Liquid chromatography-tandem mass spectrometry analysis of 2-amino-1-methyl-6-(4-hydroxyphenyl)imidazo[4,5-b]pyridine in cooked meats

Several cooked meats such as beef (fried, coated-fried), pork (fried, coated-fried), and chicken (fried, griddled, coated-fried, roasted) were analyzed for the heterocyclic amine 2-amino-1-methyl-6-(4-hydroxyphenyl)imidazo[4,5- b]pyridine (4'-OH-PhIP) not commonly determined in food and 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP). The highest content of 4'-OH-PhIP was found in fried and griddled chicken breast, the concentration being 43.7 and 13.4 ng/g, respectively, whereas the corresponding PhIP concentrations were 19.2 and 5.8 ng/g. The estimated concentration of both pyridines in fried pork loin, in fried pork sausages, and in coated-fried chicken was below 2.5 ng/g. In the rest of the samples, 4'-OH-PhIP was not detected. The analyses were performed by solid-phase extraction and LC-MS/MS. The fragmentation of 4'-OH-PhIP in an ion trap mass analyzer was studied in order to provide information for the identification of 4'-OH-PhIP. Additionally, the effect of red wine marinades on the formation of 4'-OH-PhIP in fried chicken was examined, finding a notable reduction (69%) in the amine's occurrence.     

Chitosan nanoparticles enhance developmental competence of in vitro-matured porcine oocytes

Oxidative stress is inevitable as it is derived from the handling, culturing, inherent metabolic activities and medium supplementation of embryos. This study was performed to investigate the protective effect of chitosan nanoparticles (CNPs) on oxidative damage in porcine oocytes. For this purpose, cumulus–oocyte complexes (COCs) derived from porcine slaughterhouse ovaries were exposed to different concentrations of CNPs (0, 10, 25 and 50 µg/ml) during in vitro maturation (IVM). Oocytes treated with 25 µg/ml CNPs showed significantly higher levels of GSH, along with a significant reduction in ROS levels compared to control, CNPs10 and CNPs50 groups. In parthenogenetic embryo production, the maturation rate was significantly higher in the CNPs25 group than that in the control and all other treated groups. In addition, when compared to the CNPs50 and control groups, CNPs25-treated oocytes showed significantly higher cleavage and blastocyst development rates. The highest concentration of CNPs reduced the total cell number and ratio of ICM: TE cells in parthenogenetic embryos, suggesting that there is a threshold where benefits are lost if exceeded. In cloned embryos, the CNPs25 group, as compared to all other treated groups, showed significantly higher maturation and cleavage rates. Furthermore, the blastocyst development rate in the CNPs25-treated group was significantly higher than that in the CNPs50-treated group, as was the total cell number. Moreover, we found that cloned embryos derived from the CNPs25-treated group showed significantly higher expression levels of Pou5f1, Dppa2, and Ndp52il genes, compared with those of the control and other treated groups. Our results demonstrated that 25 µg/ml CNPs treatment during IVM improves the developmental competence of porcine oocytes by reducing oxidative stress.     

Quantitative structure-activity study of herbicidal N-aryl-3,4,5,6-tetrahydrophthalimides and related cyclic imides

The quantitative relationship between structure and inhibitory activity against the root growth of sawa millet of 30 N-aryl-3,4,5,6-tetrahydrophthalimides and 88 related N-aryl cyclic imide compounds (3-aryl-1,5-tetramethylenehydantoins, 4-aryl-1,2-tetramethylenetriazolidine-3,5-diones, and their thiocarbonyl analogs) was analyzed by means of physicochemical parameters of the aryl substituents and regression analysis. The results showed that, in each series of these classes of compounds, the position-specific steric effects of aromatic substituents, as represented by Verloop's STERIMOL values, are highly important in determining the inhibitory potency of the compound.     

Characterization of an EMS-induced soybean mutant with an increased content of Af saponin and a new component Ab-δ in the seed hypocotyl

Saponins are one of the components present in the soybean seeds that have various functional properties. The chemical structures and concentration of soyasaponins affect the taste of the processed soyfood, thereby limiting its industrial applications. Therefore, it is important to understand saponin biosynthesis to explore natural and artificial variation in the saponin components, which can be modified to suit its application. The objective of the present study was to identify and characterize an EMS-induced soybean mutant with an altered saponin composition from a pool of 892 M4 lines. The mutant PE1905 showed an increased content of saponin Af (336.0%). The content of saponin Ab, DDMP-αg, and DDMP-βg was decreased in the mutant PE1905 by 89.3, 24.8, and 63.1%, respectively compared to the wild-type Pungsannamul. Additionally, four new components were detected in the mutant PE1905 that were absent in the wild type. Of these, the compound 4 (designated as Ab-δ) had the highest concentration, and therefore it was further characterized by HPLC and LC-PDA/MS/MS analysis to know the chemical structure, and molecular weight and formula. Considering these details, along with the alterations in the saponin Af and Ab concentrations, it was presumed that the Ab-δ acts as a precursor for the synthesis of saponin Af and Ab. Thus, we predicted a biosynthetic pathway from the Ab-δ to Ab saponin. The inheritance analysis showed that the concentration of saponin Ab-δ is controlled by a single recessive gene in the mutant PE1905. The results from the present study would be helpful in understanding the mechanisms behind altered seed saponin composition in soybeans.     

Quantitative trait locus analysis for soybean (Glycine max) seed protein and oil concentrations using selected breeding populations

Soybean seed protein and oil concentrations are important traits that directly affect the quality of soyfoods. Many studies and breeding programmes have been conducted to find major quantitative trait loci (QTL) that regulate protein and oil concentrations and to develop soybean cultivars with high protein and/or oil content. The purpose of this study was to identify these QTL using a selected breeding population. The population was tested in field conditions over a period of 3 years. Seed protein and oil concentrations were measured each year. Single‐nucleotide polymorphisms (SNPs) were used to construct genetic map using a 180K SoyaSNP array, which identified 1,570 SNPs. We identified 12 QTL for seed protein, 11 for seed oil concentration and four for both traits. Among these, 17 QTL were closely mapped to previously reported QTL, whereas ten sites were novel. Several QTL were detected across at least two experimental years. These loci are good candidate QTL for optimal seed protein and oil concentrations. Our results demonstrate that favourable target QTL can be successfully identified using selected breeding populations.     

Re-examination of the standard cultivation practices of rice in response to climate change in Korea

The impact of climate change has been simulated or estimated to bring about yield decline and quality deterioration of rice in Korea, a temperate country. To cope with these negative impacts, here we propose likely changes in the standard cultivation practices of rice in consideration of the current information on the progress/prediction of climate change and related physiological aspects of rice. Shifting grain-filling period could be a promising choice to optimize temperature for rice grain-filling, a crucial growth stage to productivity, hence providing rice plants with sufficient duration and solar radiation for the period. This choice, however, shortens days to flowering by the additive combined effects of high temperature by delayed transplanting, temperature rise itself by climate change, and accelerated phase change of rice by high temperature and short day-length. These combined effects may delay the time of transplanting in a great extent, hence reducing biomass accumulation of rice before flowering. In these conditions, recovering decreased biomass production is the key concern for rice productivity. This includes raising healthy seedlings by omitting the covering process, dense planting, and nutrient management to enhance plant’s uptake activity. Current standard water management, especially mid-term drainage and intermittent irrigation, would have more importance in the future since they mitigate methane emission from the paddy. Field monitoring for weeds and pests would be an important first step to identify newly developing or thriving species for establishing subsequent controlling strategies. Earlier weed control should be emphasized through adjusting time of herbicide application.     

Evaluation of waterlogging tolerance with the degree of foliar senescence at early vegetative stage of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

Maize is highly susceptible to waterlogging, which is becoming one of worldwide abiotic threats in many agricultural areas. This study was evaluated to establish the screening method and to find tolerant maize genotypes. Six Korean maize inbred lines were subjected to waterlogging at V3 for 15 days using a big size pot with single maize plant (big pot method) and a box containing 31 maize plants at a time (box method). The degree of foliar senescence and the number of senescent leaves were better indicators for selecting waterlogging tolerant maize genotypes than SPAD value and plant height. The degree of foliar senescence revealed that KS124, KS140, and KS141 are tolerant, and KS85 is susceptible to waterlogging at the early growth stage. These responses of foliar senescence were in clear accordance with those of plant grain yield, which was supported by stress tolerance index for grain yield. The box method also showed the similar response of foliar senescence to the big pot method. Therefore, this box method based on foliar senescence may be simple and efficient for large-scale screening of maize germplasm against waterlogging stress. It was concluded that foliar senescence can be a good indicator for selecting tolerant maize genotypes against waterlogging at the early growth stage.     

Quantitative real-time PCR effectively detects and quantifies colonization of sclerotia of Sclerotinia sclerotiorum by Trichoderma spp.

Some members of the fungal genus Trichoderma are able to colonize and destroy sclerotia, the thick-walled resting structures of the soilborne plant pathogenic fungus Sclerotinia sclerotiorum, thus providing a potential means of biological disease control. However, current methods to detect and quantify colonization of sclerotia are labor-intensive, and generally qualitative rather than quantitative in nature. Our objective was to develop quantitative real-time PCR (polymerase chain reaction) methods to detect and measure colonization of sclerotia by Trichoderma spp. Specific PCR primer/probe sets were developed for Trichoderma spp. and for S. sclerotiorum. A total of 180 ITS1 (internal transcribed spacer) and ITS2 sequences from different species in the genus Trichoderma were aligned, and consensus sequences were determined. Six candidate primer sets were based on conserved regions of the consensus sequence, and the specificity of each nucleotide sequence was examined using BLAST (Basic Local Alignment Search Tool; NCBI) software. Each candidate primer set was tested on genomic DNA of T. harzianum strain ThzID1-M3, as well as six different Trichoderma isolates from soil, and on genomic DNA of S. sclerotiorum. The optimum primer/probe set selected, TGP4, successfully amplified genomic DNA of all Trichoderma isolates tested, and showed high precision and reproducibility over a linear range of eight orders of magnitude, from 85 ng to 8.5 fg of T. harzianum genomic DNA. TGP4 did not amplify S. sclerotiorum DNA. A specific PCR primer/probe set (TMSCL2) was developed for S. sclerotiorum, based on the calmodulin gene sequence. TMSCL2 did not amplify Trichoderma DNA. Quantitative real-time PCR with the primers then was evaluated in experiments to test differential effects of two soil moisture levels (−50 kPa, −1000 kPa matric potential) on biocontrol of S. sclerotiorum by indigenous Trichoderma spp. Periodically over 40 days, Trichoderma and S. sclerotiorum DNA in sclerotia were quantified by PCR with appropriate primers. Over 90% of the sclerotia were colonized by indigenous Trichoderma spp. at −1000 kPa, over the 40-day period, compared to only 60% at −50 kPa. In addition to determining incidence of colonization, the PCR method allowed measurement of the extent of sclerotial colonization, which also was significantly greater in the drier soil. Quantitative real-time PCR with the TGP4 primer/probe set provides a sensitive detection and measurement tool to evaluate colonization of sclerotia by Trichoderma spp.