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High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

  • Kim, Jong Youn;Adhikari, Prakash Babu;Ahn, Chang Ho;Kim, Dong Hwi;Kim, Young Chang;Han, Jung Yeon;Kondeti, Subramanyam;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.38-48
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    • 2019
  • Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

Developing a Scientific Creativity Test to Explore the Relationship between Elementary Students' Creative Process and Product - Focusing on Biology - (초등학생의 창의 과정과 산물의 관계를 탐색하기 위한 과학 창의성 검사 도구 개발 - 생명 영역을 중심으로 -)

  • Kim, Minju;Lim, Chaeseong
    • Journal of Korean Elementary Science Education
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    • v.40 no.4
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    • pp.520-544
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    • 2021
  • This study aims to develop a scientific creativity test for exploring the relationship between elementary students' creative process and product. For this, the researcher reviewed the literatures of scientific creativity and developed the items based on the constructs of creative process and product. After a review conducted by nine science education specialists, a pilot test, and additional revision and supplementation of observation test, the test, consisting of two sets-"animals" and "plants"-was finally conducted on 105 fifth-grade students. The test results were analyzed by using statistical analysis software. WinSteps, SPSS, and AMOS. The main findings from this study are as follows. First, when it comes to scientific creativity, creative process consists of science knowledge, inquiry skills, and creative thinking skills (divergent, convergent, and associative thinking skills). Creative product in science is a new and scientifically useful idea realized in a certain form. Second, observation, which was selected as a representative inquiry skill in this research, should not be related to creative thinking skills. Third, among the rest of the items, usefulness had the lowest averages, as it was, perhaps, difficult to satisfy the teachers' criteria for the scientific validity and usefulness. Fourth, the Spearman correlation coefficients between the items of "animals" and "plants" to find out the parallel-form reliability were significant, except for the item of originality. Fifth, the test was satisfactory with regard to the three aspects of construct validity-convergent, discriminant, and nomological. This study concludes by discussing the usefulness of this test, which has the possibility of exploring the relationship between creative process and product and of playing a role as an authentic evaluation tool in school.

First Report of Tomato Spotted Wilt Virus in Oxypetalum coeruleum in Korea (옥시페탈룸에서 발생한 토마토반점위조바이러스 국내 첫 보고)

  • Eseul, Baek;Peter, Palukaitis;Ju-Yeon, Yoon
    • Research in Plant Disease
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    • v.28 no.4
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    • pp.231-236
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    • 2022
  • Oxypetalum coeruleum, commonly known as Tweedia, is a perennial herbaceous plant of the Apocynaceae family native to southern Brazil and Uruguay. Tweedia plants are grown as one of the most popular ornamental flowers for floral arrangement in Korea. In May 2021, several tweedia plants in a single greenhouse in Gimje, Jeollabuk-do were found to show virus-like symptoms including necrotic rings, vein-clearing, chlorotic mottle, and mosaic on the leaves, and necrosis on the stems. Here, we have identified tomato spotted wilt virus (TSWV) in symptomatic tweedia leaves by applying high-throughput RNA sequencing. In the result, a single infection by TSWV was verified without mixed infections of different virus species. To confirm the presence of TSWV, a reverse transcription polymerase chain reaction was performed with a specific primer set to the N gene of TSWV. The complete genomic sequence of L, M, and S segments of TSWV 'Oxy' isolate were determined and deposited in GenBank under accession numbers LC671525, LC671638, and LC671639, respectively. In the phylogenetic tree analysis by maximum likelihood method, 'Oxy' isolate showed a high relationship with TSWV 'Gumi' isolate from Gerbera jamesonii in Gyeongsangbuk-do, Korea; for all three RNA segments. To our knowledge, this is the first report of TSWV infection of O. coeruleum in Korea.

Improvement of Pre-harvest Sprouting Resistance in Korean japonica Varieties through a Precision Marker-based Breeding

  • Kamal Bhattarai;Patricia Izabelle Lopez;Sherry Lou Hechanova;Ji-Ung Jeung;Hyun-Sook Lee;Eok-Keun Ahn;Ung-Jo Hyun;Jong-Hee Lee;So-Myeong Lee;Jose E. Hernandez;Sung-Ryul Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.269-269
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    • 2022
  • Pre-harvest sprouting (PHS) on rice panicles is getting problematic in recent several years in Korea due to climate changes such as high temperature and more frequent typhoons during harvesting season. PHS negatively affects grain quality severely and also yield. Genetic improvement of Korean varieties (Oryza sativa ssp. japonica) through a marker assisted-backcross breeding (MAB) with the known PHS resistant genes must be one of ideal solutions. However, the final breeding products of MAB occasionally exhibit unwanted traits, especially the cross between genetically distant parents. This might be caused by linkage drag and/or presence of the gene-unlinked donor introgressions, resulting that the final products could not be released to the farmers. The major PHS resistance gene, Sdr4 (Seed dormancy 4) originated from an indica cultivar, Kasalath was selected as a donor gene. In order to avoid unexpected phenotypes in the breeding products, we performed a precision marker-based breeding (PMBB) consisting of foreground, recombinant, and background selections (FS, RS, and BS) which aim to develop 'single small introgression lines' (~100 kb introgression). Korean varieties (Ilpum and Gopum) were crossed with Kasalath. We developed Sdr4-allele specific markers for FS and a set of polymorphic flanking markers near the Sdr4 (-350kb and +420kb) for RS. To minimize linkage drag, the small introgression (< 125kb) containing Sdr4 was selected in Ilpum background (BC2F4) through 1st RS with ~1,200 F2 or BC1F2 plants (one side trimmed) and then 2nd RS with ~1,000 progenies from the 1st RS selected plants (another side trimmed). After RS, the selected lines were genotyped by using Infinium 7K SNP chip to detect other donor introgressions and the lines were backcrossed. Currently BS is on-going from the backcross-derived progenies with BS markers to remove residual introgressions. During the PMBB process, genetic effect of Sdr-4-Kasalath allele was confirmed in Ilpum and Gopum backgrounds by PHS phenotyping using the segregating BC2F3 or BC1F4 materials. The Sdr4 PMBB lines in Ilpum background (< 125kb introgression) will be valuable genetic resources to improve PHS resistance in modem popular temperate japonica varieties.

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Interactions between Oxidative Pentose Phosphate Pathway and Enzymes of Nitrate Assimilation "Nitrate Reductase, Nitrite Reductase, Glutamine Synthetase$_1$" and Ammonium Reassimilation "Glutamine Synthetase$_2$" as affected by $No_3$-Concentration ($No_3$-수준이 Oxidative Pentose Phosphate Pathway와 질산동화작용 효소"Nitrate Reductase, Nitrite Reductase, Glutamine Synthetase$_1$" 및 암모늄재동화작용 주요효소"Glutamine Synthetase$_2$"활성도의 상호관계에 미치는 영향)

  • Sohn, Sang-Mok;Michael James Emes
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.5
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    • pp.468-475
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    • 1992
  • In order to understand more clearly the integration between N-assmilation and C-metabolism in relation to N fertilization, a pot experiment with 5 different level of N fertilization(0, 5, 10, 25, 50 mM NO$_3$$_{[-10]}$ ) was conducted in Manchester, U.K. The peas (Pisum sativum L., cv. Early Onward) were sown in vermiculate (5 cm depth) and cultivated for 6 days under temperature controlled dark room conditions ($25^{\circ}C$). The plants received frequent irrigation with a nutrient solution: it was fertilized every 2 days, 3 times a day at 10h, 13h, 16h respectively. Elevated NO$_3$$^{[-10]}$ concentration, the activity levels of NR, NiR, total GS(crude extract), GS$_2$(plastid) in both root and shoot were increased and reached the peak in 5~25 mM, except NiR specific activity which increased its activity continually until 50 mM NO$_3$$^{[-10]}$ treatment. Total activities of GS (crude extract) in both root and shoot became higher than those of GS$_2$(Plastid), and the activity ratios of total GS in the crude extract and GS$_2$ in the plastids were 3.0 to 4.3 in root, but 3.2 to 10.6 in shoot. It was concluded that the reductants and A TP from OPPP itself should be enough to achieve the high rate of NR, NiR, GS$_1$, GS$_2$ in plant root and shoot for reduction or assimilation of nitrogen, but these enzyme activities might be inhibited by an excess of NO$_3$$^{[-10]}$ influx over the reduction capacity.

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The Brassica rapa Tissue-specific EST Database (배추의 조직 특이적 발현유전자 데이터베이스)

  • Yu, Hee-Ju;Park, Sin-Gi;Oh, Mi-Jin;Hwang, Hyun-Ju;Kim, Nam-Shin;Chung, Hee;Sohn, Seong-Han;Park, Beom-Seok;Mun, Jeong-Hwan
    • Horticultural Science & Technology
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    • v.29 no.6
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    • pp.633-640
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    • 2011
  • Brassica rapa is an A genome model species for Brassica crop genetics, genomics, and breeding. With the completion of sequencing the B. rapa genome, functional analysis of the genome is forthcoming issue. The expressed sequence tags are fundamental resources supporting annotation and functional analysis of the genome including identification of tissue-specific genes and promoters. As of July 2011, 147,217 ESTs from 39 cDNA libraries of B. rapa are reported in the public database. However, little information can be retrieved from the sequences due to lack of organized databases. To leverage the sequence information and to maximize the use of publicly-available EST collections, the Brassica rapa tissue-specific EST database (BrTED) is developed. BrTED includes sequence information of 23,962 unigenes assembled by StackPack program. The unigene set is used as a query unit for various analyses such as BLAST against TAIR gene model, functional annotation using MIPS and UniProt, gene ontology analysis, and prediction of tissue-specific unigene sets based on statistics test. The database is composed of two main units, EST sequence processing and information retrieving unit and tissue-specific expression profile analysis unit. Information and data in both units are tightly inter-connected to each other using a web based browsing system. RT-PCR evaluation of 29 selected unigene sets successfully amplified amplicons from the target tissues of B. rapa. BrTED provided here allows the user to identify and analyze the expression of genes of interest and aid efforts to interpret the B. rapa genome through functional genomics. In addition, it can be used as a public resource in providing reference information to study the genus Brassica and other closely related crop crucifer plants.

Characteristics of Light Harvesting Chlorophyll-Protein Complex and Singlet Oxygen ($^1O_2$) Quenching in Leaf-burning Disease from Panax ginseng C. A. Meyer (인삼 Light Harvesting Chlorophyll Protein의 특성 및 엽소병에서 Singlet Oxygen($^1O_2$) Quenching)

  • 양덕조;이성택
    • Journal of Ginseng Research
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    • v.13 no.2
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    • pp.158-164
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    • 1989
  • In order to determine the relationships between the lea(-burning disease and the light harvesting chlorophyll-protein (LHCP) complex in Panax ginseng C. A. Meyer, we investigated the chlorophyll-protein (CP) complex of the thylakoid membrane and its characteristics. In P. ginseng four Cp-complex bands determined by non-denaturing SDS-PAGE were identified CP I'(containing reaction center of photosystem I and LHCP I antennae), CP I (reaction center of photosystem I) LHCP II** (oligoform of LHCP II), and LHCP II (photosystem II antennae, CP 26 and CP 29) by Bassis and Dunahay's procedures. Under our experimental condition, the CP I band was only observed in P. ginseng and the band intensity of LHCP II** in P ginseng was higher than in spinach and soybean. There were differences in the absorption and fluorescence spectra and chlorophyll a/b ratio of the CP-complex bands between P. ginseng and other Plants. The Polypeptidr content of P. ginseng thylakoid was lower than in spinach and soybean thylakoid, and the Polypeptide profiles of P. ginseng was low band intensity, especially about 29-35 kD, 55 kD, and 60 kD, compared to spinach and soybean. The inhibitory effects of 2,5-dimethylfuran, specific singlet oxygen ($^1O_2$) quencher, showed that singlet oxygen destroyed 60% of chl.a, 90% of chl.b and 70% of carotenoid in bleaching P. ginseng with leaf-burning disease.

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Development of Molecular Marker for the authentication of Patriniae Radix by the analysis of DNA barcodes (DNA 바코드 분석을 통한 패장 기원종 감별용 분자 마커 개발)

  • Kim, Wook Jin;Ji, Yunui;Lee, Young Mi;Kang, Young Min;Choi, Goya;Kim, Ho Kyoung;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.45-53
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    • 2014
  • Objectives : Due to the morphological similarity of in the roots of herbal medicine, the official herbal medicine is very difficult to authenticate between the original plants of Patriniae Radix and two adulterant Patrinia species. Therefore, we introduced DNA barcode analysis to establish a powerful tool for the authentication of Patriniae Radix from its adulterants. Methods : To analyze DNA barcode regions, genomic DNA was extracted from twenty-nine specimens of Patrinia scabiosaefolia, Patrinia villosa, Patrinia saniculifolia, and Patrinia rupestris, and internal transcribed spacer 2(ITS2), matK and rbcL genes were amplified. For identification of species specific sequences, a comparative analysis was performed by the ClastalW based on entire sequences of ITS2, matK and rbcL genes, respectively. Results : In comparison of three DNA barcode sequences, we identified 22, 22, and 12 species-specific nucleotides enough to distinguish each four species from ITS2, matK and rbcL gene, respectively. The sequence differences at the corresponding positions were available genetic marker nucleotides to discriminate the correct species among analyzed four species. These results indicated that comparative analysis of ITS2, matK and rbcL genes were useful genetic markers to authenticate Patriniae Radix. Conclusions : The marker nucleotides enough to distinguish P. scabiosaefolia, P. villosa, P. saniculifolia, and P. rupestris, were obtained at 22 SNP marker nucleotides from ITS2 and matK DNA barcode sequences, but they were confirmed at 12 SNP marker nucleotides from rbcL. These differences could be used to authenticate Patriniae Radix from its adulterants as well as discriminating each four species.

Case Analysis on High Concentration of SO2 and Review on Its Reduction Policy in the Ulsan Metropolitan Area since 2001 (울산 지역에서 2001년 이후 이산화황(SO2)의 고농도 사례 분석과 저감 정책 방안의 검토)

  • Moon, Yun-Seob
    • Journal of Environmental Science International
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    • v.17 no.4
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    • pp.423-437
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    • 2008
  • Until comparatively lately, the annual time series of the $SO_2$ concentration had been shown in a decreasing trend in Ulsan as well as other Korean cities. However, the high concentration of $SO_2$ was frequently found in the specific countermeasure region including the national industrial complex such as Mipo and Onsan in the Ulsan city for the period of $2001{\sim}2004$. There are many conditions that can influence the high concentration of $SO_2$ at monitoring sites in Ulsan, such as: First, annual usage of the fuel including sulfur increased in comparison with the year before in spite of the fuel conversion policy which wants to use low sulfur oil less than 3% and LNG. Second, point source, such as the power plants and the petroleum and chemistry stacks, was the biggest contributor in $SO_2$ emission, as a analyzed result of both the air quality modeling and the stack tole-monitoring system (TMS) data. And third, the air pollutants that occurred in processes of homing and manufacturing of the fuel including sulfur were transported slow into a special monitoring site by accumulating along the frontal area of see-breeze. It was concluded that Ulsan's current environmental policy together with control methods should be changed into the regulation on total amount of emission, including a market-based emission trading with calculating of atmospheric environmental critical loads, for the $SO_2$ reduction like the specific countermeasure for the $O_3$ and PM10 reduction in the Seoul metropolitan area. And this change should be started in the big point sources of $1{\sim}3$ species because they are big contributors of Ulsan's $SO_2$ pollution. Especially it is necessary to revitalize of the self-regulation environmental management. Other control methods for sustaining the $SO_2$ reduction are as follows: maintenance of the fuel conversion policy, reinforcement of the regional stationary source emission standard, and enlargement of the stack TMS.

Expressional Patterns of Anther-specific Genes from Chinese Cabbage during the Flower Development (배추 약 발달 시기별 유전자의 발현 양상)

  • Kim, Hyun Uk;Chung, Kyu Hwan
    • Horticultural Science & Technology
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    • v.17 no.1
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    • pp.7-10
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    • 1999
  • This study was carried out with the purpose of looking into the transcriptionally regulated genes related to the anther development, characterizing them, and applying their promoters to induce male-sterile plants and restore their fertility. Fifteen anther-specific clones were isolated from the anther cDNA library of Chinese cabbage through the differential screening and sequenced partially at both ends. These partial sequence data showed that cDNA clones BAN52, 84, 101, and 229 are very similar to polygalacturonase, ascorbate oxidase, $H^+-translocating$ ATPase, and pectin esterase genes respectively. However, the other clones have not been matched to any of gene sequences in data bank. In northern dot blot analysis, the transcripts of cDNA clone BAN5, 10, 33, 52, 57, 102, 103, 215, 229 appeared in the flower bud of 2.1 mm in length and their amounts were gradually increased along with the anther development. Transcription of cDNA clone BAN32, 54, 62, 84, 101 began in flower bud of 3.9 mm, which is the late stage in anther development. However, the transcription of BAN87 was very small, but its transcript was detected in all anther developmental stages.

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