• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.724-730
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• 2006

A sequence characterized amplified region (SCAR) marker, which was tightly linked with the A1 mating type locus in Phytophthora infestans, was developed. During the random amplified polymorphic DNA-based phylogenic studies of 33 isolates of P infestans collected from year 2002 to 2004, we found an A1 mating type-specific DNA fragment. This 573-bp DNA fragment was generated only in the genomic DNA of the A1 mating types, when OPC-5 primer was used. Based on the specific DNA sequence, we designed the primer sets for generating the A1 mating type-specific 569-bp DNA fragment. When 33 genomic DNAs of P. infestans were subjected to PCR amplification using different primer combinations, the A1 mating type-specific DNA was amplified, when LB-1F and LB-2R primers were used. The specific 569-bp DNA fragment was generated only from all 18 A1 strains, but not from 15 A2 mating type strains. These results corresponded to the mating type discriminating bioassay of 33 isolates of P. infestans. Therefore, the primer combination of LB-1F/LB2R was chosen as a SCAR marker. Overall, this study indicates that the SCAR marker could be developed into a useful tool for mating type determination of P. infestans.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1520-1522
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• 2001

Relationship among Tibetan cashmere goats, Inner Mongolia cashmere goats and Liaoning cashmere goats was studied using the technique of random amplified polymorphic DNA (RAPD). One primer and four primer combinations were screened. With the five primers and primer combinations, DNA fragments were amplified from the three breeds. Each breed has 28 samples. According to their RAPD fingerprint maps, the Nei's (1972) standard genetic distance was: 0.0876 between Tibetan cashmere goats and Inner Mongolia cashmere goats, 0.1601 between Tibetan cashmere goats and Liaoning cashmere goats, 0.0803 between the Inner Mongolia cashmere goats and Liaoning cashmere goats. It coincides with their geographic location. The genetic heterogeneity of Tibetan cashmere goats, Inner Mongolia cashmere goats and Liaoning cashmere goats is 0.3266, 0.2622 and 0.2475 respectively. It is also consistent with their development history.

• Korean Journal of Breeding Science
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• v.43 no.5
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• pp.399-404
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• 2011

Genetic diversity among 28 Cymbidium varieties was evaluated by using a sequence-related amplified polymorphism (SRAP) marker system. The SRAP marker which was based on the open reading frames (ORFs) regions was developed primarily for Brassica species, but has been applied to various crops. A total of 30 SRAP primer combinations were initially screened. Twenty-eight SRAP primer combinations showed high polymorphism among the 28 Cymbidium varieties, which were consisted of breeding varieties and their parents in National Institute of Horticultural & Herbal Science (NIHHS). The amplified DNA fragments were separated by denaturing acrylamide gels and detected silver staining method. One hundred ninety six polymorphic bands (7 per primer) were generated and ranged from 0.3 to 1.0 kb in size. Polymorphic fragments were scored for calculating simple matching coefficient of genetic similarity and cluster analysis with multi-variate statistical package (MVSP) 3.1. The mean genetic similarity coefficient value was 0.588. The results showed that the correlation between $F_1$ varieties and their parents was high. These studied SRAP markers will be useful tools for genotype identification, germplasm conservation, genetic relationships in Cymbidium.

• Molecules and Cells
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• v.25 no.2
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• pp.196-204
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• 2008

Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum 'CM334' and C. annuum 'Chilsung-cho'. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to $F_2$ genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.4
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• pp.322-328
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• 2011

AFLP markers were employed to detect genetic diversity and genetic relationship among 26 accessions of foxtail millet collected from Korea. Analysis of 26 accessions of foxtail millet with nine AFLP primer set combinations detected a total of 170 bands, of which 145 (85.3%) showed polymorphic at the species level. The phenotypic diversity (Hs) calculated for the nine primer combinations ranged from 1.84 to 6.8, with an average of 3.85. The average phenotypic diversity values were 3.39 and 2.99 for accessions collected from Gangwon province (Group 1), and accessions collected from the other areas including Gyeonggi province (Group 2), respectively. In the cluster analysis of 26 accessions, two major groups were recognized at 73% genetic similarity. Group I includes 13 accessions, which were collected in Gangwon province, and 1 accession, which was collected in Gyeonggi province, with genetic similarity of 76.8%. Group II includes two accessions, which were collected in Gangwon province, and 10 accessions, which were collected in the other areas with genetic similarity of 78.9%. The presenting data on the assessment of genetic diversity and genetic relationships among Korean accessions of foxtail millet will be helpful for efficient collection or conservation strategy of foxtail millet germplasm in Korea.

• BMB Reports
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• v.28 no.6
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• pp.538-545
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• 1995

When DNA replication of simian virus 40 (SV40) is initiated on the replication origin, the regions containing the initiation sites of DNA primase, which participates in the transient RNA primer synthesis for formation of Okazaki fragments in the lagging strand, were chosen as the target sites of antisense RNA for studies of the inhibition of SV40 DNA replication. Four recombinant transcription vectors, pUC-PrI, pUC-PrII, pGEM-PrBS, and pGEM-PrSN, coding antisense RNA, were constructed. Four antisense RNAs (named as I, II, BS, and SN) having the size of 18, 19,58, and 123 nts, respectively, were made from the transcription vectors by in vitro transcription. And then, antisense RNA in the concentration of 2${\mu}m$ were added to COS cells transfected with pATSV-W which is a recombinant plasmid containing the SV40 origin of replication. The inhibitory extent of DNA replication was measured by DpnI resistance and was confirmed by measurement of transient RNA primer synthesis. The result shows that six combinations of antisense RNA (I, II, BS, SN, I+SN, and BS+SN) lead to the inhibition of SV40 DNA replication by up to 85%.

• Mycobiology
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• v.29 no.3
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• pp.135-141
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• 2001

Differential display of reverse transcription(DDRT)-PCR was conducted to have a profile of the differentially expressed genes during the formation of fruiting body of Lentinus edodes. The lines of L. edodes(ImHyup-1) employed were cultivated in the artificial blocks of sawdust, and the fruiting body was induced from the mycelia or the mass protruded from the brown surface of the sawdust blocks. RNAs were prepared from the four different developmental stages; mycelial, primordial, and stipes and pileus of fruiting body. The fragments of cDNA were synthesized from the combinations of the arbitrary primers and 3' one anchored Oligo-dT primer. Twelve combinations using the primers have been tested, and among them nineteen bands were identified as differentially expressed. Those genes were further analyzed by DNA sequencing and followed by homology search. Characterization of one clone was conducted as a preliminary data and more are under investigation.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.9
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• pp.1217-1220
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• 2005

Random amplified polymorphic DNA (RAPD-PCR) analysis of 56 animals of four different genetic groups of dwarf cattle in Kerala was done as a single step analysis. Bandsharing (BS) values were calculated for animals of each group and between groups as an analytical tool to find out genetic variation among animals. The different factors affecting BS values were estimated using Harvey''s Least squares analysis. The effects of genetic group, Guanine-cytosine (GC) content of primer and gel on BS values were found significant. Bandsharing values of Kasargode-Highrange dwarf animals were significantly different from Vechur, Vatakara and their combinations. The Vechur, Vatakara and Vechur-Vatakara combinations were found to be more uniform (high BS value) compared with other combinations. The bandsharing value was lowest with primers of GC content 90% and highest with 80% GC content. The effect of gel on BS value points to the need of adjustments of gel factor for calculation of BS values.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.3
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• pp.259-268
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• 2006

Miniature inverted transposable elements (MITEs) are abundant genomic components in plant including rice. MITE-transposon display (MITE-TD) is an Amplified Fragment Length Polymorphism (AFLP)-related technique based on MITE sequence. In this study, we used the MITE-AFLP for the analysis of diversity and relation-ship of the 114 japonica accessions. Of the several MITEs, the mPing family was applied to detect polymorphisms based on PCR amplification. The BfaI adaptor primer and the specific primer derived from mPing terminal inverted repeat (TIR) region were used to PCR amplification of 114 accessions. Nine primer pairs produced a total of 160 polymorphic bands. PIC values of the polymorphic bands generated by nine primer pairs ranged from 0.269 (BfaI + ACT) to 0.426 (BfaI + T). Each accession revealed a distinct fingerprint with two primer combinations, BfaI + G and BfaI + C. Cluster analysis using marker-based genetic similarity classified 114 accessions into five groups. MITE-AFLP markers were genetically mapped using a population of 80 BILs (BC1F7) derived from a cross between the rice accessions, Milyang 23 and Hapcheonaengmi 3. Eight of the markers produced with the primer pair BfaI + 0 were mapped on chromosomes 1, 2, 4, 5, 7, and 9. Considering that one MITE-AFLP marker on chromosome 7 was tightly linked to the Rc gene, the MITE-AFLP markers will be useful for gene tagging and molecular cloning.

• Research in Plant Disease
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• v.23 no.2
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• pp.193-201
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• 2017

Major tomato viruses in Korea are Tomato chlorosis virus (ToCV), Tomato spotted wilt virus (TSWV), Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), and Tomato mosaic virus (ToMV). RT-PCR conditions for the viruses were examined, especially in primer set and RT-PCR mixture. Total 46 primer sets from the unique sequence of the viruses were tested for nonspecific background products in a RT-PCR mixture without template. Among them 16 primer sets were applied to healthy tomato RNA, resulting the compatibility between RT-PCR mixture and primer set influenced RT-PCR to reduce nonspecific background products. Based on the combinations among cDNA synthesis parameters and RT-PCR mixtures, two reaction mixtures were finally selected for ToCV detection. The condition allowed to determine more specific primer sets; C029 (ToCV), C072 (TSWV), C070 (CMV), C048 (PepMoV), and C065 (ToMV). These primer sets are expected to be of use to specific detection of the major viruses in tomato plants.