Responses of Dodonaea viscosa growth and soil biological properties to nitrogen and phosphorus additions in Yuanmou dry-hot valley

Nitrogen(N) and phosphorus(P) are limited nutrients in terrestrial ecosystems, and their limitation patterns are being changed by the increase in N deposition. However, little information concerns the plant growth and the soil biological responses to N and P additions among different soils simultaneously, and these responses may contribute to understand plant-soil interaction and predict plant performance under global change. Thus, this study aimed to explore how N and P limitation changes in different soil types, and reveal the relationship between plant and soil biological responses to nutrient additions. We planted Dodonaea viscosa, a globally distributed species in three soil types(Lixisols, Regosols and Luvisols) in Yuanmou dry-hot valley in Southwest China and fertilized them factorially with N and P. The growth and biomass characters of D. viscosa, soil organic matter, available N, P contents and soil carbon(C), N, P-related enzyme activities were quantified. N addition promoted the growth and leaf N concentration of D. viscosa in Lixisols; N limitation in Lixisols was demonstrated by lower soil available N with higher urease activity. P addition promoted the growth and leaf P concentration of D. viscosa in Luvisols; severe P limitation in Luvisols was demonstrated by a higher soil available N: P ratio with higher phosphatase activity. Urease activity was negatively correlated with soil available N in Nlimited Lixisols, and phosphatase activity was negatively correlated with soil available P in P-limited Luvisols. Besides, the aboveground biomass and leaf N concentration of D. viscosa were positively correlated with soil available N in Lixisols, but the aboveground biomass was negatively correlated with soil available P. Our results show similar nutrient limitation patterns between plant and soil microorganism in the condition of enough C, and the nutrient limitations differ across soil types. With the continued N deposition, N limitation of the Lixisols in dry hot valleys is expected to be alleviated, while P limitation of the Luvisols in the mountaintop may be worse in the future, which should be considered when restoring vegetation.     

Changes of Biogenic Elements in <Emphasis Type="Italic">Phragmites australis</Emphasis> and <Emphasis Type="Italic">Suaeda salsa</Emphasis> from Salt Marshes in Yellow River Delta,China

Little information is available on biogenic elements (carbon, nitrogen, phosphorus and sulfur) and the ecological stoichiometric characteristics of plants in coastal wetlands. To investigate the contents of carbon, nitrogen, phosphorus and sulfur of plants, and their ecological stoichiometric characteristics in the Yellow (Huanghe) River Delta, plant samples were collected from two typical salt marshes (Suaeda salsa and Phragmites australis wetlands) during the period of from August to October in 2007, and the ratios of C/N, C/P, N/P, C/N/P and C/N/P/S were calculated. Results showed that during the studying period, plant C, N and P were lower than the global average values, and plant N and P were lower than the China’s average values. Leaf C and S in Suaeda salsa were significantly lower than those in Phragmites australis (P < 0.05), and leaf N and P in Suaeda salsa and Phragmites australis showed no significant differences (P > 0.05). Average C/N ratios were 23.75 in leaf, 73.36 in stem, 65.67 in root of Suaeda salsa, and 33.77 in leaf, 121.68 in stem, 97.13 in root of Phragmites australis. Average C/N ratios of Suaeda salsa and Phragmites australis were all great than 25, indicating the salt marsh in the Yellow River Delta is an N limitation system. Average C/P ratios were 276.78 in leaf, 709.28 in stem and 1031.32 in root of Suaeda salsa, and 536.94 in leaf, 768.13 in stem and 875.22 in root of Phragmites australis. The average N/P ratios of Suaeda salsa were 12.92 in leaf, 10.77 in stem and 10.91 in root, and the average N/P ratios of Phragmites australis were 16.40 in leaf, 7.40 in stem and 6.92 in root, indicating the Suaeda salsa wetlands were N limited and Phragmites australis wetlands were N limited in August and P limited in October in 2007. The average C/N, C/P and C/N/P ratios in Suaeda salsa and Pragmites australis were higher than the global average values, indicating the lower quality of organic matter provided by wetland plants in the Yellow River delta.     

Effects of nitrogen and phosphorus on the growth of <Emphasis Type="Italic">Levanderina fissa</Emphasis>: How it blooms in Pearl River Estuary

Effects of nitrogen (N) and phosphorus (P) from different sources and at different concentrations on the growth of Levanderina fissa (= Gyrodinium instriatum) were studied in laboratory conditions. The findings might explain the recurrent blooms of this species in Pearl River Estuary, China. Results showed that nutrient limitation significantly inhibited the growth of L. fissa. The values of specific growth rate (μ _max) and half-saturation nutrient concentration (K _S) were 0.37 divisions/d and 8.49 μmol L^?1 for N, and 0.39 divisions/d and 1.99 μmol L^?1 for P, respectively. Based on K _S values, dissolved inorganic N level in PRE was sufficient to support the high proliferation of L. fissa, while dissolved inorganic P concentration was far lower than the minimum requirement for its effective growth. L. fissa was not able to utilize dissolved organic N (DON) compounds such as urea, amino acids, and uric acid. However, it grew well by using a wide variety of dissolved organic P (DOP) sources like nucleotides, glycerophosphate, and 4-nitrophenylphosphate. The results from this study suggested that the ability in DOP utilization of L. fissa offers this species a competitive advantage in phytoplankton communities. The high level and continuous supply of DIN, enrichment of DOP, together with warm climate and low salinity in the Pearl River Estuary provided a suitable nutrient niche for the growth of L. fissa, and resulted in the recurrent blooms in the estuary.     

Effects of different phosphorus concentrations and N/P ratios on the growth and photosynthetic characteristics of <Emphasis Type="Italic">Skeletonema costatum</Emphasis> and <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis>

The effects of different phosphorus (P) concentrations (0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen (N)/P ratios on the growth and photosynthetic characteristics of Skeletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P (HP) concentration group showed the greatest algal density and highest specific growth rate. Changes in the maximum efficiency of photosystem II (F _v /F _m ) were monitored under the various P and N/P conditions. The largest decrease in F _v /F _m was in the low-P (LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields (Φ _PSII) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinoflagellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ _{PS II} values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang (Yangtze) River estuary from a photosynthesis perspective.     

Leaf litter decomposition dynamics in unmanaged <Emphasis Type="Italic">Phyllostachys pubescens</Emphasis> stands at high elevations in the Daiyun Mountain National Nature Reserve

Due to its rapid growth and high rate of spread, Phyllostachys pubescens can suppress other species, eventually leading to pure P. pubescens forests. Research on the patterns of litter decomposition and nutrient release from P. pubescens across stand mixtures are helpful to understand the decomposition and nutrient dynamics during the invasion process. However, little is known for unmanaged conditions at high elevations. Therefore, we studied the decomposition of four litter compositions comprised of different combinations of P. pubescens and Cunninghamia lanceolata foliage for one year to explore the pattern of litter decomposition and nutrient release at the Daiyun Mountain National Nature Reserve. The results show that: (1) the litter decomposed faster with a higher proportion of P. pubescens based on Olson’s decay model; (2) the remaining litter compositions followed a trigonometric function model better than Olson’s decay model and fluctuated periodically in tandem with changing temperature and precipitation; (3) the litter mixture decompositions did not support the biomass-ratio hypothesis test; and (4) there were significant binomial, power and exponential relationships between initial concentration and final amounts remaining after 1 year decomposition for C, N and P. The correlations between litter decomposition and nutrient releases were significant. The results may be used to explain the rapid spread of P. pubescens and to guide the natural management of plantations.     

Effects of <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> YC459 and soil types on seed germination and seedling growth in rock slope restoration

We carried out experiments with various concentrations of Trichoderma harzianum YC459 in different soil types (forest soil, mixed soil, merchantable soil, and leaf mold soil) to evaluate its effect on seed germination and seedling establishment of four species (Festuca arundinacea Schreb., Dianthus barbatus var. asiaticus Nakai, Lespedeza cyrtobotrya Miq., and Parthenocissus tricuspidata Planch) for rock slope restoration. We also investigated the use of drilled slanted holes on the rock slopes for seedling establishment. The results showed that T. harzianum concentration had significant effects on seed germination, seedling growth, and seedling survival for all the species with different soil types. Seed germination and survival rates peaked at 5% T. harzianum concentration with leaf mold soil and decreased as T. harzianum concentration increased from 5% to 10%. Seedling survival rates of all four species were generally lowest at 0% T. harzianum concentration in all soil types. The height of F. arundinacea and L. cyrtobotrya peaked at 5% T. harzianum concentration whereas that of D. barbatus and P. tricuspidata peaked at 10% T. harzianum concentration. We concluded that 5% T. harzianum concentration with leaf mold soil is appropriate for seed germination and seedling survival rates of most species, thus enhancing seedling establishment. Practical application of the findings of this study will contribute in the vegetation restoration of steep rocks in mountain environments     

Identification and bioinformatics analysis of microRNAs from the sporophyte and gametophyte of <Emphasis Type="Italic">Pyropia haitanensis</Emphasis>

Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata (Porphyra haitanensis) is an economically important genus that is cultured widely in China. P. haitanensis is cultured on a larger scale than Pyropia yezoensis, making up an important part of the total production of cultivated Pyropia in China. However, the majority of molecular mechanisms underlying the physiological processes of P. haitanensis remain unknown. P. haitanensis could utilize inorganic carbon and the sporophytes of P. haitanensis might possess a PCK-type C₄-like carbon-fixation pathway. To identify microRNAs and their probable roles in sporophyte and gametophyte development, we constructed and sequenced small RNA libraries from sporophytes and gametophytes of P. haitanensis. Five microRNAs were identified that shared no sequence homology with known microRNAs. Our results indicated that P. haitanensis might posses a complex sRNA processing system in which the novel microRNAs act as important regulators of the development of different generations of P. haitanensis.     

Effects of wetland vegetation on soil microbial composition: A case study in Tumen River Basin,Northeast China

Hydrology plays a dominant role in wetland plant distribution and microbial composition, but few studies explicitly attempted to relate the linkage between wetland vegetation and microbial community. The present study consisted of five wetland plant communities along three adjacent flood gradients zones (zone 1 dominated by Carex appendiculat, zone 2 dominated by Eleocharis ovate, and zone 3 dominated by Phragmites australis/Bidens pilosa/Calamagrostis angustifolia, which formed separate, monoculture patches). Gram negative and arbuscular mycorrhizal fungal phospholipid fatty acid (PLFA) are more abundant in the site with short flooding period (zone 3) than in the site with long flooding period (zone 1), and they are also different in the P. australis, B. spilosa and C. angustifolia of zone 3. Principle Component Analysis (PCA) showed that the flooding period could explain 92.4% of variance in microbial composition. Redundancy Analysis (RDA) showed that available nitrogen (AN), total nitrogen (TN) and soil organic matter (SOM) could explain the 79.5% of variance in microbial composition among E. ovata, P. australis, B. pilosa and C. angustifolia. Results demonstrated that flooding period was the main factor in driving the microbial composition and plant-derived resources could influence soil microbial composition in the seasonally flooded zones.     

Loss of preferred habitat and pollen limitation threatens reproduction in a rare mountain <Emphasis Type="Italic">Paeonia delavayi</Emphasis>

Paeonia delavayi is a wild tree peony species endemic to high-altitude regions in southwestern China. Recent agricultural land expansion, however, not only causes its dwindling population size, but also poses a severe threat to its long-term persistence. Since our knowledge of the reproductive biology of P. delavayi is very limited, and some management misconceptions have further exacerbated the already worrisome situation, the aim of the present study is to provide some scientific evidence regarding habitat preference and breeding system of P. delavayi, and to help correct some management misconceptions. Specifically, morphological traits of P. delavayi populations growing in two different habitats (i.e., the open area versus forest understory), including basal stem diameter, plant height and flower numbers per plant, were measured and compared with each other. A range of floral manipulation experiments was further conducted to study the extent of self-compatibility and pollen limitation. The results showed that P. delavayi preferred the open habitat over forest understory as it generally grew bigger with thicker stems, and produced a higher number of flowers in the former habitat. Therefore, the open habitat should receive immediate priority for conservation action instead of being converted to agricultural land; also, P. delavayi is pollen limited, as evidenced by the fact that experimental flowers receiving supplemental non-self pollen generally displayed elevated seed production than flowers subjected to other treatments (e.g., non-supplemental self pollen, non-supplemental non-self pollen and supplemental self pollen), which is consistent with reported observations that alpine plants are more likely to be pollen-limited than lowland plants. We suggest that human intervention might be necessary to guarantee the long-term persistence of P. delavayi as harsh alpine environment, intense competition for pollinators and different anthropogenic perturbations co-limit its reproductive success.     

Equality testing for soil grid unit resolutions to polygon unit scales with DNDC modeling of regional SOC pools

Matching soil grid unit resolutions with polygon unit map scales is important to minimize the uncertainty of regional soil organic carbon(SOC) pool simulation due to their strong influences on the modeling.A series of soil grid units at varying cell sizes was derived from soil polygon units at six map scales,namely,1:50 000(C5),1:200 000(D2),1:500 000(P5),1:1 000 000(N1),1:4 000 000(N4) and 1:14 000 000(N14),in the Taihu Region of China.Both soil unit formats were used for regional SOC pool simulation with a De Nitrification-DeC omposition(DNDC) process-based model,which spans the time period from 1982 to 2000 at the six map scales.Four indices,namely,soil type number(STN),area(AREA),average SOC density(ASOCD) and total SOC stocks(SOCS) of surface paddy soils that were simulated by the DNDC,were distinguished from all these soil polygon and grid units.Subjecting to the four index values(IV) from the parent polygon units,the variations in an index value(VIV,%) from the grid units were used to assess its dataset accuracy and redundancy,which reflects the uncertainty in the simulation of SOC pools.Optimal soil grid unit resolutions were generated and suggested for the DNDC simulation of regional SOC pools,matching their respective soil polygon unit map scales.With these optimal raster resolutions,the soil grid units datasets can have the same accuracy as their parent polygon units datasets without any redundancy,when VIV 1% was assumed to be a criterion for all four indices.A quadratic curve regression model,namely,y = – 0.80 × 10~(–6)x~2 + 0.0228 x + 0.0211(R~2 = 0.9994,P 0.05),and a power function model R? = 10.394?~(0.2153)(R~2 = 0.9759,P 0.05) were revealed,which describe the relationship between the optimal soil grid unit resolution(y,km) and soil polygon unit map scale(1:10 000x),the ratio(R?,%) of the optimal soil grid size to average polygon patch size(?,km~2) and the ?,with the highest R~2 among different mathematical regressions,respectively.This knowledge may facilitate the grid partitioning of regions during the investigation and simulation of SOC pool dynamics at a certain map scale,and be referenced to other landscape polygon patches' mesh partition.     

Age-related changes of leaf traits and stoichiometry in an alpine shrub (<Emphasis Type="Italic">Rhododendron agglutinatum</Emphasis>) along altitudinal gradient

Leaf morphological and stoichiometric characteristics are considered to represent both the interior inheritable characters in the plant and its adaptations to specific exterior environments. Rhododendron agglutinatum, an evergreen alpine shrub species, occupies a wide range of habitats above timberline in the Miyaluo Natural Reserve, southwestern China. Along an altitudinal gradient ranging from 3700 to 4150 m, we measured leaf morphological characters including leaf dry matter content (LDMC), leaf dry mass per unit area (LMA), and one leaf area (OLA), as well as carbon (C) and nutrient (N, P) contents in leaves of three different age groups (juvenile leaves, mature leaves and senescent leaves). We also calculated the stoichiometric relationships among carbon and nutrients (C/N, C/P and N/P). Results showed that both age and altitude affected the leaf morphological and stoichiometric properties of R. agglutinatum. Mature leaves possessed the highest LDMC, LMA and C contents both on a dry mass basis and on a unit area basis. Younger leaves possessed higher contents of nutrients. OLA as well as ratios between carbon and nutrients (C/N, C/P) increased with ages. Juvenile leaves possessed lowest ratio between nitrogen and phosphorus. In juvenile leaves, nutrients increased with altitudinal elevation, whereas other traits decreased. In mature leaves, nutrients and their ratios with carbon showed consistent trends with juvenile leaves along increasing altitude, whereas LMA and carbon on a unit area basis showed opposite trends with juvenile leaves along increasing altitude. In senescent leaves, only content of phosphorus on a unit area basis and N/P were found linearly correlated with altitude. Our results demonstrated a clear pattern of nutrient distribution with aging process in leaves and indicated that a high possibility of N limitation in this region. We also concluded that younger leaves could be more sensitive to climate changes due to a greater altitudinal influence on the leaf traits in younger leaves than those in elder leaves.     

Interspecific competition and allelopathic interaction between <Emphasis Type="Italic">Karenia mikimotoi</Emphasis> and <Emphasis Type="Italic">Dunaliella salina</Emphasis> in laboratory culture

Algal allelopathy is a manifold ecological/physiological phenomenon that is focused on chemical interactions and autotoxicity. We investigated the allelopathic interactions between Karenia mikimotoi and Dunaliella salina in laboratory cultures based on diff erent temperature (15°C, 20°C, and 25°C) and lighting (40, 80, and 160 μmol/(m²·s)) conditions. The growth of D. salina in bi-algae culture (1:1 size/density) was significantly restrained. The results of cell-free filtrate culture indicate that direct cell-tocell contact was not necessary in interspecific competition. Further experimental results demonstrated that allelochemicals released from K. mikimotoi were markedly influenced by both temperature (P =0.013) and irradiance (P =0.003), resulting in diff erent growth characteristics of D. salina in filtrate mediums. Compared with the plateau period, K. mikimotoi exudates in the exponential phase had a stronger short-term inhibition effect on D. salina in normal conditions. A clear concentration-dependent relationship was observed in the effect of allelochemicals released from K. mikimotoi with low-promoting and high-repressing effects on D. Salina in a short time-scale. In addition, allelopathic substances remain stable and effective under high temperature and pressure stress. Many flocculent sediments adhering with D. salina cells were observed in all filtrate mediums, while the quantity and color depended on the original culture conditions.     

Heavy metals in sediments and their bioaccumulation in <Emphasis Type="Italic">Phragmites australis</Emphasis> in the Anzali wetland of Iran

Accumulation of metals in both sediments and Phragmites australis organs was studied. Samples were collected from seven stations located in Anzali wetland, Iran. The samples were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). The results showed that concentration of the studied metals (except As and Cd) were higher in sediments than in P. australis organs. Metal accumulation was found to be significantly (P <0.05) higher in roots than in above-ground organs of P. australis. The bioaccumulation factor (BAF) and the transfer factor (TF) also verified the highest rate of metal accumulation in roots and their reduced mobility from roots to the above-ground organs. Pearson correlation coefficient showed significant relationships between metal concentrations in sediments and those in plant organs. It should be pointed out that sediment and plant samples exhibited higher metal concentrations in eastern and central parts than in western and southern parts of the wetland. The mean concentrations of all studied elements (except for Fe, V and Al) were higher in these sediment samples than in the Earth’s crust and shale. High accumulation of metals in P. australis organs (roots and shoots) is indicative of their high bioavailability in sediments of the wetland. The correlation between metal concentrations in sediments and in P. australis indicates that plant organs are good bioindicators of metal pollution in sediments of Anzali wetland.     

Soil nutrients in relation to vertical roots distribution in the riparian zone of Three Gorges Reservoir,China

Since the impoundment of the Three Gorges Reservoir (TGR), the riparian zone has been subjected to numerous environmental changes. This study was conducted to recognize the distribution of grass roots and its impacts on soil nutrients in the water level fluctuation zone of TGR. Roots of four predominant herbaceous plants in the study area, specifically, Cynodon dactylon, Hemarthria altissima, Hemarthria compressa, and Paspalum paspaloides, and their corresponding relation with soil nutrient contents were investigated. Root surface area density was determined with WinRHIZO, and the relationships of root distribution with soil depths and soil nutrient contents were studied. The results indicates that most roots are distributed in the top soil layer of 0–10 cm. Estimated root surface area density for the selected grass species ranges from 0.16 to 13.44 cm2/cm3, and decreases exponentially with an increase in soil depth. Soil organic matter and total nitrogen contents are significantly lower on bare control area than the corresponding values on the grasslands. Total nutrient contents on grasslands of C. dactylon and H. compressa are higher than those of other grass areas. Root length density and root surface area density are significantly correlated with soil organic matter and total nitrogen content for the four grasslands. The present results suggests that plant roots have significant effects on the distribution of soil nutrients in soil profiles in the riparian zone along the TGR. Nevertheless, additional investigations are needed to reveal the specific interactions between plant roots distribution, soil nutrients and water level fluctuations.     

Lowland pasture in Himalayan highland: edaphic properties and species composition

This study was carried out to investigate if on-farm cut vs. grazed pastures differed in soil fertility and species density in the temperate Himalayan highlands. Soil fertility and species relative density were measured from 30 cut and 30 grazed dairy pastures. In both types of pasture, soil pH, available phosphorus (P) and exchangeable potassium (K) were negatively correlated with pasture age and slope. In cut pasture, cocksfoot (Dactylis glomerata) and pasture age were positively correlated, whereas in grazed pasture, they were negatively correlated. In grazed pasture, unsown species and pasture age were positively correlated. Soil available P was significantly greater in cut pastures whereas soil exchangeable K was significantly greater in grazed pastures. In terms of species density, cut pasture had greater densities of cocksfoot and Italian ryegrass (Lolium multiflorum), whereas grazed pasture showed greater densities of white clover, sedge and local grass. Our study suggests that, if there are no improvements in the current method of pasture management, the cut pastures in the future are likely to have a simple pasture mixture constituting only cocksfoot and white clover (Trifolium repens). Whereas in grazed pasture, the pasture mixture is likely to be comprised of white clover and unsown species such as sedge, local grass and broadleaf weeds.     

Comparison of artificial neural networks,geographically weighted regression and Cokriging methods for predicting the spatial distribution of soil macronutrients (N,P, and K)

Soil macronutrients(i.e. nitrogen(N), phosphorus(P), and potassium(K)) are important soils components and knowing the spatial distribution of these parameters are necessary at precision agriculture. The purpose of this study was to evaluate the feasibility of different methods such as artificial neural networks(ANN) and two geostatistical methods(geographically weighted regression(GWR) and cokriging(CK)) to estimate N, P and K contents. For this purpose, soil samples were taken from topsoil(0–30 cm) at 106 points and analyzed for their chemical and physical parameters. These data were divided into calibration(n = 84) and validation(n = 22). Chemical and physical variables including clay, p H and organic carbon(OC) were used as auxiliary soil variables to estimate the N, P and K contents. Results showed that the ANN model(with coefficient of determination R~2 = 0.922 and root mean square error RMSE = 0.0079%) was more accurate compared to the CK model(with R~2 = 0.612 and RMSE = 0.0094%), and the GWR model(with R~2 = 0.872 and RMSE = 0.0089%) to estimate the N variable. The ANN model estimated the P with the RMSE of 3.630 ppm, which was respectively 28.93% and 20.00% less than the RMSE of 4.680 ppm and 4.357 ppm from the CK and GWR models. The estimated K by CK, GWR and ANN models have the RMSE of 76.794 ppm, 75.790 ppm and 52.484 ppm. Results indicated that the performance of the CK model for estimation of macro nutrients(N, P and K) was slightly lower than the GWR model. Also, the accuracy of the ANN model was higher than CK and GWR models, which proved to be more effective and reliable methods for estimating macro nutrients.     

Trophic Interaction in a <Emphasis Type="Italic">Portunus rituberculatus</Emphasis> Polyculture Ecosystem Based on Carbon and Nitrogen Stable Isotope Analysis

Trophic interaction among various biomass groups in a swimming crab Portunus trituberculatus polyculture pond was investigated using carbon and nitrogen stable isotope analysis. The polycultured animal species also included white shrimp Litopenaeus vannamei, short-necked clam Ruditapes philippinarum, and redlip mullet Liza haematochila. The mean δ¹³C value for all the biomass groups in polyculture ecosystem ranged from ?25.61‰ to ?16.60‰, and the mean δ¹⁵N value ranged from 6.80‰ to 13.09‰. Significant difference in the δ¹³C value was found between particulate organic matter (POM) and sediment organic matter (SOM) (P < 0.05), indicating that these two organic matter pools have different material sources. Assuming that a ¹³C-enrichment factor of 1.00‰ and a ¹⁵N-enrichment factor of 2.70‰ existed between consumer and prey, diets of the four cultured animals were estimated using a stable isotope mixing model. The estimated model results indicated that P. trituberculatus mainly feed on Aloidis laevis; L. vannamei mainly feed on shrimp feed; while A. laevis, R. philippinarum and L. haematochelia mainly feed on POM. Shrimp feed was also an important food source of R. philippinarum and L. haematochelia. The diets of P. trituberculatus, L. vannamei, R. philippinarum, and L. haematochila showed complementary effects in this polyculture ecosystem. Our finding indicated that the polyculture of these four organisms with suitable farming density could make an effective use of most of the food sources, which can make a highly efficient polyculture ecosystem.     

Screening and isolation of the algicidal compounds from marine green alga <Emphasis Type="Italic">Ulva intestinalis</Emphasis>

Twenty species of seaweed were collected from the coast of Zhejiang, China, extracted with ethanol, and screened for algicidal activity against red tide microalgae Heterosigma akashiwo and Prorocentrum micans. Inhibitory effects of fresh and dried tißsues of green alga Ulva intestinalis were assessed and the main algicidal compounds were isolated, purified, and identified. Five seaweed species, U. intestinalis, U. fasciata, Grateloupia romosissima, Chondria crassicaulis, and Gracilariopsis lemaneiformis, were investigated for their algicidal activities. Fresh tissues of 8.0 and 16.0 mg/mL of U. intestinalis dissolved in media significantly inhibited growth of H. akashiwo and P. micans, respectively. Dried tissue and ethyl acetate (EtOAc) extracts of U. intestinalis at greater than 1.2 and 0.04 mg/mL, respectively, were fatal to H. akashiwo, while its water and EtOAc extracts in excess of 0.96 and 0.32 mg/mL, respectively, were lethal to P. micans. Three algicidal compounds in the EtOAc extracts were identified as 15-ethoxy-(6z,9z,12z)-hexadecatrienoic acid (I), (6E,9E,12E)-(2-acetoxy-β-D-glucose)-octadecatrienoic acid ester (II) and hexadecanoic acid (III). Of these, compound II displayed the most potent algicidal activity with IC₅₀ values of 4.9 and 14.1 µg/mL for H. akashiwo and P. micans, respectively. Compound I showed moderate algicidal activity with IC₅₀ values of 13.4 and 24.7 µg/mL for H. akashiwo and P. micans, respectively. These findings suggested that certain macroalgae or products therefrom could be used as effective biological control agents against red tide algae.     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.