• Title/Summary/Keyword: 저항성 유전자

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GUS Gene expression and plant regeneration via somatic embryogenesis in cucumber (Cucumis sativus L.) (오이에서 체세포배 발생을 통한 GUS유전자의 발현 및 식물체 재생)

  • Kim, Hyun-A;Lee, Boo-Youn;Jeon, Jin-Jung;Choi, Dong-Woog;Choi, Pil-Son;Utomo, Setyo Dwi;Lee, Jae-Hyoek;Kang, Tong-Ho;Lee, Young-Jin
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.275-280
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    • 2008
  • One of the limitation for Agrobacterium-mediated transformation via organogenesis from cotyledon explants routinely in cucumber is the production of chimeric plants. To overcome the limitation, Agrobacterium-mediated transformation system via somatic embryogenesis from hypocotyl explants of cucumber (c.v., Eunsung) on the selection medium with paromomycin as antibiotics was developed. The hypocotyl explants were inoculated with Agrobacterium tumefaciens strain EHA101 carrying binary vector pPTN290; then were subsequently cultured on the following media: co-cultivation medium for 2 days, selection medium for $5{\times}14$ days, and regeneration medium. The T-DNA of the vector (pPTN290) carried two cassettes, Ubi promoter-gus gene as reporter and 35S promoter-nptll gene conferring resistance to paromomycin as selectable agent. The confirmation of stable transformation and the efficiency of transformation was based on the resistance to paromomycin indicated by the growth of putative transgenic calli on selection medium amended with 100mg/L paromomycin, and GUS gene expression. Forty eight clones (5.2%) with GUS gene expressed of 56 callus clones with resistance to paromomycin were independently obtained from 928 explants inoculated. Of 48 clones, transgenic plants were only regenerated from 5 clones (0.5%) at low frequency. The histochemical GUS assay in the transgenic seeds ($T_1$) also revealed that the gus gene was successfully integrated and segregated into each genome of transgenic cucumber.

Detection of LM canola with tolerance to glyphosate and glufosinate-ammonium via the Environmental monitoring in South Korea (자연생태계 모니터링을 통한 glyphosate와 glufosinate-ammonium에 저항성을 가지는 유전자변형 캐놀라의 발견)

  • Shin, Su Young;Jo, Beom-Ho;Moon, Jeong Chan;Lee, Jung Ro;Choi, Wonkyun;Seol, Min-A;Kim, Mi-Jeong;Song, Hae-Ryong
    • Journal of Plant Biotechnology
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    • v.43 no.4
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    • pp.479-485
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    • 2016
  • Living modified (LM) crops are imported each year to South Korea as food and feeds, LM canola being one of the imported crops. The cultivation of LM crops is not permitted in South Korea but the import of these crops is increasing. In this study, we surveyed the environmental risk of imported LM canola at 9 provinces, from March 2009 to June 2013. Monitoring of canola was conducted around feed factories, roadsides, harbors, farmhouses, and flower festival regions. From the total of 595 canola samples collected from 1850 monitoring sites, we identified 6 LM canola samples. The LM canola samples were subjected to protein and DNA based analysis. PCR analyses using approved 5 single event primers (T45, MS8, RT73, Rf3 and Topas 19-2) revealed that two crops were glyphosate-resistant LM canolas, and four were glufosinate-resistant LM canolas. This study suggested that environmental monitoring is a useful research tool to manage LM crops unintentionally introduced into the environment in South Korea. This result can be used as a basis for future post-management of canola crops.

Characterization of a Drought-Tolerance Gene, BrDSR, in Chinese Cabbage (배추의 건조 저항성 유전자, BrDSR의 기능 검정)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.102-111
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    • 2016
  • The goal of this study was to characterize the BrDSR (Drought Stress Resistance in B. rapa) gene and to identify the expression network of drought-inducible genes in Chinese cabbage under drought stress. Agrobacterium-mediated transformation was conducted using a B. rapa inbred line ('CT001') and the pSL100 vector containing the BrDSR full length CDS (438 bp open reading frame). Four transgenic plants were selected by PCR and the expression level of BrDSR was approximately 1.9-3.4-fold greater than that in the wild-type control under drought stress. Phenotypic characteristics showed that BrDSR over-expressing plants were resistant to drought stress and showed normal growth habit. To construct a co-expression network of drought-responsive genes, B. rapa 135K cDNA microarray data was analyzed to identify genes associated with BrDSR. BrDSR was directly linked to DARK INDUCIBLE 2 (DIN2, AT3G60140) and AUTOPHAGY 8H (ATG8H, AT3G06420) previously reported to be leaf senescence and autophagy-related genes in plants. Taken together, the results of this study indicated that BrDSR plays a significant role in enhancement of tolerance to drought conditions.

Characterization of a New Gene Resistant to Alkylating Agents and 3-Aminobenzamide When Knocked Out in Fission Yeast (분열형 효모에서 유전자 결실에 의해 알킬화제와 3-AMINOBENZAMIDE에 저항성을 나타내는 새로운 유전자의 특성 분석)

  • 박종군;차재영;황성진;박세근;김미영;백성민;최인순;이정섭
    • Journal of Life Science
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    • v.12 no.2
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    • pp.219-225
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    • 2002
  • The organization of eukayotic chromatin into specific conformation that are associated with transcription, replication, reapir and other nuclear processes are achieved via a series of DNA-protein interaction. These interactions are mediated by a range of DNA-binding domains such as SAP domain et at. By searching S. pombe genomic DNA database, we have found a gene named SAPuvs (SAP UV Sensitive) whose amino acid sequence is in part similar to SAP domain of Arabidopsis poly (ADP-ribose) polymerase and Ku7O. Knock-out cell of S. pombe SAPuvs gene was constructed using Ura4 as a selection marker. Survival analysis of knock-out cell indicated that treatment with UV significantly reduces the survival compared to wild type cell. Potentiation of MMS-induced cytotoxicity by 3AB post-treatment was observed in wild type cells, but not in knock-out cells. These data suggested that the protein encoded by SAPuvs gene is associated with chromatin reorganization during DNA repair.

Transgenic Rice Expressing Snowdrop Lectin (Galanthus nivalis agglutinin; GNA) Shows Resistance to Rice Brown Planthopper (Nilaparvata lugens Stål) (Snowdrop lectin (Galanthus nivalis agglutinin: GNA) 유전자 도입에 의한 벼멸구 저항성 형질전환 벼 개발)

  • Lee, Soo In;Yoon, In Sun;Kim, Jin A;Hong, Joon Ki;Park, Beom-Seok;Lee, Yeon-Hee
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1614-1620
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    • 2012
  • Transgenic rice plants with increased resistance to rice brown planthopper (Nilaparvata lugens St${\aa}$l) were generated by particle bombardment-mediated transformation of plants with a gene encoding snowdrop lectin (Galanthus nivalis agglutinin; GNA) under control of the rice Rubisco small subunit (rbcS) promoter.. A large number of transgenic rice plants containing the GNA gene were generated. The integration, expression, and inheritance of this gene in the $R_1$ and $R_2$ generations were demonstrated by Southern and western blot analyses. The plants contained one to five copies of the transgene. The GNA protein comprised approximately 0.01-2.0% of total soluble protein in the $R_1$ and $R_2$ transgenic plants. Insect bioassays and feeding studies showed that the GNA protein expressed in the $R_2$ transgenic rice plants reduced the survival of brown planthoppers. The introduction of GNA into rice plants therefore can help to control insect pests.

Responses of Resistant Genes to Barley Yellow Mosaic Virus (BaYMV) Strains in Korea (국내 분포 보리호위축바이러스(Barley Yellow Mosaic Virus) strain에 대한 저항성 유전자 반응)

  • Park, Jong-Chul;Noh, Tae-Hwan;Park, Chul-Soo;Kang, Chun-Sik;Kang, Mi-Hyung;Lee, Eun-Sook;Lee, Jun-Hee;Lee, Jung-Joon;Kim, Tae-Soo
    • Research in Plant Disease
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    • v.15 no.2
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    • pp.72-76
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    • 2009
  • It was investigated the responses of BaYMV resistant genes to Korean BaYMV(Barley yellow mosaic virus) strains. BaYMV was distributed dominantly with about 51% detection ratio among the three investigated virus such as BaYMV, BaMMV(Barley mild mosaic virus) and SBWMV(Soil-borne wheat mosaic virus) in ELISA test. Double infection with BaYMV and BaMMV was detected also higher as 38.8%, however, BaMMV sole infection ratio was lower with only 1.4%. The 11 BaYMV resistant genes were tested their responses to four Korean BaYMV strains, BaYMV-N, H, I and M. Generally, rym 3 genes showed resistant to Korean BaYMV strains and rym 4m and 5a also was better. Three genes, rym 1+5(Mokusekko-3), rym 3(Ea 52, Baitori) and rym 5a(Solan) showed resistant responses to BaYMV-N type. In -H strain test, seven genes that rym 2(Mihori Hadaka 3), rym 3(Ea 52, Haganemugi, Baitori), rym 4m(Diana, Franka), rym 5a(Solan), rym 7(Hor 3365), rym 9(Bulgarian 347), rym 12(Jochiwon Covered 2) were considered as resistant. The three genes that rym 1+5, rym 3 and rym 5a was effective to -I strain, and rym 3, rym 4m and rym 5a showed resistant to -M strain.

Isolation and Functional Identification of BrDSR, a New Gene Related to Drought Tolerance Derived from Brassica rapa (배추 유래 신규 건조 저항성 관련 유전자, BrDSR의 분리 및 기능 검정)

  • Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.575-584
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    • 2015
  • Drought stress is a crucial environmental factor determining crop survival and productivity. The goal of this study was to clearly identify a new drought stress-tolerance gene in Brassica rapa. From KBGP-24K microarray data with the B. rapa ssp. pekinensis inbred line 'Chiifu' under drought stress treatment, a gene which was named BrDSR (B. rapa Drought Stress Resistance) was chosen among 738 drought-responsive unigenes. BrDSR function has yet to be determined, but its expression was induced over 6-fold by drought. To characterize BrDSR, the gene was isolated from B. rapa inbred line 'CT001' and found to contain a 438-bp open reading frame encoding a 145 amino acid protein. The full-length cDNA of BrDSR was used to construct an over-expression vector, 'pSL100'. Tobacco transformation was then conducted to analyze whether the BrDSR gene can increase drought tolerance in plants. The BrDSR expression level in T1 transgenic tobacco plants selected via PCR and DNA blot analyses was up to 2.6-fold higher than non-transgenic tobacco. Analysis of phenotype clearly showed that BrDSR-expressing tobacco plants exhibited more tolerance than wild type under 10 d drought stress. Taking all of these findings together, we expect that BrDSR functions effectively in plant growth and survival of drought stress conditions.

Isolation of Superoxide Dismutase cDNAS from an Weedy Rice Variety and Transformation of a Cultivated Rice Variety (잡초성벼의 superoxide dismutase cDNA cloning과 재배벼로의 형질전환)

  • Park, Sang-Gyu;Park, Jong-Suk;Lee, Seung-In;Suh, Suk-Chul;Kim, Byung-Keuk;Jo, Youl-Lae;Suh, Hak-Soo
    • Korean Journal of Environmental Agriculture
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    • v.21 no.2
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    • pp.156-161
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    • 2002
  • Two different cDNA clones for superoxide dismutase (SOD) were isolated from an weedy rice variety (Oryza sativa, cv. Bhutan14Ad) and were introduced into a cultivated rice variety (Oryza sativa, cv. Nakdong) in order to develop the environmental stress-resistant rice plants. Sequence analysis of the cloned cDNAS indicated that the deduced amino acid sequence of SOD-A is 88.4% identical to that of SOD-B. Furthermore, the nucleotide sequence of SOD-A is 99.3% identical to that of a Cu/Zn SOD gene of Oryza sativa (GenBank accession No. L36320). The nueleotide sequence of SOD-B was identical to that of the previously published SOD gene (Accession No. D01000). A cultivated rice variety, Nakdong-byeo, was transformed with chimeric SOD genes containing a actin promoter of rice and pin2 terminator using a particle bombardment technique. Transformed calli were selected on an selection medium containing phosphinothricin (PPT). Transgenic rice plants were regenerated from the PPT-resistant calli. PCR analysis with genomic DNAs from transgenic plants revealed that transgenes are introduced into rice genome.

Localized Induced Resistance to Erysiphe graminis f. sp. hordei in Near-Isogenic Barley Lines. (근 동질유전자 보리계통에서 보리흰가루병에 대한 국부적 유도저항성)

  • Cho Baik Ho;Peterson V. Smedegaard
    • Korean Journal Plant Pathology
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    • v.1 no.1
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    • pp.22-27
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    • 1985
  • Localized resistance against a virulent race of Erysiphe graminis f. sp. hordei by prior inoculation of a virulent or an avirulent race of the same fungus was induced on the near-isogenic barley leaves. Induced resistance could be detected within one hour following challenge inoculation with a virulent or an avirulent inducer race, hut the resistance increased greatly as the interval between the two inoculations was increased, showing the highest level by 6-9 hours of exposure to the inducer race. The level of the induced resistance was proportional to the amount of inducer inoculum applied. The resistances elicited by virulent or avirulent inducer races were similar with respect to the level of resistance and the time needed for its induction.

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Characterization of PR-10 gene derived from highly resistant '93-3-98' pear inoculated with scab (Venturia nashicola) (배 검은별무늬병(Venturia nashicola) 고도 저항성 '93-3-98' 유래 PR-10 유전자의 특성)

  • Chun, Jae An;Kim, Se Hee;Cho, Kang Hee;Kim, Dae Hyun;Choi, In Myong;Shin, Il Sheob
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.25-33
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    • 2015
  • A PyrcpPR-10 gene with differentially expressed was isolated by using the suppression subtractive hybridization assay between '93-3-98' (highly resistant against scab caused by Venturia nashicola) and 'Sweat Skin'(highly susceptible) and analyzed the expression pattern according to organs and cultivars. The full length of PyrcpPR-10 was cloned as 743bp with 480bp's ORP, and was determined to encode a protein of 159 amino acid residues. On analyzing PyrcpPR-10 gene sequence compared with resistant and susceptible cultivars, 'Hwangsilri' (resistant), 'Gamcheonbae' (moderately resistant), 'Wonhwang' (moderately susceptible), 'Niitaka' (highly susceptible), and 'Sweat Skin' (highly susceptible) had identical gene sequence but 'Bartlett' (highly resistant) showed partly different sequences. The deduced amino acid sequence showed 64 ~ 98% homology and had the GXGGXG motif to known amino acid of other plants PR-10 by the BLAST X analysis. Among several organs or tissues, petal was showed highest expression level of PyrcpPR-10 gene followed by leaf, floral axis, bud, and bark. The expression level of PyrcpPR-10 gene was dramatically increased at 24 hr after inoculation in all cultivars and also up-regulated in accordance with resistant degree of cultivars. While resistant cultivars ('Bartlett', '93-3-98', and 'Hwangsilri') induced relatively high expression level of PyrcpPR-10 gene, susceptible cultivars ('Niitaka', and 'Sweat Skin') showed low expression level. PyrcpPR-10 gene is assumed that it is directly connected with defense mechanisms to pear scab.