Single and double inoculation with Azospirillum/Trichoderma: the effects on dry bean and wheat

A plant growth-promoting rhizobacterium (Azospirillum brasilense Sp7) and a bio-control fungus, which can solubilize insoluble phosphorus (Trichoderma harzianum Rifai 1295-22), were evaluated for their single and combined effects on dry bean (Phaseolus vulgaris) and wheat (Triticum aestivum L.) grown in soil. A pot experiment with bean and a field experiment with both bean and wheat were established. In contrast to single inoculation of Trichoderma, the single inoculation of Azospirillum and the double inoculation did not significantly (P >0.05) increase nodule numbers and nodule mass at 45 days after planting in pot grown beans. However, the Azospirillum inoculation with supplementary phosphorus significantly (P <0.05) increased nodule mass. There were no significant (P >0.05) differences among the inoculation treatments for plant dry weight, total plant nitrogen, and total plant phosphorus at 45 days after planting in both pot and field experiments with bean. However, the combined inoculation and rock phosphate application at 1 Mg ha^–1 significantly (P <0.05) increased bean seed yield, total seed nitrogen and phosphorus in the bean field trial. This treatment more than doubled the mentioned properties compared to the control. The microbial inoculations, with the exception of the combined inoculation, significantly (P <0.05) increased total seed nitrogen, but never affected seed yield in the wheat field trial (P >0.05). The combined inoculation improves many plant and yield parameters and, therefore, has some advantages over single inoculation provided that rock phosphate was supplied at an amount not exceeding 1 Mg ha^–1. Higher rock phosphate application rates decreased many plant and yield parameters in our study.This work was carried out at Gaziosmanpaa University, Tokat, Turkey.     

A simple extraction and chromatographic system for the simultaneous analysis of anthocyanins and stilbenes of Vitis species

A separation system has been developed to permit the simultaneous analysis of major anthocyanins and stilbenes from berries of Vitis species in <1 h. The system makes use of separation by HPLC and detection by UV-visible absorption and fluorescence, for anthocyanins and stilbenes, respectively, with the two detection systems linked in series. Monitoring the absorption at 520 nm permits ready identification and quantification of major anthocyanins. Chromatograms derived from fluorescence (330 nm excitation/374 nm emission) produce less clear data for the piceids and resveratrols, which are present in much lower amounts; peaks for these compounds can be verified by means of external or internal standards.     

Insecticidal activity of Arum maculatum tuber lectin and its binding to the glycosylated insect gut receptors

Mannose binding approximately 50 kDa homotetrameric lectin, purified from edible Arum maculatum tuber, was analyzed through SDS-PAGE and studied for its agglutination property using rabbit erythrocytes. Cross reactivity of the purified lectin was verified through western blot using Colocasia esculantum(Family, Araceae) tuber lectin antibody. The insecticidal activity of Arum maculatum tuber lectin (ATL) was tested against two economically important sucking pests, Lipaphis erysimi and Aphis craccivora, in an artificial diet. The LC(50) values for L. erysimi and A. craccivora were determined to be 21 microg/mL and 16 microg/mL, respectively. Addition of alpha-d-mannose in ATL-supplemented diet reduced the aphid mortality. Two major receptor proteins of ATL (approximately 40 kDa and approximately 35 kDa) were detected from the brush border membrane vesicle (BBMV) protein of L. erysimi and A. craccivora guts, respectively, using ligand-binding assay. Alpha-d-Mannose was found to be a deterrent to such binding of ATL to the BBMV receptors.     

Seasonal variations of oil residues in water of Shatt al-Arab river,Iraq

The distribution and seasonal variations of petroleum residues in the Shatt al-Arab water column have been determined spectrofluorometrically. Their concentrations were found to vary between 1.7 to 35.4 μg L^?1 Kuwait crude oil equivalents. The results suggested that petroleum hydrocarbons present in this river originated from diverse sources. Hydrocarbon amounts tend to be highest in winter (averaged 17.4 μg L^?1) and lowest in summer (averaged 3.1 μg L^?1).     

Studies of Wet Deposition and Dustfall at Pune, India

Rain water and dustfall deposition samples were collected at Pune, an urban site (1992–98) and at Sinhagad, a rural site (1992–94). The samples were collected with wet-only and bulk collectors at Pune and with bulk collector at Sinhagad. The samples were analyzed for major ions, pH and conductivity. The study showed that the rain water at both places is alkaline (pH > 5.6). The average pH at Pune was 6.1. Neutralising components, indicated by Ca and non sea salt (nss) Mg have higher concentrations than the acidifying components SO₄ and NO₃. The wet deposition fluxes of all the ionic components were higher than the dustfall fluxes. Relative contribution from dustfall was largest for K, Ca, Mg and NO₃. Dustfall was greater at Pune, compared to Sinhagad for all components and up to double for Ca.     

Genetic polymorphism of plastid DNA in Tunisian date-palm germplasm (Phoenix dactylifera L.) detected with PCR-RFLP

Fifteen plastid fragments were amplified from a set of Tunisian date-palm accessions by PCR with consensus primers and analysed by RFLP. Polymorphic DNA bands were obtained as reliable molecular markers to estimate genetic distances among the accessions and to examine their phylogenetic relationships. The ctDNA PCR-RFLP method permitted the identification of two haplotypes that differ in the presence or absence of the HinfI restriction site. Phenetic groups composed of cultivars clustered together but does not constitute monophyletic groups. This typology agrees with the haplotypes' organization and provides a common genetic background within the implied varieties.     

Suppression of Meloidogyne javanica by Pseudomonas aeruginosa IE-6S in tomato: the influence of NaCl, oxygen and iron levels

Understanding the environmental factors that influence the suppression of disease-suppressive strains of Pseudomonas fluorescens is an essential step toward improving the level and reliability of their biocontrol activity. A 0.8 M NaCl concentration was optimal for in vitro survival and growth of IE-6S⁺ while, nematicidal activity by IE-6S⁺ was maximal when the bacterium was exposed to 0.4 M NaCl. The bacterium was highly sensitive to high (1.6 M) NaCl concentration. Culture filtrate of the bacterium resulting from the medium supplemented with 0.2 or 0.4 M NaCl showed the presence of secondary metabolite, hydrogen cyanide (HCN). Soil amendment with IE-6S⁺ alone or in conjunction with up to 0.8 M NaCl enhanced bacterial efficacy towards Meloidogyne javanica, the root-knot nematode. Soil amendment with NaCl up to 0.8 M also resulted in enhanced bacterial rhizosphere colonization and growth of tomato seedlings. Protein content of the shoot was reduced when soil was amended with 1.6 M NaCl. Inner root establishment of the bacterium was greatly affected in the soils treated with 1.6 M NaCl. Under in vitro conditions, IE-6S⁺ showed enhanced growth when kept at ambient oxygen conditions while the growth of bacterium affected when incubated at low oxygen conditions. Culture filtrate of the bacterium resulting from low oxygen level caused greater mortality of M. javanica juveniles in vitro compared with the filtrates obtained from ambient oxygen conditions. Culture filtrate from low oxygen conditions also showed the presence of hydrogen cyanide while those from ambient oxygen condition did not. Under glasshouse conditions, regardless of bacterial application, nematode penetration rate was greater when the pots were watered from the top; nematode penetration was lowered in bacterized pots compared with non-bacterized controls. IE-6S⁺ applied in the pots either watered from the top or bottom had no significant impact on growth of tomato but protein contents of the leaves increased after treatment with the bacterium. Rhizosphere and inner root colonization of the bacterium increased when the pots were watered from the top. Under in vitro conditions, with an increased iron concentration in the form of FeEDDHA, growth of IE-6S⁺ and its nematicidal activity increased. Culture filtrate of IE-6S⁺ obtained from liquid King's B medium supplemented with 0.5 or 1.0 mM FeEDDHA showed the presence of HCN. Under glasshouse conditions, soil treated with FeEDDHA alone did not reduce nematode penetration rates but did reduce greatly when applied in conjunction with IE-6S⁺. FeEDDHA applied at 0.5 mg/kg of soil in combination with IE-6S⁺ significantly enhanced plant growth and leaf protein contents. FeEDDHA at 1 mg/kg of soil increased bacterial populations both in the rhizosphere and inner root tissues of tomato.     

Determination of N-methylcarbamate pesticides in liver by liquid chromatography

A liquid chromatographic (LC) method using a 2-step purification technique for the simultaneous determination of 10 carbamates in bovine, swine, and duck livers has been developed. Carbamates are extracted from liver samples with methylene chloride. After evaporation, the residues from the extract are dissolved in methylene chloride-cyclohexane (1 + 1) and cleaned up by gel permeation chromatography. The eluate containing carbamate residues is evaporated to dryness, reconstituted in methylene chloride, further purified by passing it through an aminopropyl Bond Elut extraction cartridge, and analyzed by liquid chromatography using post-column derivatization with orthophthalaldehyde and fluorescence detection. Excitation and emission are set at 340 and 418 nm, respectively. Liver samples for beef, pork, and duck were fortified with 5, 10, and 20 ppb of mixed carbamate standards. The average of 10 recoveries of 10 carbamates at all 3 levels of fortification was greater than 80% with coefficients of variation less than 17%.     

Monitoring of the fermentation media of citric acid by the trimethylsilyl derivatives of the organic acids formed

In this approach, a derivatization method is described for monitoring of organic acids in fermentation media without any separation step. The aqueous phase of fermentation media was evaporated and heated in a silylation reagent to form trimethylsilyl (TMS) derivatives. The silylated compounds are analyzed by 29Si nuclear magnetic resonance (29Si NMR) and gas chromatography-mass spectrometry (GC-MS). 29Si NMR can qualitatively monitor the components produced in the Krebs cycle. Quantification of these compounds is investigated by using selected ion monitoring mode of mass spectrometry. In this mode, mass to charge (m/z) values of their [M - 15]+ ions, which are 465, 275, 247, 221, 335, 251, and 313 of TMS derivatives of citric, alpha-ketoglutaric, succinic, fumaric, l-malic, oxaloacetic, and palmitic (as an internal standard), acids, respectively, are used. The limit of detection and the linear working range for derivatized citric acid were found to be 0.1 mg L(-1) and 10-3 x 10(4) mg L(-1). The relative standard deviation of the method for five replicates was 2.1%. The average recovery efficiency for citric acid added to culture media was approximately 97.2%. Quantitative results of GC-MS are compared with those obtained by an ultraviolet-visible method.