Seed viability of five wild Saudi Arabian species by germination and X-ray tests

Our objective was to evaluate the usefulness of the germination vs. the X-ray test in determining the initial viability of seeds of five wild species (Moringa peregrina, Abrus precatorius, Arthrocnemum macrostachyum, Acacia ehrenbergiana and Acacia tortilis) from Saudi Arabia. Usually several days were required to determine the viability of all five species via germination tests. However, X-ray test will give immediate results on filled/viable seeds. Seeds of all species, except Acacia ehrenbergiana and Acacia tortilis showed high viability in both germination (96–72% at 25/15 °C, 94–70% at 35/25 °C) and X-ray (100–80%) test. Furthermore, there was a general agreement between the germination (19%, 14% at 25/15 °C and 17% and 12% at 35/25 °C) and X-ray (8%, 4%) tests in which seed viability of Acacia ehrenbergiana and Acacia tortilis was very low due to insect damaged embryo as shown in X-ray analysis. Seeds of Abruspreca torius have physical dormancy, which was broken by scarification in concentrated sulfuric acid (10 min), and they exhibited high viability in both the germination (83% at 25/15 °C and 81% at 35/25 °C) and X-ray (96%) tests. Most of the nongerminated seeds of the five species except those of Acacia ehrenbergiana and Acacia tortilis, were alive as judged by the tetrazolium test (TZ). Thus, for the five species examined, the X-ray test was proved to be a good and rapid predictor of seed viability.     

Germination response and viability of an endangered tropical conifer Widdringtonia whytei seeds to temperature and light

The tropical conifer Widdringtonia whytei Rendle is an endangered species endemic to Mulanje Mountain in Malawi. A study was conducted for the first time under controlled conditions in order to assess the effects of temperature and light on germination and viability of W. whytei seeds. Seeds incubated at a constant temperature of 20 °C attained the highest cumulative germination percentage (100%) followed by 87% germination under fluctuating temperatures of 15 °C night/25 °C day. No seed germination occurred at temperatures below 15 °C. Seeds that failed to germinate at temperatures below 15 °C showed the highest (> 90%) viability compared to the seeds incubated at 25 °C (60%). Across temperature regimes, germination was significantly higher under light (44.7%) than dark (35.6%) conditions. It is concluded that temperature is one of the critical factors for germination of W. whytei seed. The ability of W. whytei seeds to germinate both in light and darkness implies that the species would unlikely form a persistent soil seed bank, an attribute which is common in species that survive in habitats frequently disturbed by fires.     

Temperature requirements for dormancy break and seed germination vary greatly among 14 wetland Carex species

We evaluated dormancy loss in seeds of 14 Carex species (C. atherodes, C. brevior, C. comosa, C. cristatella, C. cryptolepis, C. granularis, C. hystericina, C. lacustris, C. pellita, C. scoparia, C. stipata, C. stricta, C. utriculata, C. vulpinoidea) under growing season and stratification conditions and determined the temperature requirements for germination. Seeds were germinated for 1 year at a diel temperature regime (5/1 °C, 14/1 °C, 22/8 °C, or 27/15 °C) or a seasonal regime (seeds moved among the four diel regimes to mimic seasonal temperatures). All species had conditionally dormant seeds at maturity. The optimal temperature for germination of most species was 27/15 °C. The 14 species were grouped by their seed viability, dormancy, and germination with a Seed Regeneration Index (SRI; range 0–1) using the results of this study and a previously published paper on stratification effects on Carex seed dormancy and germination. The eight species that had an SRI value >0.5 (C. brevior, C. comosa, C. cristatella, C. cryptolepis, C. hystericina, C. scoparia, C. stipata, C. vulpinoidea) had high seed viability (>60%) and required little to no stratification to germinate readily over a broad range of temperatures. The six species with an SRI value <0.5 (C. atherodes, C. granularis, C. lacustris, C. pellita, C. stricta, C. utriculata) generally had low seed viability (<50% and often <1%) and required stratification or particular temperatures (35/30 °C or 5/1 °C for C. stricta; 35/30 °C for C. utriculata; 27/15 °C for C. atherodes, C. lacustris, C. pellita; 5/1 °C for C. granularis) for germination ≥50%. These six species will require more attention from restoration practitioners to ensure that there are sufficient viable seeds to meet revegetation goals, that dormancy break is achieved, and that seeds are sown when temperatures are optimal for germination. The different seed germination syndromes that we found for these Carex species likely contribute to variable seed bank formation and emergence patterns, and species coexistence.     

Seed storage and germination in Kumara plicatilis,a tree aloe endemic to mountain fynbos in the Boland,south-western Cape,South Africa

Seed storage under appropriate conditions is a relatively inexpensive means of safeguarding plant genetic material for ex situ conservation. Post-storage germination trials are used to determine the viability of stored seeds, and hence the efficacy of the particular storage treatment. Kumara plicatilis (= Aloe plicatilis) is a tree aloe endemic to mountain fynbos in the Boland, south-western Cape. The viability and germination behaviour of K. plicatilis seeds were assessed for seeds stored for four and nine months at − 80 °C, 4 °C, 25 °C and under ambient conditions in a laboratory. Seeds were germinated under controlled conditions and germination rates and percentages determined. Ungerminated seeds were tested for viability using tetrazolium salt. Seed viability was not significantly reduced during storage. Seeds stored at − 80 °C for four and nine months exhibited the fastest germination rate overall (both 5.9 ± 0.3 weeks, mean ± S.E.), and slowest was for seeds stored under ambient conditions for four and nine months (both 7.8 ± 0.4 weeks). All seed lots showed similar percentage germination after four months of storage (78.0–90.4%). The highest percentage germination overall was for seeds stored at − 80 °C for four months (90.4%) and the lowest was for seeds kept at 4 °C and − 80 °C for nine months (39.2 and 39.6%, respectively). Respective percentage viability for ungerminated seeds in these two treatments was 82% and 87%, respectively, indicating the induction of secondary dormancy. Induced dormancy triggered by protracted cold temperatures may be an adaptation that enables seeds to survive prolonged extreme conditions that are unfavourable for germination. Further research on the long-term storage of aloe seeds would be beneficial for developing long-term seed storage and germination testing protocols for ex situ conservation.     

Germination characteristics of the grass weed Digitaria nuda (Schumach.)

The effect of various pre-treatments and their interaction with temperature on cumulative percentage and the rate of germination were evaluated for Digitaria nuda. Stored and fresh seeds were pre-treated with either 0.02 M KNO₃, soaked in water for 24 h (priming), sterilized with 0.5% NaOCl or heat treated at 60 °C. Seeds were germinated at constant temperatures of 25 and 30 °C and fluctuating temperature regimes of 25/10 and 30/15 °C. The effect of pre-chilling on germination of stored and fresh seed was evaluated at 30/15 °C, and seed emergence in two soil types at different burial depths (0, 0.5, 1, 2, 3, 4, 5 and 6 cm) was also determined. The pre-treatment of stored seed with KNO₃ resulted in the highest germination percentage (100%), whereas the pre-treatment of fresh seed with water for 24 h gave the best germination (99%), at constant temperatures of 25 and 30 °C. Pre-chilling of seed increased germination by more than 30%. Emergence from clay loam soil was greater compared with the emergence from sandy loam soil. Total seedling emergence decreased exponentially with increasing burial depths with only 5% of seed germinating from a burial depth of 6 cm. Results from this study showed that germination requirements are species specific and knowledge of factors influencing germination and emergence of grass weed seed can assist in predicting flushes in emergence allowing producers to implement control practices more effectively.     

Smoke-water effect on the germination of Amazonian tree species

Smoke stimulates seed germination of a range of species from ecosystems that may or may not be fire prone. We evaluated the effects of smoke-water on germination of ten tree species of economic value in the Amazon region. Two materials were burnt to produce smoke-water: germination paper and the wood of Cecropia palmata Willd. Seven dilutions of the solutions were tested. Seeds of nine forest trees were germinated under controlled laboratory conditions (25 °C ± 2 °C) in the laboratory. Bertholletia excelsa Humb. & Bonpl., was tested in the nursery (approximately 25–36 °C) because of its large seeds. Irrespective of the material burned, smoke-water significantly increased seed germination of three species: Cordia goeldiana Hub., Ochroma pyramidale (Cav. ex Lam.) Urb. and Jacaranda copaia (Aubl.) D. Don. and there was a significant inhibitory effect on Swietenia macrophylla King. Germination was accelerated by smoke in J. copaia, B. excelsa and Bellucia grossularioides (L.) Triana. The most pronounced effect was observed in B. excelsa, as the mean germination time of 108 d (control) was reduced to 76 d with smoke-water made from germination paper (dilution of 1:25) and to 61 d with the one from Cecropia wood (dilution of 1:250). For five of the ten species studied, smoke-water either increased or accelerated seed germination, irrespective of the materials used for its production. Seeds with low vigour and prolonged germination time seemed to be more receptive to smoke.     

Effect of yak rumen content treatments on seed germination of 11 alpine meadow species on the Qinghai-Tibetan Plateau

The Qinghai-Tibetan Plateau is located in the ‘Third Pole’ of the world, characterized by a harsh environment. Despite this, the alpine meadow ecosystem have developed over a wide area but serious grassland degradation is threatening the ecological environment on the Plateau. Recruitment of new plants to the population, via germination and establishment, is vital to plant community survival. Previous work on the seeds in this area has mainly focused on community-wide germination strategies, seed germination characteristics and their correlations with seed size and seed mass. However, there have been no studies on the effects of soaking in rumen contents on the plant seed germination characteristics of alpine meadow species. The present study had two main objectives: (i) to determine the effect of fresh rumen content from yaks on seed germination characteristics and seedling growth of species common to the eastern Tibetan Plateau alpine meadow, and (ii) to develop an effective method to enhance seed germination. Seeds of 11 common species were collected together with fresh rumen content from three yaks that grazed there. Seed germination tests were conducted after they had been soaked in rumen content for one of six soaking periods (12, 24, 36, 48, 60 or 72 h). The seeds were incubated under natural light conditions of 8 h light at 25 °C and 16 h darkness at 15 °C, for the germination period of 34 days. The results showed that seed germination and seedling growth were affected by soaking time, seed coat completeness and seed type. After soaking in rumen content, the germination percentages of scarified (peeled or with the seed coat cut through) seeds of some species (true seeds Oxytropis ochrocephala and Medicago ruthenia var inschanica, nutlet Carex enervis, achenes Anemone rivularis and Polygonum sibiricum) and complete seeds of C. enervis, and A. rivularis were improved but the duration of soaking was also important. Seed germination of caryopsis Achnatherum inebrians (a toxic grass) was significantly inhibited by any exposure to rumen fluids. Scarified seeds generally had higher germination percentages than complete ones after treatment, but with the increase in soaking time, germination percentages declined and scarified seeds were more sensitive to the treatment than the complete seeds. After soaking in yak rumen content, the germination indices of scarified M. ruthenia at 12 h treatment, O. ochrocephala and achene Rumex acetosa at 12–24 h treatment, nutlet Kobresia humilis at 24 h treatment, P. sibiricum at 24–48 h treatment, C. enervis at 12–48 h treatment and A. rivularis at 12–60 h treatment were significantly higher than the control (P < 0.05), while the germination indices of complete C. enervis seeds at 12 h and 36 h treatment, and A. rivularis at 12–60 h treatment were significantly higher compared with the control. The germination indices of other species gradually decreased with the increase in soaking time. We concluded that yak rumen digestion could enhance, inhibit or not affect seed germination and seedling growth of the alpine meadow species, which might influence seedling recruitment, interspecific competition, and the plant community structure of the eastern Tibetan Plateau alpine meadow. Overall, yak digestion has a positive effect on alpine meadow seed germination and seed dispersal.     

Temperature requirements for seed germination of Pereskia aculeata and Pereskia grandifolia

Pereskia aculeata and Pereskia grandifolia have been studied widely due to their high nutritional and therapeutic values. However, little is known about the biological requirements of their seeds for the various germination factors. Thus, this experiment aimed to evaluate the thermal effects on the germination of these species at the temperatures of 24 °C, 27 °C, 30 °C, 33 °C and 36 °C. After verification of the existence of differences in the performance of germination, a non-linear regression was carried out, relating the germination to temperature and identifying its point of maximum efficiency. We found that the lowest synchronization indexes of germination were observed close to 30 °C. The best germination response of the P. aculeata and P. grandifolia was observed at 30 °C and 33 °C, respectively, with greater germination strength and fewer days to attain 63.21% of germinations. The results obtained from the germination of P. aculeata and P. grandifolia can be described by the Weindull distribution model with three parameters, as proposed by Carneiro and Guedes (1992).     

Impact of temperature on olive (Olea europaea L.) pollen performance in relation to relative humidity and genotype

Olive varieties ‘Koroneiki’, ‘Kalamata’, ‘Mastoidis’ and ‘Amigdalolia’ were employed in two experiments for 3 years to assess the effect of temperature on olive pollen germination and tube growth in relation to relative humidity and genotype. Pollen samples were subjected to pre-incubation at 10, 20, 30 or 40 °C in combination with decreased air relative humidity – 80, 40, 30 or 20%, respectively – for 24 h to simulate temperature stress that is observed during pollen dispersal; and subsequently in vitro cultured. In the second experiment, pollen was exposed at 15, 20, 25 and 30 °C for 24 h in vitro to evaluate pollen response in conditions of water and nutrients availability and to determine the optimum pollen germination and tube growth temperatures for each cultivar. The highest pre-incubation temperature treatment (40 °C) prevented pollen germination in ‘Koroneiki’ and ‘Mastoidis’, with the less affected varieties (‘Amigdalolia’ and ‘Kalamata’) having average germination percentages of only 7.6 and 2%, respectively. Pre-incubation at 30 °C had a negative impact on pollen germination in ‘Koroneiki’ (?65%), ‘Kalamata’ (?20%) and ‘Amigdalolia’ (?72%) compared to the control (20 °C). Pollen pre-incubation at 40 °C decreased significantly the pollen tube length in ‘Kalamata’ (?50%) and ‘Amigdalolia’ (?52%). In the second experiment, in vitro pollen germination increased after incubation at 25 °C for ‘Koroneiki’ (+6%), ‘Mastoidis’ (+52%), ‘Kalamata’ (+10%) and ‘Amigdalolia’ (+10%) compared to the control (20 °C). At 30 °C germination percentages for ‘Mastoidis’, ‘Kalamata’ and ‘Amigdalolia’ were 8, 6 and 14% higher, respectively, compared to the control (20 °C). Pollen tube length also increased with incubation temperature for all of the studied cultivars. Based on the cumulative stress response index (CSRI) that was calculated for high temperature stress the varieties were classified: ‘Mastoidis’ and ‘Kalamata’ as tolerant and ‘Koroneiki’ and ‘Amigdalolia’ as intermediate at 30 °C while all studied cultivars were sensitive at 40 °C. The observed strong genotype-differentiated response in high and low temperature stress could be exploited by plant breeders towards producing new tolerant olive varieties.     

Effects of different temperatures on the life history of Evania appendigaster L. (Hymenoptera: Evaniidae), a solitary oothecal parasitoid of Periplaneta americana L. (Dictyoptera: Blattidae)

The influence of temperatures on the life parameters of the solitary oothecal parasitoid Evania appendigaster, was investigated in the laboratory. Parasitized oothecae of Periplaneta americana were left to develop under seven constant temperatures: 15, 17, 20, 25, 30, 35, and 40 °C. At the end, we found that: (i) E. appendigaster was able to complete development within the temperature range of 17–34 °C; (ii) mean adult longevity decreased as temperature increased, with the temperature of 40 °C being fatal in a matter of hours; (iii) males lived longer than females between 15 and 30 °C; (iv) adult emergence rate was the highest at 25 °C, and (v) no wasps emerged at 15 or 40 °C. Non-emerged oothecae contained either unhatched eggs or dead larvae. We determined the theoretical lower developmental threshold and thermal constant for the complete development as 12.9 °C and 584.8 day-degrees for males, and 13.1 °C and 588.2 day-degrees for females, respectively. A good balance between faster development, maximum adult longevity and good egg viability was obtained between 25–30 °C, and that would be the best temperature range for rearing E. appendigaster.     

Effects of rabbit gut passage on seed retrieval and germination of three shrub species

It has been known for a few decades that European rabbits consume seeds and fleshy fruits of native woody plants, but relevant factors in the endozoochory processes such as seed predation (chewing and digestion), sexual differentiation, or the temporal pattern of seed recovery have been little evaluated until now. In this study, we examined seed dispersal of three Mediterranean shrub species by wild rabbits through monitoring of seed retrieval and germination after gut passage. Twelve adult wild rabbits (Oryctolagus cuniculus; six males and six females) of similar size and age were fed seeds of three shrub species with fleshy fruits (Crataegus monogyna, Myrtus communis and Pistacia lentiscus). After ingestion of fruits, seeds were retrieved from dung every 12 h for a day and a half. The viability and germination of retrieved seeds were tested along with that of uneaten seeds. Between 5% and 76% of ingested seeds were retrieved from dung, with significant differences between species and sex. Most M. communis seeds were retrieved with 12–24 h after ingestion; almost all C. monogyna seeds were recovered with 0–12 h after ingestion; no seeds of P. lentiscus were recovered. Only in the case of M. communis seed was the recovery rate greater in female than in male. Passage through the rabbit gut significantly increased seed germination in M. communis, and decreased it in C. monogyna. In conclusion, wild rabbits acted in this study as predators of C. monogyna and P. lentiscus seeds and potential dispersers of M. communis seeds.     

Role of calcium in alleviating effect of salinity on germination of Phragmites karka seeds

Phragmites karka (Retz.) Trin, ex. steud, a perennial reed with creeping rhizome from the family Poaceae, is distributed as pure population in brackish water swamps. Populations primarily propagate using ramets but also produce numerous seeds which form part of the seed bank after dispersal and are exposed to extremes of temperature, drought, and salinity stress. Seeds were germinated under a range of salinity (0, 100, 200, 300, 400, 500 mM NaCl) and temperature (10/20 °C, 15/25 °C, 20/30 °C, 25/35 °C, night/day) regimes in 12 h light:12 h dark photoperiod or in complete darkness with 0, 5, 10, 25 mM CaCl₂. Salinity, absence of light and high temperature (25/35 °C) reduced germination while calcium generally reversed this effect, more so at cooler temperature regimes. Calcareous soil around Karachi would help alleviate the salinity effect on the germination of P. karka and facilitate its survival.     

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.     

Biology and thermal requirements of Chrysoperla genanigra (Neuroptera: Chrysopidae) reared on Sitotroga cerealella (Lepidoptera: Gelechiidae) eggs

Chrysoperla genanigra Freitas is a common green lacewing associated with melon pests in the Northeastern Brazil. All life stages of this recently described species were studied under a range of constant temperature conditions (17, 21, 25, 29, 33, 35 and 37 °C), a photoperiod of 12 h:12 h (L:D) and 70 ± 10% relative humidity. Adults of C. genanigra were fed on a diet consisting of a 1:1 (v/v) mixture of brewer’s yeast and honey, while larvae were provided with eggs of Sitotroga cerealella (Olivier) ad libitum. The duration of preimaginal development of the species was inversely proportional to temperature and ranged from approximately 63 days at 17 °C to 15 days at 35 °C. The percentage of adult emergence varied from 6.7% at 17 °C to 76.7% at 25 °C, although no larvae were able to complete development at 37 °C. The lower thermal threshold for total preimaginal development was approximately 10.8 °C and the thermal requirement was 336.7 degree-days. Egg production, along with the longevity of both males and females, were significantly affected by temperature. It is concluded that the best temperature for rearing C. genanigra is 25 °C, with the lowest preimaginal mortality and the highest egg production (992.7 eggs/female).     

The germination characteristics of Alexandrium minutum (Dinophyceae), a toxic dinoflagellate from the Fal estuary (UK)

The germination characteristics of Alexandrium minutum cysts from the Fal estuary were studied at different conditions of temperature (4–24 °C) and salinity (15–35‰) and in the dark and low light intensity (2 μmol^?2 s^?1). Sediment sub-samples were directly cultured and processed at the end of the experiment for counts of non-germinated cysts. A decrease in the number of cysts was interpreted as germination that was calculated by comparison of the number of cysts over time with that of initial counts. The 50% germination time (time at which 50% of the total initial number of cysts had germinated) was calculated for each condition. A. minutum did not germinate in the dark but it germinated under all other conditions studied. Highest germination occurred at salinities of 30 psu and 35 psu and temperatures from 8 °C to 24 °C (germination rate—expressed as the inverse of the 50% germination time: 1.1–1.2). Lowest germination occurred at 15 psu and 4 °C and 24 °C (germination rate: 3.9–3.8). However, little variation in germination rates occurred across the conditions studied. As these conditions represent those likely in the estuary it is probable that A. minutum cysts on the surface of the sediments represent a constant source of cells to the water column and sediment disturbance (revealing buried cysts) could rapidly inoculate the water column with vegetative cells. This data was used to develop a model for Alexandrium germination from coastal sediments.     

Effect of variable dew temperatures on infection of green foxtail by Pyricularia setariae,Drechslera gigantea,and Exserohilum rostratum

A thermogradient apparatus was used to investigate the effect of variable dew temperatures on infection of green foxtail by the indigenous pathogen Pyricularia setariae (Ps) and the exotic pathogens Drechslera gigantea (Dg), and Exserohilum rostratum (Er) from the southern USA that showed bioherbicide potential against several grassy weeds. This device is capable of creating multiple diurnal temperature cycles, mimicking daily temperature fluctuations that occur under field conditions. Seven temperature regimes, i.e., 15/10 °C, 20/5 °C, 20/15 °C, 25/10 °C, 25/20 °C, 30/15 °C, and 30/25 °C (maximum/minimum), were used with temperature cycling from maximum to minimum and then back up to maximum in a 24 h period. Ps and Dg were much more virulent than Er on green foxtail, resulting in higher levels of disease and weed control. Dg was little affected by the dew temperatures in terms of plant infection and was more efficacious than Ps under cooler dew temperatures (15/10 °C and 20/5 °C), causing twice as much disease. This greater amount of disease coincided with higher conidial germination, appressorial formation and infection-hypha frequency by Dg at the lower temperatures. The efficacy of Ps improved as dew temperature increased, accompanied by a higher percentage of germination and more frequent appressorial production. Dg caused severe disease 2 d after inoculation whereas Ps required 4 d to initiate disease symptoms. These observations suggest that Dg is a superior candidate than Ps for green foxtail control on the Canadian prairies.     

Effects of various salts on the germination of three perennial salt marsh species

We studied the germination responses of Arthrocnemum macrostachyum, Juncus acutus and Schoenus nigricans to saline stress caused by different salt types. The germination percentage and mean time to germination data were obtained by incubating seeds for 30 d in 1, 2, 3, 4 or 5% saline solutions of NaCl, MgCl₂, MgSO₄ and Na₂SO₄ at 30/20 °C and with a 12 h photoperiod. A. macrostachyum was the most tolerant species to salinity during the germination (65% in 2% NaCl). S. nigricans showed the lowest germination (none germinated in salt and only 26% in distilled water). J. acutus showed intermediate behaviour between the two above species, its germination being inhibited by high salt concentrations. The sulphates had less inhibitory effect than the equivalent chloride concentrations.     

Effects of temperature,salinity and their interaction on growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a seafood poisoning that is widespread in tropical, subtropical and temperate regions of the world. In the main island of Japan, distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3, have been reported. To discuss the bloom dynamics of these Japanese species/phylotypes of Gambierdiscus, first we tested six culture media to optimize growth conditions and then clarified the effects of temperature and salinity and temperature–salinity interactions on growth. All strains of the species/phylotypes tested showed the highest cell yields when they were cultivated in IMK/2 medium. G. australes, G. scabrosus and Gambierdiscus sp. type 2 grew in the range 17.5–30 °C, whereas Gambierdiscus sp. type 3 grew in 15–25 °C. The semi-optimal temperature ranges (≥80% of the maximal growth rate) of the former three species/phylotypes were 19–28 °C, 24–31 °C and 21–28 °C, respectively, whereas that of the latter phylotype was 22–25 °C. Hence, Gambierdiscus sp. type 3 may be adapted to relatively lower water temperatures of ≤25 °C. In contrast, G. australes, G. scabrosus and Gambierdiscus sp. type 2 presumably possess adaptability to relatively high water temperatures. The optimal temperature for G. scabrosus was 30 °C, whereas the optimal temperature for the others was 25 °C. G. australes and Gambierdiscus sp. type 3 grew in a salinity range of 25–40 whereas G. scabrosus and Gambierdiscus sp. type 2 grew in salinity 20–40. Furthermore, the semi-optimal salinity range of G. australes, G. scabrosus, Gambierdiscus spp. type 2 and type 3 were salinity 27–38, 24–36, 22–36 and 29–37, respectively. Among the species/phylotypes, G. scabrosus and Gambierdiscus sp. type 2 grew even at salinity 20 where the others did not grow, thus possessing adaptability to low salinity waters. Our results clearly demonstrate that the optimal and tolerable temperature–salinity conditions differ among Japanese Gambierdiscus species/phylotypes. Considering these results, temperature–salinity interactions may play an important role in bloom dynamics and the distribution of the Gambierdiscus species/phylotypes in Japanese coastal waters.     

Effects of salt,alkali and salt–alkali mixed stresses on seed germination of the halophyte Salsola ferganica (Chenopodiaceae)

Salsola ferganica L. (Chenopodianceae) is an annual halophytic species. Experiments were carried out in laboratory to determine the effects of temperature, perianths and various types of salinity on seed germination and germination recovery. Seeds were germinated at 6 levels of temperature with perianths, plus perianths and removed perianths in complete darkness for 9 days. The germination responses of the seeds without perianths at 25 °C were determined over a wide range of NaCl, NaHCO₃ or NaCl–NaHCO₃ mixed stress for 13 days. Perianths seriously affected germination as a barrier for seed germination and the optimal temperature was at 25 °C. Highest germination percentage was obtained under control and seed germination was progressively inhibited with the increase of salinity concentration. The negative effect of NaHCO₃ at the same concentration on germination was stronger than that of NaCl and NaCl–NaHCO₃ mixed. When substrate salinity was removed, seeds exposed to a high NaCl concentration (400–800 mM), NaHCO₃ (50–200 mM) and NaCl–NaHCO₃ mixed (100–400 mM) germinated well. Final germination of Salsola ferganica seeds was significantly affected by types of salt at the low salinity (?200 mM) and with increased salinity it was influenced mainly by salinity concentration for various proportion of salt–alkali mixed stress.     

Overwintering temperatures affect freezing temperatures of turions of aquatic plants

The effect of different overwintering temperatures (2.5 ± 1 °C in a refrigerator or outdoor natural overwintering on wet topsoil with weak frosts) on the freezing temperature and survival rate of turions of 10 aquatic plant species with different ecological traits (free-floating habit or bottom rooting) was studied using mini thermocouples. Dormant, non-hardened turions of 9 species exhibited freezing within a narrow temperature range of ?7.0 to ?10.2 °C, while Hydrocharis morsus-ranae froze at ?3.6 °C. The survival rate of the turions after the measurements was, however, very low (0–38%). In several species, the freezing temperature of turions at the beginning of germination was not significantly different (at p < 0.05) from the dormant ones. The mean freezing temperature of outdoor hardened turions of 6 species was within a very narrow range of ?2.8 to ?3.3 °C and was thus significantly higher by 4–7 °C (p < 0.0002) than that for the non-hardened turions. It is assumed that the freezing temperatures indicate freezing of the extracellular water. The hardened turions of all 7 species were able to survive mild winter frosts under the topsoil conditions at a rate of 76–100%. These characteristics suggest that the turions of aquatic species can be hardened by weak frosts and that their frost hardiness is based on the shift from frost avoidance in non-hardened turions to frost tolerance.