• Title/Summary/Keyword: Gene characterization

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Functional characterization of ABA signaling components using transient gene expression in rice protoplasts

  • Song, In-Sik;Moon, Seok-Jun;Kim, Jin-Ae;Yoon, Insun;Kwon, Taek-Ryoun;Kim, Beom-Gi
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.109-109
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    • 2017

  • The core components of ABA-dependent gene expression signaling have been identified in Arabidopsis and rice. This signaling pathway consists of four major components; group A OsbZIPs, SAPKs, subclass A OsPP2Cs and OsPYL/RCARs in rice. These might be able to make thousands of combinations through interaction networks resulting in diverse signaling responses. We tried to characterize those gene functions using transient gene expression for rice protoplasts (TGERP) because it is instantaneous and convenient system. Firstly, in order to monitor the ABA signaling output, we developed reporter system named pRab16A-fLUC which consists of Rab16A promoter of rice and luciferase gene. It responses more rapidly and sensitively to ABA than pABRC3-fLUC that consists of ABRC3 of HVA1 promoter in TGERP. We screened the reporter responses for over-expression of each signaling components from group A OsbZIPs to OsPYL/RCARs with or without ABA in TGERP. OsbZIP46 induced reporter most strongly among OsbZIPs tested in the presence of ABA. SAPKs could activate the OsbZIP46 even in the ABA independence. Subclass A OsPP2C6 and -8 almost completely inhibited the OsbZIP46 activity in the different degree through the SAPK9. Lastly, OsPYL/RCAR2 and -5 rescued the OsbZIP46 activity in the presence of SAPK9 and OsPP2C6 dependent on ABA concentration and expression level. By using TGERP, we could characterize successfully the effects of ABA dependent gene expression signaling components in rice. In conclusion, TGERP represents very useful technology to study systemic functional genomics in rice or other monocots.

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Sequence and Characterization of the Genomic Clone of the FVFD16 and FVFD30 Gene Isolated from Flammulina velutipes (팽이버섯에서 분리된 FVFD16과 FVFD30 유전자의 게놈클론의 염기서열 및 특성)

  • Kim, Dool-Yi;Azuma, Tomo-Nori
    • The Korean Journal of Mycology
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    • v.28 no.1
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    • pp.26-31
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    • 2000

  • We isolated genomic clone of FVFD16 and FVFD30 gene specifically expressed during fruit body formation of Flammulina velutipes [(Curt: Fr.) Sing] and determinated the sequences. The FVFD16 gene is including two introns in open reading frame, and FVFD30 gene is including four introns. The introns were matched GT/AG rule. The FVFD16 and FVFD30 genes contained CAAT box with similarity arrange and TATA box. CT-rich region was presented before the transcription start point. FVFD30 gene is investigated that expected the most activity of CCACC arrange. The result of FVFD16 gene analysis showed 80% homology by cDNA clone that is gene family. From the results of genomic southern blot analysis, we presumed more than two copy number gene family of FVFD16 and FVFD30 gene.

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Identification and Molecular Characterization of Methionine Sulfoxide Reductase B Gene in Rice Blast Fungus, Magnaporthe oryzae (벼도열병균에서의 methionine sulfoxide reductase B 유전자의 분자적 특성)

  • Kim, Jeong-Hwan;Kim, Jin-Soo;Jeong, Mi-Yeon;Choi, Woo-Bong
    • Journal of Life Science
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    • v.19 no.3
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    • pp.343-348
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    • 2009

  • Magnaporthe oryzae, a major cause of rice blast, is one of the most destructive plant fungal pathogens. Secretion of reactive oxygen species (ROS) during the infection phase of plant pathogenic fungus plays a key role in the defense mechanism of a plant. ROS causes oxidative damage and functional modification to the proteins in a pathogenic fungus. Methionine, especially, is a major target of ROS, which oxidizes it to methionine sulfoxide. To survive from the attack of ROS, plant pathogenic fungus has antioxidative systems - one example would be methionine sulfoxide reductase B (MSRB), which reverses the oxidative alteration of methionine to methionine sulfoxide. In the present study, identification and molecular characterization of the MSRB gene in M. oryzae KJ201 were investigated. The MSRB gene was amplified by PCR from the M. oryzae KJ201 genomic DNA. The copy number of MSRB in the genome of M. oryzae KJ201 was identified by Southern blot analysis, which revealed that the gene exists as a single copy. To study the molecular function of an MSRB gene, the expression level of the MSRB gene was assayed with hydrogen peroxide treatment by Northern blot analysis and RT-PCR. The expression of the MSRB gene was increased by treatment of hydrogen peroxide, without significant correlation to hydrogen peroxide concentrations. These results indicate that the MSRB gene in M. oryzae KJ201 could contribute to protection against plant defense compounds such as ROS and offer a novel strategy for the control of rice blast.

  • https://doi.org/10.5352/JLS.2009.19.3.343 인용 PDF KSCI