• Title/Summary/Keyword: Seed purity

Search Result 72, Processing Time 0.033 seconds

Antifungal and Plant Growth Promotion Activities of Recombinant Defensin Proteins from the Seed of Korean Radish (Raphanus sativus L.)

  • Hwang, Cher-Won
    • Korean Journal of Environmental Agriculture
    • /
    • v.28 no.4
    • /
    • pp.435-441
    • /
    • 2009
  • In the present study, we analyzed the defensin protein deduced from Korean radish (Raphanus sativus L.) seeds.To express the genes in E. coli, we constructed a recombinant expression vector with a defensin gene, named rKRs-AFP gene isolated from Korean radish seeds. Over expressed rKRs-AFP proteins was separated by SDS-PAGE to determine the purity, and protein concentration was determined by the Bradford method. Antifungal activity was assessed by disk assay method against the tested fungi. As a result, when 500 mL of cell culture were disrupted by sonicator, 32.5 mg total proteins were obtained. The purified protein showed a single band on SDS-PAGE with estimated molecular weight about 6 KDa, consistent with the molecular mass calculated from the deduced amino acid sequence. The purified rKRs-AFP protein showed remarkable antifungal activities against several fungi including Aspergillus niger, Botrytis cinerea causing the gray mold disease, and Candida albicans. In field tests using the purified rKRs-AFP protein, the protein showed the reducing activity of disease spot and the mitigating effect of spreading of disease like agrichemicals. The immuno-assay of rKRs-AFP protein showed that the purified protein entirely accumulated at B. cinerea cytoplasm through the hyphal septa shown by fluorescence imaging. There was no fluorescence inside the cell, when the hypha was incubated without the protein. These all results indicate that the recombinant rKRs-AFP proteins can be utilized as a potential antifungal drug to control harmful plant fungal pathogens.

Purification and Serology of Potato Virus S (감자 바이러스 S의 순화와 항혈청제조)

  • Lee Soon Hyung;Lee Key Woon;Chung Bong Jo
    • Korean journal of applied entomology
    • /
    • v.16 no.3 s.32
    • /
    • pp.145-148
    • /
    • 1977
  • he study was conducted to produce an antiserum of potato virus S for identification and screening of seed-potatoes. Potato virus S was isolated from infected plants and identified by means of indicator plants and electro microscopy. Isolated potato virus S was multiplied in Nicotiana deebneyii and the virus was purified by a modified method that was developed through this study. The purity of potato virus S was 1.18mg/ml. Purified potato virus S was injected into rabbit intravenously once a week for 5 weeks. Antiserum was collected 10 days after the last injection. The produced antiserum was determined to have a titer of, 1/2048 by means of microprecipitin tests.

  • PDF

Titanium Dioxide Recovery from Soda-roasted Spent SCR Catalysts through Sulphuric Acid Leaching and Hydrolysis Precipitation (소다배소 처리된 탈질 폐촉매로부터 황산침출과 가수분해 침전반응에 의한 TiO2의 회수)

  • Kim, Seunghyun;Trinh, Ha Bich;Lee, Jaeryeong
    • Resources Recycling
    • /
    • v.29 no.5
    • /
    • pp.48-54
    • /
    • 2020
  • Sulphuric acid (H2SO4) leaching and hydrolysis were experimented for the recovery of titanum dioxide (TiO2) from the water-leached residue followed by soda-roasting spent SCR catalysts. Sulphuric acid leaching of Ti was carried out with leachate concentration (4~8 M) and the others were fixed (temp.: 70 ℃, leaching time: 3 hrs, slurry density: 100 g/L, stirring speed: 500 rpm). For recovering of Ti from the leaching solution, hydrolysis precipitation was conducted at 100 ℃ for 2 hours in various mixing ratio (leached solution:distilled water) of 1:9 to 5:5. The maximum leachability was reached to 95.2 % in 6 M H2SO4 leachate. on the other hand, the leachability of Si decreased dramatically 91.7 to 3.0 % with an increase of H2SO4 concentration. Hydrolysis precipitation of Ti was proceeded with leaching solution of 8 M H2SO4 with the lowest content of Si. The yield of precipitation increased proportionally with a dilution ratio of leaching solution. Moreover, it increased generally by adding 0.2 g TiO2 as a precipitation seed to the diluted leaching solution. Ultimately, 99.8 % of TiO2 can be recovered with the purity of 99.46 % from the 1:9 diluted solution.

Development of highly uniform variety for processing using SSR markers in radish (Raphanus sativus L) (분자표지를 활용한 고품질 가공용 고순도 무 품종 육성)

  • Jung, Un-Hwa;Oh, Jong-Hyuck;Kim, Young-Gyu;Ahn, Chun-Hee;Lee, Kwang-Sik;Choi, Su-Ryun;Lim, Yong-Pyo;Park, Su-Hyoung;Choi, Ki-Young;Lee, Yong-Beom
    • Journal of Plant Biotechnology
    • /
    • v.41 no.1
    • /
    • pp.56-63
    • /
    • 2014
  • Using commercial radish varieties for processing, about 30% of radish was discarded due to the root shape and low purity. To raise the processing ability, we tried to develop a new variety producing H-shaped root. As another characteristic required in variety for processing is high purity, we tried to raise purity using simple sequence repeats (SSR) markers for testing seed purity in every segregating generation. To develop Male-sterile (MS) seeding parent, we crossed commercial variety of 'Gwan dong spring' and 'Gyeo ryong spring'. One elite inbred was selected as recurrent parent for the MS plant. The major horticultural traits of selected inbred line were disease resistance, late bolting, heat resistance and bright green root top color. To develop pollen parent, we crossed commercial variety of 'Tae sang king' and 'Seoul spring'. We used individual selection method to develop H-shaped hard root and disease resistant inbred. In each segregating generation, we selected one plant based on phenotype and the uniformity of selected plant was tested by SSR markers using self-pollinated seeds. In the first segregating generation, 64.6% of sib plants shared the same band in PCR amplification using ACMP-490 primer and 66.7% using cnu-316 primer. The uniformity of segregating generations using ACMP-490 and cnu-316 raised in second generation to 68.8%, 70.8%, respectively; in third generation to 93.8%, 100%; in fourth generation to 93.8%, 100%; in fifth generation to 95.8%, 100%; in sixth generation to 100%, 100%. A novel cross was made using selected MS parent and pollen parent. When we checker the horticultural traits using autumn cultivation, the novel cross variety produced H-shaped root comparing other commercial varieties and produced highly uniform radish. Thus we registered this novel cross variety as 'YR ORE' at 2013 (Registration No. 4550).

The Aspect of Natural Regeneration for Major Tree Species in the Natural Deciduous Forest (천연문엽수임내(天然聞葉樹林內) 주요(主要) 구성(構成) 수종(樹種)의 천연경신(天然更新) 양상(樣相))

  • Kim, Ji Hong;Yang, Hee Moon;Jin, Guang Ze;Lee, Won Sup;Kang, Sung Kee
    • Journal of Forest and Environmental Science
    • /
    • v.17 no.1
    • /
    • pp.1-17
    • /
    • 2001
  • Forming a part of "Cooperative Practical Study for the Modernization of the Management of National Forest", this study was conducted to provide overall ecological information for the natural regeneration of major tree species on the basis of community structural attributes in the deciduous forest ecosystem. Followings are summarized characteristics of the natural regeneration for the selected tree species. Betula costata : Although large number of seeds are dispersed by wind, they require mineral soils to germinate. Thick litter layer could be an obstacle to germinate. After germination, the seedling requires large amount of light for successful establishment. Acer mono : Characterized by high shade tolerance and weak drought resistance, the seedling should be overcasted with more than 50% of canopy coverage. High stand density should be maintained to produce good quality of timber. The potential of coppice may be high. Ulmus laciniata : Since this species needs high rate of troll moisture and light, around 60% of canopy coverage should be maintained to retain moisture and incoming light. The competition with other vegetation should be removed for the favor of successful seedling establishment. Fraxinus mandshurica : This species requires moist mineral soils to germinate. After germination, the seedling needs large amount of light and moisture for successful establishment. Site preparation should be applied to reduce competition with weedy vegetation. Fraxinus rhynchophylla : Interval of large seed crops may be highly varied. Thick litter layer could be an obstacle to germinate. Site preparation should be applied to reduce competition with weedy vegetation so as to achieve successful seedling establishment. Quercus mongolica : Including the difficulty of seed supply by the consumption, thick litter layer and mountain bamboo cover could be the obstacle to germinate. More than 50% of relative light intensity is necessary to achieve successful seedling establishment. Kalopanax pictus : Thick litter layer could be an obstacle to germinate. The seedling needs large amount of light and moisture for successful establishment. Abies holophylla : In spite of high shade tolerance, the growth rate in sapling stage may be extremely slow. Cornus controversa : Seeds (drups) are consumed and dispersed by animals, tending to be not sufficient in seed supply. This species requires large amount of light for successful germination and seedling establishment. Tilia amurensis : The difficulty of seed supply might be expected with low seed purity and double dormancy. Since thick litter layer could be an obstacle to germinate, the species requires moist mineral soils for successful germination. The potential of coppice may be extremely high.

  • PDF

Purification of p-Dioxanone from p-Dioxanone and Diethylene Glycol Mixture by a Layer Melt Crystallization (경막형 용융결정화에 의한 파라디옥사논과 디에틸렌글리콜 혼합물로부터 파라디옥사논의 정제)

  • Kim, Sung-Il;Kim, Chul-Ung;Park, So-Jin
    • Korean Chemical Engineering Research
    • /
    • v.43 no.5
    • /
    • pp.595-602
    • /
    • 2005
  • In order to purify diethylene glycol as main impurity included in p-dioxanone, SLE (solid-liquid equilibria) and mixture density on two components system of p-dioxanone and diethylene glycol were measured and a layered melt crystallization with seed has been applied. The SLE of p-dioxanone and diethylene glycol were a simple eutectic system and the temperature and PDX concentration at eutectic point were 0.08 and 246 K, respectively. Densities of their binary mixtures were well fitted by the best correlation equation, ${\rho}_l=0.405+1.361x+0.002T-0.004xT$. In the melt crystallization, the growth rate (G) was proportional to the 1.5th power of the subcooling degree. The effective distribution coefficient ($K_{eff}$) as the degree of impurity removal was observed to increase with increasing the growth rate and initial p-dioxanone concentration. And also, $K_{eff}$ was correlated with Z function using Wintermantel's model such as $K_{eef}=-0.0604+6.392{\times}Z$. Finally, PDX purity through the optimization of this process can be obtained over 99%.

Use of Microsatellite Markers Derived from Genomic and Expressed Sequence Tag (EST) Data to Identify Commercial Watermelon Cultivars (수박 시판 품종의 식별을 위한 Genomic과 Expressed Sequence Tag (EST)에서 유래된 Microsatellite Marker의 이용)

  • Kwon, Yong-Sham;Hong, Jee-Hwa;Kim, Du-Hyun;Kim, Do-Hoon
    • Horticultural Science & Technology
    • /
    • v.33 no.5
    • /
    • pp.737-750
    • /
    • 2015
  • This study was carried out to construct a DNA profile database for 102 watermelon cultivars through the comparison of polymorphism level and genetic relatedness using genomic microsatellite (gMS) and expressed sequence tag (EST)-microsatellite (eMS) markers. Sixteen gMS and 10 eMS primers showed hyper-variability and were able to represent the genetic variation within 102 watermelon cultivars. With gMS markers, an average of 3.63 alleles per marker were detected with a polymorphism information content (PIC) value of 0.479, whereas with eMS markers, the average number of alleles per marker was 2.50 and the PIC value was 0.425, indicating that eMS detects a lower polymorphism level compared to gMS. Cluster analysis and Jaccard's genetic distance coefficients using the unweighted pair group method with arithmetic average (UPGMA) based on the gMS, eMS, and combined data sets showed that 102 commercial watermelon cultivars could be categorized into 6 to 8 major groups corresponding to phenotypic traits. Moreover, this method was sufficient to identify 78 out of 102 cultivars. Correlation analysis with Mantel tests for those clusters using 3 data sets showed high correlation ($r{\geq}0.80$). Therefore, the microsatellite markers used in this study may serve as a useful tool for germplasm evaluation, genetic purity assessment, and fingerprinting of watermelon cultivars.

Development of SNP marker set for marker-assisted backcrossing (MABC) in cultivating tomato varieties

  • Park, GiRim;Jang, Hyun A;Jo, Sung-Hwan;Park, Younghoon;Oh, Sang-Keun;Nam, Moon
    • Korean Journal of Agricultural Science
    • /
    • v.45 no.3
    • /
    • pp.385-400
    • /
    • 2018
  • Marker-assisted backcrossing (MABC) is useful for selecting offspring with a highly recovered genetic background for a recurrent parent at early generation unlike rice and other field crops. Molecular marker sets applicable to practical MABC are scarce in vegetable crops including tomatoes. In this study, we used the National Center for Biotechnology Information- short read archive (NCBI-SRA) database that provided the whole genome sequences of 234 tomato accessions and selected 27,680 tag-single nucleotide polymorphisms (tag-SNPs) that can identify haplotypes in the tomato genome. From this SNP dataset, a total of 143 tag-SNPs that have a high polymorphism information content (PIC) value (> 0.3) and are physically evenly distributed on each chromosome were selected as a MABC marker set. This marker set was tested for its polymorphism in each pairwise cross combination constructed with 124 of the 234 tomato accessions, and a relatively high number of SNP markers polymorphic for the cross combination was observed. The reliability of the MABC SNP set was assessed by converting 18 SNPs into Luna probe-based high-resolution melting (HRM) markers and genotyping nine tomato accessions. The results show that the SNP information and HRM marker genotype matched in 98.6% of the experiment data points, indicating that our sequence analysis pipeline for SNP mining worked successfully. The tag-SNP set for the MABC developed in this study can be useful for not only a practical backcrossing program but also for cultivar identification and F1 seed purity test in tomatoes.

DNA fingerprinting analysis for soybean (Glycine max) varieties in Korea using a core set of microsatellite marker (핵심 Microsatellite 마커를 이용한 한국 콩 품종에 대한 Fingerprinting 분석)

  • Kwon, Yong-Sham
    • Journal of Plant Biotechnology
    • /
    • v.43 no.4
    • /
    • pp.457-465
    • /
    • 2016
  • Microsatellites are one of the most suitable markers for identification of variety, as they have the capability to discriminate between narrow genetic variations. The polymorphism level between 120 microsatellite primer pairs and 148 soybean varieties was investigated through the fluorescence based automatic detection system. A set of 16 primer pairs showed highly reproducible polymorphism in these varieties. A total of 204 alleles were detected using the 16 microsatellite markers. The number of alleles per locus ranged from 6 to 28, with an average of 12.75 alleles per locus. The average polymorphism information content (PIC) was 0.86, ranging from 0.75 to 0.95. The unweighted pair group method using the arithmetic averages (UPGMA) cluster analysis for 148 varieties were divided into five distinctive groups, reflecting the varietal types and pedigree information. All the varieties were perfectly discriminated by marker genotypes. These markers may be useful to complement a morphological assessment of candidate varieties in the DUS (distinctness, uniformity and stability) test, intervening of seed disputes relating to variety authentication, and testing of genetic purity in soybean varieties.

Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

  • Park, Kyunghyuk;Frost, Jennifer M.;Adair, Adam James;Kim, Dong Min;Yun, Hyein;Brooks, Janie S.;Fischer, Robert L.;Choi, Yeonhee
    • Molecules and Cells
    • /
    • v.39 no.10
    • /
    • pp.768-775
    • /
    • 2016
  • The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.