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Genome Wide Association Study for Phytophthora sojae Resistance with the Two Races Collected from Main Soybean Production Area in Korea with 210 Soybean Natural Population

  • Beom-Kyu Kang;Su-Vin Heo;Ji-Hee Park;Jeong-Hyun Seo;Man-Soo Choi;Jun-Hoi Kim;Jae-Bok Hwang;Ji-Yeon Ko;Yun-Woo Jang;Young-Nam Yun;Choon-Song Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.202-202
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    • 2022
  • Recently days, soybean production in paddy field is increasing, from 4,422 ha in 2016 to 10,658 ha in 2021 in Korea. It is easy for Phytophthora stem and root rot (PSR) occurring in paddy field condition, when it is poorly drained soils with a high clay content, and temporary flooding and ponding. Therefore PSR resistant soybean cultivar is required. The objective of this study is to identify QTL region and candidate genes relating to PSR resistance of the race in main soybean cultivation area in Korea. 210 soybean materials including cultivars and germplasm were used for inoculation and genome-wide association study (GWAS). Inoculation was conducted using stem-scar method with 2 replications in 2-year for the race 3053 from Kimje and 3617 from Andong. 210 materials were genotyped with Soya SNP 180K chip, and structure analysis and association mapping were conducted with QTLMAX V2. The results of inoculation showed that survival ratio ranged from 0% to 96.7% and mean 9.7% for 3053 and ranged from 0% to 100% and mean 7.6% for 3617. Structure analysis showed linkage disequillibrium (LD) was decayed below r2=0.5 at 335kb of SNP distance. Significant SNPs (LOD>7.0) were identified in Chr 1, 2, 3, 4, 5, 11, 14, 15 for 3053 and Chr 1, 2, 3, 7, 10, 14 for 3617. Especially, LD blocks (AX-90455181;15,056,628bp~AX-90475572;15,298,872bp) in Chr 2 for 3053 and 3067 were duplicated. 29 genes were identified on these genetic regions including Glyma.02gl47000 relating to ribosome recycling factor and defense response to fungus in Soybase.

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QTL Identification for Slow Wilting and High Moisture Contents in Soybean (Glycine max [L.]) and Arduino-Based High-Throughput Phenotyping for Drought Tolerance

  • Hakyung Kwon;Jae Ah Choi;Moon Young Kim;Suk-Ha Lee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.25-25
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    • 2022
  • Drought becomes frequent and severe because of continuous global warming, leading to a significant loss of crop yield. In soybean (Glycine max [L.]), most of quantitative trait loci (QTLs) analyses for drought tolerance have conducted by investigating yield changes under water-restricted conditions at the reproductive stages. More recently, the necessity of QTL studies to use physiological indices responding to drought at the early growth stages besides the reproductive ones has arisen due to the unpredictable and prevalent occurrence of drought throughout the soybean growing season. In this study, we thus identified QTLs conferring wilting scores and moisture contents of soybean subjected to drought stress in the early vegetative stage using an recombinant inbred line (RIL) population derived from a cross between Taekwang (drought-sensitive) and SS2-2 (drought-tolerant). For the two traits, the same major QTL was located on chromosome 10, accounting for up to 11.5% of phenotypic variance explained with LOD score of 12.5. This QTL overlaps with a reported QTL for the limited transpiration trait in soybean and harbors an ortholog of the Arabidopsis ABA and drought-induced RING-D UF1117 gene. Meanwhile, one of important features of plant drought tolerance is their ability to limit transpiration rates under high vapor pressure deficiency in response to mitigate water loss. However, monitoring their transpiration rates is time-consuming and laborious. Therefore, only a few population-level studies regarding transpiration rates under the drought condition have been reported so far. Via employing an Arduino-based platform, for the reasons addressed, we are measuring and recording total pot weights of soybean plants every hour from the 1st day after water restriction to the days when the half of the RILs exhibited permanent tissue damage in at least one trifoliate. Gradual decrease in moisture of soil in pots as time passes refers increase in the severity of drought stress. By tracking changes in the total pot weights of soybean plants, we will infer transpiration rates of the mapping parents and their RILs according to different levels of VPD and drought stress. The profile of transpiration rates from different levels of severity in the stresses facilitates a better understanding of relationship between transpiration-related features, such as limited maximum transpiration rates, to water saving performances, as well as those to other drought-responsive phenotypes. Our findings will provide primary insights on drought tolerance mechanisms in soybean and useful resources for improvement of soybean varieties tolerant to drought stress.

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Development of Ceramide NP Analysis Method in Cosmetic Formulations Using Liquid Chromatography (액체크로마토그래피를 이용한 화장품 제형 내 세라마이드엔피 분석법 확립)

  • Ye Ji Lee;Young Eun Kim;Jae Yong Seo;Hyun Dae Cho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.4
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    • pp.291-298
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    • 2023
  • In this study, a quantitative analysis method was developed using high-performance liquid chromatography (HPLC) to analyze the content of ceramide NP in lotion, cream, and cleanser formulations in cosmetics. The analysis was performed using a C18 column, and the mobile phase was set at a ratio of 70 : 30 for acetonitrile and methanol, the flow rate was set to 0.8 mL/min, and the column temperature was set to 20 ℃. The method was verified by analyzing specificity, linearity, limit of detection, limit of quantitation, accuracy, and precision in accordance with the ICH guidelines. As a result of validating the method, the linearity of the calibration curve was excellent (R2 = 0.99984). The accuracy of the lotion, cream, and cleanser formulations was confirmed with a recovery rate ranging from 95.11% to 100.48%. The precision analysis showed a low relative standard deviation (RSD) of less than 0.26%. The limit of detection was 0.902 ㎍/mL, and the limit of quantitation was 2.733 ㎍/mL. Through this quantitative analysis method of ceramide NP applied in cosmetics, it is expected to assist in the quality control of products by enabling measurement even when it is difficult to separate the main peak due to the influence of interfering substances.

Development of Simultaneous Analysis for Multiple Agricultural Pesticides in Raw Milk Products using GC-MS/MS (GC-MS/MS를 이용한 원유 원료 중 농약 동시분석법 확립)

  • Young Nae Choi;Yoon ho Shin;Hwangeui Cho;Jung Bok Kim
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.420-429
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    • 2023
  • GC-MS/MS using liquid-liquid extraction (LLE) and C18 cartridges was used to identify and quantify levels of chlorpyrifos, chlorpyrifos-methyl, cypermethrin, deltamethrin and permethrin in bulk raw milk. A calibration curve spanning 10 ng/mL to 200 ng/mL was obtained with a satisfactory correlation coefficient of 0.99. The limits of detection (LOD) and limits of quantitation (LOQ) for chlorpyrifos, chlorpyrifos-methyl, cypermethrin, deltamethrin, and permethrin in the matrix ranged from 0.06 to 1.81 ng/mL and 0.19 to 6.04 ng/mL, respectively. The recoveries of 5 pesticides from spiked samples at 37.5-125 ng/mL ranged from 86.1 to 102.1%. The measurement of uncertainty of the GC-MS/MS method for these five pesticides was developed based on the analytical process and quantification. An analysis method that is easier and faster than the method specified in the Korean food standards codes for analyzing these five pesticides in raw material milk was developed. Moreover, the analytical method for chlorpyrifos, chlorpyrifos-methyl, cypermethrin, deltamethrin, and permethrin in bulk raw milk by GC-MS/MS was established.

Simultaneous Analysis of Cold Medicine Component by High-Performance Liquid Chromatography(HPLC) (고성능 액체크로마토그래피(HPLC)를 이용한 Cold Medicine 성분의 동시 분석)

  • Wonju Lee;Seung-Tae Choi;Keun-Sik Shin;Jin-Young Park;Jae-Ho Sim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.6
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    • pp.867-873
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    • 2023
  • In this study, for the purpose of standardized quality control of a cold medicine, we simultaneous analyzed four main chemical components of a cold medicine: acetaminophen, caffeine, methyl paraben, and propyl paraben. The sample was subjected to quantitative analysis using high performance liquid chromatography (HPLC), after pretreatment of four components. The experiment was carried out by using Isocratic elution at wavelength of 270nm. Acetonitrile and water (H2O) were used as a mobile phase at a flow rate of 1.0mL/min in a commercial C18 reversed-phase column. A volume of 10uL cold medicine were injected into the column with column oven temperature at 35℃. As a result of the experiment, the values of Resolution were 4.983, 1.596, 5.519, and 1.678 respectively-well over Rs >1.5, which indicates that the separation of four components were efficient. In addition, value of symmetry factor of the components was 1.056, 1.069, 1.032, and 1.133 respectively, to show its symmetrical stability. The calibration curve of all four components exhibits good linearity with R2 >0.9995 to 0.9999. Furthermore, the limit of detection(LOD) were between 0.0118 to 1.5973 mg/mL, while the limit of quantification (LOQ) were between 0.0353 to 4.7919 ㎍/mL with the recovery rate of 79.6% ~ 120.5%. The results of this study showed an efficient quality evaluation of a simultaneous analysis method for cold medicine components.

Determination of Domoic Acid in Seafood Matrices using HPLC-UV with Solid Phase Extraction Cleanup (고체상 추출 전처리 및 HPLC-UV를 이용한 수산물 중 domoic acid의 분석)

  • Si Eun Kim;Sang Yoo Lee;Ji Eun Park;Hyunjin Jung;Hyang Sook Chun
    • Journal of Food Hygiene and Safety
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    • v.38 no.5
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    • pp.297-304
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    • 2023
  • Domoic acid (DA), a neurotoxin produced naturally by diatoms, is responsible for incidents of amnesic shellfish poisoning. In this study, a modified analytical method was established to determine domoic acid in seafood using solid phase extraction cleanup and optimizing the amount of sample and extraction solvent to reduce interference effects. The modified method using high-performance liquid chromatography with ultraviolet detection was validated using three seafood matrices (mussel, red snow crab, and anchovy) at three concentrations (1, 2, and 4 mg/kg) and compared to the Food Code method. Compared to the Food Code method, the modified method showed better performance in terms of linearity (R2>0.999), detection limit (0.02-0.03 mg/kg), quantification limit (0.05-0.09 mg/kg), intra-/inter-day accuracy (86.2-100.4%), and intra-/inter-day precision (0.2-4.0%). Furthermore, the method was successfully applied for the analysis of 87 seafood samples marketed in Korea, and DA was detected at a low concentration of 140 ㎍/kg in one anchovy sample. These results suggest that the modified method can be used for routine determination of DA in seafood.

Residual Characteristics and Processing Factors of Difenoconazole in Fresh Ginseng and Processed Ginseng Products (인삼 및 가공품 중 difenoconazole의 잔류특성 및 가공계수)

  • Noh, Hyun-Ho;Lee, Kwang-Hun;Lee, Jae-Yun;Lee, Eun-Young;Park, Young-Soon;Park, Hyo-Kyoung;Oh, Jae-Ho;Im, Moo-Hyeog;Lee, Yong-Jae;Baeg, In-Ho;Kyung, Kee-Sung
    • The Korean Journal of Pesticide Science
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    • v.16 no.1
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    • pp.35-42
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    • 2012
  • This study was carried out to elucidate the residual characteristics and calculate processing factors of difenoconazole in ginseng and its processed products, such as dried ginseng, red ginseng and their water and alcohol extracts. The pesticide was sprayed onto the ginseng according to its pre-harvest intervals in 2009 (four-year-old ginseng) and 2010 (five-year-old ginseng). Harvested ginseng was processed to dried ginseng, red ginseng and their extracts according to the commercially well-qualified conventional methods provided by the Korea Ginseng Corporation. Limit of detection (LOD) and limit of quantitation (LOQ) of difenoconazole in fresh ginseng were 0.001 and 0.003 mg/kg, respectively. In case of processed ginseng products, their levels were 0.002 and 0.007 mg/kg, respectively. Concentration of difenoconazole in both fresh ginseng and its processed products increased with the experimental period. Processing factors, calculated as a ratio of difenoconazole concentration in processed products to fresh ginseng were found to be 1.71 to 2.17 and 1.62 to 2.03 in case of dried and red ginseng, respectively, while those for their extracts ranged from 1.76 to 2.98. In case of five-year-old dried ginseng and red ginseng as well as their extracts, the ranges of processing factor of difenoconazole were found to be 2.9 to 3.1, 1.9 to 2.2 and 2.4 to 4.7, respectively.

Determination of Residue Levels of Ethyl Carbamate in Alcoholic Beverages by Gas Chromatography/Tandem Mass Spectrometry (GC/MS/MS) (가스크로마토그래피/질량분석기(GC/MS/MS)를 이용한 주류 중 에틸카바메이트 잔류량 조사)

  • Kim, Dong-Ho;Jang, Han-Sub;Choi, Gu-Il;Kim, Hyun-Jung;Kim, Ho-Jin;Kim, Hyo-Lin;Kim, Keun-Sung
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.63-68
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    • 2013
  • Ethyl carbamate (EC) is a contaminant generated in the fermentation processes of various fermented foods. In this study, residue levels of EC in 95 alcoholic beverage samples were determined by using Gas Chromatography/Tandem Mass Spectrometry (GC/MS/MS). All the samples were purified by a liquid-liquid extraction (LLE) method using dichloromethane. The LLE method enables an improvement in time and cost to detection and specificity over the conventional extraction methods. The limits of detection and quantification (LOD and LOQ) to analyze EC were 1.3 and 4.0 ng/mL, respectively. The recovery rates of EC were ranged from 90.0 to 97.5% at the levels of 50, 100, and 500 ug/L. Among traditional grain-based alcoholic beverage samples (n = 34), the average residue levels of EC in takju, yakju, and cheongju were 0.63, 7.01, and 14.11 ug/L, respectively. Among fruit-based alcoholic beverage samples (n = 48), those of EC in japanese apricot spirits, bokbunjaju, grape wines, and other fruit wines were 79.18, 1.66, 2.64, and 2.39 ug/L, respectively. Among distilled or diluted alcoholic beverage samples (n = 13), those of EC in soju (distilled or diluted), general distillates, liquors, and brandies were 0, 3.30, 8.20, and 8.52 ug/L, respectively. Therefore, this study reports that the residue levels of EC in the alcoholic beverages, distributed in the current domestic markets, did not reach its maximum allowed levels of 30 and 400 ug/L established for grape and fruit wines in Canada, respectively.

Development and Validation of an Official Analytical Method for Determination of Ipfencarbazone in Agricultural Products using GC-ECD (GC-ECD를 이용한 농산물 중 Ipfencarbazone의 신규분석법 개발 및 검증)

  • Jang, Jin;Kim, Heejung;Lee, Eun-Hyang;Ko, Ah-Young;Ju, Yunji;Kim, Sooyeon;Chang, Moon-Ik;Rhee, Gyu-Seek
    • The Korean Journal of Pesticide Science
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    • v.19 no.3
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    • pp.210-217
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    • 2015
  • Ipfencarbazone is a herbicide of the tetrazolinone class, and is believed to be an inhibitor of very long chain fatty acids (VLCFAs), which control cell division in weeds. The objective of this study was to develop and validate an official analytical method for ipfencarbazone determination in agricultural products. The ipfencarbazone residues in agricultural products were extracted with acetone, partitioned with n-hexane, and then purified through silica SPE cartridge. Finally, the analyte was quantified by gas chromatograph-electron capture detector (GC-ECD) and confirmed with gas chromatograph/mass spectrometer(GC/MS). The linear range of ipfencarbazone was 0.01 to 1.0 mg/L with the coefficient of determination ($r^2$) of 0.9999. The limit of detection (LOD) and quantification (LOQ) was 0.003 and 0.01 mg/kg, respectively. In addition, average recoveries of ipfencarbazone ranged from 80.6% to 112.3% at the different concentration levels LOQ, 10LOQ and 50LOQ, while the relative standard deviation was 2.2-8.6%. All values were consistent with the criteria ranges requested in the CODEX guidelines. Furthermore, and inter-laboratory study was conducted to validate the method. This proposed method for determination of ipfencarbazone residues in agricultural products can be used as an official analytical method.

Multi-residue Pesticide Analysis in Cereal using Modified QuEChERS Samloe Preparation Method (곡물류 중 잔류농약 다성분 분석을 위한 개선된 QuEChERS 시료 정제법의 개발)

  • Yang, In-Cheol;Hong, Su-Myeong;Kwon, Hye-Young;Kim, Taek-Kyum;Kim, Doo-Ho
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.314-334
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    • 2013
  • This study explored an efficient modified Quick, Easy, Cheap, Effective, Rugged and safe (QuEChERS) method combined with liquid chromatography-electrospray ionization with tandem mass spectrometric detection for the analysis of residues of 76 pesticides in brown rice, barley and corn including acidic sulfonylurea herbicides. Formic acid (1%) acid in acetonitrile and dispersive solid phase extractions used for extraction of pesticides and clean-up of the extract respectively. Two fortified spikes at 50 and 200 ng $g^{-1}$ levels were performed for recovery test. Mean recoveries of majority of pesticides at two spike levels ranged from 73.2 to 132.2, 80.9 to 136.8, 66.6 to 143.5 for brown rice, barley and corn respectively with standard error (CV) less than 10%. Good linearity of calibration curves were achieved with $R^2$ > 0.9907 within the observed concentration ranged. The modified method also provided satisfactory results for sulfonylurea herbicides. The method was applied to the determination of residues of target pesticides in real samples. A total of 26 pesticides in 36 out of 98 tasted samples were observed. The highest concentration was observed for tricyclazole at 1.17 mg $kg^{-1}$ in brown rice. This pesticide in two brown rice samples exceeded their MRLs regulated for rice in republic of Korea. Except tricyclazole none of the observed pesticides' concentration was higher than their MRLs. The results reveal that the method is effectively applicable to routine analysis of residues of target pesticides in brown rice, barley and corn.