• Title/Summary/Keyword: Genetic Marker

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Validity Test for Molecular Markers Associated with Resistance to Phytophthora Root Rot in Chili Pepper (Capsicum annuum L.) (고추의 역병 저항성과 연관된 분자표지의 효용성 검정)

  • Lee, Won-Phil;Lee, Jun-Dae;Han, Jung-Heon;Kang, Byoung-Cheorl;Yoon, Jae-Bok
    • Horticultural Science & Technology
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    • v.30 no.1
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    • pp.64-72
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    • 2012
  • Phytophthora root rot has been causing a serious yield loss in pepper production. Since 2004, the year in which commercial cultivars resistant to the disease were firstly commercialized, it has been necessary to introduce the resistance into domestic pepper cultivars for dried red pepper. Therefore, developing molecular markers linked to the resistance is required for an accurate selection of resistant plants and increasing breeding efficiency. Until now, several markers associated with the major dominant gene resistant to Phytophthora root rot have been reported but they have some serious limitations for their usage. In this study, we aimed to develop molecular markers linked to the major dominant gene that can be used for almost of all genetic resources resistant to Phytophthora root rot. Two segregating $F_2$ populations derived from a 'Subicho' ${\times}$ 'CM334' combination and a commercial cultivar 'Dokyacheongcheong' were used to develop molecular markers associated with the resistance. After screening 1,024 AFLP primer combinations with bulked segregant analysis, three AFLP (AFLP1, AFLP2, and AFLP3) markers were identified and converted into three CAPS markers (M1-CAPS, M2-CAPS, and M3-CAPS), respectively. Among them, M3-CAPS marker was further studied in ten resistants, fourteen susceptibles, five hybrids and 53 commercial cultivars. As a result, M3-CAPS marker was more fitted to identify Phytophthora resistance than previously reported P5-SNAP and Phyto5.2-SCAR markers. The result indicated that the M3-CAPS marker will be useful for resistance breeding to Phytophthora root rot in chili pepper.

Skewed Inheritance of EST-SSR Alleles in Reciprocal Crosses of Cut Roses (절화장미 품종간 정역교배에 있어서 EST-SSR 마커의 유전)

  • Kim, Jin-Ki;Ahn, Dong-Chun;Oh, Hye-Jeong;Kim, Kwang-Hwan;Choi, Young-Mi;Oh, Seung-Yong;Kang, Nam-Jun;Jeong, Byoung-Ryong;Kim, Zhoo-Hyeon;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.618-626
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    • 2010
  • Matroclinal inheritance of morphological characters in interspecific crosses of Rosa spp. can be influenced by cytoplasmic inheritance, apomixis, and asynaptic heterogamy. In asynaptic heterogamy, which is often observed from interspecific crosses of Rosa sect. $Caninae$, the polyploidy of the seed parent (especially for 5x=35) is recovered in the progeny through the pollens that include only a set of bivalents (x=7) and egg cells that contain a set of bivalents (x=7) and other univalents (3x=21). In this study, we investigated the causes of matroclinal offsprings observed from reciprocal crosses of tetraploid cut rose cultivars ($Rosa$ $hybrida$ L.) by analyzing EST-SSR marker distribution in the progeny populations. From EST-SSR marker analysis of eight offsprings per six reciprocal crosses among six cultivars, cases of cytoplasmic inheritance were not observed. Apomixis was also very rare as compared to the reports on interspecific crosses of sect. $Caninae$; only one apomitic plant was identified from the cross 'Redtem' ${\times}$ 'Red Sandra'. Although a clear-cut pattern of asynaptic heterogamy was not found, cultivar-specific marker transmission skewed to seed parent in four cultivars implied that genetic inheritance can be highly influenced by the seed parent depending on crosses among cut rose cultivars; especially, 10 out of 11 alleles specific to 'Yellow King' distributed in progenies at higher ratios when the cultivars were crossed as the seed parent.

Identification of Quantitative Trait Loci Associated with Anthracnose Resistance in Chili Pepper (Capsicum spp.) (고추 탄저병 저항성 관련 양적형질 유전자좌 분석)

  • Kim, Su;Kim, Ki-Taek;Kim, Dong-Hwi;Yang, Eun-Young;Cho, Myeong-Cheoul;Jamal, Arshad;Chae, Young;Pae, Do-Ham;Oh, Dae-Geun;Hwang, Ju-Kwang
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1014-1024
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    • 2010
  • Pepper ($Capsicum$ spp.) anthracnose caused by $Colletotrichum$ $acutatum$ is a destructive disease susceptible to areas where chili peppers are grown. $Capsicum$ $baccatum$ var. $pendulum$ (Cbp) is resistant to anthracnose and has actively been used for interspecific hybridization for the introgression of resistance gene(s) into cultivated chili peppers. The goals of this study were to determine the inheritance of resistance to anthracnose within $Capsicum$ $baccatum$ and to map quantitative trait loci (QTLs) for the anthracnose resistance. A genetic mapping population consisting of 126 $F_2$ plants derived from a cross between $Capsicum$ $baccatum$ var. $pendulum$ (resistant) and $Capsicum$ $baccatum$ 'Golden-aji' (susceptible) was used for linkage mapping. The linkage map was constructed with 52 SSRs, 175 AFLPs, and 100 SRAPs covering 1,896cM, with an average interval marker distance of 4.0cM. Based on this map, the number, location, and effect of QTLs for anthracnose resistance were studied using plants inoculated in the laboratory and field. A total of 19 quantitative trait loci (2 major QTLs and 16 minor QTLs) were detected. Two QTLs ($An8.1$, $An9.1$) showed 16.4% phenotypic variations for anthracnose resistance after wounding inoculation. In addition, five minor QTL loci ($An7.3$, $An7.4$, $An4.1$, $An3.1$, $An3.2$) showed a total of 60.73% phenotypic variations of anthracnose resistance in the field test. Several significant QTLs were also detected and their reproducibility was confirmed under different inoculation conditions. These QTLs are now being confirmed with different breeding populations. Markers tightly linked to the QTLs that are reliable under different environmental conditions will help to determine the success of marker-assisted selection for anthracnose -resistant breeding programs in chili pepper.

Noninvasive Prenatal Diagnosis using Cell-Free Fetal DNA in Maternal Plasma: Clinical Applications

  • Yang, Young-Ho;Han, Sung-Hee;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.1-16
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    • 2011
  • Owing to the risk of fetal loss associated with prenatal diagnostic procedures (amniocentesis, chorionic villus sampling), noninvasive prenatal diagnosis (NIPD) is ultimate goal of prenatal diagnosis. The discovery of circulating cell-free fetal DNA (cffDNA) in maternal plasma in 1997 has opened up new probabilities for NIPD by Dr. Lo et al. The last decade has seen great development in NIPD. Fetal sex and fetal RhD status determination by cffDNA analysis is already in clinical use in certain countries. For routine use, this test is limited by the amount of cell-free maternal DNA in blood sample, the lack of universal fetal markers, and appropriate reference materials. To improve the accuracy of detection of fetal specific sequences in maternal plasma, internal positive controls to confirm to presence of fetal DNA should be analyzed. We have developed strategies for noninvasive determination of fetal gender, and fetal RhD genotyping using cffDNA in maternal plasma, using real-time quantitative polymerase chain reaction (RT-PCR) including RASSF1A epigenetic fetal DNA marker (gender-independent) as internal positive controls, which is to be first successful study of this kind in Korea. In our study, accurate detection of fetal gender through gestational age, and fetal RhD genotyping in RhD-negative pregnant women was achieved. In this assay, we show that the assay is sensitive, easy, fast, and reliable. These developments improve the reliability of the applications of circulating fetal DNA when used in clinical practice to manage sex-linked disorders (e.g., hemophilia, Duchenne muscular dystrophy), congenital adrenal hyperplasia (CAH), RhD incompatibility, and the other noninvasive pregnant diagnostic tests on the coming soon. The study was the first successful case in Korea using cffDNA in maternal plasma, which has created a new avenue for clinical applications of NIPD.

Physical Characteristics and Microsatellite Polymorphisms in Miryang Native Dogs (밀양지방 토종개의 형태학적 특징 및 유전적 다양성 연구)

  • Cho, Byung-Wook;Cho, Gil-Jae
    • Journal of Life Science
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    • v.16 no.4
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    • pp.626-631
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    • 2006
  • This study was carried out to investigate the physical characteristics (height, body lenght, chest depth, head type, ear type, body color, eye type and tail type) and genetic diversity using 15 microsatellite DNA markers (PEZ 1, 5, 8, 10, 11, 12, 13, 15, 16, 17, 20, 21, FHC 2010, FHC 2054 and FHC 2079) in 44 random Miryang native dogs(6 months${\sim}$12 years old). The height, body lenght, and chest depth of Miryang native dogs were 43-55 cm(mean 49.5 cm), 45-60 cm(mean 54.3 cm), and 50-64 cm(mean 57.9 cm), respectively. Miryang native dog was medium sized. The head and eye type were reverse-triangle(100%), triangle (90.9%) and newborn moon(9.1%), respectively. Most of body color had white coat color(93.2%), light pink tongue color(100%), light black anal color(90,9%) and pink claw color(100%). The ear type showed erect ear(100%), and half-curled(56.8%), upward(34.1%), curled(9.1%) in tail type, respectively. Number of alleles observed at a single locus ranged from 2 (PEZ 21 and FHC 2010) to 14 (PEZ 13), with average number of alleles per locus of 6.13. The expected heterozygosities of 15 microsatellite loci were estimated based on gene frequencies. The highest expected heterozygosity, 0.863 was estimated in PEZ 13 locus and the lowest, 0.455 in PEZ 21 and FHC 2010 locus. And the mean expected heterozygosity of 15 microsatellite markers was calculated as 0.635. Polymorphic information content (PIC) values were ranged from 0.348 (PEZ 21 and FHC 2010) to 0.837 (PEZ 13), and the mean PIC value was calculated as 0.570. Of the 15 markers, PEZ 10, PEZ 13, PEZ 17 and FHC 2054 loci have relatively high PIC value (> 0.7) in Miryang native dog. In order to determine the efficieney of parentage control, exclusion probabilities (PE) were calculated for each allele. The highest PE 1 and PE 2 in PEZ 13 locus was caculated to 0.548 and 0.710, respectively. And the total exclusion power in PE 1 and PE 2 was calculated to 0.9895 and 0.9996, respectively. These results can give basic information for perservation and research in Miryang native dog, and phylogenetic relationships of the Korean native dog and Asian dog breeds.

Validation of QF-PCR for Rapid Prenatal Diagnosis of Common Chromosomal Aneuploidies in Korea

  • Han, Sung-Hee;Ryu, Jae-Song;An, Jeong-Wook;Park, Ok-Kyoung;Yoon, Hye-Ryoung;Yang, Young-Ho;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.7 no.1
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    • pp.59-66
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    • 2010
  • Purpose: Quantitative fluorescent polymerase chain reaction (QF-PCR) allows for the rapid prenatal diagnosis of common aneuploidies. The main advantages of this assay are its low cost, speed, and automation, allowing for large-scale application. However, despite these advantages, it is not a routine method for prenatal aneuploidy screening in Korea. Our objective in the present study was to validate the performance of QF-PCR using short tandem repeat (STR) markers in a Korean population as a means for rapid prenatal diagnosis. Material and Methods: A QF-PCR assay using an Elucigene kit (Gen-Probe, Abingdon, UK), containing 20 STR markers located on chromosomes 13, 18, 21, X and Y, was performed on 847 amniotic fluid (AF) samples for prenatal aneuploidy screening referred for prenatal aneuploidy screening from 2007 to 2009. The results were then compared to those obtained using conventional cytogenetic analysis. To evaluate the informativity of STR markers, the heterozygosity index of each marker was determined in all the samples. Results: Three autosomes (13, 18, and 21) and X and Y chromosome aneuploidies were detected in 19 cases (2.2%, 19/847) after QF-PCR analysis of the 847 AF samples. Their results are identical to those of conventional cytogenetic analysis, with 100% positive predictive value. However, after cytogenetic analysis, 7 cases (0.8%, 7/847) were found to have 5 balanced and 2 unbalanced chromosomal abnormalities that were not detected by QF-PCR. The STR markers had a slightly low heterozygosity index (average: 0.76) compared to those reported in Caucasians (average: 0.80). Submicroscopic duplication of D13S634 marker, which might be a unique finding in Koreans, was detected in 1.4% (12/847) of the samples in the present study. Conclusion: A QF-PCR assay for prenatal aneuploidy screening was validated in our institution and proved to be efficient and reliable. However, we suggest that each laboratory must perform an independent validation test for each STR marker in order to develop interpretation guidelines of the results and must integrate QF-PCR into the routine cytogenetic laboratory workflow.

Current status and prospects of molecular marker development for systematic breeding program in citrus (감귤 분자육종을 위한 분자표지 개발 현황 및 전망)

  • Kim, Ho Bang;Kim, Jae Joon;Oh, Chang Jae;Yun, Su-Hyun;Song, Kwan Jeong
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.261-271
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    • 2016
  • Citrus is an economically important fruit crop widely growing worldwide. However, citrus production largely depends on natural hybrid selection and bud sport mutation. Unique botanical features including long juvenility, polyembryony, and QTL that controls major agronomic traits can hinder the development of superior variety by conventional breeding. Diverse factors including drastic changes of citrus production environment due to global warming and changes in market trends require systematic molecular breeding program for early selection of elite candidates with target traits, sustainable production of high quality fruits, cultivar diversification, and cost-effective breeding. Since the construction of the first genetic linkage map using isozymes, citrus scientists have constructed linkage maps using various DNA-based markers and developed molecular markers related to biotic and abiotic stresses, polyembryony, fruit coloration, seedlessness, male sterility, acidless, morphology, fruit quality, seed number, yield, early fruit setting traits, and QTL mapping on genetic maps. Genes closely related to CTV resistance and flesh color have been cloned. SSR markers for identifying zygotic and nucellar individuals will contribute to cost-effective breeding. The two high quality citrus reference genomes recently released are being efficiently used for genomics-based molecular breeding such as construction of reference linkage/physical maps and comparative genome mapping. In the near future, the development of DNA molecular markers tightly linked to various agronomic traits and the cloning of useful and/or variant genes will be accelerated through comparative genome analysis using citrus core collection and genome-wide approaches such as genotyping-by-sequencing and genome wide association study.

Quality and genetic relationship on dehulling characteristic in oat variety (국내 귀리품종 탈부특성에 따른 품질 및 유연관계 분석)

  • Kim, Kyeong-Min;Shin, Dongjin;Kim, Kyeong-Hoon;Kwon, Yu-Ri;Park, Hyo-Jin;Hyun, Jong-Nae;Kim, Young-Jin;Oh, Myung-Kyu;Park, Young-Hoon
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.375-381
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    • 2018
  • The consumption of oats (Avena sativa L.) with high nutritional utility is accelerating due to the increased consumers' demand for functional foods. In Korea, naked oats are used as food, while covered oats are used for animal feed. However, it is difficult to distinguish naked oats from covered oats when the husk is removed from the grains by a special process. The present study was carried out to investigate experimental methods that would be beneficial in the segregation of different types of oats after husk removal. Grain quality-related biochemical compounds were analyzed in a bid to differentiate the oat dehulling characteristics. In addition, 61 SSR markers were examined for genetic relationship and variety identification of oats using five naked and seven covered oat varieties. Results showed that, the contents of protein, lipid, and ${\beta}-glucan$ were not significantly different among the oat varieties and this could not be used as an index for distinguishing oats husk character. However, in the fatty acid composition ratio,, naked oats had a higher ratio of stearic acid (C18:0) and oleic acid (C18:1) than covered oats, and covered oats had a higher ratio of linoleic acid (C18:2) and linoleic acid (C18:3) than naked oats. The assessment of SSR marker genotype revealed that 33 polymorphic bands among 12 oat varieties and 1 variety could be distinguished through the combination of polymorphic markers thus indicating the usability of these markers for variety identification in oats.

Comparative Analysis of Freshwater Fish Species in Civilian Control Zone in South Korea: A Comparison between Direct Survey Results and Indirect Assessment via eDNA (우리나라 민간인통제구역 내 수계 어류에 대한 비교분석: 직접조사 결과와 eDNA를 통한 간접조사 결과 비교)

  • Soon-Jae Eum;Naeyoung Kim;Min-A Seol;Ji Young Kim
    • Korean Journal of Ichthyology
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    • v.35 no.4
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    • pp.224-235
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    • 2023
  • South Korea is the only divided nation globally, marked by a military demarcation line establishing demilitarized and civilian control zones, ensuring national security. Consequently, these areas exhibit relatively minimal ecological disruption compared to other regions. However, the threat to safety persists due to the presence of unexploded ordnances and landmines, imposing significant constraints on ecological research. To address this, we conducted a comparative study utilizing eDNA analysis as a supplementary and alternative approach within three points of the "Road of Peace" - Inje, Yanggu, and Hwacheon courses, located within the civilian control zone. Direct surveys and indirect eDNA sampling were carried out in May, July, and September of 2022. Genetic material obtained from the samples underwent amplification, library preparation, MiSeq sequencing, and subsequent ASV generation for indirect analysis. These results were then compared with the findings of direct surveys. Our findings revealed the detection of eDNA for both observed species at the Yanggu-1 point, and for two out of four species at Yanggu-2. Hwacheon-1 displayed the detection of eDNA for one out of one observed species, whereas Hwacheon-2 yielded seven out of twelve, Hwacheon-3 showed four out of six, and all one observed species at Hwacheon-4 exhibited eDNA detection. Consequently, approximately 69% of the fish species identified through direct surveys were confirmed by indirect eDNA analysis. It is necessary to verify if certain fish species, such as the continental trout and catfish, have genetic information registered in the NCBI database. Additionally, it is believed that further marker development research utilizing different genetic sequences is essential. Given the limitations imposed by the hazardous nature of the surveyed civilian control zone, eDNA analysis proves to be a suitable supplement for fish research in the area.

Effect of the Fatty Acid Synthase Gene for Beef Quantity Traits in Hanwoo Breeding Stock (한우 Fatty Acid Synthase (FASN) 유전자 반수체형의 후대검정우 육량 및 육질에 미치는 영향)

  • Kim, Sang-Wook;Lee, Jun-Heon;Kim, Jin-Ho;Won, You-Seog;Kim, Nae-Soo;Kim, Kwan-Suk
    • Journal of Animal Science and Technology
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    • v.52 no.1
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    • pp.9-16
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    • 2010
  • A previous study has shown that the g.17924G>A polymorphism of fatty acid synthase (FASN) is associated with unsaturated fatty acid composition in the Hanwoo beef, hence this study was conducted to evaluate the effect of single nucleotide polymorphisms (SNPs) within FASN gene on the selection phenotypes of Hanwoo breeding stock. A total of 925 progeny test steers were used to genotype g.11280G>A, g.13125T>C, and g.17924G>A polymorphisms and significant associations were found among g.11280G>A, g.17924G>A, and carcass traits, such as carcass weight, backfat thickness, and beef quantity index. No significant association was found between g.13125T>C and carcass traits. Although the degree of linkage disequilibrium (LD) was not strong among g.11280G>A, g.13125T>C, and g.17924G>A in the LD analysis, four major haplotype classes were formed with the genotypic information within the FASN gene; the frequencies of the halpotypeswere -GCG-[0.378], -ATG-[0.301], -GTA-[0.191], and -ACG-[0.063], respectively. Phenotypic association was performed among these haploptypes, and the haplotype 2 (-ATG-)was significantly associated with higher carcass weight when compared to the other haplotypes, i.e. haplotype 1 (-GCG-) and haplotype 3 (-GTA-). A copy number of the FASN haplotype 3 (-GTA-) had also a significant association with carcass weight of subjects. In conclusion, it was observed that two polymorphisms (g.11280G>A and g.17924G>A) and their haplotypes within the FASN gene are consistently associated with carcass traits. Therefore, it is desirable to use the FASN polymorphisms for pre-selection program as genetic marker with improved carcass yield and beef quality of the Hanwoo sire at the Hanwoo Improvement Center as well as for commercial Hanwoo producers, the FASN genotypic information can be used for a part of selecting Hanwoo dam for superior calf production.