Chromosome diversity in Dasypyrum villosum, an important genetic and trait resource for hexaploid wheat engineering.

BACKGROUND AND AIMS: Dasypyrum villosum (2n = 2x = 14) harbours potentially beneficial genes for hexaploid and tetraploid wheat improvement. Highly diversified chromosome variation exists among and within accessions due to its open-pollination nature. The wheat-D. villosum T6VS·6AL translocation was widely used in breeding mainly because gene Pm21 in the 6VS segment conferred high and lasting powdery mildew resistance. However, the widespread use of this translocation may narrow the genetic base of wheat. A better solution is to utilize diversified D. villosum accessions as the genetic source for wheat breeding. Analysis of cytological and genetic polymorphisms among D. villosum accessions also provides genetic evolution information on the species. Using cytogenetic and molecular tools we analysed genetic polymorphisms among D. villosum accessions and developed consensus karyotypes to assist the introgression of beneficial genes from D. villosum into wheat. METHODS: A multiplex probe of repeats for FISH, GISH and molecular markers were used to detect chromosome polymorphisms among D. villosum accessions. Polymorphic signal block types, chromosome heterogeneity and heterozygosity, and chromosome polymorphic information content were used in genetic diversity analysis. KEY RESULTS: Consensus karyotypes of D. villosum were developed, and the homoeologous statuses of individual D. villosum chromosomes relative to wheat were determined. Tandem repeat probes of pSc119.2, (GAA)10 and the AFA family produced high-resolution signals and not only showed different signal patterns in D. villosum chromosomes but also revealed the varied distribution of tandem repeats among chromosomes and accessions. A total of 106 polymorphic chromosomes were identified from 13 D. villosum accessions and high levels of chromosomal heterozygosity and heterogeneity were observed. A subset of 56 polymorphic chromosomes was transferred into durum wheat through wide crosses, and seven polymorphic chromosomes are described in two newly developed durum-D. villosum amphidiploids. CONCLUSIONS: Consensus karyotypes of D. villosum and oligonucleotide FISH facilitated identification of polymorphic signal blocks and a high level of chromosomal heterozygosity and heterogeneity among D. villosum accessions, seen in newly developed amphiploids. The abundant genetic diversity of D. villosum and range of alleles, exploitable through interploid crosses, backcrosses and recombination (chromosome engineering), allow introduction of biotic and abiotic stress resistances into wheat, translating into increasing yield, end-use quality and crop sustainability.

Surface-enhanced Raman spectroscopy aptasensor for simultaneous determination of ochratoxin A and zearalenone using Au@Ag core-shell nanoparticles and gold nanorods.

The design and fabrication of a surface-enhanced Raman scattering (SERS) aptasensor for simultaneous detection of zearalenone (ZEN) and ochratoxin A (OTA) in wheat and corn samples is described. The capture and reporter probes were SH-cDNA-modified gold nanorods and SH-Apt-modified Au@Ag core-shell nanoparticles, respectively. After recognizing OTA and ZEN aptamers and complementary strands (SH-cDNA), the reporter probe generated a strong SERS signal. The preferred binding of OTA and ZEN aptamers to OTA and ZEN, respectively, caused reporter probes to release the capture probes, resulting in a linear decrease in SERS intensity. The detection of OTA showed good linearity with an R2 value of 0.986, which could be maintained across a wide concentration range (0.01 to 100 ng/mL), with the limit of detection of 0.018 ng/mL. For detection of ZEN, good linearity with an R2 value of 0.987 could be maintained across a wide concentration range (0.05 to 500 ng/mL), with 0.054 ng/mL as the limit of detection. Good accuracy (relative standard deviation < 4.2%) during mycotoxin determination as well as excellent quantitative recoveries (96.0-110.7%) during the analysis of spiked real samples was achieved. The proposed SERS aptasensor exhibited excellent performance in the detection of OTA and ZEN in real food samples. Hence, by simply changing the aptamer, this new model can be applied to the detection of multiple mycotoxins in the food industry.

(Fe,Co)@nitrogen-doped graphitic carbon nanocubes derived from polydopamine-encapsulated metal-organic frameworks as a highly stable and selective non-precious oxygen reduction electrocatalyst.

A facile approach is reported to synthesize (Fe,Co)@nitrogen-doped graphitic carbon (NGC) nanocubes (NCs) via the pyrolysis of polydopamine-encapsulated Fe3[Co(CN)6]2 NCs at 700 °C. Besides the comparable catalytic activity for oxygen reduction reaction (ORR) to the Pt/C catalyst, it showed much more outstanding catalytic selectivity and superior durability.

In-syringe demulsified dispersive liquid-liquid microextraction and high performance liquid chromatography-mass spectrometry for the determination of trace fungicides in environmental water samples.

An in-syringe demulsified dispersive liquid-liquid microextraction (ISD-DLLME) technique was developed using low-density extraction solvents for the highly sensitive determination of the three trace fungicides (azoxystrobin, diethofencarb and pyrimethanil) in water samples by high performance liquid chromatography-mass spectrometry chromatography-diode array detector/electrospray ionisation mass spectrometry. In the proposed technique, a 5-mL syringe was used as an extraction, separation and preconcentration container. The emulsion was obtained after the mixture of toluene (extraction solvent) and methanol (dispersive solvent) was injected into the aqueous bulk of the syringe. The obtained emulsion cleared into two phases without centrifugation, when an aliquot of methanol was introduced as a demulsifier. The separated floating organic extraction solvent was impelled and collected into a pipette tip fitted to the tip of the syringe. Under the optimal conditions, the enrichment factors for azoxystrobin, diethofencarb and pyrimethanil were 239, 200, 195, respectively. The limits of detection, calculated as three times the signal-to-noise ratio (SN(-1)), were 0.026 µg L(-1) for azoxystrobin, 0.071 µg L(-1) for diethofencarb and 0.040 µg L(-1) for pyrimethanil. The repeatability study was carried out by extracting the spiked water samples at concentration levels of 0.02 µg mL(-1) for all the three fungicides. The relative standard deviations varied between 4.9 and 8.2% (n=5). The recoveries of all the three fungicides from tap, lake and rain water samples at spiking levels of 0.2, 1, 5 µg L(-1) were in the range of 90.0-105.0%, 86.0-114.0% and 88.6-110.0%, respectively. The proposed ISD-DLLME technique was demonstrated to be simple, practical and efficient for the determination of different kinds of fungicide residues in real water samples.

Ultrasound-enhanced surfactant-assisted dispersive liquid-liquid microextraction and high-performance liquid chromatography for determination of ketoconazole and econazole nitrate in human blood.

A simple and efficient method, based on ultrasound-enhanced surfactant-assisted dispersive liquid-liquid microextraction (UESA-DLLME) followed by high-performance liquid chromatography (HPLC) has been developed for extraction and determination of ketoconazole and econazole nitrate in human blood samples. In this method, a common cationic surfactant, cetyltrimethylammonium bromide (CTAB), was used as dispersant. Chloroform (40 µL) as extraction solvent was added rapidly to 5 mL blood containing 0.068 mg mL(-1) CTAB. The mixture was then sonicated for 2 min to disperse the organic chloroform phase. After the extraction procedure, the mixture was centrifuged to sediment the organic chloroform phase, which was collected for HPLC analysis. Several conditions, including type and volume of extraction solvent, type and concentration of the surfactant, ultrasound time, extraction temperature, pH, and ionic strength were studied and optimized. Under the optimum conditions, linear calibration curves were obtained in the ranges 4-5000 µg L(-1) for ketoconazole and 8-5000 µg L(-1) for econazole nitrate, with linear correlation coefficients for both >0.99. The limits of detection (LODs, S/N = 3) and enrichment factors (EFs) were 1.1 and 2.3 µg L(-1), and 129 and 140 for ketoconazole and econazole nitrate, respectively. Reproducibility and recovery were good. The method was successfully applied to the determination of ketoconazole and econazole nitrate in human blood samples.

Application of an ultrasound-assisted surfactant-enhanced emulsification microextraction method for the analysis of diethofencarb and pyrimethanil fungicides in water and fruit juice samples.

In this work, a simple, practical and environmentally friendly sample pre-treatment method, ultrasound-assisted surfactant-enhanced emulsification microextraction coupled with high performance liquid chromatography-diode array detector/electrospray ionisation mass spectrometry, was developed to determine diethofencarb and pyrimethanil residues in water and fruit juice samples. Tween 80 was used as an emulsifier and carbon tetrachloride was chosen as the extraction solvent, and no dispersive organic solvent was needed, which is typically required in common dispersive liquid-liquid microextraction methods. Several variables, such as the type and volume of extraction solvent and surfactant, extraction temperature and ultrasound extraction time were investigated and optimised. Under optimal conditions, the enrichment factors were 265 and 253 for diethofencarb and pyrimethanil, respectively. The limits of detection (LODs), calculated as three times the signal-to-noise ratio (S/N), were 0.01 µg L(-1) for both diethofencarb and pyrimethanil. The linearity of the method was obtained in the range of 0.05-2000 µg L(-1), with correlation coefficients of 0.9994-0.9998. The water (at fortified levels of 0.1 and 1.0 µg L(-1)) and fruit juice samples (at fortified levels of 0.1 and 1.0 µg L(-1)) were successfully analysed using the proposed method, and the relative recoveries were in the range of 88-114%, 93-111%, 86-117% and 94-101%, respectively.