• Mycobiology
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• v.45 no.4
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• pp.401-408
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• 2017

Colletotrichum gloeosporioides is an economically important fungal pathogen causing substantial yield losses indifferent host plants. To understand the genetic diversity and molecular epidemiology of this fungus, we have developed a novel, high-resolution multi-locus microsatellite typing (MLMT) method. Bioinformatic analysis of C. gloeosporioides unannotated genome sequence yielded eight potential microsatellite loci, of which five, CG1 $(GT)_n$, CG2 $(GT1)_n$, CG3 $(TC)_n$, CG4 $(CT)_n$, and CG5 $(CT1)_n$ were selected for further study based on their universal amplification potential, reproducibility, and repeat number polymorphism. The selected microsatellites were used to analyze 31 strains of C. gloeosporioides isolated from 20 different host plants from India. All microsatellite loci were found to be polymorphic, and the approximate fragment sizes of microsatellite loci CG1, CG2, CG3, CG4, and CG5 were in ranges of 213-241, 197-227, 231-265, 209-275, and 132-188, respectively. Among the 31 isolates, 55 different genotypes were identified. The Simpson's index of diversity (D) values for the individual locus ranged from 0.79 to 0.92, with the D value of all combined five microsatellite loci being 0.99. Microsatellite data analysis revealed that isolates from Ocimum sanctum, Capsicum annuum (chili pepper), and Mangifera indica (mango) formed distinct clusters, therefore exhibited some level of correlation between certain genotypes and host. The developed MLMT method would be a powerful tool for studying the genetic diversity and any possible genotype-host correlation in C. gloeosporioides.

• The Plant Pathology Journal
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• v.31 no.4
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• pp.402-413
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• 2015

Yellow rust (stripe rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive foliar diseases of wheat in Egypt and worldwide. In order to identify wheat genotypes resistant to yellow rust and develop molecular markers associated with the resistance, fifty F8 recombinant inbred lines (RILs) derived from a cross between resistant and susceptible bread wheat landraces were obtained. Artificial infection of Puccinia striiformis was performed under greenhouse conditions during two growing seasons and relative resistance index (RRI) was calculated. Two Egyptian bread wheat cultivars i.e. Giza-168 (resistant) and Sakha-69 (susceptible) were also evaluated. RRI values of two-year trial showed that 10 RILs responded with RRI value >6 <9 with an average of 7.29, which exceeded the Egyptian bread wheat cultivar Giza-168 (5.58). Thirty three RILs were included among the acceptable range having RRI value >2 <6. However, only 7 RILs showed RRI value <2. Five RILs expressed hypersensitive type of resistance (R) against the pathogen and showed the lowest Average Coefficient of Infection (ACI). Bulked segregant analysis (BSA) with eight simple sequence repeat (SSR), eight sequence-related amplified polymorphism (SRAP) and sixteen random amplified polymorphic DNA (RAPD) markers revealed that three SSR, three SRAP and six RAPD markers were found to be associated with the resistance to yellow rust. However, further molecular analyses would be performed to confirm markers associated with the resistance and suitable for marker-assisted selection. Resistant RILs identified in the study could be efficiently used to improve the resistance to yellow rust in wheat.

• The Plant Pathology Journal
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• v.26 no.4
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• pp.306-312
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• 2010

The pathotypes of Cercospora beticola, causal agent of sugar beet leaf spot disease, were identified by application of pathogenicity test using 100 isolates obtained from the provinces with intensive sugar beet cultivation. For the identification of pathotypes, five sugar beet cultivars were used each with different resistance factors. Cultivar reactions were determined by inoculation of cultivars with the isolates under controlled conditions and measuring disease severity on the $15^{th}$ day according to the 1-9 KWS Scale. Based on the reactions of the five cultivars, a total of 15 pathotypes were detected. All employed sugar beet cultivars were resistant to Pathotype no:1 comprising most of the isolates. Genetic diversity of the causal agent was characterized by AFLP reaction. The products acquired at the end of AFLP reaction were detected by means of Beckman CEQ 8800 DNA Capillary Series Analysis and the results obtained were evaluated according to the similarity index UPGMA. For the genetic analysis of C. beticola isolates, 9874 polymorphic fragments of sizes between 100 and 500 bp were analysed which were generated by nine primers. The dendrogram derived from AFLP analysis depicted the existence of five different subgroups. The polymorphism rate among isolates was 91.13% and the dendrogram distribution of the pathotypes obtained by pathogenicity indicated that pathotypes were not discriminated and did not compose any groups.

• Journal of Plant Biotechnology
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• v.46 no.1
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• pp.1-8
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• 2019

The main goal of this study was to determine the genetic diversity among 36 grape cultivars grown in Palestine by using ISSR-polymerase chain reaction (PCR) fingerprints. Among the tested primers, 17 produced reasonable amplification products with high intensity and pattern stability. A total of 57 DNA fragments (loci) separated by electrophoresis on agarose gels were detected and they ranged in size, from 150 to 900 bp. Out of these fragments, 55 (88%) were polymorphic and 2 (3.5%) monomorphic. Our results also revealed an average of 3.1 loci per primer. A minimum of 1 and maximum of 10 DNA fragments were obtained (S-17, #820 and #841) and (S-31) primers, respectively. Therefore, the later primer (S-31) is considered to be the most powerful primer among the tested ones. The genetic distance matrix showed an average distance range of between 0.05 and 0.76. The maximum genetic distance value of 0.76 (24% similarity) was exhibited between the (Shami and Marawi.Hamadani.Adi) as well as (Bairuti and Marawi.Hamadani.Adi) genotypes. On the other hand, the lowest genetic distance of 0.05 (95% similarity) was exhibited between (Jandali.Tawel.Mofarad and Jandali. Kurawi.Mlzlz) along with (Shami.Aswad and Shami.mtartash. mlwn) genotypes. Furthermore, the UPGMA dendrogram generally clusters the grape cultivars into eight major clusters in addition to an isolated genotype. Based on these figures, the cultivars tested in this study could be characterized by large divergence at the DNA level. This is taking the assumption that our region has a very rich and varied clonal grape genetic structure.

• Journal of Korean Society of Forest Science
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• v.90 no.1
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• pp.43-54
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• 2001

Genetic differentiation of populations is resulted from the environmental and the genetic effects, and the interactions between them. Whereas, the major factors influencing to the genetic differentiation within populations are the gene flow induced by seed or pollen dispersial, the microsite heterogeneity, and the density-dependent distribution of individuals. For the purpose of studying spatial genetic structure and the distribution pattern of Korean pines(Pinus koraiensis), we set up one $100{\times}100m$ plot at a Korean pine stand in Quercus mongolica community on Mt. Jumbong in Korea. To estimate the coefficient of spatial autocorrelation as Moran's index and an analogue, simple block distance, isozyme markers were analyzed in 325 Korean pines. For 11 polymorphic loci observed in 9 enzyme systems, the average percentage of polymorphic loci, the observed and expected heterozygocity were 72.2% 0.200, and 0.251, respectively. It was revealed the excess of homozygotes was observed in the plot, which suggests that here may be more number of consanguineous trees than expected. On the basis of isozyme genotypes observed in this study, 325 trees were classified into 147 groups in which the maximum number of trees for one group was 34. From the distance class of 24-32m, the genetic heterogeneity began to increase. The variation of simple block distance against the growth performance by tree height and diameter also showed the same trend at 24~32m class. According to high fixation index(F=0.204), the spatial genetic structure within a stand, the analysis of the growth performance, and the distribution patterns of identical genotypes, we inferred that the genetic structure of a Korean pine stand in Mt. Jumbong has been maintained rather density-dependent mechanism than the gene flow, such as the pollen dispersial or the heavy input of seeds following the forest gaps. The genetic patchy size was determined between 24~32m, which suggests that the selection of individuals for the ex situ conservation of Korean pine in Mt. Jumbong may be desirable to be made with the spatial distance over 37 meters between trees.

• Korean Journal of Microbiology
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• v.41 no.1
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• pp.60-66
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• 2005

To select the rapid and efficient molecular subtyping method for epidemiologic monitoring of Staphylococcus aureus (S. aureus) strains at clinical laboratory levels, a total 116 of S. aureus and MRSA (methicillin-resistant S. aureus) strains from diverse animal species [Korean cattle, goat, pig, dog, chicken, mouse] and also humans were analyzed. To evaluate the discriminatory ability (DA) of individual PCR methods, random amplified polymorphic of DNA [RAPD; 4M & RA primer], repetitive extragenic palindromic sequences PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) methods were conducted and then compared on their Simpson's index of diversity (SID) values based on the dendrogram patterns, which was produced by software program (BiolD2+ & GelCompar II). In first, RAPD using the 4M primer (SID 0.915) was expressed more higher SID value than that of RA primer (SID 0.874). 4M primer was expressed more powerful DA than RA. Both REP-PCR (SID 0.930) and ERIC-PCR (SID 0.929) methods showed much more higher DA than that of RAPD. According to the present results, both REP-PCR and ERIC-PCR among the tested analysis methods were found as the most reliable and discriminative molecular subtyping method, because they expressed the highest DA for the present S. aureus and MRSA strains.

• Korean Journal of Plant Resources
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• v.25 no.5
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• pp.517-522
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• 2012

This study was conducted to investigate the morphological and anatomical characteristics of needle and the genetic diversity of Pinus pumila Regel which is a unique and the southern peripheral population in South Korea. ANOVA test showed that there were significant differences among individuals within population in all 8 needle characteristics. Average characteristics of 66 individuals were 53.59 mm in needle length, 0.78 mm in needle width, 68.98 in needle index, 0.65 mm in needle thickness, 4.56 ea. in maximum stomata row, 3.80 ea. in minimum stomata row, 8.36 ea. in total stomata row and 1.71 ea. in resin canals, respectively. Resin canal per needle of this species ranged from one to three, depending on external type. Especially, arrangement types were 69.47% in two resin canals and 30.45% in a single resin canal. A total of 78 bands was generated from 9 selected I-SSR primers. The estimates of genetic variation were 61.5% in proportion of polymorphic bands (P), 1.698 in effective number of alleles ($A_e$), 0.388 in expected heterozygosity ($H_e$) and 0.567 in Shannon's information index (S.I.), respectively.

• Korean Journal of Breeding Science
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• v.43 no.5
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• pp.422-429
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• 2011

This study was conducted to investigate the genetic diversity and to develop a technique for cultivar identification using SSR markers in grapevine. Thirty Korean bred and introduced grapevine cultivars were evaluated by 28 SSR markers. A total of 143 alleles were produced ranging from 2 to 8 alleles with an average of 5.1 alleles per locus. Polymorphic information contents (PIC) were ranged from 0.666 (VVIp02) to 0.975 (VVIn33 and VVIn62) with an average of 0.882. UPGMA (unweighted pair-group method arithmetic average) clustering analysis based on genetic distances using 143 alleles classified 30 grapevine cultivars into 7 clusters by similarity index of 0.685. Similarity values among the tested grapevine cultivars ranged from 0.575 to 1.00, and the average similarity value was 0.661. The similarity index was the highest (1.00) between 'Jinok' and 'Campbell Early', and the lowest (0.575) between 'Alden' and 'Narsha'. The genetic relationships among the 30 studied grapevine cultivars were basically consistent with the known pedigree. The three SSR markers sets (VVIn61, VVIt60, and VVIu20) selected from 28 primers were differentiated all grapevine cultivars except for 'Jinok' and 'Campbell Early'. Five cultivars ('Narsha, 'Alden', 'Dutchess', 'Pione', and 'Muscat Hamburg') were identified by VVIn61 at the first step. Then 21 cultivars including 'Hongsodam' by VVIt60 at the second step and 2 cultivars ('Heukbosuck' and 'Suok') by VVIu20 at the third step were identified. These markers could be used as a reliable tool for the identification of Korean grapevine cultivars.

• Journal of Preventive Medicine and Public Health
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• v.55 no.3
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• pp.280-288
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• 2022

Objectives: One of the most widely used pesticides today is chlorpyrifos (CPF). Cytochrome P450 (CYP)2B6, the most prominent catalyst in CPF bioactivation, is highly polymorphic. The objective of our study was to evaluate the role of CYP2B6^*6, which contains both 516G>T and 785A>G polymorphisms, in CPF toxicity, as represented by the concentration of 3,5,6-trichloro-2-pyridinol (TCPy), among vegetable farmers in Central Java, Indonesia, where CPF has been commonly used. Methods: A cross-sectional study was conducted among 132 vegetable farmers. Individual socio-demographic and occupational characteristics, as determinants of TCPy levels, were obtained using a structured interviewer-administered questionnaire and subsequently used to estimate the cumulative exposure level (CEL). TCPy levels were detected with liquid chromatography-mass spectrometry. CYP2B6^*6 gene polymorphisms were analyzed using a TaqMan^® SNP Genotyping Assay and Sanger sequencing. Linear regression analysis was performed to analyze the association between TCPy, as a biomarker of CPF exposure, and its determinants. Results: The prevalence of CYP2B6^*6 polymorphisms was 31% for ^*1/^*1, 51% for ^*1/^*6, and 18% for ^*6/^*6. TCPy concentrations were higher among participants with CYP2B6^*1/^*1 than among those with ^*1/^*6 or ^*6/^*6 genotypes. CYP2B6^*6 gene polymorphisms, smoking, CEL, body mass index, and spraying time were retained in the final linear regression model as determinants of TCPy. Conclusions: The results suggest that CYP2B6^*6 gene polymorphisms may play an important role in influencing susceptibility to CPF exposure. CYP2B6^*6 gene polymorphisms together with CEL, smoking habits, body mass index, and spraying time were the determinants of urinary TCPy concentrations, as a biomarker of CPF toxicity.

• Clinical and Experimental Pediatrics
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• v.52 no.3
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• pp.356-363
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• 2009

Purpose : A polymorphism in the IGF-I gene promoter region is known to be associated with serum IGF-I levels, birth weight, and body length, suggesting that IGF-I gene polymorphism might influence postnatal growth. The present study aimed to investigate the role of this polymorphic cytosine-adenine (CA) repeat of the IGF-I gene in children with idiopathic short stature. Methods : The study involved 131 children (72 boys and 59 girls) diagnosed with idiopathic short stature, aged 715 years. Genomic DNA was extracted from anticoagulated peripheral whole blood. The primers were designed to cover the promoter region containing the polymorphic CA repeat. Data were analyzed using GeneMapper software. The correlations between age and serum IGF-I levels were analyzed using Spearmans correlation coefficient. Results : The CA repeat sequences ranged from 15 to 22, with 19 CA repeats the most common with an allele frequency of 40.6%. Homozygous for 19 CA repeat was 13.0%, heterozygous for 19 CA repeat was 56.5%, and 19 CA non-carrier was 30.5%. The three different genotype groups showed no significant differences in height, body weight and body mass index, and serum IGF-I levels. The serum IGF-I level and age according to the IGF-I genotypes were significantly correlated in the entire group, 19 CA repeat carrier group, and the non-carrier group. The three groups also showed no significant differences in the first year responsiveness to GH treatment. Conclusion : There were no significant different correlations between 19 CA repeat polymorphism and serum IGF-I levels according to genotype. Our results suggest that the IGF-I 19 CA repeat gene polymorphism is not functional in children with idiopathic short stature.