Precise mapping of QTL for Hessian fly resistance in the hard winter wheat cultivar 'Overland'.

KEY MESSAGE: A major QTL for Hessian fly resistance was precisely mapped to a 2.32 Mb region on chromosome 3B of the US hard winter wheat cultivar 'Overland'. The Hessian fly (HF, Mayetiola destructor) is a destructive insect pest of wheat in the USA and worldwide. Deploying HF-resistant cultivars is the most effective and economical approach to control this insect pest. A population of 186 recombinant inbred lines (RILs) was developed from 'Overland' × 'Overley' and phenotyped for responses to HF attack using the HF biotype 'Great Plains'. A high-density genetic linkage map was constructed using 1,576 single nucleotide polymorphism (SNP) markers generated by genotyping-by-sequencing (GBS). Two quantitative trait loci (QTLs) with a significant epistatic effect on HF resistance were mapped to chromosomes 3B (QHf.hwwg-3B) and 7A (QHf.hwwg-7A) in Overland, which are located in similar chromosome regions as found for H35 and H36 in the cultivar 'SD06165', respectively. QHf.hwwg-3B showed a much larger effect on HF resistance than QHf.hwwg-7A. Five and four GBS-SNPs, respectively, in the QHf.hwwg-3B and QHf.hwwg-7A QTL intervals were converted into Kompetitive allele specific polymerase chain reaction (KASP) markers. QHf.hwwg-3B was precisely mapped to a 2.32 Mb interval (2,479,314-4,799,538 bp) using near-isogenic lines (NILs) and RILs that have recombination within the QTL interval. The US winter wheat accessions carrying contrasting alleles at KASP markers KASP-3B4525164, KASP-7A47772047 and KASP-7A65090410 showed significant difference in HF resistance. The combination of the two KASP markers KASP-3B3797431 and KASP-3B4525164 is near-diagnostic for the detection of QHf.hwwg-3B in a US winter wheat panel and can be potentially used for screening the QTL in breeding programs.

Mechanisms of Cd and Cr removal and tolerance by macrofungus Pleurotus ostreatus HAU-2.

Fungi bioaccumulation is a novel and highly promising approach to remediate polluted soil. The present study revealed a high ability to tolerate Cd and Cr in the fungus Pleurotus ostreatus HAU-2. However, high concentrations of Cd and Cr can suppress fungal growth and result in a variation of hypha micromorphology. Batch experiments were performed to investigate Cd and Cr stress effects on the amount of active oxygen in fungi, activity of antioxidant enzyme, as well as the removal efficiency of Cd and Cr. The results revealed that Cd and Cr caused increasing active oxygen and malonaldehyde (MDA) concentrations. Antioxidant enzymes play a central role in removing active oxygen, while glutathione (GSH) aids the Cd detoxification within cells. In fluid culture, fungal removal rates of Cd and Cr ranged from 44.85% to 80.36% and 14.49% to 45.55%, respectively. Intracellular accumulation and extracellular adsorption were the major removal approaches. Bag cultivation testing indicated that the fungus absorbed Cd and Cr contained within soil. In particular, the accumulation ability of Cd (15.6mgkg-1) was higher compared to that of Cr (8.9mgkg-1). These results successfully establish P. ostreatus HAU-2 as promising candidate for the remediation of heavy-metal polluted soils.

UPLC-QTOF analysis reveals metabolomic changes in the flag leaf of wheat (Triticum aestivum L.) under low-nitrogen stress.

Wheat is one of the most important grain crop plants worldwide. Nitrogen (N) is an essential macronutrient for the growth and development of wheat and exerts a marked influence on its metabolites. To investigate the influence of low nitrogen stress on various metabolites of the flag leaf of wheat (Triticum aestivum L.), a metabolomic analysis of two wheat cultivars under different induced nitrogen levels was conducted during two important growth periods based on large-scale untargeted metabolomic analysis using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF). Multivariate analyses-such as principle components analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA)-were used for data analysis. PCA yielded distinctive clustering information among the samples, classifying the wheat flag samples into two categories: those under normal N treatment and low N treatment. By processing OPLS-DA, eleven secondary metabolites were shown to be responsible for classifying the two groups. The secondary metabolites may be considered potential biomarkers of low nitrogen stress. Chemical analyses showed that most of the identified secondary metabolites were flavonoids and their related derivatives, such as iso-vitexin, iso-orientin and methylisoorientin-2″-O-rhamnoside, etc. This study confirmed the effect of low nitrogen stress on the metabolism of wheat, and revealed that the accumulation of secondary metabolites is a response to abiotic stresses. Meanwhile, we aimed to identify markers which could be used to monitor the nitrogen status of wheat crops, presumably to guide appropriate fertilization regimens. Furthermore, the UPLC-QTOF metabolic platform technology can be used to study metabolomic variations of wheat under abiotic stresses.

Improvement of tolerance to lead by filamentous fungus Pleurotus ostreatus HAU-2 and its oxidative responses.

Wastewater contaminated with heavy metals is a world-wide concern. One biological treatment strategy includes filamentous fungi capable of extracellular adsorption and intracellular bioaccumulation. Here we report that an acclimated strain of filamentous fungus Pleurotus ostreatus HAU-2 can withstand Pb up to 1500 mg L(-1) Pb, conditions in which the wildtype strain cannot grow. The acclimated strain grew in liquid culture under 500 mg L(-1) Pb without significant abnormity in biomass and morphology, and was able to remove significant amounts of heavy metals with rate of 99.1% at 200 mg L(-1) and 63.3% at 1500 mg L(-1). Intracellular bioaccumulation as well as extracellular adsorption both contributed the Pb reduction. Pb induced levels of H2O2, and its concentration reached 72.9-100.9 µmol g(-1) under 200-1000 mg L(-1) Pb. A relatively higher malonaldehyde (MDA) concentration (8.06-7.59 nmol g(-1)) was also observed at 500-1500 mg L(-1) Pb, indicating that Pb exposure resulted in oxidative damage. The fungal cells also defended against the attack of reactive oxygen species by producing antioxidants. Of the three antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), CAT was the most responsive and the maximal enzyme activity was 15.8 U mg(-1) protein. Additionally, glutathione (GSH) might also play a role (3.16-3.21 mg g(-1) protein) in detoxification under relatively low Pb concentration (100-200 mg L(-1)). Our findings suggested that filamentous fungus could be selected for increased tolerance to heavy metals and that CAT and GSH might be important components of this tolerance.

Profile of natural redox mediators production of laccase-producing fungus Pleurotus ostreatus.

Polycyclic aromatic hydrocarbons (PAHs) are highly toxic organic pollutants which are abundant and environmentally widespread. Anthracene is a simple PAH that can be oxidized by laccases, copper-containing oxidase enzymes, produced by some plants, fungi, and bacteria. In this work, the extracellular culture fluid (CF) of laccase-producing fungus Pleurotus ostreatus was separated to crude laccase (CL) and aqueous ultrafiltrate (AU) fractions. The rate of anthracene oxidation by CF was 68.7 % while oxidation by CL was only 27.8 %. The addition of AU enhanced anthracene oxidation rate by CL to 60.4 %, indicating that the natural redox-mediators were present in the CF. The laccase-catalyzed anthracene oxidation rate increased with increased AU concentration, implying that oxidation rate is positively related to the concentration of natural mediators when laccase activity is constant. The AU from fungal culture containing bran or straw enhanced laccase-catalyzed anthracene oxidation; this enhancement increased further with prolonged fungus-cultivation, implying that both bran and straw induce the natural mediators. Our findings suggest increasing natural mediator levels may be an alternative strategy to improve the biodegradability of laccase-producing fungi.

Effect of CO2 enrichment on the glucosinolate contents under different nitrogen levels in bolting stem of Chinese kale (Brassica alboglabra L.).

The effects of CO(2) enrichment on the growth and glucosinolate (GS) concentrations in the bolting stem of Chinese kale (Brassica alboglabra L.) treated with three nitrogen (N) concentrations (5, 10, and 20 mmol/L) were investigated. Height, stem thickness, and dry weights of the total aerial parts, bolting stems, and roots, as well as the root to shoot ratio, significantly increased as CO(2) concentration was elevated from 350 to 800 microl/L at each N concentration. In the edible part of the bolting stem, 11 individual GSs were identified, including 7 aliphatic and 4 indolyl GSs. GS concentration was affected by the elevated CO(2) concentration, N concentration, and CO(2)xN interaction. At 5 and 10 mmol N/L, the concentrations of aliphatic GSs and total GSs significantly increased, whereas those of indolyl GSs were not affected, by elevated atmospheric CO(2). However, at 20 mmol N/L, elevated CO(2) had no significant effects on the concentrations of total GSs and total indolyl GSs, but the concentrations of total aliphatic GSs significantly increased. Moreover, the bolting stem carbon (C) content increased, whereas the N and sulfur (S) contents decreased under elevated CO(2) concentration in the three N treatments, resulting in changes in the C/N and N/S ratios. Also the C/N ratio is not a reliable predictor of change of GS concentration, while the changes in N and S contents and the N/S ratio at the elevated CO(2) concentration may influence the GS concentration in Chinese kale bolting stems. The results demonstrate that high nitrogen supply is beneficial for the growth of Chinese kale, but not for the GS concentration in bolting stems, under elevated CO(2) condition.