Plasma free amino acid kinetics in rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) using a bolus injection of <Superscript>15</Superscript>N-labeled amino acids

To gain insight into the metabolic design of the amino acid carrier systems in fish, we injected a bolus of ¹⁵N amino acids into the dorsal aorta in mature rainbow trout (Oncorhynchus mykiss). The plasma kinetic parameters including concentration, pool size, rate of disappearance (R _d), half-life and turnover rate were determined for 15 amino acids. When corrected for metabolic rate, the R _d values obtained for trout for most amino acids were largely comparable to human values, with the exception of glutamine (which was lower) and threonine (which was higher). R _d values ranged from 0.9 μmol 100 g⁻¹ h⁻¹ (lysine) to 22.1 μmol 100 g⁻¹ h⁻¹ (threonine) with most values falling between 2 and 6 μmol 100 g⁻¹ h⁻¹. There was a significant correlation between R _d and the molar proportion of amino acids in rainbow trout whole body protein hydrolysate. Other kinetic parameters did not correlate significantly with whole body amino acid composition. This indicates that an important design feature of the plasma-free amino acids system involves proportional delivery of amino acids to tissues for protein synthesis.     

Detection Methanogens in Newly Settled Sediments from Xuanwu Lake in Nanjing, China

Sediments from Xuanwu Lake have been dredged in the past 3 years to improve the water quality, but methanogenesis should still exist in the newly settled sediment. Methane production, methanogens, and physiochemical parameters were detected in the surface sediments (0–5 cm) and/or vertical sediments (0–21 cm, segmented at interval of 3 cm). Methane flux at water–air interface varied among five detected sites. Principal component analysis showed that CH₄ flux, content of water and the concentration of total nitrogen (TN), CH₄ and organic matters (OM) weighed most heavily on the component I in surface sediments while different patterns were observed for vertical sediments. The copy number of the 16S rRNA gene for bacteria was lower in the surface sediment (0–6 cm) than that in deeper sediments (12–21 cm), while 16S rRNA genes of Archaea were almost evenly distributed in the vertical sediments. Representatives belonging to the orders Methanobacteriales, Methanomicrobiales, and Methanosarcinales were detected in all samples of the vertical sediments, except that no members of the Methanococcales were detected in the samples at 0–6 cm. The level of Methanobacteriales reached a highest density at 18.1 × 10⁴ copies g⁻¹ dry weight (dw) at 6–9 cm; for Methanosarcinales (76.89 × 10⁶ copies g⁻¹ dw) and Methanococcales (82.70 × 10³ copies g⁻¹ dw) at 12–15 cm, whereas for Methanomicrobiales (43.37 × 10⁶ copies g⁻¹ dw) at 9–12 cm. Methanosarcinaceae and Methanosaetaceae reached to their highest densities at 6–9 and 9–12 cm, respectively. These data provided useful information for better understanding the methanogenesis in the newly settled sediments of a recently dredged lake.     

Nutrient content of tropical edible seaweeds, Eucheuma cottonii, Caulerpa lentillifera and Sargassum polycystum

The proximate composition, vitamin C, α-tocopherol, dietary fibers, minerals, fatty acid and amino acid profiles of three tropical edible seaweeds, Eucheuma cottonii (Rhodophyta), Caulerpa lentillifera (Chlorophyta) and Sargassum polycystum (Phaeophyta) were studied. The seaweeds were high in ash (37.15–46.19%) and dietary fibers (25.05–39.67%) and low in lipid content (0.29–1.11%) on dry weight (DW) basis. These seaweeds contained 12.01–15.53% macro-minerals (Na, K, Ca and Mg) and 7.53–71.53 mg.100 g⁻¹ trace minerals (Fe, Zn, Cu, Se and I). The crude protein content of E. cottonii (9.76% DW) and C. lentillifera (10.41% DW) were higher than that of S. polycystum (5.4% DW), and protein chemical scores are between 20 and 67%. The PUFA content of E. cottonii was 51.55%, C. lentillifera 16.76% and S. polycystum 20.34%. Eicosapentaenoic acid (EPA), accounted for 24.98% of all fatty acids in E. cottonii. These seaweeds have significant vitamin C (∼35 mg.100 g⁻¹) and α-tocopherol (5.85–11.29 mg.100 g⁻¹) contents.     

Optimization of probiotic and lactic acid production by Lactobacillus plantarum in submerged bioreactor systems

Biomass and lactic acid production by a Lactobacillus plantarum strain isolated from Serrano cheese, a microorganism traditionally used in foods and recognized as a potent probiotic, was optimized. Optimization procedures were carried out in submerged batch bioreactors using cheese whey as the main carbon source. Sequential experimental Plackett–Burman designs followed by central composite design (CCD) were used to assess the influence of temperature, pH, stirring, aeration rate, and concentrations of lactose, peptone, and yeast extract on biomass and lactic acid production. Results showed that temperature, pH, aeration rate, lactose, and peptone were the most influential variables for biomass formation. Under optimized conditions, the CCD for temperature and aeration rate showed that the model predicted maximal biomass production of 14.30 g l⁻¹ (dw) of L. plantarum. At the central point of the CCD, a biomass of 10.2 g l⁻¹ (dw), with conversion rates of 0.10 g of cell g⁻¹ lactose and 1.08 g lactic acid g⁻¹ lactose (w/w), was obtained. These results provide useful information about the optimal cultivation conditions for growing L. plantarum in batch bioreactors in order to boost biomass to be used as industrial probiotic and to obtain high yields of conversion of lactose to lactic acid.     

Seasonal changes in growth rate, carrageenan yield and lectin content in the red alga Kappaphycus alvarezii cultivated in Camranh Bay, Vietnam

The three color morphotypes of the red alga Kappaphycus alvarezii (brown, red and green) were cultured in Camranh Bay, Vietnam, using the fixed off-bottom monoline culture method to evaluate the growth rate, carrageenan yield, 3,6-anhydrogalactose, gel strength and lectin content. The brown morphotype was cultivated over a 12-month period; the red and green morphotypes were over a 6-month period. At the 60-day culture timepoint, the brown morphotype showed a higher growth rate (3.5–4.6% day⁻¹) from September to February, and lower growth rate (1.6–2.8% day⁻¹) from March to August. Significant (P < 0.05) differences in growth rate between culture months were found with the brown morphotype. High growth rates for the red (3.6–4.4% day⁻¹) and green (3.7–4.2% day⁻¹) morphotypes were obtained from September to February. The carrageenan yield, 3,6-anhydrogalactose and gel strength of the three morphotypes showed little variation, with the highest values obtained in November–December. At the 30-day sampling point, the brown morphotype had a higher lectin content (167–302 μg g⁻¹ dry alga) from August to March and a lower lectin content (23–104 μg g⁻¹ dry alga) from April to July. High lectin contents were recorded for the red (139–338 μg g⁻¹ dry alga) and green (124–259 μg g⁻¹ dry alga) morphotypes from September to February. This study shows that the different morphotypes of K. alvarezii can be grown in the tropical waters of the Camranh during the northeast monsoon, and part of the southwest monsoon, especially the brown morphotype, which can be grown during any season.     

Hormone Profiling by LC-QToF-MS/MS in Dormant <Emphasis Type="Italic">Macadamia integrifolia</Emphasis>: Correlations with Abnormal Vertical Growth

A method for analyzing multiple plant hormone groups in small samples with a complex matrix was developed to initiate a study of the physiology of abnormal vertical growth (AVG) in Macadamia integrifolia (cv. HAES344). Cytokinins (CKs), gibberellins (GAs), abscisic acid (ABA), and auxins were detected in xylem sap and apical and lateral buds using high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QToF-MS/MS). The extraction method separated compounds with high sensitivity in positive (CKs) and negative (ABA, auxins, GAs) modes of QToF-MS/MS. CK profiles differed in xylem sap and apical and lateral buds irrespective of AVG symptoms. Trans-zeatin riboside (t-ZR) was dominant in sap of normal and AVG trees (∼4 and 6 pmol g⁻¹ FW, respectively). In apical buds isopentenyl adenine (iP) (∼30 pmol g⁻¹ FW) was the most abundant CK, and in lateral buds trans-zeatin (t-Z) (22–24 pmol g⁻¹ FW) and iP (24–30 pmol g⁻¹ FW) were the most abundant. t-Z levels of AVG trees were higher in apical buds (13.88 vs. 6.6 pmol g⁻¹ FW, p < 0.05) and lower in sap (0.16 vs. 0.51 pmol ml⁻¹, p < 0.005) compared to normal trees. ABA in lateral buds was 1.9 times higher (p < 0.001) in AVG. IAA was below quantification, whereas indole-3-butyric acid (IBA) was consistently present. GA₇ was the dominant GA in apical and lateral buds of all trees (100–150 pmol g⁻¹ FW). GA_{3, 4, & 9} were consistently present at low concentrations (<12 pmol g⁻¹ FW) in buds. GAs_{1, 3, & 9} were detected in xylem sap at low concentrations (<0.5 pmol g⁻¹ FW). Differences in sap amino acids (AA) were also assessed. In sap from AVG trees, asparagine and glutamine increased significantly (p < 0.05) in their contribution to total AA. Potential AVG hormone correlations are discussed.     

Phenylpropanoid production in callus and cell suspension cultures of <Emphasis Type="Italic">Buddleja cordata</Emphasis> Kunth

Plant tissue cultures represent a potential source for producing secondary metabolites. In this work, Buddleja cordata tissue cultures were established in order to produce phenylpropanoids (verbascoside, linarin and hydroxycinnamic acids), as these metabolites are credited with therapeutic properties. Highest callus induction (76.4–84.3%) was obtained in five treatments containing 2,4-Dichlorophenoxyacetic acid (2,4-d: 0.45–9.05 μM) with Kinetin (KIN: 2.32, 4.65 μM), whereas highest root induction (79.6%) corresponded to the α-Naphthaleneacetic acid (9.05 μM) with KIN (2.32 μM) treatment. Verbascoside was the major phenylpropanoid produced in in vitro cultures (root, white and green callus) [66.24–86.26 mg g⁻¹ dry weight (DW)], while linarin and hydroxycinnamic acid production was low (0.95–3.01 mg g⁻¹ DW). Verbascoside and linarin production were improved in cell suspension culture (116 mg g⁻¹ DW and 8.12 mg g⁻¹ DW, respectively).     

Changes in fatty acids and sterols during batch growth of <Emphasis Type="Italic">Pavlova viridis</Emphasis> in photobioreactor

Changes in the composition of fatty acids and sterols of Pavlova viridis cultured in an air-lift photobioreactor were studied using gas chromatography-mass spectrometry (GC-MS). The results show that radical changes in fatty acid and sterol contents and compositions occurred during growth phase transitions: the total lipid increased along with the culture age, from 166.4 mg g⁻¹ (late exponential phase) to 232.7 mg g⁻¹ (linear phase), and increased further to be 235.1 mg g⁻¹ in the stationary phase. Polyunsaturated fatty acids (PUFAs), especially eicosapentaenoic acid (EPA), decreased along with the culture time, PUFAs, and EPA contents maximized in the late exponential phase to become 46.2 mg g⁻¹ and 22.1 mg g⁻¹ respectively; there was no significant change in docosahexaenoic acid (DHA) content during the whole growth phase, although it reached the peak in the linear phase with 3.5 mg g⁻¹. As for the sterols, two unique sterols with two hydroxyl groups, termed pavlovols, were observed. 4α,24-Dimethylcholestan-3β,4β-diol, one of the pavlovols, increased almost 2-fold from the late exponential phase (2.5 mg g⁻¹) to the stationary phase (4.3 mg g⁻¹). On the contrary, the contents of stigmasterol and sitosterol decreased with culture age, with the maximum content of 2.4 mg g⁻¹ and 3.1 mg g⁻¹, both obtained in the late exponential phase, respectively. The results indicate that growth phase control could be used as a methodology to optimize the total lipid, EPA, PUFA, and sterol contents with the potential for both aquaculture feeds and nutraceutical applications, especially for further research into unique pavlovols.     

Effect of light intensity on the proximate biochemical and fatty acid composition of Isochrysis sp. and Nannochloropsis oculata for use in tropical aquaculture

The total protein, carbohydrate, lipid and ash compositions, and fatty acid contents of two species of marine microalgae, the eustigmatophyte Nannochloropsis oculata (formerly ‘Chlorella sp., Japan’) and the chrysophyte Isochrysis sp. (Tahitian) used in tropical Australian mariculture, were studied. The microalgae were grown under a range of culture conditions (41 and 601 laboratory culture, 3001 bag culture, and 80001 outdoor culture) and four light regimes (100 to 107 μ E m⁻² s⁻¹, 240 to 390 μ E m⁻² s⁻¹, 340 to 620 μ E m⁻² s⁻¹, and 1100 to 1200 μE m⁻² s⁻¹ respectively) to determine the effect of light intensity on the chemical composition of large scale outdoor cultures. Laboratory and bag cultures were axenic and cultured in Walne medium while outdoor cultures were grown in a commercial medium designed for optimum nutrition in tropical outdoor aquaculture operations. Change in growth medium and photon flux density produced only small changes in the proximate biochemical composition of both algae. N. oculata and Isochrysis sp. both showed a trend towards slightly lower carbohydrate and higher chlorophyll a in shaded outdoor culture. Isochrysis sp. showed significant concentrations of the essential polyunsaturated fatty acid 22:6(n−3) (docosahexaenoic acid) from 5.3 to 10.3% of total fatty acid, and 20:5(n−3) (eicosapentaenoic acid) ranged from 0.6 to 4.1%. In contrast, N. oculata had high concentrations of 20:5(n−3) (17.8 to 39.9%) and only traces of 22:6(n−3). The fatty acid composition of Isochrysis sp. grown at high photon flux density (1100–1200 μE m⁻² s⁻¹) under outdoor culture showed a decrease in the percentage of several highly unsaturated fatty acids, including 20:5(n−3), and an increase in 22:6(n−3). N. oculata showed a similar decrease in the percentage of 20:5(n−3). High light intensity caused a decrease in the ratio of total C₁₆ unsaturated fatty acids to saturated 16:0 in N. oculata, and a decrease in the ratio of total C₁₈ unsaturated fatty acids to saturated 18:0 together with a decrease in the ratio of total unsaturated fatty acids to total saturated fatty acids in both microalgae.     

The biosynthetic pathway of carotenoids in the astaxanthin-producing green alga <Emphasis Type="Italic">Chlorella zofingiensis</Emphasis>

The carotenoid composition of the astaxanthin-producing green alga Chlorella zofingiensis was investigated using high-performance liquid chromatography. Astaxanthin, adonixanthin, and zeaxanthin are the major carotenoids in this alga. The pigment pattern was characterized during the accumulation period, and in response to diphenylamine (DPA), an inhibitor of carotenoid biosynthesis. An increase in zeaxanthin followed by a decrease in xanthophyll was seen after the induction of astaxanthin biosynthesis by glucose. This biphasic kinetics of zeaxanthin was parallel to the marked increase in adonixanthin (from 0 mg g⁻¹ to 0.21 mg g⁻¹) and astaxanthin (from 0.05 mg g⁻¹ to 0.35 mg g⁻¹) and decrease of β-carotene (from 0.30 mg g⁻¹ to 0.03 mg g⁻¹). More importantly, unlike the Haematococcus alga, in which there was a high β-carotene accumulation after DPA treatment, C. zofingiensis showed an accumulation of extra zeaxanthin instead of β-carotene after treatment of the cells with DPA. All these results observed in vivo studies corroborate the observations in vitro studies at the enzyme level that zeaxanthin is a substrate for the carotenoid oxygenase in C. zofingiensis. It is suggested that zeaxanthin might be an important intermediate and not an end product of the biosynthetic pathway of astaxanthin. Therefore, a new pathway for astaxanthin formation by C. zofingiensis, which is different from that of the other astaxanthin-producing microorganisms, is proposed. An understanding of the astaxanthin biosynthetic pathway may yield important information toward the optimization of astaxanthin production, especially for the improvement of astaxanthin through genetic manipulations.     

Cadmium Accumulation in Relation to Organic Acids and Nonprotein Thiols in Leaves of the Recently Found Cd Hyperaccumulator <Emphasis Type="Italic">Rorippa globosa</Emphasis> and the Cd-accumulating Plant <Emphasis Type="Italic">Rorippa islandica</Emphasis>

It has been proposed that organic acid and nonprotein thiol are involved in the hyperaccumulation of metals. In this study, Cd accumulation, organic acid, and nonprotein thiol production and their relationships in the leaves of Cd-hyperaccumulator Rorippa globosa were examined and compared with a closely related species, Rorippa islandica. The results showed that there was no reduction in biomass of R. globosa when treated with 25 μg Cd g⁻¹ (T₂), despite Cd accumulation in the leaves was up to 158.2 μg g⁻¹ DW. On the other hand, the growth of Cd-treated R. islandica was obviously inhibited as it accumulated more than 100 μg g⁻¹ DW in the leaves. Therefore, R. islandica behaved as a Cd-accumulating plant. The Cd treatments could significantly induce the synthesis of acetic acid in both species, suggesting that acetic acid, as the most abundant organic acid, might be related to the Cd accumulation. Significant positive correlations between Cd concentrations and both tartaric and malic acid concentrations in the leaves of R. globosa were observed. There was a significant positive correlation between Cd concentrations and acetic acid concentrations in the leaves of R. islandica. This trend of tartaric and malic acids in the leaves of R. globosa and acetic acid in the leaves of R. islandica might be related to Cd accumulation. In addition, a quadratic relationship was obviously observed for NP-SH contents and total Cd concentrations in the leaves of R. globosa, indicating that NP-SH was significantly related to Cd accumulation and tolerance.     

Hydrophobic response of the fungus <Emphasis Type="Italic">Rhinocladiella similis</Emphasis> in the biofiltration with volatile organic compounds with different polarity

Rhinocladiella similis biodegraded volatile organic compounds (VOCs) of different polarity in gas-phase biofilters. Elimination capacities, (EC) of 74 g_hexane m⁻³ h⁻¹, 230 g_ethanol m⁻³ h⁻¹, 85 g_toluene m⁻³ h⁻¹ and 30 g_phenol m⁻³ h⁻¹ were obtained. EC values correlated with the solubility of the VOCs. R. similis grown with n-hexane or ethanol in biofilters packed with Perlite showed that the surface hydrophobicity was higher with n-hexane than ethanol. The hydrophobin-like proteins extracted from the mycelium produced with n-hexane (15 kDa) were different from those in the ethanol biofilter (8.5 kDa and 7 kDa).     

Mercury contamination and life history traits of Allis shad <Emphasis Type="Italic">Alosa alosa</Emphasis> (Linnaeus, 1758) and Twaite shad <Emphasis Type="Italic">Alosa fallax</Emphasis> (Lacépède, 1803) in the Gironde estuary (South West France)

Mercury concentration [Hg] was assessed in 20 adult Allis shad Alosa alosa (54–59 cm) and 20 adult Twaite shad Alosa fallax (36–44 cm) collected during their spawning migration in the Dordogne and the Garonne rivers (France). [Hg] was measured in the gills, dorsal muscle, liver and kidney. Twaite shad exhibited higher [Hg] than Allis shad. Median [Hg] values were [Hg]_Gills = 0.33 μg g⁻¹ (dw), [Hg]_Muscle = 1.22 μg g⁻¹, [Hg]_Liver = 1.99 μg g⁻¹, [Hg]_Kidney = 1.93 μg g⁻¹ for Twaite shad and [Hg]_Gills = 0.06 μg g⁻¹, [Hg]_Muscle = 0.20 μg g⁻¹, [Hg]_Liver = 1.18 μg g⁻¹, [Hg]_Kidney = 1.08 μg g⁻¹ for Allis shad. In order to understand such differences, we investigated some life history traits of the two species: migratory history, age (3–6 years), size at age (an expression of growth) and the number of spawning events (0–2 events). The difference in estuarine residence time between the juveniles of both species, which was assumed to influence [Hg], was investigated using otolith Sr:Ca ratio. The microchemical analysis revealed a significant difference in the residence time of juveniles in the estuary (medians are 21 d and 10 d for Twaite shad and Allis shad, respectively) but this residence time seems too short to influence [Hg] (Allis shad: r _spearman < 0.128; Twaite shad: r _spearman < −0.340). As both species show the same age structure, the influence of age on [Hg] was negligible. The literature shows that the differences in growth and in the number of spawning events reported in our study are in favour of a higher [Hg] for Twaite shad than for Allis shad. Although trophic status was not investigated here, the literature reveals that it is another factor that could produce higher [Hg] in Twaite shad, since its diet includes higher trophic levels than Allis shad. Guest editors: S. Dufour, E. Prévost, E. Rochard & P. Williot Fish and diadromy in Europe (ecology, management, conservation)     

Cadmium accumulation in relation to organic acids in leaves of Solanum nigrum L. as a newly found cadmium hyperaccumulator

The influence of various cadmium concentrations on organic acid levels in leaves of the Cd hyperaccumulator, Solanum nigrum L. and a closely related species, Solanum melongena L., were investigated. In particular, the relationship of organic acids with Cd accumulation in the two plants was investigated. The results showed that Cd accumulation in the shoots of S. nigrum was significantly higher than that of S. melongena. The accumulation of Cd in the leaves of S. nigrum ranged from 2.0 to 167.8 μg g⁻¹ dry weight (DW), but only from 1.2 to 64.0 μg g⁻¹ DW in S. melongena. Solanum melongena was considerably less tolerant to Cd than S. nigrum. Approximately 20% of the total Cd in S. nigrum leaves was water-soluble, suggesting that some accumulated Cd was associated with water-soluble compounds such as organic acids. Malic acid in the leaves of S. nigrum was the most abundant organic acid [up to 115.6–145.7 μmol g⁻¹ fresh weight (FW)], but this acid was not significantly affected by the Cd concentration in soil. However, the level of malic acid in S. melongena plants was much lower, only 16.3–75.4 μmol g⁻¹ FW. The significant positive correlations between total Cd and water-soluble Cd concentrations and both acetic and citric acid concentrations in the leaves of S. nigrum were observed. In contrast, there was no correlation between concentrations of the two acids and Cd concentrations in the leaves of S. melongena. These results indicated that acetic and citric acids in the leaves of S. nigrum might be related to its Cd hyperaccumulation.     

Nutrient and hormone levels in cotton ovules during embryony

In vitro zygotic and somatic embryogenesis protocols rely on nutrient and hormone levels from media to satisfy the physiological and developmental requirements of embryony. To better understand these requirements for cotton, we quantified levels of major and minor elements, carbohydrates, NH₄ ⁺, free amino acids and six hormones in whole cotton ovules (with fibers removed), nucelli (ovules with integuments removed), or ovule fluid (extracted from the endosperm region). Samples were collected from field-grown cotton at 1–18 days-past-anthesis (DPA) during each of three growing seasons. Replication across 2 years was obtained for carbohydrates, NH₄ ⁺, free amino acids and hormones from nucellus samples. The year effect was large primarily for hormones only. The most abundant minerals across tissue types and years were K, P, Mg and S. Potassium was the most abundant at 260, 600 and 1,660 mmol kg⁻¹ dry mass (DM) in nucelli, whole ovules and ovule fluid, respectively. Magnesium, Ca, Zn and Mn levels were 2–8-fold higher in ovule fluid compared to whole ovules or nucelli. In the free amino acid plus NH₄ ⁺ category, NH₄ ⁺, alanine, serine, glycine, asparagine (plus aspartic acid), glutamine (plus glutamic acid), leucine, threonine and arginine predominated in nucelli and ovule fluid, and levels tended to be higher in the older samples across years and tissue types. Fructose and glucose levels also increased with age with very high levels being found in late DPA ovule fluid. Arabinose, inositol and melibiose were also prominent sugars. Indole-3-acetic acid levels were similar between nucelli and ovule fluid and ranged from 10 to 80 μmol kg⁻¹ DM. An abscisic acid spike, from 15 to 400 μmol kg⁻¹ DM, occurred in nucelli and whole ovules from 2 to 8 DPA. Thereafter, abscisic acid levels remained between 5 and 10 μmol kg⁻¹ DM. Zeatin and zeatin riboside were the most abundant cytokinins, and levels of these hormones fluctuated between 1 and 4 μmol kg⁻¹ DM in both nucelli and ovule fluid.     

<Emphasis Type="Italic">Fusarium</Emphasis> species (section <Emphasis Type="Italic">Liseola</Emphasis>) occurrence and natural incidence of beauvericin,fusaproliferin and fumonisins in maize hybrids harvested in Mexico

Fusarium species can produce fumonisins (FBs), fusaric acid, beauvericin (BEA), fusaproliferin (FUS) and moniliformin. Data on the natural occurrence of FBs have been widely reported, but information on BEA and FUS in maize is limited. The aims of this study were to establish the occurrence of Fusarium species in different maize hybrids in Mexico, to determine the ability of Fusarium spp. isolates to produce BEA, FUS and FBs and their natural occurrence in maize. Twenty-eight samples corresponding to seven different maize hybrids were analyzed for mycobiota and natural mycotoxin contamination by LC. Fusarium verticillioides was the dominant species (44–80%) followed by F. subglutinans (13–37%) and F. proliferatum (2–16%). Beauvericin was detected in three different hybrids with levels ranging from 300 to 400 ng g⁻¹, while only one hybrid was contaminated with FUS (200 ng g⁻¹). All samples were positive for FB₁ and FB₂ contamination showing levels up to 606 and 277 ng g⁻¹, respectively. All F. verticillioides isolates were able to produce FB₁ (13.8–4,860 μg g⁻¹) and some also produced FB₂ and FUS. Beauvericin, FUS, FB₁ and FB₂ were produced by several isolates including F. proliferatum and F. subglutinans and co-production was observed. This is the first report on the co-occurrence of these toxins in maize samples from Mexico. The analysis of the presence of multiple mycotoxins in this substrate is necessary to understand the significance of these compounds in the human and animal food chains.     

Identification of QTL underlying isoflavone contents in soybean seeds among multiple environments

Soybean isoflavones are valued in certain medicines, cosmetics, foods and feeds. Selection for high-isoflavone content in seeds along with agronomic traits is a goal of many soybean breeders. The aim of the study was to identify the quantitative trait loci (QTL) underlying seed isoflavone content in soybean among seven environments in China. A cross was made between ‘Zhongdou 27’, a soybean cultivar with higher mean isoflavone content in the seven environments (daidzein, DZ, 1,865 μg g⁻¹; genistein, GT, 1,614 μg g⁻¹; glycitein, GC, 311 μg g⁻¹ and total isoflavone, TI, 3,791 μg g⁻¹) and ‘Jiunong 20’, a soybean cultivar with lower isoflavone content (DZ, 844 μg g⁻¹; GT, 1,046 μg g⁻¹; GC, 193 μg g⁻¹ and TI, 2,061 μg g⁻¹). Through single-seed-descent, 130 F₅-derived F₆ recombinant inbred lines were advanced. A total of 99 simple-sequence repeat markers were used to construct a genetic linkage map. Seed isoflavone contents were analyzed using high-performance liquid chromatography for multiple years and locations (Harbin in 2005, 2006 and 2007, Hulan in 2006 and 2007, and Suihua in 2006 and 2007). Three QTL were associated with DZ content, four with GT content, three with GC content, and five with TI content. For all QTL detected the beneficial allele was from Zhongdou 27. QTL were located on three (DZ), three (GC), four (GT) and five (TI) molecular linkage groups (LG). A novel QTL was detected with marker Satt144 on LG F that was associated with DZ (0.0014 > P > 0.0001, 5% < R ² < 11%; 254 < DZ < 552 μg g⁻¹), GT (0.0027 > P > 0.0001; 4% < R ² < 9%; 262 < GT < 391 μg g⁻¹), and TI (0.0011 > P > 0.0001; 4% < R ² < 15%; 195 < TI < 871 μg g⁻¹) across the various environments. A previously reported QTL on LG M detected by Satt540 was associated with TI across four environments and TI mean (0.0022 > P > 0.0001; 3% < R ² < 8%; 182 < TI < 334 μg g⁻¹) in China. Because both beneficial alleles were from Zhongdou 27, it was concluded that these two QTL would have the greatest potential value for marker-assisted selection for high-isoflavone content in soybean seed in China. G. Zeng, D. Li and Y. Han have equal contributions to the paper.     

Drought,warming and soil fertilization effects on leaf volatile terpene concentrations in <Emphasis Type="Italic">Pinus halepensis</Emphasis> and <Emphasis Type="Italic">Quercus ilex</Emphasis>

The changes in foliar concentrations of volatile terpenes in response to water stress, fertilization and temperature were analyzed in Pinus halepensis and Quercus ilex. The most abundant terpenes found in both species were α-pinene and Δ³-carene. β-Pinene and myrcene were also abundant in both species. P. halepensis concentrations were much greater than those of Q. ilex in agreement with the lack of storage in the latter species (15205.60 ± 1140.04 vs. 0.54 ± 0.08 μg g⁻¹ [d.m.]). The drought treatment (reduction to 1/3 of full watering) significantly increased the total terpene concentrations in both species (54% in P. halepensis and 119% in Q. ilex). The fertilization treatment (addition of either 250 kg N ha⁻¹ or 250 kg P ha⁻¹ or both) had no significant effects on terpene foliar concentrations. The terpene concentrations increased from 0.25 μg g⁻¹ [d.m.] at 30°C to 0.70 μg g⁻¹ [d.m.] at 40°C in Q. ilex (the non-storing species) and from 2,240 μg g⁻¹ [d.m.] at 30°C to 15,621 μg g⁻¹ [d.m.] at 40°C in P. halepensis (the storing species). Both species presented negative relationship between terpene concentrations and relative water contents (RWC). The results of this study show that higher terpene concentrations can be expected in the warmer and drier conditions predicted for the next decades in the Mediterranean region.     

Effect of nitrogen limitation on oleic acid biosynthesis in <Emphasis Type="Italic">Botryococcus braunii</Emphasis>

The influence of nitrogen (N) deficiency on the cell growth and intracellular lipid production of the alga Botryococcus braunii UTEX 572 was investigated. Biomass concentration and lipid content of B. braunii cultivated in modified Chu-13 medium containing 0.04, 0.37, and 3.66 mM nitrate were 0.23–0.38 g L⁻¹ and 36–63% of dry cell weight, respectively. The specific growth rate of B. braunii reached a constant of 0.185 day⁻¹ during cultivation with an initial nitrate feed of 3.66 mM. The maximum lipid content of B. braunii was 63% with 0.04 mM nitrate. However, the maximum lipid productivity of 0.019 g L⁻¹ day⁻¹ was achieved with 0.37 mM nitrate. The level of oleic acid, an important component of biodiesel, was higher at 86% of the total fatty acids under N-limited conditions (0.04 mM nitrate) compared to 69% under N-sufficient conditions (3.66 mM nitrate). Furthermore, expression of the stearoyl-ACP desaturase gene (sad) encoding a stearoyl-ACP desaturase involved in the synthesis of oleic acid was 2.6-fold higher under N-limited conditions than under N-sufficient conditions.     

Large-scale cultivation of adventitious roots of Echinacea purpurea in airlift bioreactors for the production of chichoric acid, chlorogenic acid and caftaric acid

Adventitious roots of Echinacea purpurea were cultured in airlift bioreactors (20 l, 500 l balloon-type, bubble bioreactors and 1,000 l drum-type bubble bioreactor) using Murashige and Skoog (MS) medium with 2 mg indole butyric acid l⁻¹ and 50 g sucrose l⁻¹ for the production of chichoric acid, chlorogenic acid and caftaric acid. In the 20 l bioreactor (containing 14 l MS medium) a maximum yield of 11 g dry biomass l⁻¹ was achieved after 60 days. However, the amount of total phenolics (57 mg g⁻¹ DW), flavonoids (34 mg g⁻¹ DW) and caffeic acid derivatives (38 mg g⁻¹ DW) were highest after 50 days. Based on these studies, pilot-scale cultures were established and 3.6 kg and 5.1 kg dry biomass were achieved in the 500 l and 1,000 l bioreactors, respectively. The accumulation of 5 mg chlorogenic acid g⁻¹ DW, 22 mg chichoric acid g⁻¹ DW and 4 mg caftaric acids g⁻¹ DW were achieved with adventitious roots grown in 1,000 l bioreactors.