• Title/Summary/Keyword: PCR. Rice(Oryza sativa L.)

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Black Rice (Oryza sativa L. var. japonica) Hydrolyzed Peptides Induce Expression of Hyaluronan Synthase 2 Gene in HaCaT Keratinocytes

  • Sim, Gwan-Sub;Lee, Dong-Hwan;Kim, Jin-Hwa;An, Sung-Kwan;Choe, Tae-Boo;Kwon, Tae-Jong;Pyo, Hyeong-Bae;Lee, Bum-Chun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.271-279
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    • 2007
  • Black rice (Oryza sativa L. var. japonica) has been used in folk medicine in Asia. To understand the effects of black rice hydrolyzed peptides (BRP) from germinated black rice, we assessed the expression levels of about 20,000 transcripts in BRP-treated HaCaT keratinocytes using human 1A oligo microarray analysis. As a result, the BRP treatment showed a differential expression ratio of more than 2-fold: 745 were activated and 1,011 were repressed. One of the most interesting findings was a 2-fold increase in hyaluronan synthase 2 (HAS2) gene expression by BRP. Semiquantitative RT-PCR showed that BRP increased HAS2 mRNA in dose-dependent manners. ELISA showed that BRP effectively increased hyaluronan (HA) production in HaCaT keratinocytes.

Transformation of Rice (Oryza sativa L.) with Phosphate Transporter cDNA from Tobacco (Nicotiana tabacum L.) (담배 인산수송자 유전자를 이용한 벼의 형질전환)

  • 유남희;윤성중
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.441-445
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    • 2000
  • In order to improve phosphate use efficiency of rice using phosphate transporter (PT), transgenic rice plants containing a tobacco PT gene were developed. Calli from Dongjinbyeo (Oryza sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring PT gene. Multiplied calli were transferred to MS medium supplemented with 50 mg/L hygromycin, 500 mg/L carbenicillin, 2 mg/L kinetin, 0.1 mg/L NAA. After 2 weeks, hygromycin resistant shoots were obtained from the calli on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 250 mg/L cabenicillin. Plant regeneration rate from the calli was about 52%. Stable incorporation of the tobacco PT gene into rice genomic DNA was confirmed by PCR and Southern blot analysis.

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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.

Transcriptome and Small RNAome Analyses Reveal the Association of pre-harvest Sprouting and Heat Stress Response in Rice (Oryza sativa L.)

  • Minsu Park;Woochang Choi;Sang-Yoon Shin;Yujin Kweon;Jihyun Eom;Minsun Oh;Chanseok Shin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2023.04a
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    • pp.157-157
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    • 2023
  • Pre-harvest sprouting (PHS) in rice (Oryza sativa L.) is one of the main problems associated with seed dormancy. PHS causes yield loss and reduction of grain quality under unpredictable humid conditions at the ripening stage, thus affecting the economic value of the rice crop. To resolve this issue, it is important to understand the molecular mechanism underlying seed dormancy in rice. Recent studies have shown that seed dormancy is affected by a large number of genes associated with plant hormones. However, the effect of heat stress on seed dormancy and plant hormones is not well understood. In this study, we compared the PHS rate as well as the transcriptome and small RNAome of the seed embryo and endosperm of two different accessions of rice, PHS-susceptible rice (low dormancy) and PHS-resistant rice (high dormancy) under three different maturation stages. We identified and verified the candidate genes associated with seed dormancy and heat stress-related responses in rice using quantitative real-time PCR. We newly discovered hormone-related genes, heat shock protein-related genes, and miRNAs potentially involved in PHS. These findings provide a foundation for understanding the dynamics of transcriptome and small RNAome of hormone- and heat stress-related genes, which affect PHS during seed maturation.

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Transformation of Rice (Oryza sativa L.) with Sucrose Transporter cDNA from Potato (Solanum tuberosum L.) (감자 Sucrose Transporter 유전자의 벼 Genome 내로의 도입)

  • 백소현;유남희;윤성중
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.97-101
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    • 2001
  • The transport and allocation of photosynthetic assimilate is an important regulatory factor in plant productivity, In order to modify assimilate partitioning in rice, transgenic plants containing a potato sucrose transporter (SuT) gene were developed. Calli derived from rice seeds (Oryza sativa L. cv Dongjin) were cocultured with A. tumefaciens LBA 4404 harboring the SuT gene. Calli were transferred to MS medium supplemented with 50 mg/L hygromycin, 500 mg/L carbenicillin, 2 mg/L kinetin, 0.1 mg/L NAA. After 2 weeks, hygromycin resistant shoots were obtained from the calli on the selection medium. Roots were induced from the putative transgenic shoots on rooting MS medium supplemented with 250 mg/L cabenicillin. Plant regeneration rate from the calli was about 150%. Stable incorporation of the potato SuT gene into rice genomic DNA was confirmed by PCR and Southern blot analysis.

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Molecular Cloning and Nucleotide Sequencing of a DNA Clone Encoding Arginine Decarboxylase in Rice (Oryza sativa L.) (벼의 arginine decarboxylase DNA clone의 재조합 및 염기서열 분석)

  • Hong, Sung-Hoi;Jeung, Ji-Ung;Ok, Sung-Han;Shin, Jeong-Sheop
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.112-117
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    • 1996
  • Arginine decarboxylase (ADC) is the first enzyme in one of the two pathways of diamine putrescine biosynthesis in plants. The genes encoding ADC have previously been cloned from Escherichia coli, oat and tomato genome. Two degenerate oligonucleotides (17-mer) corresponding to two conserved regions of ADC were used as primers in polymerase chain reaction of rice (Oryza sativa L.) genomic DNA, and an approximately 1.0 kbp fragment was obtained. This amplified PCR product showed an open reading frame which contains 1,022 bp of nucleotide sequences. This PCR product was cloned into pGEM-originated T vector and the short 500 bp PstI digested fragment was subcloned into pGEM-3zf(+/-) vectors to facilitate sequencing. The nucleotide sequence of this PCR product showed about 74% and 70% identity with the same regions of the oat and tomato ADC cDNA sequences, respectively. The predicted amino acid sequence exhibited 45% and 62% identity with oat and tomato ADC polypeptide fragments, respectively. The sequence similarities of 34%, 47% and 38% were previously reported in oat and E. coli, tomato and oat, and tomato and E. coli ADC amino acids, respectively. Therefore, similarities and identities between rice and oat or tomato are remarkably higher than those others of the previous reports. In the highly conserved regions in both the amino acid sequence and spacing regions among the sequences of these three, rice ADC open reading frame also has the exactly same regions with the striking similarity. RNA blot analysis showed that hnc is expressed as a transcript of approximately 2.5 kbP in the rice seedling leaf tissues.

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Production of miraculin protein in suspension cell lines of transgenic rice using Agrobacterium (Agrobacterium을 이용한 형질전환 벼 현탁 세포주에서 miraculin 단백질의 생산)

  • Kim, Hee Kyoung;Go, Ji Yun;Park, So-Young;Kang, Kwon Kyoo;Jung, Yu Jin
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.227-234
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    • 2020
  • To produce the miraculin protein in suspension cultures, rice (Oryza sativa L.) was transformed with Agrobacterium tumefacience EHA105 containing the miraculin AB512278 gene. The cell suspension cultures were established using cell lines selected from transgenic rice callus. The integration of the miraculin gene into the rice chromosome was confirmed using genomic PCR analysis. In addition, RT-PCR analysis indicated that the miraculin gene is expressed in the selected suspension cell lines. Thus, the recombinant miraculin was expressed in the transgenic suspension cell line, HK-2. Therefore, we have successfully developed a HK-2 line that produces miraculin. These results demonstrate that transformed cell suspension cultures can be used to produce a taste-modifying protein such as miraculin.

Responses of Photosynthetic Efficiency and Ascorbate Peroxidase Induced by Salt Stress in Rice (Oryza sativa L.) (벼의 salt stress에 의해 유도된 산화 stress에 대한 ascorbate peroxidase 반응)

  • Koo, Jeung-Suk;Im, Kyoung-Nam;Chun, Hyun-Sik;Lee, Chin-Bum
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1173-1180
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    • 2010
  • We investigated changes in photosynthesis and activity of ascorbate peroxidase (APX) that scavenges ROS as responses to oxidative stress induced by salinity in rice (Oryza sativa L.). Photosynthetic efficiency of rice leaves, monitored in terms of Fv/Fm, declined with the increase of salt concentration (100-300 mM NaCl). Salinity caused an increase of $H_2O_2$ in leaves of rice, with an increase of APX activity. Among total APX isoforms, an isoform of stromal-APX 1 in leaves of rice was completely inactivated by 300 mM NaCl, but was not affected by chilling or drought. The results suggest that salt stress acts in quite a different mechanism in relation to the activity of stromal-APX from that of other stresses such chilling and drought. We carried out RT-PCR for analysis of genes expression of APX isoforms as affected by salt stress. The expression of cytosolic APX/thylakoid-bound APX genes in leaves of rice exposed to salt stress was increased, while stromal APX gene expression rapidly declined.

Structure and Expression of OsUBP6, an Ubiquitin-Specific Protease 6 Homolog in Rice (Oryza sativa L.)

  • Moon, Yea Kyung;Hong, Jong-Pil;Cho, Young-Chan;Yang, Sae-Jun;An, Gynheung;Kim, Woo Taek
    • Molecules and Cells
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    • v.28 no.5
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    • pp.463-472
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    • 2009
  • Although the possible cellular roles of several ubiquitin-specific proteases (UBPs) were identified in Arabidopsis, almost nothing is known about UBP homologs in rice, a monocot model plant. In this report, we searched the rice genome database (http://signal.salk.edu/cgi-bin/RiceGE) and identified 21 putative UBP family members (OsUBPs) in the rice genome. These OsUBP genes each contain a ubiquitin carboxyl-terminal hydrolase (UCH) domain with highly conserved Cys and His boxes and were subdivided into 9 groups based on their sequence identities and domain structures. RT-PCR analysis indicated that rice OsUBP genes are expressed at varying degrees in different rice tissues. We isolated a full-length cDNA clone for OsUBP6, which possesses not only a UCH domain, but also an N-terminal ubiquitin motif. Bacterially expressed OsUBP6 was capable of dismantling K48-linked tetra-ubiquitin chains in vitro. Quantitative real-time RT-PCR indicated that OsUBP6 is constitutively expressed in different tissues of rice plants. An in vivo targeting experiment showed that OsUBP6 is predominantly localized to the nucleus in onion epidermal cells. We also examined how knock-out of OsUBP6 affects developmental growth of rice plants. Although homozygous T3 osubp6 T-DNA insertion mutant seedlings displayed slower growth relative to wild type seedlings, mature mutant plants appeared to be normal. These results raise the possibility that loss of OsUBP6 is functionally compensated for by an as-yet unknown OsUBP homolog during later stages of development in rice plants.