Ochratoxin A production in aniseed-based media by selected fungal strains and in anise fruits (<Emphasis Type="Italic">Pimpinella anisum</Emphasis> L.)

The growth conditions and ochratoxin A (OTA) production of Aspergillus strains were studied in aniseed (Pimpinella anisum L.)-based media. The results showed that methanol/NaHCO₃ (50:50, v/v) mixture for extraction and competitive direct ELISA analytical method are capable of detecting low OTA concentrations in this raw material, which were confirmed by HPLC with fluorescence detection (R ² = 0.994). In aniseed meal extract agar artificially contaminated with selected fungi, the higher OTA values obtained were 283.8 ± 28.1 μg L^-1 for A. carbonarius and between 1.7 ± 0.1 μg L^-1 and 16.5 ± 12.8 μg L^-1 for A. steynii strains. While the optimal conditions of growth for A. carbonarius and A. steynii are 28°C and 0.98 a _w, the optimal production of OTA was observed at 0.99 a _w for both A. carbonarius and A. steynii but at 22°C and 28°C, respectively. Except in one sample, all the aniseed samples analysed were negative for OTA natural contamination. This study demonstrates that aniseed can be a matrix capable to contamination with OTA, at least produced by A. carbonarius and A. steynii strains, regardless of the antimicrobial properties of aniseed essential oil.     

A <Emphasis Type="Italic">FRUITFULL-like</Emphasis> gene is associated with genetic variation for fruit flesh firmness in apple (<Emphasis Type="Italic">Malus domestica</Emphasis> Borkh.)

The FRUITFULL (FUL) and SHATTERPROOF (SHP) genes are involved in regulating fruit development and dehiscence in Arabidopsis. We tested the hypothesis that this class of genes are also involved in regulating the development of fleshy fruits, by exploring genetic and phenotypic variation within the apple (Malus domestica) gene pool. We isolated and characterised the genomic sequences of two candidate orthologous FUL-like genes, MdMADS2.1 and MdMADS2.2. These were mapped using the reference population ‘Prima x Fiesta’ to loci on Malus linkage groups LG14 and LG06, respectively. An additional MADS-box gene, MdMADS14, shares high amino acid identity with the Arabidopsis SHATTERPROOF1/2 genes and was mapped to Malus linkage group LG09. Association analysis between quantitative fruit flesh firmness estimates of ‘Prima x Fiesta’ progeny and the MdMADS2.1, MdMADS2.2 and MdMADS14 loci was carried out using a mixed model analysis of variance. This revealed a significant association (P < 0.01) between MdMADS2.1 and fruit flesh firmness. Further evidence for the association between MdMADS2.1 and fruit flesh firmness was obtained using a case–control population-based genetic association approach. For this, a polymorphic repeat, (AT)_n, in the 3′ UTR of MdMADS2.1 was used as a locus-specific marker to screen 168 apple accessions for which historical assessments of fruit texture attributes were available. This analysis revealed a significant association between the MdMADS2.1 and fruit flesh firmness at both allelic (χ ² = 34, df = 9, P < 0.001) and genotypic (χ ² = 57, df = 32, P < 0.01) levels.     

Loci underlying resistance to Race 3 of soybean cyst nematode in Glycine soja plant introduction 468916

Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is an important soybean [Glycine max (L.) Merr.] pest in the U.S. and throughout the world. Genetic resistance is the primary method for controlling SCN and there is a need to identify new resistance genes. Glycine soja Sieb. and Zucc. is the wild ancestor of domesticated soybean and is a potential source of new SCN resistance genes. The goal of this research was to map quantitative trait loci (QTLs) that provide resistance to SCN Race 3 from the G. soja plant introduction (PI) 468916. Fifty seven F₂-derived lines from a cross between the G. soja PI 468916 and the G. max experimental line A81-356022 were tested for resistance to an SCN population with a Race-3 phenotype. These lines were also genotyped with 1,004 genetic markers and resistance genes were mapped by composite interval mapping with the computer program QTL-Cartographer. In the F₂ population, three significant (LOD > 3.0) QTLs were detected that explained from 5% to 27% of the variation for Race-3 resistance. The two most significant QTLs identified in the F₂ population were tested in a population of 100 BC1F₂ plants developed by crossing A81-356022 to a line from the F₂ population that carried the two resistance QTLs from G. soja. In the backcross population, both Race-3 resistance QTLs were significant, which confirms the existence of these QTLs. The QTLs identified in this experiment map to positions where SCN resistance genes have not been previously identified, suggesting that these are novel genes that could be useful for diversifying the resistance genes currently used in cultivar development. Received: 7 August 2000 / Accepted: 4 December 2000     

Highly efficient plant regeneration and <Emphasis Type="Italic">in vitro</Emphasis> polyploid induction using hypocotyl explants from diploid mulberry (<Emphasis Type="Italic">Morus multicaulis</Emphasis> Poir.)

Efficient plant regeneration is essential for successful transformation and in vitro polyploidy induction in mulberry. A high frequency (80%) of plant regeneration from hypocotyls occurred under in vitro conditions in mulberry (Morus multicaulis Poir.). We identified three key factors for enhancing successful regeneration based on earlier work: (1) hypocotyl position, (2) the combination and concentration of growth regulators, and (3) the addition of AgNO₃. The highest frequency of shoot regeneration was achieved using hypocotyl segments, which are proximal to apical meristems, and the optimal culture conditions were Murashige and Skoog’s (MS) (Murashige and Skoog, 1962) basal medium supplemented with 3.0 mg l⁻¹ 6-benzylamino purine, 0.3 mg l⁻¹ indole-3-acetic acid, 0.1% polyvinypyrrolidone, and 1.0 mg/l silver nitrate (AgNO₃) under subdued light at 25 ± 2°C. Treating the shoots with 0.2% colchicine (dipping for 72 h) resulted in a 14% tetraploid frequency, whereas a 20% tetraploid frequency resulted from using a 0.25% colchicine (dripping for 5 d) treatment, as determined by chromosome number counts. The diploid plant chromosome number was 28 (2n = 2x = 28) and that of tetraploid plants was 56 (2n = 4x = 56). Regenerated shoots rooted easily in 8–10 d using half-strength basal MS medium with 0.5 mg l⁻¹ indole-3-butyric acid and were successfully established in the soil.     

Photoinhibitory damage is modulated by the rate of photosynthesis and by the photosystem II light-harvesting chlorophyll antenna size

We investigated the effect of photosynthetic electron transport and of the photosystem II (PSII) chlorophyll (Chl) antenna size on the rate of PSII photoinhibitory damage. To modulate the rate of photosynthesis and the light-harvesting capacity in the unicellular chlorophyte Dunaliella salina Teod., we varied the amount of inorganic carbon in the culture medium. Cells were grown under high irradiance either with a limiting supply of inorganic carbon, provided by an initial concentration of 25 mM NaHCO₃, or with supplemental CO₂ bubbled in the form of 3% CO₂ in air. The NaHCO₃-grown cells displayed slow rates of photosynthesis and had a small PSII light-harvesting Chl antenna size (60 Chl molecules). The half-time of PSII photodamage was 40 min. When switched to supplemental CO₂ conditions, the rate of photodamage was retarded to a t_1/2 = 70 min. Conversely, CO₂-supplemented cells displayed faster rates of photosynthesis and a larger PSII light-harvesting Chl antenna size (500 Chl molecules). They also showed a rate of photodamage with t_1/2 = 40 min. When depleted of CO₂, the rate of photodamage was accelerated (t_1/2  = 20 min). These results indicate that the in-vivo susceptibility to photodamage is modulated by the rate of forward electron transport through PSII. Moreover, a large Chl antenna size enhances the rate of light absorption and photodamage and, therefore, counters the mitigating effect of forward electron transport. We propose that under steady-state photosynthesis, the rate of light absorption (determined by incident light intensity and PS Chl antenna size) and the rate of forward electron transport (determined by CO₂ availability) modulate the oxidation/reduction state of the primary PSII acceptor Q_A, which in turn defines the low/high probability for photodamage in the PSII reaction center. Received: 14 August 1997 / Accepted: 26 September 1997     

Evaluation of laboratory-based measures of soil mineral nitrogen and potentially mineralizable nitrogen as predictors of field-based indices of soil nitrogen supply in potato production

Accurate estimation of soil nitrogen (N) supply in the field is required to optimize fertilizer N management and to minimize environmental N losses in humid environments. Laboratory-based measures of N availability were evaluated as predictors of field-based indices of soil N supply within potato production systems. Pre-plant soil samples (0–15 cm) were collected from a series of forty treatments in established potato trials located in Atlantic Canada and Maine, USA. Total plant N uptake at topkill with no fertilizer N applied (PNU_0N), PNU_0N plus soil mineral N to 30 cm depth at harvest and relative yield were considered as field-based indices of soil N supply. The potentially mineralizable N (N₀) was determined by aerobic incubation at 25°C and periodic leaching for 24 weeks. A series of laboratory-based measures of soil N supply were measured in soil samples. Pre-plant soil nitrate or total mineral N at 0–30 cm depth was the best single predictor of PNU_0N (r = 0.67 and 0.71, respectively) and relative yield (r = 0.58 and 0.61). The ultraviolet absorbance of 0.01 M NaHCO₃ extract at 205 nm (NaHCO₃-205) was suitable as a predictor of PNU_0N and relative yield in each growing season, however, the relationship between this parameter and PNU_0N or relative yield varied somewhat among years. A combination of pre-plant mineral N plus N mineralized in the first 2 weeks period of incubation after re-wetting is proposed as a more robust measure of N availability compared with use of mineral N alone.     

Over-expressing GsGST14 from Glycine soja enhances alkaline tolerance of transgenic Medicago sativa

Glutathione-S-transferases (GSTs) are ubiquitous enzymes that play a key role in stress tolerance and cellular detoxification. The GST gene GsGST14 selected from the gene expression profiles of Glycine soja under alkaline stress was transformed into alfalfa (Medicago sativa L.). Transgenic alfalfa plants showed 1.73–1.99 times higher GST activity than wild-type plants. Transgenic alfalfa grew well in the presence of 100 mM NaHCO₃, while wild-type plants exhibited chlorosis and stunted growth, even death. There were marked changes in malondialdehyde content and relative membrane permeability caused by alkaline stress in non-transgenic lines compared to transgenic lines. The results indicate that the gene GsGST14 could enhance alkaline resistance in transgenic alfalfa.     

Effects of <Emphasis Type="SmallCaps">l</Emphasis>-Canavanine and ozone on vascular reactivity in septicemic rats

Septicemia leads to oxidative stress with overproduction of reactive-oxygen species (ROS) and consumption of endogenous antioxidant enzymes. We tested a twofold hypothesis: (1) does oxidative stress (OxS) induced by sepsis acting alone or in concert with augmented inflammatory processes contributes to sepsis-related vascular dysfunction, and, (2) whether ozone (O₃) and l-canavanine (CAV) mitigate the negative impact of the aforementioned phenomena. We investigated the relative impact of treatment with CAV and/or O₃ on vascular OxS associated vascular functional changes in septicemic rats. For this study, 60 male Sprague–Dawley rats were used and divided into six experimental groups (n = 10): control group (C), sham-operated (Sham), septicemic rats (S), S rats treated with CAV (100 mg/kg. i.p; S + CAV), S rats treated with O₃ (1.2 mg/kg, i.p.; S + O₃) and S rats treated with both O₃ and CAV (S + O₃ + CAV). After 22 h, the mean arterial blood pressure (MAP), the aortic ring vascular reactivity to phenylephrine, abdominal aortic blood flow (AABF), serum tumor necrosis factor-α (TNF-α) and plasma nitrite/nitrate (NOx) concentration were measured. In addition, hepatic antioxidant enzyme activities sodium dismutase (SOD) and glutathione peroxidase (GSH-Px) were estimated. Septicemia caused significant elevation of serum TNF-α (p < 0.001) and plasma NOx (p < 0.001) and significant (p < 0.001) reduction of AABF (p < 0.001), aortic vascular response to phenylephrine (p < 0.001), MAP (p < 0.001) and hepatic SOD and GSH-Px activity (p < 0.001) compared with the C group, while treatment with O₃ and/or CAV induced significant amelioration of all those increases. Abnormalities were attenuated to a similar extent with treatment with both O₃ and CAV. These results suggested that concomitant administration of O₃ and CAV alleviated the compromised vascular reactivity in septicemic conditions and prevent its progression into septic shock compared with each alone.