Temperature acclimation and heat tolerance of photosynthesis in Norwegian Saccharina latissima (Laminariales,Phaeophyceae)

Kelps, seaweeds and seagrasses provide important ecosystem services in coastal areas, and loss of these macrophytes is a global concern. Recent surveys have documented severe declines in populations of the dominant kelp species, Saccharina latissima, along the south coast of Norway. S. latissima is a cold‐temperate species, and increasing seawater temperature has been suggested as one of the major causes of the decline. Several studies have shown that S. latissima can acclimate to a wide range of temperatures. However, local adaptations may render the extrapolation of existing results inappropriate. We investigated the potential for thermal acclimation and heat tolerance in S. latissima collected from three locations along the south coast of Norway. Plants were kept in laboratory cultures at three different growth temperatures (10, 15, and 20°C) for 4–6 weeks, after which their photosynthetic performance, fluorescence parameters, and pigment concentrations were measured. S. latissima obtained almost identical photosynthetic characteristics when grown at 10 and 15°C, indicating thermal acclimation at these temperatures. In contrast, plants grown at 20°C suffered substantial tissue deterioration, and showed reduced net photosynthetic capacity caused by a combination of elevated respiration and reduced gross photosynthesis due to lowered pigment concentrations, altered pigment composition, and reduced functionality of Photo‐system II. Our results support the hypothesis that extraordinarily high temperatures, as observed in 1997, 2002, and 2006, may have initiated the declines in S. latissima populations along the south coast of Norway. However, observations of high mortality in years with low summer temperatures suggest that reduced population resilience or other factors may have contributed to the losses.     

Temperature and salinity responses of drifting specimens of Kappaphycus alvarezii (Gigartinales,Rhodophyta) farmed on the Brazilian tropical coast

Bioinvasion events causing serious environmental damage have been a concern with the mariculture of Kappaphycus alvarezii (Doty) Doty ex P.C. Silva, suggesting the importance of studying the biological aspects of drifting specimens of K. alvarezii for monitoring programs. The present study aims to evaluate the tolerance and growth of drifting color variants of K. alvarezii under different temperatures and salinities to determine their physiological capacity for growing outside cultivation rafts. Drifting color variants were collected in Paraíba State, Brazil, in November 2011(dry month) and August 2012 (rainy month), and cultivated in the laboratory under different temperatures (20, 24, 28, and 32 °C) and salinities (15, 25, 35, 45, and 55 psu). Growth rates as well as pigment and protein contents were determined. Results showed that drifting specimens collected in the dry month showed higher tolerance to variation in temperature (20 to 28 °C) and salinity (25 to 35 psu) than drifting specimens collected in the rainy month. Higher growth rates occurred in samples cultured at 20 and 24 °C (2.8–3 % day^?1) and 25 to 35 psu (3.4–3.5 % day^?1), suggesting temperature and salinity optima. Higher phycobiliprotein levels were observed in the red and brown variants under hypersaline conditions (45 and 55 psu). Higher chlorophyll a contents were associated with samples cultivated at 20–24 °C and 24–35 psu. Based on the results of the present study, drifting specimens collected in dry month are more tolerant to temperature and salinity variations, suggesting that the drifting K. alvarezii should be monitored especially during this period to prevent its establishment outside the cultivation rafts and dispersion along the northeastern coast of Brazil.     

Comparing salinity tolerance in early stages of the sporophytes of a non-indigenous kelp (Undaria pinnatifida) and a native kelp (Saccharina latissima)

The short-term effects of low salinities on the survival of germlings of an introduced kelp Undaria pinnatifida and a native kelp Saccharina latissima were assessed under laboratory conditions. This experiment was designed to compare the differential stress tolerance to salinity of the early life history stages of sporophytes of these two kelps that co-occur on European Atlantic coasts. Germlings (young sporophytes) of both species were exposed for 4 days to salinities ranging from 31 (control) to 26, 21, 16, 11, and 6 psu. Afterwards, they were post-cultured in control seawater (31 psu) for another 4 days to corroborate the viability of injured germlings. Results showed that germlings of the introduced kelp were less resistant to low salinity, surviving to as low as 16 psu; whereas the germlings of the native kelp survived in salinities as low as 11 psu. Despite the observed differences, both species are relatively tolerant to low salinity. Our observations also indicated that, at least in a short term, gametophytes of both species were able to survive in salinities as low as 6 psu. The significance of low-salinity tolerance to the distribution of these kelps and for their offshore cultivation is discussed.     

Temperature requirements for the growth of young sporophytes of Undaria pinnatifida and Undaria undarioides (Laminariales, Phaeophyceae)

The relative growth rate of young sporophytes of Undaria pinnatifida (Harvey) Suringar and Undaria undarioides (Yendo) Okamura was examined in order to understand the difference in distribution of these two species around the coast of Japan. The optimal temperature for growth of both species was similar at 20°C and the upper critical temperature for growth was also similar, at 27°C for U. pinnatifida and 26°C for U. undarioides. Therefore, the optimal and upper critical temperatures for growth of the young sporophytes are not the main factors determining the distribution of each species. Next, the lower critical temperatures for growth were examined. For the young sporophytes of U. pinnatifida, the lower limit was less than 5°C while for those of U. undarioides it was 15°C. Thus, the difference in the lower critical temperature for growth between the two species was approximately 10°C. During the period of young sporophyte growth in the field, the temperature at the mouth of Ise Bay, Japan, where U. pinnatifida occurs, ranges from 12.7°C in December to 13.1°C in April, with a minimum of 7.9°C in February. Our experiments indicate that young sporophytes are able to grow throughout this period. The temperature off Hamajima, Japan, where U. undarioides occurs, ranges from 19.1°C to 14.8°C during the same time period. Again, young sporophytes are able to growth throughout this period, although minimum winter temperatures are only just high enough for growth. These natural temperature ranges during the growth season of the sporophytes agree well with the experimentally determined temperature requirements for growth of each species. Therefore, the difference between the two species in the critical temperature required for growth of the young sporophytes, especially in the low temperature range, is one of the major factors determining the distribution pattern of each species.     

RESPONSE TO SALINITY AND HEAT STRESS IN TWO HALOTOLERANT CHLOROPHYTE ALGAE1

Two new isolates of halotolerant chlorophyte algae from the Salt Plains National Wildlife Refuge in Oklahoma, USA, tentatively identified as Dunaliella sp. Teodoresco and Nannochloris sp. Naumann, were characterized with respect to interaction between growth salinity and short‐term heat tolerance. Cells were cultured at 23–25° C over a wide range of salinity. In both species, salinity alone had little effect on maximum photochemical yield (measured by pulse modulated fluorescence) and integrity of the light harvesting system (77 K fluorescence emission spectra). In contrast, Nannochloris exhibited decreasing growth rate (μ), light‐saturated photosynthetic capacity (P^cell_max), respiration (R_d), light‐harvesting efficiency (α^cell), and chl content with increasing salinity. Cultures were heated for 2 h near their upper temperature limits (41.5° C for Dunaliella and 45° C for Nannochloris grown at 50 psu). Dunaliella was progressively more heat‐tolerant with increasing salinity. Photochemical yield of cells at 100 and 50 psu was inhibited by about 15% and 40%, respectively, and largely recovered within 30 min after return to 23° C. Thermal inhibition of photochemical yield in Nannochloris was about 45% at both 50 and 100 psu, but recovery was slower at 100 psu. At 20 psu, both species were almost 90% inhibited by high temperature and required more than a day to recover. In both species, 2 h of heating increased the PSI:PSII fluorescence emission ratio (714:690 nm) at all salinities. This ratio largely recovered within 24 h in Dunaliella at 50 and 100 psu and partially recovered in Nannochloris at 100 psu, but cells of both species heated at 20 psu were chlorotic the next day.     

Tolerance to salinity and thermal stress by larvae and adults of the serpulid annelid Ficopomatus enigmaticus

The serpulid annelid Ficopomatus enigmaticus is a widely distributed invader of shallow‐water, brackish habitats in subtropical and temperate regions, where it has numerous damaging ecological and economic effects. Its distributional pattern suggests that temperature and salinity play important roles in limiting its distribution, but because other factors often covary with these, drawing strong conclusions from these patterns is difficult. In an effort to more clearly identify the effects of these factors, we examined tolerance to acute thermal (16–28°C) and salinity (0–35 psu) stress by larvae (5‐day exposure, unfed) and adults (14‐day exposure, unfed) of F. enigmaticus in the laboratory experiments. Larvae showed higher mortality at the highest temperature tested 28°C; adult survival was unaffected by temperature. Neither larvae nor adults survived exposure to pure freshwater (0 psu), but survived well at salinities ranging 3.5–35 psu. In addition, high salinity did not slow tube growth in adults. These results suggest that salinity stress, in particular, does not directly limit the distribution of F. enigmaticus to low‐salinity habitats. Experimental work on the distribution of F. enigmaticus is uncommon in the literature, but is likely needed to identify the abiotic or biotic factors that limit the distribution of this frequently invasive species.     

Photosynthetic and respiratory responses of Gracilaria parvispora from the southeastern Gulf of California

Photosynthetic and respiratory responses (P–E curves) of Gracilaria parvispora from the southeast Gulf of California were studied at four temperatures (20, 25, 30, 35 °C) and salinity (25, 30, 35, 40 psu) combinations. The alga showed acclimation in its photosynthetic and respiratory responses to tropical temperature as well as to oceanic salinity. A positive effect of temperature on photosynthetic rate (P _max) was observed for all salinities. Photosynthetic rates for treatments at 20 and 25 °C were lower (<9.2 mg O₂?g dry weight (dw)^?1?h^?1) than for treatments at 30 and 35 °C (>12 mg O₂ g dw^?1?h^?1). G. parvispora showed limited tolerance to low salinities (25 psu) and low temperatures (20 °C) and the interaction between temperature and salinity was significant (analysis of variance, P?<?0.05). Responses to salinity indicated adaptation to oceanic salinity. Photosynthetic responses were lower at 25 psu than at higher salinities. The lowest P _max values (6.2–8.2 mg O₂?g dw^?1?h^?1) were observed at the lowest salinity (25 psu) regardless of temperature. Compensation and saturation irradiances (26–170 and 57–149 μmol photons m^?2?s^?1, respectively) indicate adaptation to lower irradiances in shallow (1–2 m depth) habitats, where turbidity can be high, and the capacity of shade adaptation has been developed. Results suggest distribution of this species is mainly related to salinity or temperature. The potential mariculture efforts of G. parvispora would be limited by low temperatures in winter, and indicate that this species will probably not be able to spread further due to low temperatures (<15 °C) in the upper part of the Gulf of California.     

Effects of temperature on growth of Vallisneria americana in a sub-tropical estuarine environment

The submersed aquatic vegetation (SAV) species Vallisneria americana Michx. (tape grass) is a valuable resource in the Caloosahatchee estuary and in many other aquatic systems. Given the variable nature of freshwater inflows and environmental conditions in the Caloosahatchee, it is necessary to understand how tape grass will respond to high and low salinity conditions at different light and temperature levels. Specifically, quantitative information is needed as input to modeling tools that can be applied to predict growth and survival of tape grass under a range of environmental conditions present in the estuary. We determined growth rates for small and medium sized tape grass plants obtained from the Caloosahatchee estuary, southwest coastal Florida, USA in freshwater (0.5 psu) under high (331 μE m^?2 s^?1) and low light (42 μE m^?2 s^?1) and at 10 psu under high light conditions. We ran six treatments at five temperatures spanning 13–32 °C for 8–9 weeks. The optimum temperature for growth was roughly 28 °C, with a minimum threshold temperature of 13 °C and a maximum threshold temperature of 38 °C. Plants grew fastest in freshwater, at high light and temperatures greater than 20 °C. The slowest growth rates were observed at 13 °C regardless of salinity, light or plant size. Our results suggest that tape grass growth is strongly influenced by water temperature and that additional stressors such as low light and elevated salinity can reduce the range of temperature tolerance, especially at colder water temperatures.     

The effect of temperature,light-spectrum,desiccation and salinity gradients on the photosynthetic performance of a subtidal brown alga,Sargassum macrocarpum,from Japan

The effect of temperature, light-spectrum, desiccation and salinity gradients on the photosynthesis of a Japanese subtidal brown alga, Sargassum macrocarpum (Fucales), was determined using a pulse amplitude modulation-chlorophyll fluorometer and dissolved oxygen sensors. Temperature responses of the maximum (F_v/F_m in darkness) and effective (ΔF/F_m′ at 50 μmol photons m⁻² s⁻¹; = Φ_PSII) quantum yields during 6-day culture (4–36°C) remained high at 12–28°C, but decreased at higher temperatures. Nevertheless, ΔF/F_m′ also dropped at temperatures below 8°C, suggesting light sensitivity under chilling temperatures because F_v/F_m remained high. Photosynthesis–irradiance responses at 24°C under red (660 nm), green (525 nm), blue (450 nm) and white light (metal halide lamp) showed that maximum net photosynthesis under blue and white light was greater than under red and green light, indicating the sensitivity and photosynthetic availability of blue light in the subtidal light environment. In the desiccation experiment, samples under aerial exposure of up to 8 h under dim-light at 24°C and 50% humidity showed that ΔF/F_m′ quickly declined after more than 45 min of emersion; furthermore, ΔF/F_m′ also failed to recover to initial levels even after 1 day of rehydration in seawater. Under the emersion state, the ΔF/F_m′ remained high when the relative water content (RWC) was greater than 50%; in contrast, it quickly dropped when the RWC was less than 50%. When the RWC was reduced below 50%, ΔF/F_m′ did not return to initial levels, regardless of subsequent re-hydration, suggesting a low capacity of photosynthesis to recover from desiccation. The stenohaline response of photosynthesis under 3-day culture is evident, given that ΔF/F_m′ declined when salinity was beyond 20–40 psu. Adaptation to subtidal environments in temperate waters of Japan can be linked to these traits.     

Combined effects of temperature and salinity on functional responses of haemocytes and survival in air of the clam Ruditapes philippinarum

The combined effects of temperature and salinity on both immune responses and survival in air of the clam, Ruditapes philippinarum, were evaluated for the first time. The animals were kept for 7 days at three differing temperature (5 °C, 15 °C, 30 °C) and salinity values (18 psu, 28 psu, 38 psu), and effects of the resulting 9 experimental conditions on total haemocyte count (THC), Neutral Red uptake (NRU), haemolymph protein concentration, and lysozyme activity in both haemocyte lysate (HL) and cell-free haemolymph (CFH) were evaluated. The survival-in-air test was also performed. Two-way ANOVA analysis revealed that temperature influenced significantly THC and NRU, whereas salinity and temperature/salinity interaction affected NRU only. Temperature and salinity did not influence significantly HL and CFH lysozyme activity, as well as haemolymph total protein content. Survival-in-air test is widely used to evaluate general stress conditions in clams. In the present study, temperature and salinity were shown to influence the resistance to air exposure of R. philippinarum. The highest LT₅₀ (air exposure time resulting in 50% mortality) value was recorded in clams kept at 18 psu and 15 °C, whereas the lowest value was observed in clams kept at 28 psu and 30 °C. Overall, results obtained demonstrated that temperature and salinity can affect some functional responses of haemocytes from R. philippinarum, and suggested a better physiological condition for animals kept at 15 °C temperature and 18 psu salinity.     

Effects of temperature and nutrients on microscopic stages of the bull kelp (Nereocystis luetkeana,Phaeophyceae)

Warming ocean temperatures have been linked to kelp forest declines worldwide, and elevated temperatures can act synergistically with other local stressors to exacerbate kelp loss. The bull kelp Nereocystis luetkeana is the primary canopy-forming kelp species in the Salish Sea, where it is declining in areas with elevated summer water temperatures and low nutrient concentrations. To determine the interactive effects of these two stressors on microscopic stages of N. luetkeana, we cultured gametophytes and microscopic sporophytes from seven different Salish Sea populations across seven different temperatures (10–22°C) and two nitrogen concentrations. The thermal tolerance of microscopic gametophytes and sporophytes was similar across populations, and high temperatures were more stressful than low nitrogen levels. Additional nitrogen did not improve gametophyte or sporophyte survival at high temperatures. Gametophyte densities were highest between 10 and 16°C and declined sharply at 18°C, and temperatures of 20 and 22°C were lethal. The window for successful sporophyte production was narrower, peaking at 10–14°C. Across all populations, the warmest temperature at which sporophytes were produced was 16 or 18°C, but sporophyte densities were 78% lower at 16°C and 95% lower at 18°C compared to cooler temperatures. In the field, bottom temperatures revealed that the thermal limits of gametophyte growth (18°C) and sporophyte production (16–18°C) were reached during the summer at multiple sites. Prolonged exposure of bull kelp gametophytes to temperatures of 16°C and above could limit reproduction, and therefore recruitment, of adult kelp sporophytes.     

The effects of temperature and salinity on vital biological functions of the Antarctic crustacean Serolis polita

Effects of temperature rise (from 0 to +5°C) and salinity decline (from 34 to 30 psu) on vital biological functions of the Antarctic isopod Serolis polita were studied in laboratory experiments. Behavioural reactions to food odour, as well as righting responses and reburying in the sediments, were measured. Both temperature increase and salinity decline impaired the ability of S. polita to perform these biological functions critical for their long-term survival, by lowering the number of isopods able to right and rebury in the sediment, increasing time-to-right, reducing locomotory activity and weakening isopod reaction to food odour. Significant interactive effects between temperature and salinity on time-to-right and time spent swimming were observed, with isopods being more vulnerable to lower salinities when exposed to higher temperatures. Some biological functions (righting, reburying) were more sensitive to temperature and salinity changes than others (swimming). In conclusion, our findings strongly suggest that Antarctic isopods are vulnerable to environmental changes, and their ability to cope with them is limited.     

EXCYSTMENT AND GROWTH OF CHRYSOPHYTES AND DINOFLAGELLATES AT LOW TEMPERATURES AND HIGH SALINITIES IN ANTARCTIC SEA-ICE1

Extreme environmental conditions have been thought to limit algal growth in the upper sea-ice. In McMurdo Sound, Antarctica, chrysophyte statocysts (stomatocysts) and dinoflagellate hypnozygotes (resting cysts) overwinter in first- and second-year land-fast sea-ice exposed to temperatures of -20° C or lower. In early November, when temperatures in the upper ice are < ?8°C and brine salinities are >126 psu, dinoflagellate cysts activate and shortly thereafter excyst. During early November, chrysophyte statocysts also begin to excyst. Net daily primary production occurs in the sea-ice brine at temperatures as low as ?7.1° C, at brine salinities as high as 129 psu, and at average photon flux densities as low as 5 μmol photons.m^?2.s^?1. Dinoflagellate densities were >10⁶ vegetative cells.L^?1 of ice while temperatures in the upper ice were between ?6.8 and ?5.8° C and brine salinities were ～100 psu. Chrysophyte densities reached >10⁶.L^?1 of ice by early December. High densities of physiologically active clyo- and halotolerant algae can occur in the upper land-fast sea-ice under extreme conditions of temperature and salinity.     

Differential response of varying salinity and temperature on zoospore induction, regeneration and daily growth rate in Ulva fasciata (Chlorophyta, Ulvales)

Seaweed cultivation is imperative to augment increasing industrial demand. Ulva fasciata Delile is a potential seaweed for cultivation with applications in food industries. There is a renewed interest in large-scale aquaculture of this species in India due to its envisaged demand in snack food products. In the present study, we have successfully demonstrated the possibility of inducing zoospores in vegetative tissue, effective regeneration and improved growth in this seaweed by manipulating salinity (from 15 to 30 psu) and temperature (from 15 to 35°C). The optimum salinity and temperature requirement for zoospores induction were found to be 15 psu and 25°C, respectively. The quadriflagellate zoospores showed negative phototaxis and the settlement and germination pattern similar to several other green seaweeds. The optimum regeneration (78.53?±?10.05%) was recorded at 25°C and 30 psu salinity. The maximum daily growth rate (16.1?±?0.28%) was at 25°C and 30 psu salinity which corresponded to the field conditions. This method could be further refined at nursery culture to achieve artificial seeding essential for the success of commercial cultivation of this seaweed.     

Plasma osmolality and oxygen consumption of perch Perca fluviatilis in response to different salinities and temperatures

The present study determined the blood plasma osmolality and oxygen consumption of the perch Perca fluviatilis at different salinities (0, 10 and 15) and temperatures (5, 10 and 20° C). Blood plasma osmolality increased with salinity at all temperatures. Standard metabolic rate (SMR) increased with salinity at 10 and 20° C. Maximum metabolic rate (MMR) and aerobic scope was lowest at salinity of 15 at 5° C, yet at 20° C, they were lowest at a salinity of 0. A cost of osmoregulation (SMR at a salinity of 0 and 15 compared with SMR at a salinity of 10) could only be detected at a salinity of 15 at 20° C, where it was 28%. The results show that P. fluviatilis have capacity to osmoregulate in hyper‐osmotic environments. This contradicts previous studies and indicates intraspecific variability in osmoregulatory capabilities among P. fluviatilis populations or habitat origins. An apparent cost of osmoregulation (28%) at a salinity of 15 at 20° C indicates that the cost of osmoregulation in P. fluviatilis increases with temperature under hyperosmotic conditions and a power analysis showed that the cost of osmoregulation could be lower than 12·5% under other environmental conditions. The effect of salinity on MMR is possibly due to a reduction in gill permeability, initiated to reduce osmotic stress. An interaction between salinity and temperature on aerobic scope shows that high salinity habitats are energetically beneficial during warm periods (summer), whereas low salinity habitats are energetically beneficial during cold periods (winter). It is suggested, therefore, that the seasonal migrations of P. fluviatilis between brackish and fresh water is to select an environment that is optimal for metabolism and aerobic scope.     

Effects of temperature,salinity and body size on routine metabolism of coastal largemouth bass Micropterus salmoides

Routine metabolism (i.e. standard metabolism plus a low level of activity) of coastal largemouth bass Micropterus salmoides from Mobile‐Tensaw Delta, AL, U.S.A. was examined as a function of temperature (15, 20, 25 and 30° C), salinity (0, 4, 8 and 12) and body mass (range 24–886 g) using flow‐through respirometry. Functionally, a cubic relationship best described the effect of salinity on respiration; the magnitude of these effects increased with temperature and body mass. The best model predicted that specific respiration (mg O₂ g^?1 h^?1) at temperatures >20° C was lowest at salinities of 0·0 and 9·7, and elevated at 3·2 and 12·0; salinity had little to no effect at temperatures ≤20° C. Respiration increased exponentially with temperature, but when compared with previously published respiration rates for M. salmoides from northern latitudes, predicted respiration was higher at cool temperatures and lower at high temperatures. The reduced energetic cost near the isosmotic level (i.e. c. 9) may be an adaptive mechanism to tolerate periods of moderate salinity levels and may help explain why M. salmoides do not flee an area in response to increased salinity. Further, these results suggest that salinity has high energetic costs for coastal populations of M. salmoides and may contribute to the observed slow growth and small maximum size within coastal systems relative to inland freshwater populations.     

Effect of temperature and photon fluence rate on gametophytes and young sporophytes of Laminaria ochroleuca Pylaie

We analysed the effects of temperature and photon fluence rate on meiospore germination, growth and fertility of gametophytes, and growth of young sporophytes of Laminaria ochroleuca. Maximum percentages of germination (91–98%) were obtained at 15°C and 18°C, independent of photon fluence rate. Optimal development of female gametophyte and maximum fecundity and reproductive success of gametophytes occurred at 15°C and 18°C and at 20 and 40 μmol m^–2 s^–1. Maximum relative growth rate of young sporophytes after 2 weeks of culture was achieved under the same conditions. L. ochroleuca gametophytes cannot reproduce and growth of its sporophytes is not competitive at temperatures close to 10°C. Received in revised form: 31 August 2001 Electronic Publication     

Growth of Ecklonia cava (Laminariales, Phaeophyta) sporophytes transplanted to a locality with different temperature conditions

Transplanting experiments were carried out to determine whether the small type sporophytes with short stipe of Ecklonia cava Kjellman (Laminariales, Phaeophyta) growing in a locality with warm temperatures, change into larger type with a long stipe when transplanted to a locality with cooler temperatures. Juvenile E. cava sporophytes, having a stipe shorter than 5 cm long were collected from Tei in Tosa Bay (southern Japan) (seawater temperature 15–29°C) and transplanted to Nabeta Bay (central Japan) (seawater temperature 13–25°C), where larger type E. cava sporophytes characterized by long stipe (ca 1 m) grow. They were attached to artificial reefs at the sea bottom (9 m depth) in Nabeta Bay to monitor their growth. For comparison, juvenile E. cava sporophytes of almost similar size growing in Nabeta Bay were also transplanted in the same way to the same experimental site. Observations of growth of sporophytes from Tei and Nabeta were carried out monthly for 2 years from November 1995 to October 1997. The transplanted Tei and Nabeta sporophytes showed an increase in stipe length and diameter from winter to spring, whereas almost no increase was observed during summer and autumn. At the end of the study period, the stipe of Nabeta sporophytes reached 25.6 cm in length and 17.0 mm in diameter, whereas that of Tei sporophytes reached 11.1 cm in length and 11.2 mm in diameter. The primary blade length was 16.0 cm in Nabeta sporophytes, whereas it was 5.5 cm in Tei sporophytes. Thus, Tei sporophytes still remained smaller than Nabeta sporophytes even under the same environmental conditions.