessed, on a per cell basis, almost identical amounts of all photosynthetic pigments but, under nitrogen fixing conditions, photosynthetic activity (oxygen evolution) was detected only after new proteins had been synthesized, before a functional heterocyst was developed and while total nitrogen remained constant. With energy provided by aerobic respiration, a wide range of intracellular amino acids characteristic of proteins was utilised to sustain the new protein synthesis. The end of this biosynthetic activity coincided with the timing of the first cell division. From this stage on, no changes in protein concentration occurred until mature heterocysts were developed. In the presence of nitrate, no significant changes in the major germination events were observed.     

Perianth colour dimorphism is related to germination properties and salinity tolerance in Salsola vermiculata in the Arabian deserts

We investigated the dimorphic perianth colour of Salsola vermiculata and its association with seed germination percentage, interactions with temperature, light, salinity and recovery from prior salinity exposure. Seeds with and without pink and yellow perianth were incubated at three thermal regimes, two photoperiods, and five salinity levels. Germination recovery after salinity exposure was observed on seeds that failed to germinate during the salinity study. The germination percentage and rate were signi?cantly related to the perianth colour, the presence of perianth wings, thermal regimes and photoperiod. The presence of a perianth wing significantly reduced germination percentage and germination rate in both the pink and the yellow morph, but the yellow morph exhibited a higher germination percentage. Perianth wing removal increased germination in saline conditions. With the perianth removed, germination recovery was higher for the pink morph than for the yellow one. We suggest that by providing two different strategies for balancing germination with dormancy during favourable conditions, the presence of two morphs makes S. vermiculata more successful in highly unpredictable desert environments.     

Germination behaviour in populations ofCapsella bursa-pastoris (Cruciferae)

Germination behaviour of variousCapsella bursa-pastoris populations collected from Scandinavia, Middle Europe and the Alps, was tested in unheated, non-illuminated greenhouses (46 populations) and in growth chambers using 5–7 alternating temperature regimes (16 populations). For all populations, the influence of temperature on germination rate is straightforward: the higher the temperature, the greater the germination. Germination capacity, however, may depend on the geographical region. There is also a strong seed age effect on both, rate and capacity of germination. Once dormancy was broken, seeds from all populations were able to germinate over the entire range of temperatures. Some populations revealed a more or less pronounced temperature optimum for germination capacity, others germinated equally well over the entire temperature range. This indicates genetic heterogeneity between populations. However, no correlation between germinability and any environmental pattern was detected. The data indicate thatCapsella bursa-pastoris has adopted a germination strategy which includes a broad temperature tolerance. Germination of wildCapsella plants seems to be regulated by the factors contributing to the inception and breaking of dormancy which depend on pre- and postharvest conditions. Adaptation in germination behaviour inCapsella bursa-pastoris is different from that in other life history traits (flowering behaviour, growth form parameters).