• Title/Summary/Keyword: Random Amplification of Polymorphic DNA

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Fine Mapping of the Rice Bph1 Gene, which Confers Resistance to the Brown Planthopper (Nilaparvata lugens Stal), and Development of STS Markers for Marker-assisted Selection

  • Cha, Young-Soon;Ji, Hyeonso;Yun, Doh-Won;Ahn, Byoung-Ohg;Lee, Myung Chul;Suh, Seok-Cheol;Lee, Chun Seok;Ahn, Eok Keun;Jeon, Yong-Hee;Jin, Il-Doo;Sohn, Jae-Keun;Koh, Hee-Jong;Eun, Moo-Young
    • 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.

Mband characterization of a cultivar "DanBi 5Ho" with a long shelf life (큰느타리버섯 단핵균주 간 교잡에 의한 저장우수형 품종 "단비5호" 육성 및 특성)

  • Lee, Young-Han;Kim, Ya-El;Seuk, Su-Won;Jeong, Jeong-Min;Ryu, Jae-San;Heo, Jae-Young;Kim, Hee-Dae;Choe, Yong-Jo;Kim, Min-Keun
    • 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.

Multiple Confirmation and RAPD-genotyping of Enterobacter sakazakii Isolated from Sunsik (선식에서 분리한 Enterobacter sakazakii의 복합동정 및 RAPD를 이용한 genotyping)

  • Choi, Jae-Won;Kim, Yun-Ji;Lee, Jong-Kyung;Kim, Young-Ho;Kwon, Ki-Sung;Hwang, In-Gyun;Oh, Se-Wook
    • 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.

Discrimination of Korean Apple Cultivars Using Combination of RAPD-SCAR Markers (RAPD-SCAR 마커 조합을 이용한 국내 육성 사과 품종 판별)

  • Cho, Kang-Hee;Heo, Seong;Kim, Hyun-Ran;Kim, Jeong-Hee;Shin, Il-Sheob;Han, Sang-Eun;Kim, Se-Hee;Kim, Dae-Hyun
    • 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.

Changes of Efficacy of Antioxidant, Antidyslipidemic, Antidiabetic and Microbiological Characteristics in Fermented and Salt-treated Fermented Codonopsis lanceolata (발효 더덕 및 소금 처리 발효 더덕의 미생물 특성과 항산화, 항비만, 항당뇨 효능 변화)

  • Seong, Eun-Hak;Lee, Myeong-Jong;Kim, Hojun;Shin, Na Rae
    • Journal of Korean Medicine for Obesity Research
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    • v.18 no.2
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    • pp.106-114
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    • 2018
  • Objectives: We investigated about the microbial properties and changes in the efficacy of the Codonopsis lanceolata (CL) by natural fermentation. Methods: CL was fermented for four weeks in a well-ventilated place with 2.5% salt. pH, total sugar, total polyphenol, and total flavonoid were measured to determine fermentation characteristics according to fermentation period and salt treatment. Polymerase chain reaction denaturing gradient gel electrophoresis and random amplification of polymorphic DNA-polymerase chain reaction were carried out for microbial analysis during fermentation. In addition, HepG2 cell was cultured to check the lipid accumulation through oil red O staining and the glucose uptake was analyzed by measuring the 2-NBDG at C2C12 cell. Results: The pH level and the total sugar decreased with the CL fermentation. Total polyphenol and flavonoid increased after CL fermentation. It was confirmed that Leuconostoc mesenteroides were maintained continuously during fermentation. In the salt treatment CL, there was a sharp increase in Rahnella aquatilis. Lactobacillus plantarum matrix was observed in fermented CL. In addition, Lactococcus lactis, Weissella koreensis, R. aquatilis, L. plantarum, Leu. mesenteroides have been added to the salt treatment. Glucose uptake were significantly increased after fermentation with salt for four weeks. Lipid accumulation in the HepG2 cells was observed that there was difference (P<0.01) between free fatty acid group (100%) and decreased 4 weeks after fermentation (90.38%) at $800{\mu}g/mL$. Conclusions: Total polyphenol and flavonoid were increased after CL fermentation. Especially, percentage of the glucose uptake and lipid accumulation inhibition increased in CL fermentation with salt. It is expected that fermentation of salt treated CL will be more effective in diabetes and fatty liver.

Spread of Bacterial Canker of Kiwifruit by Secondary Infection of Pseudomonas syringae pv. actinidiae Biovar 3 in Gyeongnam in 2016 (2016년 경남지역 Pseudomonas syringae pv. actinidiae Biovar 3의 2차감염에 의한 키위 궤양병의 확산)

  • Kim, Gyoung Hee;Choi, Eu Ddeum;Lee, Young Sun;Jung, Jae Sung;Koh, Young Jin
    • 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.