• Horticultural Science & Technology
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• v.33 no.5
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• pp.722-729
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• 2015

Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.648-655
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• 2015

Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

• Journal of Practical Agriculture & Fisheries Research
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• v.3 no.1
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• pp.48-69
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• 2001

A lot of clones of the genus Dioscorea have been introduced from some tropical and subtropical regions since 1997. In 33 clones of water yams (Dioscorea alata L.), some morphological characteristics were investigated at the field. Variation ranges of the total weight and tuber number per stump were within the ranges from 90 to 2,147 g with an average of 610 g ; and 1.3-4.7 with an average of 2.8, respectively. The color tones observed on the tuber-flesh were sorted into 3 color-categories, i.e., white, pale brown and pale purple, and those on leaves were sorted into 3 color-categories, i.e., green, heavy green and purplish green. Intraspecific genetic relationship of 19 variation types of the Yam classified by their external morphological characteristics such as leaf and tuber shape was assessed by DNA using random and specific primers. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)4 sequence]) had been used in PCR-amplification. Only 12 primers, however, were successful in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0%), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90.

• Journal of Mushroom
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• v.16 no.3
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• pp.140-146
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• 2018

Pleurotus eryngii is one of the most commercially important mushrooms cultivated in Korea. However, the shelf-life of the fruiting body is short, limiting its export. A new hybrid strain H17 of P. eryngii was developed to extend the shelf-life by mono-mono crossing between monokaryotic strains derived from DanBi and KNR2774. Although the cultivation period of H17 was slightly longer than that of the reference cultivar Kenneutari No.2, the quality did not change and remained normal after a period of 65.0 days at $4^{\circ}C$. This result was significantly different from that of the reference cultivar Kenneutari No.2. Analysis of the genetic characteristics of the new hybrid strain H17 revealed a different profile from that of the parental and reference cultivars when random amplification of polymorphic DNA (RAPD) primers was used. These results demonstrate that H17 is a new cultivar with improved storability after harvesting.

• The Korean Journal of Mycology
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• v.26 no.4 s.87
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• pp.466-477
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• 1998

Twenty-two Phellinus strains were characterized using colony morphologies and polymerase chain reaction (PCR) to divide into Phellinus linteus. There were some differences in mycelial growth and colony shapes among the strains when they were grown on various media such as PDA, MCM, MEA and YM. Phellinus linteus was slowly growing, formed golden-yellow colony, and produced blue pigment on PDA media. When the regions of internal transcribed spacer (ITS) were amplified from ribosomal RNA (rRNA) coding genes of P. igniarius and P. linteus strains by means of PCR, two types of band (700 bp and 800 bp) were appeared, respectively. For the amplified intergenic region I (IGRI), P. igniarius strains showed a different band among 500, 600, 700 and 800 bp according to the strains, whereas P. linteus strains did one specific band of 700 bp. By polymorphism analysis after digesting the amplified products with 6 different restriction enzymes, a band specific to P. linteus was generated when the products for ITS region were digested with HaeIII, suggesting that the enzyme digestion could provide effective method to distinguish between P. igniarius and P. linteus. And also, the analysis of genetic relationship showed that the genetic similarities were 89% and 95% in P. igniarius and P. linteus strains, respectively. Random amplification polymorphic DNA (RAPD) analysis using multiple primer sets and arbitrarily primed PCR (AP-PCR) with ITS3 primer could also result in a reproducible way to identify P. linteus strains.

• Korean Journal of Food Science and Technology
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• v.40 no.1
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• pp.101-105
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• 2008

Enterobacter sakazakii is implicated in severe forms of neonatal infections such as meningitis and sepsis. This organism has been isolated from a wide range of foods, including cheese, vegetables, grains, herbs, and spices, but its primary environment is still unknown. Generally, dried infant milk formula has been epidemiologically identified as the source of E. sakazakii. Sunsik (a powdered mixture of roasted grains and other foodstuffs) is widely consumed in Korea as a side dish or energy supplement. Sunsik is consumed without heat treatment; thus, lacking an additional opportunity to inactivate foodborne pathogens. Therefore, its microbiological safety should be guaranteed. In this study, the prevalence of E. sakazakii was monitored in 23 different sunsik component flours, using FDA recommended methods; but E. sakazakii medium (Neogen) and Chromogenic E. sakazakii medium (Oxoid) were used as the selective media. In total, presumptive E. sakazakii strains were isolated from 8 different sunsik powders. Subsequently, an API 20E test was conducted, and 15 strains from 5 different sunsik flours (sea tangle, brown rice, non-glutinous rice, cheonggukjang, dried anchovy) were confirmed as E. sakazakii. Fifteen strains were again confirmed by PCR amplification, using three different primer sets (tDNA sequence, ITS sequence, 16S rRNA sequence), and compared to ATCC strains (12868, 29004, 29544, 51329). They were once again confirmed by their enzyme production profiles using an API ZYM kit. Finally, RAPD (random amplified polymorphic DNA)-genotyping was carried out as a monitoring tool to determine the contamination route of E. sakazakii during processing.

• Horticultural Science & Technology
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• v.28 no.5
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• pp.828-835
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• 2010

Conventional methods for identification of apple cultivars are based on the evaluation of sets of morphological characteristics, however, closely related cultivars often cannot be distinguished by morphological traits. This study was conducted to develop DNA markers for discrimination of the apple cultivars bred in Korea. Thirty random primers generated eighty-three random amplified polymorphic DNA (RAPD) markers from thirty-one Korean bred and introduced apple cultivars. Fifty-two RAPD fragments were cloned and sequenced for conversion into sequence characterized amplified region (SCAR) markers. Among them only seventeen SCAR markers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Several combinations of six (AN11_433, AN08_566, A408_592, AK17_653, AO04_711, AO04_779 or AW15_368, AN11_433, A408_592, AK17_653, AO04_711, AO04_779, or AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AO04_779) to seven (AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AM16_708, AO04_779 or A330_424, AN11_433, AG14_502, AN08_566, A408_592, AK17_653, AO04_779 or A330_424, AN11_433, AK14_564, A408_592, AK17_653, AM16_708, AT14_789) SCAR markers provided enough polymorphism to identify sixteen Korean apple cultivars among thirty-one tested cultivars. Therefore, application of the seventeen SCAR markers was sufficient to identify the thirty-one tested apple cultivars. These markers could be utilized as a reliable tool for cultivar discrimination of Korean apples.

• Molecules and Cells
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• v.26 no.2
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• pp.146-151
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• 2008

The brown planthopper (BPH) is a major insect pest in rice, and damages these plants by sucking phloem-sap and transmitting viral diseases. Many BPH resistance genes have been identified in indica varieties and wild rice accessions, but none has yet been cloned. In the present study we report fine mapping of the region containing the Bph1 locus, which enabled us to perform marker-aided selection (MAS). We used 273 F8 recombinant inbred lines (RILs) derived from a cross between Cheongcheongbyeo, an indica type variety harboring Bph1 from Mudgo, and Hwayeongbyeo, a BPH susceptible japonica variety. By random amplification of polymorphic DNA (RAPD) analysis using 656 random 10-mer primers, three RAPD markers (OPH09, OPA10 and OPA15) linked to Bph1 were identified and converted to SCAR (sequence characterized amplified region) markers. These markers were found to be contained in two BAC clones derived from chromosome 12: OPH09 on OSJNBa0011B18, and both OPA10 and OPA15 on OSJNBa0040E10. By sequence analysis of ten additional BAC clones evenly distributed between OSJNBa0011B18 and OSJNBa0040E10, we developed 15 STS markers. Of these, pBPH4 and pBPH14 flanked Bph1 at distances of 0.2 cM and 0.8 cM, respectively. The STS markers pBPH9, pBPH19, pBPH20, and pBPH21 co-segregated with Bph1. These markers were shown to be very useful for marker-assisted selection (MAS) in breeding populations of 32 F6 RILs from a cross between Andabyeo and IR71190, and 32 F5 RILs from a cross between Andabyeo and Suwon452.

• Journal of Mushroom
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• v.14 no.2
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• pp.64-69
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• 2016

Pleurotus eryngii is one of the most widely cultivated and important edible mushrooms. However, the shelf life of the fruiting body is short, which. To solve this problem, a new cultivar, DanBi 5Ho, of Pleurotus eryngii was developed by mono-mono crossing between monokaryotic strains derived from KNR2598 and KNR2610. The optimum temperature formycelial growth was 25 and that forfruiting body development was 15-16. The quality did not change after a period of 40.0 d at 4. This result was significantly different compared to that ofthe control strain Knneuari 2 Ho. Analysis of genetic characteristics of the new hybrid strain DanBi 5Ho revealeda different profile thanthat of the parental and control strains when random amplification of polymorphic DNA (RAPD) primers we used. The results obtained from this study show that DanBi 5Ho is a new hybrid strain, characterized by improved storability after harvesting.

• Research in Plant Disease
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• v.22 no.4
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• pp.276-283
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• 2016

Bacterial canker caused by Pseudomonas syringae pv. actinidiae (Psa) occurred at 202 kiwifruit orchards for the survey period of 2013-2016, of which Psa biovar 2 (Psa2) and Psa biovar 3 (Psa3) were detected at 73 and 129 kiwifruit orchards, respectively. The number of kiwifruit orchards infected by Psa3 in 2016 increased nearly two times compared to 2015. Psa3 was detected from all the kiwifruit cultivars except some kiwiberry cultivars growing in Korea. Yellow-fleshed cultivars Hort16A and Jecy-gold and red-fleshed cultivar Hongyang were highly susceptible to Psa3. Our epidemiological and random amplification of polymorphic DNA analyses indicated that the first Psa3 incidence on Hongyang orchard in Sacheon, Gyoungnam might result from an introduction of Psa3-contaminated pollens from China for artificial pollination in 2014 and recent outbreaks of Psa3 in Sacheon and Goseong, Gyoungnam in 2016 might be due to rapid spread of bacterial canker by secondary infection of Psa3 from Hongyang orchard to neighboring Jecy-gold and Hayward orchards.