• Title/Summary/Keyword: Random Amplified Polymorphic DNA(RAPD)

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Determination of paraquat-resistant biotype on Conyza canadensis and the resistant mechanism (Paraquat 저항성 생태형 망초의 선발과 저항성 기작)

  • Kim, Sung-Eun;Kim, Seung-Yong;Ahn, Sul-Hwa;Chun, Jae-Chul
    • The Korean Journal of Pesticide Science
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    • v.9 no.1
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    • pp.88-96
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    • 2005
  • Paraquat-resistant biotype of Conyza canadensis (L.) Cronq. was determined by chlorophyll loss and random amplified polymorphic DNA (RAPD) analysis and the resistant mechanism was investigated with respect to absorption, translocation, and binding constant. RAPD analysis for paraquat resistant (R) and susceptible (S) biotypes found in a pear orchard revealed that the biotypes possessed remote genetic relationship. Chlorophyll loss, as an indication of paraquat toxicity, of S biotype was 7.8-fold greater than that of R biotype. There were no differences in contents of epicuticular wax and cuticle and amounts of [14C]paraquat penetrating the cuticle between the two biotypes. Little translocation of the herbicide out of the treated leaf was observed in either biotype. Binding constants of paraquat to the cell wall and thylakoid membrane were 7.4-fold and 16.9-fold, respectively, higher in R biotype than in S biotype. The results suggest that the resistance mechanism of C. canadensis biotype is due partly to high binding affinity of paraquat to the cell wall and thylakoid membrane.

Studies on Microbial Ecology of Actinomycetes in Tideland Soils. (서해 아암도 갯벌토양 미생물의 개체군 분석 및 RAPD 분석에 의한 방선균의 생태학적 연구)

  • 조영주;김정한;전은수;이상미;박동진;이재찬;이향범;김창진
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.79-85
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    • 2002
  • Ecological characteristics of microorganisms in tideland soils were studied by investigation of microbial diversity and population. Twenty soil samples were taken at surface, 10, 20 and 30 cm depth each. Bacteria, actinomycetes and fungi were isolated on each selective isolation medium containing different concentration of NaCl. Actinomycetes were the most isolated from soil samples taken at 10 cm depth and isolated by humic acid-vitamin (HV) medium without sea water or salt. Twenty nine strains of actinomycetes were isolated at surface soil and 74, 39, 37 strains were at 10, 20, and 30 cm depth, respectively. All these isolates were analysed and grouped by random amplified polymorphic DNA (RAPD)-PCR analysis. Many of the isolates were clustered into Microtetraspora and Pseudonocardia. Fungal isolates were highly distributed at the surface soil and isolated well on potato dextrose agar (PDA) medium with sea water. Bacterial isolates were higly distributed at surface soil and isolated well by nutrient medium without sea water or salt. Soil samples taken at 10 cm depth showed the highest microbial diversity and population.

Characteristics and breeding of a new cultivar Pleurotus eryngii var. ferulae, 'Beesan No.1' (아위느타리 신품종 '비산1호'의 육성 및 자실체 특성)

  • Shin, Pyung-Gyun;Yoo, Young-Bok;Kong, Won-Sik;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.12 no.1
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    • pp.52-57
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    • 2014
  • To develop a new cultivar of King oyster mushroom white variety (Pleurotus ferulae), GW10-95 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2850-24 derived from ASI 2850 and dikaryotic strain ASI 2803. The GW10-95(ASI 2803 ${\times}$ ASI 2850-24) was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Beesan No.1'. The 'Beesan No.1' was formed incompatibility line distinctly in the confrontation growth of parental strains ASI 2803 and ASI 2850. Analysis of the genetic characteristics of the new cultivar 'Beesan No.1' showed a different DNA profile as that of the control strains, ASI 2803 and ASI 2850, when RAPD(Random Amplified Polymorphic DNA) primer URP6 was used. The optimum temperature and pH arrange for mycelial growth were $25^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about 245 g using demonstration farms. And also the stipe was thick and long. This new cultivar 'Beesan No.1' of Pleurotus ferulae was characterized by fast fruitbody formation, the stipe was thick and long and high quality yield compared to that of other cultuvars. We therefore expect that this new strain will increase of farmer's income by construction of stabilized production system.

Breeding of the native vegetables using the biotechnology

  • Iwamoto, Yuzuri
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.106-111
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    • 2005
  • For breeding of a new rootstock for eggplant production, somatic hybrids between two species, Solanum integrifolium and S. sanitwongsei were obtained through protoplast fusion. The former species has been commonly used for rootstock for eggplant production in Japan. Eggplants on these rootstocks are more productive than ungrafted plants, but are susceptible to bacterial wilt caused Ralstonia solanacearum. While the latter species is resistant, the growth of eggplants on this rootstock is rather slow and low yield. Protoplast of both species were isolated from cotyledons, and inactivated with iodoacetamide or UV-irradiation, then fused electrically. The fused products were then cultured. Regenerated plantlets were then transplanted on soil then maintained in a green house. The plants were classified into four groups. Those in the first group showed morphological characters intermediate of the parentalspecies. The plants bore fruit with viable seeds. The plants showed a chromosome number of 2n=48, the sum of those of the parental species, and are suggested to be symmetric fusion products. While plants in the other groupswas less vigorous and showed chromosome number 2n= 68 to 72 suggesting asymmetric fusion products by genomic in situ hybridization(GISH). Isozyme pattern of shikimate dehydrogenase (SKDH; EC 1.1.1.25), isocitrate dehydrogenase (IDH; EC 1.1.1.41) and phosphoglucomutase (PGM; EC 2.7.5.1) showed that 24 regenerated plants in three groups were somatic hybrids. Analysis of random amplified polymorphic DNA (RAPD) showed that 43 S. integrifolium-specific and 57 S. sanitwongsei-specific bands were all found in 24 plants. Both somatic hybrids and its S1 plants were found to be resistant to bacterial wilt, and eggplant grafted these plants using for rootstocks were more productive than grafted mother plants. Now, S1 progenies are used for commercial eggplant production in Osaka Prefecture.

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Biochemical and Molecular Characterization of High Population Density Bacteria Isolated from Sunflower

  • Goes, Kelly Campos Guerra Pinheiro De;Fisher, Maria Luisa De Castro;Cattelan, Alexandre Jose;Nogueira, Marco Antonio;Carvalho, Claudio Guilherme Portela De;Oliveira, Andre Luiz Martinez De
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.437-447
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    • 2012
  • Natural and beneficial associations between plants and bacteria have demonstrated potential commercial application for several agricultural crops. The sunflower has acquired increasing importance in Brazilian agribusiness owing to its agronomic characteristics such as the tolerance to edaphoclimatic variations, resistance to pests and diseases, and adaptation to the implements commonly used for maize and soybean, as well as the versatility of the products and by-products obtained from its cultivation. A study of the cultivable bacteria associated with two sunflower cultivars, using classical microbiological methods, successfully obtained isolates from different plant tissues (roots, stems, florets, and rhizosphere). Out of 57 plant-growth-promoting isolates obtained, 45 were identified at the genus level and phylogenetically positioned based on 16S rRNA gene sequencing: 42 Bacillus (B. subtilis, B. cereus, B. thuringiensis, B. pumilus, B. megaterium, and Bacillus sp.) and 3 Methylobacterium komagatae. Random amplified polymorphic DNA (RAPD) analysis showed a broad diversity among the Bacillus isolates, which clustered into 2 groups with 75% similarity and 13 subgroups with 85% similarity, suggesting that the genetic distance correlated with the source of isolation. The isolates were also analyzed for certain growth-promoting activities. Auxin synthesis was widely distributed among the isolates, with values ranging from 93.34 to 1653.37 ${\mu}M$ auxin per ${\mu}g$ of protein. The phosphate solubilization index ranged from 1.25 to 3.89, and siderophore index varied from 1.15 to 5.25. From a total of 57 isolates, 3 showed an ability to biologically fix atmospheric nitrogen, and 7 showed antagonism against the pathogen Sclerotinia sclerotiorum. The results of biochemical characterization allowed identification of potential candidates for the development of biofertilizers targeted to the sunflower crop.

Molecular Identification and Effects of Temperature on Survival and Growth of Hybrids between Haliotis gigantea Gmelin (♀) and Haliotis discus hannai Reeve (♂)

  • An, Hye Suck;Han, Jong Won;Hwang, Hyun-Ju;Jeon, Hancheol;Jung, Seung-Hyun;Jo, Seonmi;Choi, Tae-Young;Hyun, Young Se;Song, Ha Yeun;Whang, Ilson
    • Journal of Marine Life Science
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    • v.2 no.2
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    • pp.83-89
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    • 2017
  • In abalones, interspecific hybridization has been suggested as a possible means to increase production and desired traits for the industry. In Korea, Haliotis gigantea is considered a species with a larger size and higher temperature tolerance than H. discus hannai. However, H. discus hannai is considered the most valuable and popular fishery resource due to its better acceptance and higher market prices. Thus, viable interspecific hybrids have been produced by artificial inseminating H. gigantea eggs with H. discus hannai sperm. However, the reciprocal hybrid cross was not successful. In this study, the hybridity and the growth and thermal tolerance performance of the interspecific hybrids were examined. A combination of various assays revealed maximum growth occurrence at 21℃ and the higher growth rate in the hybrids than that of H. discus hannai parent. In addition, the growth and survival at high-temperature (28℃) of the hybrids was equivalent to that of the highly tolerant H. gigantea parent, suggesting new possibilities to overcome the mass mortality in H. discus hannai during high temperature periods of summer season in Korea. Furthermore, the induced interspecific hybrid status was confirmed by the presence of species-specific bands for each parental species of the random amplified polymorphic DNA (RAPD) profiles using universal rice primer (URP), which could be used as speciesspecific markers to distinguish the hybrids and their parental species.

Geographical Variation of the Oriental Fruit Fly, Bactrocera dorsalis, Occurring in Taiwan (오리엔탈과실파리 유전변이 - 대만 지역 집단변이)

  • Kim, Yonggyun;Kim, Hyoil;Mollah, Md. Mahi Imam;Al Baki, Md. Abdullah
    • Korean journal of applied entomology
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    • v.58 no.2
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    • pp.133-142
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    • 2019
  • This study analyzed genetic variation of the Oriental fruit fly (OFF), Bactrocera dorsalis, which is designated to be a quarantine insect pest in Korea. OFF samples endemic to Taiwan were collected at three different locations (Taipei, Taichung, and Kaohsiung) for three days from July 30 to August 1 in 2018 and assessed in their age and mitochondrial DNA sequence variations. In these places, 1,085 OFF males were collected using methyl eugenol lure while 30 males of Zeugodacus cucurbitae and one male of Bactrocera tau were collected using Cuelure. A protein diet lure attracted 6 flies including one OFF and 5 flies of Z. cucurbitae. Male heads of OFF contained pterin, which increased in contents with age from 32 to $59{\mu}g/head$. There was a local variation in pterin amounts in OFF heads, in which Kaohsiung population had lower amounts of pterin than Taipei and Taichung populations. Genetic distance among these three populations were measured by random amplified polymorphic DNA and showed that Taipei population was separated from Taichung/Kaohsiung cluster. Genetic variation was also analyzed in sequence variations in cytochrome oxidase I (CO-I) and NADH dehydrogenase I (ND-I). There was 7.8% variation in CO-I sequence (360 residues) and 6.6% variation in ND-I sequence (213 residues). These polymorphic sites are proposed to be used to develop SNP (single nucleotide polymorphism) markers characteristic to Taiwan OFF populations.

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.

A study of morphological characteristics and hybridization on Lepista nuda (민자주방망이버섯(Lepista nuda )의 형태적 특성 및 교배에 관한 연구)

  • Jung, Won Soon;Kim, Jong Bong
    • Journal of Mushroom
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    • v.11 no.1
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    • pp.1-8
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    • 2013
  • In this study, twelve of Lepista nuda were collected from various localities in Korea. Also thirteen exotic L. nuda species were collected from Japan, France, Switzerland and Portugal. Spores were isolated under optical microscope. These spores were placed on the surface of YM medium for inducing to germination. Eleven mating-groups were selected by morphological characters of fruit body such as size, color and stipe patterns. Intra-isolate crosses were made between two single-spore isolates derived from mating-groups. Also, dikaryotic crossing using the isolates from L. nuda were carried out to evaluated tetrakaryon formation. Cross-mating compatibility tests also verified its dikaryotic state by microscopic or molecular genetic observation of clamp connection and Random Amplified Polymorphic DNA (RAPD) band pattern. To analyze the growth rate of hybrids and parents mycelium in dikaryons obtained from compatible mating groups were placed on PDA medium. Intra-isolate crosses determined eleven mating-groups within L. nuda. The typical clamp connection were mostly observed in mating-groups of Korean L. nuda in $K1{\times}K2$, $K1{\times}K3$, $K1{\times}K4$, $K1{\times}K6$, $K1{\times}K5$, $K2{\times}K4$, $K2{\times}K3$, $K2{\times}K6$, $K3{\times}K4$, $K4{\times}K5$, and $K4{\times}K6$. Korean L. nuda type of dikaryon, shown to cross-incompatibility with L. sordida, it seemed that mating induce more rapidly than wild types in a view of growth rate. In conclusion, it would be useful to improve mass production with better morphological characteristics through a special mating of L. nuda.

Characterization of a New Cultivar "Dan Bi" by Mono-mono Hybridization in Pleurotus eryngii (단핵균주간 교잡에 의한 큰느타리버섯 신품종 "단비"의 특성)

  • Kim, Min-Keun;Ryu, Jae-San;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.39-43
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    • 2011
  • A new cultivar "Dan Bi" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2312 and KNR2596. The parental strains, KNR2312 and KNR2596, are characterized by the property of high quality and a small number of primordia formation, respectively. The optimum temperature of mycelial growth was 25 and that of fruiting body development was $15{\sim}16^{\circ}C$. The period of harvesting including primordia formation was 0.7~1.3 days later than that of control strain Knneutari No. 3 in the culling cultivation. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has a scale like as cobweb. The yield was $93{\pm}9.7$ g per 850 cc of plastic bottle. Analysis of the genetic characteristics of the new commercial variety "Dan Bi" showed a different profile as that of the control strain, Knneutari No. 3, when RAPD (Random Amplified Polymorphic DNA) primer #8005 was used. This new variety "Dan Bi" of Pleurotus eryngii is characterized by a small number of primordia formation after scratching.