Seagrass feeding choices and digestive strategies of the herbivorous fish Sarpa salpa

This is the first study investigating the plant–herbivore interaction between Sarpa salpa, which has overgrazed seagrass transplants in Portugal, and the seagrasses Cymodocea nodosa, Zostera marina and Zostera noltii, which have been considered for restoration. When offered the choice between the three seagrasses in outdoor tanks, adult S. salpa clearly preferred Z. noltii. Testing the seagrasses separately, mean ± s.d. feeding rates ranged from 21 ± 11 g seagrass fresh mass kg^?1 fish mass day^?1 for Z. marina to 32 ± 9 g seagrass fresh mass kg^?1 fish mass day^?1 for C. nodosa and 40 ± 11 g seagrass fresh mass kg^?1 fish mass day^?1 for Z. noltii (temperature = 16° C). Food‐processing rate in S. salpa did not differ between seagrasses, and there was no evidence of a regulation of processing rate according to food intake. Seagrasses differed substantially in nitrogen content and C:N, with C. nodosa containing the highest nitrogen content and lowest C:N (2·5 ± 0·1% and 14·0 ± 1·0), followed by Z. noltii (2·1 ± 0·1% and 17·0 ± 1·0) and Z. marina (1·4 ± 0·1% and 26·0 ± 2·0). Food‐processing rate in S. salpa and the nutritional value of the seagrasses were not correlated with the observed feeding preference and rate. The study suggests that C. nodosa and Z. marina are less at risk of overgrazing by S. salpa and might thus be preferable to Z. noltii for seagrass restoration in areas with noticeable abundances of this fish.     

Characterization and seasonal variation of individual flavonoids in Zostera marina and Zostera noltii from Norwegian coastal waters

The flavonoid content in leaves of Zostera marina and the endangered Zostera noltii, including mono- and disulphated flavonoids, from different sample localities were characterized. Seasonal variation of both individual and total flavonoid, as well as rosmarinic acid concentration were revealed. Minor amounts of luteolin 7-(6″-malonyl)glucoside (6) and apigenin7-(6″-malonyl)glucoside (11) were identified in Z. noltii for the first time. The total flavonoid content was found to be higher in Z. noltii than in Z. marina at most of the examined localities, and the qualitative flavonoid content was somewhat different in the two species. The quantitative variation of flavonoids and rosmarinic acid was found to be relatively consistent from year to year in Z. marina during a period of three years. The two species appeared though to have a different flavonoid production in the various seasons at the West coast. While Z. marina had the highest content in young leaves in May or June, with a markedly decrease from June to September and the lowest measured content in February, Z. noltii had the lowest measured flavonoid content in May/June followed by an increase from June to September and the highest measured content during wintertime in February. The observed seasonal differences may be related to the fact that Z. noltii is considered a perennial, thermophilous species, and the increasing flavonoid production during the colder seasons from September to March/April in Norway may serve as a protective function.     

Evaluation of Zostera detritus as a potential new source of zosteric acid

Detritus of the seagrasses Zostera noltii and Z. marina collected on the beaches of Arcachon Lagoon (France) over a 3-year period was screened as a new source of zosteric acid (ZA). This natural sulphated phenolic acid is a high value-added product capable of preventing settlement of marine organisms and protecting crops from fungal diseases. The seasonal variation of the ZA content was quantified in methanolic and aqueous crude extracts using high-performance liquid chromatography. The concentration found ranged from 65 to 456 μg g⁻¹ dry wt for Z. noltii and 51–692 μg g⁻¹ dry wt for Z. marina, respectively. This is the first report of ZA in Zostera noltii. Detrital leaves of Zostera have never before been screened for their bioactive substances. These results show that this low cost, very abundant and renewable, but heretofore unused, marine resource has potential as a source of a rare and naturally occurring bioactive product.     

Chromosome numbers of the European seagrasses

The chromosome numbers of the five European seagrasses have been determined in material from several sites along the coasts of the Atlantic Ocean, the North Sea and the Mediterranean:Zostera marina L., 2n = 12;Z. noltii Hornem., 2n = 12;Posidonia oceanica (L.)Delile, 2n = 20;Cymodocea nodosa (Ucria)Aschers., 2n = 14, 2n = 28;Halophila stipulacea (Forsk.)Aschers., 2n = 18. The difference in chromosome morphology betweenZ. marina andZ. noltii supports the division of the genus into two subgenera.     

Characterization of microsatellite loci in the dwarf eelgrass Zostera noltii (Zosteraceae) and cross‐reactivity with Z. japonica

Zostera noltii is an important species of eelgrass occurring along European, north African, Mediterranean, Black Sea and Azov Sea coasts. Nine microsatellite loci were developed and no linkage disequilibrium was observed. Cross‐amplification was observed for all loci (polymorphic) in Z. japonica; only four loci amplified (monomorphic) in Z. marina.     

Seasonal dynamics of <Emphasis Type="Italic">Zostera noltii</Emphasis> Hornem. in two Mediterranean lagoons

The density, biomass and shoot morphology of two populations of Zostera noltii were monitored from January 1998 to July 1999 at two shallow Mediterranean lagoons of Biguglia and Urbino, which differ in hydro-morphological conditions and nutrient loading. Monitoring included the principal biological and foliar parameters (shoot density, aboveground and belowground biomass, length, width and number of leaves, LAI and coefficient A: percentage of leaves having lost their apex), the organic matter contents of the sediment and the environmental conditions (salinity, turbidity, temperature, nutrient concentrations and dissolved oxygen levels). The two populations of Z. noltii displayed seasonal changes in density (1600–19600 m²), aboveground biomass (11–153 g. DW. m⁻²), leaf length (33–255 mm), and leaf width (0.9–1.8 mm). Temperature and turbidity were significant environmental factors influencing the temporal changes observed in the Z. noltii meadows studied. Conversely, the belowground biomass, the number of leaves per shoot and the LAI did not undergo any seasonal changes. In the Biguglia lagoon, the functioning dynamics of the Z. noltii seagrass beds are determined by the catchment area and the inputs of nutrients derived from it, whereas in the Urbino lagoon the dynamics of the Z. noltiibeds depend on low levels of water turbidity.     

Temperature tolerance and survival of intertidal populations of the seagrass <Emphasis Type="Italic">Zostera noltii</Emphasis> (Hornemann) in Southern Europe (Ria Formosa,Portugal)

The dwarf seagrass Zostera noltii is an important primary producer in Atlantic coastal ecosystems from Mauritania to southern Norway and the Mediterranean Sea. Sessile intertidal organisms existing at the interface between marine and terrestrial environments may be particularly vulnerable to environmental change. In this study, we asked how near to thermal tolerance limits natural populations of Z. noltii are in the Ria Formosa coastal lagoon system in southern Portugal. We recorded the maximum temperatures in the Ria Formosa during the 2007 summer, and conducted experiments to determine the sub-lethal temperature of Z. noltii shoots sampled at two sites located at different tidal heights. Mortality rates and photosynthetic performance were recorded within a range of heat shock temperatures between 35 and 41°C. Survival was recorded ≤37°C, while higher temperatures led to a sudden drop in photosynthetic capacity followed by mortality (shoot loss) that occurred more rapidly with increasing temperatures. At 39°C and above, the rate of shoot mortality in both sites was close to 100%, occurring between 5 and 13 days after the heat shock. Survival was ca. 95 and 90% at 35 and 37°C, respectively. From these results for Z. noltii populations in the Ria Formosa we estimated sub-lethal temperature to be approximately 38°C for Z. noltii, close to the maximum of 36°C recorded in the summer 2007. Considering predicted trajectories in the coming decades, these results raise concern as to the future viability of intertidal Z. noltii populations near the southernmost edge of their distribution. Handling editor: S. M. Thomaz     

Seasonal variations of community production,respiration and biomass of different primary producers in an intertidal <Emphasis Type="Italic">Zostera noltii</Emphasis> bed (Western English Channel,France)

Gross primary production (GPP) and respiration (R) in an intertidal Zostera noltii (Hornemann, 1832) community were estimated each month from in situ CO₂ flux measurements during the emersion period in the Bay of Morlaix (France). Simultaneously, the Chlorophyll a biomass of Z. noltii leaves, associated epiphytes and sediment within the benthic chambers were also monitored. The significant positive exponential relationship established between R and the maximum daily temperature emphasized that temperature was a good predictor of seasonal variation in the community R in Z. noltii bed. In contrast, the amplitude of the fitted curve on GPP was low and represented <25% of the annual mean value (90.9 ± 15.2 mgC m⁻² h⁻¹). The Chlorophyll a biomass of sediment was high (133.70 ± 22.96 mg m⁻²) but it was likely constituted by detritus which did not contribute much to the community GPP. The Chlorophyll a biomass of associated epiphytes was higher in winter and varied over the year inversely with Z. noltii. Therefore, the asynchronous dynamic of primary producers could support a constant rate of production at community scale over the year.     

Leaf dynamics and standing stocks of intertidal Zostera noltii Hornem. and Cymodocea nodosa (Ucria) Ascherson on the Banc d'Arguin (Mauritania)

Leaf dynamics and standing stocks of intertidal seagrasses were studied in the Baie d'Aouatif (Parc National du Banc d'Arguin, Mauritania) in April and September 1988. Standing stocks of Zostera noltii Hornem. suggest a unimodal seasonal curve similar to what is found for populations at higher latitudes. Also, leaf growth rates (0.03 cm² cm^–2 day^–1 on average) were similar to those found at higher latitudes in these months. Variation in leaf loss over tidal depth, time and different locations in the Baie d'Aouatif was larger and more often significant than variation in leaf growth. In general, Z. noltii beds in the Baie d'Aouatif had comparable leaf growth rates and standing stocks. In both months losses were almost always higher than or equal to growth.Variation in leaf loss over time was much higher in the plots that were situated high in the intertidal than in lower plots. This is explained by differences in susceptibility to sloughing, which is presumably higher in periods with low tide around noon for shallow depths.In an experiment using artificial shading nets, in situ leaf growth was affected negatively from 94% shading onwards. This shading was observed to reduce the light intensity reaching the seagrass bed to a level below the reported range of light compensation points for Z. noltii. Cymodocea nodosa (Ucria) Ascherson on average had higher leaf area and relative growth rates than Z. noltii and much lower loss rates, resulting in a positive net increase in September. Standing stocks were also higher than for Z. noltii. A mixed seagrass bed containing the above two species and Halodule wrightii Ascherson had the highest observed total biomass: 335 g m^–2 ash-free dry weight.     

Seasonal variation in rates of heterotrophic nitrogen fixation (acetylene reduction) in Zostera noltii meadows and uncolonised sediments of the Bassin d'Arcachon, south-west France

Nitrogen fixation (acetylene reduction) rates were measured over an annual cycle in meadows of the seagrass Z. noltii and uncolonised sediments of the Bassin d'Arcachon, south-west France, using both slurry and whole core techniques. Measured rates using the slurry technique in Z. noltii colonised sediments were consistently higher than those determined in isolated cores. This was probably due to the release of labile organic carbon sources during preparation of the slurries. Thus, in colonised sediments the whole core technique may provide a more accurate estimate of in situ activity. Acetylene reduction rates measured by the whole core technique in colonised sediments were 1.8 to 4-fold greater, dependent upon the season, in the light compared with those measured in the dark, indicating that organic carbon released by the plant roots during photosynthesis was an important factor regulating nitrogen fixation. In contrast acetylene reduction rates in uncolonised sediments were independent of light.Addition of sodium molybdate, a specific inhibitor of sulphate reduction inhibited acetylene reduction activity in Z. noltii colonised sediments by > 80% as measured by both slurry and whole core techniques irrespective of the light regime, throughout the year inferring that sulphate reducing bacteria (SRB) were the dominant component of the nitrogen fixing microflora. A mutualistic relationship between Z. noltii and nitrogen fixing SRB in the rhizosphere, based on the exchange of organic carbon and fixed nitrogen is proposed. In uncolonised sediments sodium molybdate initially severely inhibited acetylene reduction rates, but the level of this inhibition declined over the course of the year. These data indicate that the nitrogen fixing SRB associated with the Zostera roots and rhizomes were progressively replaced by an aerobic population of nitrogen fixers associated with the decomposition of this recalcitrant high C:N ratio organic matter.Acetylene and sulphate reduction rates in the seagrass beds showed distinct summer maxima which correlated with a reduced availability of NH ₄ ⁺ in the sediment and the growth cycle of Z. noltii in the Bassin. Overall, these data indicate that acetylene reduction (nitrogen fixation) activity in the rhizosphere of Z. noltii was regulated both by release of organic carbon from the plant roots and maintenance of low ammonium concentrations in the root zone due to efficient ammonium assimilation.Nitrogen fixation rates determined from acetylene reduction rates measured by the whole core technique ranged from 0.1 to 7.3 mg N m^–2 d^–1 in the Z. noltii beds and between 0.02 and 3.7 mg N m^–2 d^–1 in uncolonised sediments, dependent upon the season. Nitrogen fixation in the rhizosphere of Z. noltii was calculated to contribute between 0.4 and 1.1 g N m^–2 y^–1 or between 6.3 and 12% of the annual fixed nitrogen requirement of the plants. Heterotrophic nitrogen fixation therefore represents a substantial local input of fixed nitrogen to the sediments of this shallow coastal lagoon and contributes to the overall productivity of Z. noltii in this ecosystem.     

Rise in human activities on the mudflats and Brent Geese (Branta bernicla) wintering distribution in relation to Zostera spp. beds: a 30-year study

Human activities may restrict access of wintering birds to their food resources, but habitat destruction and fragmentation may interact with disturbance to reduce carrying capacity. We analysed the abundance and distribution of wintering Brent Geese (Branta bernicla) in a major French wintering site from 1970 to 2000, when they experienced increases in human foreshore activities. We used mixed-effects models to test whether spatial extent of two Zostera species, Z. noltii and Z. marina, influenced the distribution of Brent Geese. The number of geese increased significantly with the area of both Zostera species beds. However, the relationship changed in the period after the increase in human activities. More specifically, fragmentation of the most accessible food resource, Z. noltii, negatively affected Brent Goose distribution. This fragmentation was in turn increased by human activities and reduced by the protection of Z. noltii (by banning human access). This implies that direct disturbance, although not excluded, might not have been the major cause of changes in Brent Goose distribution, but rather the effects of human activities on food resources. Thus, our results show that establishment of resource protection areas are efficient as conservation measures, and they underline the need for studies of impacts of human activities on resources, in addition to disturbance effects.     

Suitable water temperature for seed storage of Zostera japonica for subtropical seagrass bed restoration

Restoration of Zostera japonica is needed. Laboratory culture experiments to know the germination characteristics might be helpful for implementation of actual restoration. As a part of germination experiments, we explored suitable water temperature for long-term storage of Z. japonica seeds. This work was based on earlier reports of Zostera marina, which presumably has similar physiological properties to Z. japonica. This study consisted of two experiments: (1) preservation experiments to investigate the fate of stored Z. japonica seeds and (2) germination experiments to investigate the germination potential of the stored seeds. The results of the preservation experiments suggested that seed condition, that is, germinated, degraded, unstable, stable, etc., showed variations between the seeds stored at 4 and 23 °C. The majority of the seeds stored at 4 °C were germinated, while those at 23 °C seemed to be degraded, presumably by bacteria and mold. The germination experiments suggested high germination potential of seeds stored at 4 °C even after 302 days had elapsed. In conclusion, including previously reported results on Z. marina, low temperature was suitable for the preservation of seeds to maintain germination potential.     

Desiccation,Moisture Content and Germination of Zostera marina L. Seed

Zostera marina is the only seagrass species whose seeds have been successfully used in large‐scale restoration. Although progress has been made in refining Z. marina restoration protocols, additional information on Z. marina seed physiology is necessary as the science of seagrass restoration evolves. We tested the germination rates of Z. marina seeds under different relative humidities and temperatures for different periods of time. Z. marina seed moisture content (MC) and germination rates were also tested when seeds were exposed to a temperature of 25°C and relative humidity of 50%. Z. marina seeds suffered higher mortality when exposed to lower relative humidity and higher temperature for longer period of exposure time. A significant negative correlation was detected between seed germination rate and MC. Z. marina seeds are sensitive to desiccation exposure and long periods of exposure to air should be prevented to minimize seed mortality when seeds are used in restoration projects.     

The effects of the polychaete Nereis diversicolor on the distribution and transplanting success of Zostera noltii

Loss of saltmarsh vegetation in south-east England is a significant problem for conservation and coastal defence. The losses of vegetation began in the 1930s with the loss of intertidal Zostera marina and have continued more recently. Some preliminary trials at re-establishing Zostera in some estuaries of Essex have not been successful. This paper addresses the hypothesis that the infauna, particularly the polychaete Nereis diversicolor, may restrict natural colonisation by Zostera and reduce the success of transplanting trials. In field experiments, Z. noltii were transplanted into areas where Nereis were common, close to an established seagrass bed and into two other estuaries. The transplants protected from the effects of the polychaetes by netting had a higher survival rate, lower index of root damage and greater biomass at the end of the experiments than those that were unprotected. In laboratory experiments, Nereis reduced the survival of Z. noltii. They were observed grasping the leaves and pulling them into their burrows. These results indicate that herbivory and disturbance by N. diversicolor is responsible, at least in part, for the restriction of the distribution of Z. noltii and may have been important in limiting the success of previous transplanting experiments. A hypothesis is proposed which states that there are two stable states on the upper mud-flats. One state is dominated by plants, including Zostera spp., which prevent colonisation by burrowing infauna, and the other is dominated by infauna which prevent colonisation by plants. Managing these two states could be the key to re-establishing the early successional stages of saltmarsh development. Received: 15 June 1999 / Received in revised form: 6 July 1999 / Accepted: 8 July 1999     

Are epiphytes a significant component of intertidal Zostera noltii beds?

The role of epiphytes in an intertidal Zostera noltii seagrass bed in Marennes-Oléron Bay was assessed in comparison with the other main benthic primary producers (Z. noltii, microphytobenthos) at two bathymetric levels and on a seasonal basis. Assemblage and biomass of epiphytes were studied using scanning electron microscopy (SEM). Z. noltii and its detrital matter followed a typical seasonal pattern: microphytobenthos was present in large quantities throughout the year representing 21% of the total biomass while detrital matter, above-ground parts and below-ground parts accounted for 65, 9 and 5%, respectively. Only two species of epiphytic diatoms, Cocconeis scutellum and Cocconeis placentula, were observed on seagrass leaves. Epiphyte biomass was very low, representing on average less than 0.001% of that of microphytobenthos or leaves. This low epiphyte biomass is linked with the absence of macroalgae and also with the low biovolume of Cocconeis, which formed a monolayer of cells on leaves. This can be explained by the severe conditions of the intertidal and the high leaf turn-over of Z. noltii leaves.     

Morphological characteristics of annual Zostera marina shoots at various germination temperatures

The timing of the transition from seed, seedlings and development into flowering is paramount importance in annual-type Zostera marina, because flowering is the first step of sexual reproduction. A majority of plants use environmental cues to regulate the transition to their developmental stages because plants must flower synchronously for successful outcrossing and must complete their sexual reproduction under favorable external conditions. The morphological characteristics (seeds and lateral shoot production, branch number, and inflorescence length) of reproductive shoots of Z. marina L. were examined in outdoor mesocosms to better understand the reproductive strategies of annual populations. Seeds in the germination experiment were divided into two groups: those exposed to cold (7 °C; vernalized group) and those left untreated (25-21 °C; non-vernalized group). All 600 seeds (300 from each group) were cultured for 2 months at 7, 10, 15, 20, and 25 °C in an indoor incubator. In the vernalized group, the germination rates were almost significantly higher than in the non-vernalized group. However, germination rates were not significantly affected by germination temperature. In outdoor mesocosms, production of vegetative shoots was observed in plants germinated at 15 and 20 °C in the vernalized group and at 10, 15 and 20 °C in the non-vernalized group. The highest number of vegetative shoots produced (35) was observed in plants germinated at 20 °C in the vernalized group, whereas seeds of either group failed to produce vegetative shoots when germinated at a low temperature (7 °C).In the flowering phase, the number of branches per shoot in the vernalized group was significantly higher than in the non-vernalized group. The total number of spadices on the 1st branches of plants in the vernalized group (germination at 20 °C) was significantly lower than that in the non-vernalized group at the same germination temperature. The total number of spadices per reproductive shoot in the vernalized group (germination at 10 °C) was also higher than in the non-vernalized group. Thus, both low temperature (vernalization) and seed germination temperature have implications for the sexual and asexual propagation of annual Z. marina populations.     

Differential anaerobic decomposition of seagrass (Zostera noltii) and macroalgal (Monostroma obscurum) biomass from Arcachon Bay (France)

Arcachon Bay is characterized by extensive meadows of the seagrass Zostera noltii. Moreover, as a consequence of eutrophication, massive proliferations of the macroalga (Monostroma obscurum) have occurred since the beginning of 1990s.This paper describes the anaerobic decomposition of biomass of both species under experimental conditions by two methods. Firstly, the dynamics of decomposition were studied in situ using litter bags. The remaining biomass and the elemental composition of the decomposing macrophytes were monitored. Secondly, degradation was studied in experimental containers under anoxic conditions in which the release of inorganic nutrients and the development of fermentative and sulfate-reducing bacterial populations were followed.The decomposition rate of total biomass was faster for macroalgae than for the vascular plants, thus corroborating previous observations. However, both in situ and laboratory experiments showed that the anaerobic decomposition of the seagrass Z. noltii resulted in rapid release of inorganic N and P, and increasing C/N and C/P ratios of the residual biomass. As a result, the recycling of inorganic nitrogen and phosphorus compounds was slightly more efficient for Z. noltii than for M. obscurum. Recycling of inorganic nutrients appears to be of a great importance to the whole ecosystem, because of the extensive spreading of Z. noltii in the bay.     

Seagrasses are negatively affected by organic matter loading and Arenicola marina activity in a laboratory experiment

When two ecosystem engineers share the same natural environment, the outcome of their interaction will be unclear if they have contrasting habitat-modifying effects (e.g., sediment stabilization vs. sediment destabilization). The outcome of the interaction may depend on local environmental conditions such as season or sediment type, which may affect the extent and type of habitat modification by the ecosystem engineers involved. We mechanistically studied the interaction between the sediment-stabilizing seagrass Zostera noltii and the bioturbating and sediment-destabilizing lugworm Arenicola marina, which sometimes co-occur for prolonged periods. We investigated (1) if the negative sediment destabilization effect of A. marina on Z. noltii might be counteracted by positive biogeochemical effects of bioirrigation (burrow flushing) by A. marina in sulfide-rich sediments, and (2) if previously observed nutrient release by A. marina bioirrigation could affect seagrasses. We tested the individual and combined effects of A. marina presence and high porewater sulfide concentrations (induced by organic matter addition) on seagrass biomass in a full factorial lab experiment. Contrary to our expectations, we did not find an effect of A. marina on porewater sulfide concentrations. A. marina activities affected the seagrass physically as well as by pumping nutrients, mainly ammonium and phosphate, from the porewater to the surface water, which promoted epiphyte growth on seagrass leaves in our experimental set-up. We conclude that A. marina bioirrigation did not alleviate sulfide stress to seagrasses. Instead, we found synergistic negative effects of the presence of A. marina and high sediment sulfide levels on seagrass biomass.     

Photosynthetic adaptation of Zostera noltii Hornem.

The study of Zostera noltii Hornem. under stressed conditions in an estuary in southern Spain gives evidence that this seagrass can tolerate high light intensities without damage to its pigment pool. Therefore, photosynthesis at high light intensities remains as intense as at saturation point and it is sustained principally by Photosystem I.The existence of a short-term pigmentary response (60 min) was evidenced and the relationship between assimilation rates and the pigment pool was established.Comparisons made with Z. marina L. and between the respective environmental characteristics of both plants suggests that the special adaptive photosynthetic performance of Z. noltii can be supported by a C₄-type metabolism.     

NADP+-dependent isocitrate dehydrogenase from a psychrophilic bacterium,Psychromonas marina

The gene encoding NADP⁺-dependent isocitrate dehydrogenase (IDH; EC 1.1.1.42) of a psychrophilic bacterium, Psychromonas marina, was cloned and sequenced. The open reading frame of the gene encoding IDH of P. marina (PmIDH) was 2229 bp in length and corresponded to a polypeptide composed of 742 amino acids. The molecular mass of IDH was calculated as 80,426 Da. The deduced amino acid sequence of PmIDH exhibited high degrees of homology with the monomeric IDH from other bacteria such as Colwellia maris (62% identity) and Azotobacter vinelandii (AvIDH) (64%). His-tagged PmIDH overexpressed in Escherichia coli cells was purified and characterized. The optimum temperature of PmIDH activity was about 35 °C; however, the enzyme lost 74% of the activity after incubation for 10 min at 30 °C, indicating that this enzyme is thermolabile. Chimeric enzymes produced through domain swapping between PmIDH and mesophilic AvIDH were constructed and their optimum temperatures and thermostability were determined. The results suggest that regions 2 and 3, especially region 3, of the two IDHs are involved in their catalytic activities and optimum temperature and thermostability for activity.