• Research in Plant Disease
• /
• v.18 no.2
• /
• pp.73-79
• /
• 2012

Phytophthora katsurae is a fungal pathogen responsible for chestnut ink disease. We designed two duplex primer sets (SOPC 1F/1R+KatI 3F/5R, SOPC 1-1F/1-1R+KatI 3F/5R) to detect P. katsurae. SOPC 1F/1R and SOPC 1-1F/1-1R primer pairs were designed for sequence characteristic amplification regions (SCAR) marker, and KatI 3F/5R primer pair was used for P. katsurae-specific primer designed from internal transcribed spacer (ITS) region. To assess the sensitivity of duplex PCR, genomic DNA was serially diluted 10-fold to make the final concentrations from 1 mg/ml to 1 ng/ml. The sensitivity for two primer sets were 1 ${\mu}g/ml$ and 100 ng/ml, respectively. To find detection limits for zoospores of P. katsurae, each zoospore suspension was serially diluted 10-fold to make the final concentrations from $1{\times}10^6$ to $1{\times}10^2$ cells/ml, and then DNA was extracted. The limits of detection for all of two primer sets were $1{\times}10^5$ cells/ml. All of two primer sets were specific to P. katsurae in PCR detection and did not produce any P. katsurae-specific PCR amplicons from other 16 Phytophthora species used as the control. This study shows that duplex PCR using two primer sets might be a useful tool for rapid and efficient detection of P. katsurae.

• Journal of Life Science
• /
• v.15 no.3 s.70
• /
• pp.474-478
• /
• 2005

This study was conducted to analyze genetic characteristics and to develop the specific marker for Cheju native horse (Coo) at the level of sequence characterized amplified regions (SCARs). We collected blood samples from Cheju native horse and Thoroughbred horse (Th) and obtained genomic DNA from the blood of 50 individuals randomly selected within the breeds. Seven hundred primers were chosen randomly and were used to examin the polymorphism and 40 kinds of primers showed polymorphic RAPD band patterns between two breeds. Thirty primers of them showed horse specific bands. With the primer MG 30, amplified band of 2.0 kb showed the specificity to Cheju native horse (Cnh). Additionally MG 53 detected the thoroughbred horse (Th) specific markers at size of 2.3 kb. As the next, 2.3 kb band from MG 53 was checked with the all individuals from all the breeds of this study, and it maintained the reproducible breed specificity to thoroughbred horse (Th). With this results, 2.3 kb band was cloned into plasmid vector and sequenced bidirectionally from both ends of the cloned fragment. With the obtained sequences 10 nucleotide extended primers including the original arbitray primer were designed as a SCARs primer. Finally, the primer with extended sequence showed the reproducible breed differentiation pattern and it was possible to identify Cheju native horse (Cnh) from other breeds. The SCARs marker 2.3 kb from MG 53 could be used to identify Cheju native horse (Cnh) for not only registration but also horse breeding programe.

• Journal of Crop Science and Biotechnology
• /
• v.10 no.2
• /
• pp.98-105
• /
• 2007

Phylogenetic relationships in Korean watermelons were evaluated by genetic similarity coefficients using 15 SSR(simple sequence repeat), 14 SCAR(sequence characterized amplified region) and 14 CAPS(sequence characterized amplified region) markers. The SSR markers were selected from previously reported melon and watermelon SSRs through testing polymorphisms within a set of commercial $F_1$ varieties. The SCAR and CAPS markers were developed from polymorphic AFLP(amplified fragment length polymorphism) markers between inbred lines 'BN4001' and 'BN4002'. From the AFLP analysis, 105 polymorphic fragments were identified between the inbred lines using 1,440 primer combinations of EcoRI+CNNN and XbaI+ANNN. Based on the sequencing data of these polymorphic fragments, we synthesized sequence specific primer pairs and detected clear and reliable polymorphisms in 27 primer pairs by indels(insertion/deletion) or RFLP(restriction fragment length polymorphism). A total of 43 sequence-based PCR markers were obtained and polymorphic information content(PIC) was analyzed to measure the informativeness of each marker in watermelon varieties. The average PIC value of SCAR markers was 0.41, which was similar to that of SSR markers. Genetic diversity was also estimated by using these markers to assess the phylogenetic relationships among commercial varieties of watermelon. These markers differentiated 26 Korean watermelon varieties into two major phylogenetic groups, but this grouping was not significantly correlated with their morphological and physiological characteristics. The mean genetic similarity was 66% within the complete set of 26 commercial varieties. In addition, these sequence-based PCR markers were reliable and useful to identify cultivars and genotypes of watermelon.

• Archives of Craniofacial Surgery
• /
• v.12 no.1
• /
• pp.71-74
• /
• 2011

Purpose: Sarcoidosis is a systemic noncaseating granulomatous disease of an unknown origin, and can involve any organ including the skin. The infiltration of sarcoid granuloma in an old cutaneous scar is an uncommon cutaneous manifestation of sarcoidosis. This paper reports a 35-year old female who presented with cutanesous nodules in previous facial scars. Methods: A 35-year-old female presented with cutaneous nodules for 2 months in previous scars of the forehead and lower lip that she had acquired in the childhood. An excisional biopsy of the lower lip mass, serologic examinations and radiologic studies were performed. Results: The excisional biopsy revealed noncaseating granulomas consistent with sarcoidosis. Chest CT revealed both hilar and paratracheal lymphadenopathy with nodular densities in both lung fields. Routine laboratory tests, serologic tests, serum angiotensin converting enzyme level, sputum for acid-fast bacilli, ophthalmoscopic examination, TB-PCR and NTM-PCR showed normal findings. Therefore, the patient was diagnosed with sarcoidosis. The patient refused fiberoptic bronchoscopy and medication with oral steroid. Currently, the clinical manifestation and progress are being monitored closely, and treatment is expected to start with oral steroid according to the progress. Conclusion: For the proper management of cutaneous sarcoidosis, particularly scar sarcoidosis, plastic surgeons should be aware of the many clinical and histopathological features of sarcoidosis and recommend a systemic evaluation for early diagnosis and proper treatment.

• The Korea Journal of Herbology
• /
• v.37 no.5
• /
• pp.9-16
• /
• 2022

Objectives : The purpose of this study is to report that applying the genetic discrimination method to Pinelliae Tuber is suitable as a countermeasure for the limitations of morphological identification announced publicly in the Ministry of Food and Drug Safety(MFDS). Methods : Randomly selected fifty samples in Pinelliae Tuber imported from China were used for morphological and genetic identification. The morphological identification was applied method announced publicly by the MFDS. The traits of morphological identification were classified as Pinellia ternata, P. tripartita, Pinellia pedatisecta, and Typhonium flagelliforme, according to the formation of tuberous root and tuber morphology. The genetic identifications were conducted by Sequence Characterized Amplified Region(SCAR) marker and DNA barcoding analysis for cross-validation, respectively. SCAR marker was verified according to the presence or absence of amplicon through PCR amplification using species-specific primers. DNA barcoding analysis used sequence information of the matK region. Results : As a result of the morphological identification, 27 out of 50 samples were identified as original species 'P. ternata' of genuine 'Pinelliae Tuber', and 23 were identified as adulterant species 'P. pedatisecta'. Unlike this, the genetic identification was identified as the original species 'P. ternata' in all 50 samples in the SCAR marker and matK regional sequence analysis. Conclusions : Pinelliae Tuber of morphological mutant that can not be classified by morphological identification is imported from China. The SCAR marker would be used as accurate and efficient assays for species identification of the morphological mutant.

• Korean Journal of Breeding Science
• /
• v.43 no.4
• /
• pp.265-272
• /
• 2011

This study is to generate SCARs markers for identification of Perilla species. A SCAR is a genomic DNA fragment at a single genetically defined locus that is identified by PCR amplification using a pair of specific oligonucleotide primers. We derived SCARs by sequencing and cloning the both ends of the amplified products of RAPD markers. Sixteen sequence-specific primers were synthesized from eight RAPD markers, which were completely sequenced. We developed the species-specific SCAR markers which could be used successfully in detecting genetic variation in four Perilla species. These markers could be used to verify species-origins of various forms of Perilla germplasms.

• Korean Journal of Medicinal Crop Science
• /
• v.12 no.1
• /
• pp.53-59
• /
• 2004

Finding a means to discriminate the commercial herb medicines when they were dried and sliced is a very important and imminent project in Korea. To differentiate plant origins and the commercial herb medicines of Atractylodes japonica and A. macrocephala, two discriminative methods using anatomical characteristics and SCAR marker were applied. It was possible to discriminate plant origins and the commercial herb medicines between A. japonica and A. macrocephala by anatomical characteristics: development of periderm, layer of stone cell, distribution of laticiferous vessels, development of xylem fiber in xylem ray, contained quantity of clustered crystals and others. While, two SCAR markers were developed from RAPD clones: SAjR2 (600 bp) from AjR2 and SAmR1 (1,200 bp) from AmR1. These two markers were enough for discrimination plant origins and the commercial herb medicines between A. japonica and A. macrocephala. The result of application of anatomical characteristics and SCAR markers to investigate current status in domestic herb market, Daegu and Kumsan herb market, it was identified to be current herb medicines of A japonica.

• Childhood Kidney Diseases
• /
• v.5 no.1
• /
• pp.43-50
• /
• 2001

Purpose : The urinary tract infection associated with vesicoureteral reflux(VUR) in children may result in serious complications such as renal scarring, hypertension, proteinuria and end stage renal disease. The purpose of this study was to evaluate the factors affecting renal scar such as age, gender, grade of VUR, and ACE gene polymorphism, and body growth in the patients with and those without renal scar associated with VUR Methods : During the period from January 1994 to July 2000, We had 93 children with urinary tract infection associated with VUR who were admitted to the Department of pediatrics of Chonbuk National University Hospital. The patients were divided into two groups according to follow up 99mTc-DMSA renal scan; patients with renal scar group and those with non-scar group. We analyzed and compared the factors associated with renal scarring between the two groups. Results : There were no significant difference in gender, causative organism, ACE gene polymorphism, height and weight at diagnosis between renal scar group and non-scar group. Fifty four patients were in renal scar group and forty seven of them had VUR. The age at diagnosis was significantly higher in renal scar group (2.48${\pm}$2.64yr) than in non renal scar group (1.26${\pm}$1.83yr). Especially, the infants who were less than 1 year of age with VUR developed relatively more renal scar compared with infants older than 1 tear of age. The incidence of renal scarring showed a direct correlation with the severity of VUR. Conclusion : The factors affecting renal scar formation were age at diagnosis, presence and grade of VUR, but the other factors such as gender, causative organism, ACE gene polymorphism were not associated with renal scarring. Therefore, further evaluation about uropathogenic E coli and foflow up study about body growth associated with severity of renal scar would be necessary. (J. Korean Soc Pediatr Nephrol 5 : 43- 50, 2001)

• The Plant Pathology Journal
• /
• v.22 no.1
• /
• pp.63-67
• /
• 2006

Apple scar skin, one of the most destructive diseases affecting apple, is caused by Apple scar skin viroid (ASSV d). Fruit dappling appeared on several cultivars in Korea and has been distributed to major cultivated areas since 2001. ASSVd was identified from infected fruits by using nucleic acid sequence-based amplification with electrochemiluminescence (NASBA-ECL). NASBA-ECL method was faster and hundredfold more sensitive than reverse transcription-polymerase chain reaction (RT-PCR) for ASSVd detection in apple leaves/ stems. ASSVd was rapidly transmitted to the entire tree in the second year after artificial inoculation. The ASSVd could be transmitted efficiently by using contaminated pruning scissors to both lignified stems (60 to $70\%$) and green shoots (20 to $40\%$) of apple tree and young plants. Dipping of contaminated scissors in $2\%$ sodium hypochlorite solution effectively prevented viroid transmission. In the ASSV d-infected fruits, the viroid was easily detected from fruit skin, seed coat, and embryo. Moreover, embryo and endosperm separately excised from the ASSVd-infected seeds were ASSVd positive in NASBA-ECL assay. Seedlings germinated from ASSVd-positive seeds showed $7.7\%$ infection rate., which indicated that ASSVd is seed-borne.

• Korean journal of applied entomology
• /
• v.55 no.3
• /
• pp.245-256
• /
• 2016

Elytra of Harmonia axyridis exhibit varied color patterns. In the present study, we deciphered the genetic basis for intraspecific diversity of elytra color patterns in H. axyridis, using amplified fragment length polymorphism (AFLP). Twenty-eight AFLP reactions were performed to generate a total of 2,741 bands. Of these, 20 bands were polymorphic for each color pattern. The polymorphic bands showed differences of genetic character among different color patterns of H. axyridis. Among them, ten candidate AFLP markers were color-linked. S1, S2, and S20 markers were detected in Succinea 1 and 2 variants of H. axyridis, whereas S3 and S5 were specifically detected in the Conspicua variant. S15, S18, and S19 were specific to the Succinea 2 variant. Polymerase chain reaction (PCR) products of these ten AFLP markers were sequenced. BLAST analysis of these sequences against the GenBank database revealed their homology to DNA fragments of unknown function. Based on the color-linked AFLP markers, sequence characterized amplified region (SCAR) markers were designed for PCR amplification of genomic DNA. Of the ten AFLP markers, five were successfully converted into SCAR markers, which could discriminate elytra color polymorphism in H. axyridis.