• Title/Summary/Keyword: 형질전환벼

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Levels of Resistance and Fitness in Glufosinate-ammonium-Resistant Transgenic Rice Plants (Glufosinate-ammonium 저항성 형질전환벼의 저항성 수준과 적응성에 관한 연구)

  • Yun, Young Beom;Kuk, Yong In
    • Weed & Turfgrass Science
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    • v.1 no.4
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    • pp.50-56
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    • 2012
  • The objectives of this research were to quantify resistance levels of transgenic rice expressing the bar gene to glutamine synthetase (GS)-inhibiting, and methionine sulfoximine and photosynthesis-inhibiting herbicide, paraquat, and compare the ammonium accumulation, chilling injury, and yield between transgenic and non-transgenic rice. The transgenic rice lines were 45-96-fold more resistant to glufosinate-ammonium than non-transgenic rice. The transgenic rice lines were also 18-fold more resistant to methionine sulfoximine, but was not resistant to paraquat, which has different target site. Glufosinate-ammonium increased the ammonium accumulation in leaves of non-transgenic rice plants, but had minimal or no effect on leaves of transgenic lines. The transgenic lines except for 258, 411, 607 and 608 were more susceptible during chilling and recovery than non-transgenic rice plants. The yield of transgenic lines 142, 144, 258 and 608 was similar or higher than that of non-transgenic rice in pot conditions.

Difference in Physiological Responses to Environmental Stress in Protox Inhibitor Herbicide-Resistant Transgenic Rice and Non-transgenic Rice (Protox 저해형 제초제 저항성 형질전환벼와 비형질전환벼의 환경스트레스에 대한 생리적 반응 차이)

  • Yun, Young-Beom;Kwon, Oh-Do;Shin, Dong-Young;Hyun, Kyu-Hwan;Lee, Do-Jin;Jung, Ha-Il;Kuk, Yong-In
    • Korean Journal of Weed Science
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    • v.32 no.1
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    • pp.35-43
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    • 2012
  • The objective of this research was to confirm the difference in physiological responses to environmental stresses such as chilling, high temperature, NaCl, and chemical stress (paraquat) in Protox inhibitor resistant-transgenic rice (MX, PX, and AP37) and its non-transgenic counterpart (WT). Transgenic and non-transgenic rice plants were exposed to a chilling temperature of $5^{\circ}C$ for 1 day or a high temperature of $45^{\circ}C$ for 4 days and allowed to recover at $25^{\circ}C$ for 6 days after the chilling treatment or 8 days after the high temperature treatment. Leaf injury, shoot fresh weight, porphyrin biosynthesis substances, and chlorophyll content were investigated in transgenic and non-transgenic rice at 6 days after 0.5% and 1% NaCl treatments or at 5 days after 0~300 ${\mu}M$ paraquat treatments. No significant difference in leaf injury and shoot fresh weight were observed between transgenic and non-transgenic rice during chilling and recovery. Plant height and shoot fresh weight were also similar between transgenic and non-transgenic rice during the high temperature and recovery period (0~5 days). However, plant height and shoot fresh weight in transgenic rice line MX and PX were lower than in non-transgenic rice at 6 days for recovery. Leaf injury, chlorophyll, and Mg-Proto IX ME contents had no significant difference between transgenic rice and non-transgenic rice after NaCl treatment, but Proto IX content for AP37 and shoot fresh weight for PX and AP37 in 0.5% NaCl treatment were significantly reduced compared with non-transgenic rice. There was no difference in leaf injury and shoot fresh weight when comparing transgenic rice and non-transgenic rice after paraquat treatment. Although transgenic rice and non-transgenic rice showed a little difference at a particular measurement period in certain environmental stresses, there was generally no difference in physiological responses between transgenic rice and non-transgenic rice.

Generation of Transgenic Rice without Antibiotic Selection Marker through Agrobacterium-mediated Co-transformation System (아그로박테리움 동시 형질전환 시스템을 통한 항생제 선발 마커가 없는 형질전환벼의 생산)

  • Park, Soo-Kwon;Kwon, Tack-Min;Lee, Jong-Hee;Shin, Dong-Jin;Hwang, Woon-Ha;Song, You-Chun;Cho, Jun-Hyun;Nam, Min-Hee;Jeon, Seung-Ho;Lee, Sang-Yeol;Park, Dong-Soo
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1152-1158
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    • 2012
  • Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.

Functional characterization of a CCCH type zinc-finger protein gene OsZF2 by ectopic overexpression of the gene in rice (과발현 형질전환벼에서 CCCH type zinc-finger protein 유전자 OsZF2 기능 분석)

  • Lee, Jung-Sook;Yoon, In-Sun;Yoon, Ung-Han;Lee, Gang-Seob;Byun, Myung-Ok;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.23-29
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    • 2009
  • We have previously isolated a CCCH type zinc-finger protein gene, OsZF2 (Oryza sativa Zinc Finger 2), from the cold-treated rice cDNA library. To investigate the potential role of OsZF2, transgenic rice lines over-expressing OsZF2 under the control of CaMV 35S promoter have been developed through Agrobacterium-mediated transformation. Elevated level of OsZF2 transcripts was confirmed by RNA gel blot analysis in transgenic rice. Under the 100 mM NaCl condition, the transgenic rice showed significantly enhanced growth rate in terms of shoot length and fresh weight, implicating that OsZF2 is likely to be involved in salt response of rice. In the field condition, however, the transgenic rice showed a dwarf phenotype and flowering time was delayed. Genome expression profiling analysis of transgenic plants using the 20K NSF rice oligonucleotide array revealed many up-regulated genes related to stress responses and signaling pathways such as chaperone protein dnaJ 72, salt stress-induced protein, PR protein, disease resistance proteins RPM1 and Cf2/Cf5 disease resistance protein, carbohydrate/ sugar transporter, OsWAK kinase, brassinosteroid LRR receptor kinase, and jasmonate O-methyltransferase. These data suggest that the CCCH type zinc-finger protein OsZF2 is a upstream transcriptional factor regulating growth and stress responsiveness of rice.

Mechanism of Growth Inhibition in Herbicide-Resistant Transgenic Rice Overexpressing Protoporphyrinogen Oxidase (Protox) Gene (Protoporphyrinogen Oxidase (Protox) 유전자 과다발현 제초제 저항성 형질전환 벼의 생육저해 기작)

  • Kuk, Yong-In;Shin, Ji-San;Yun, Young-Beom;Kwon, Oh-Do
    • Korean Journal of Weed Science
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    • v.30 no.2
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    • pp.122-134
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    • 2010
  • We investigated the levels of resistance and accumulation of terapyrroles, reactive oxygen species, lipid peroxidation, and antioxidative enzymes for reasons of growth reduction in herbicide-transgenic rice overexpressing Myxococcus xanthus, Arabidopsis thaliana, and human protoporphyrinogen oxidase (Protox) genes. The transgenic rice overexpressing M. xanthus (MX, MX1, PX), A. thaliana (AP31, AP36, AP37), and human (H45, H48, H49) Protox genes showed 43~65, 41~72 and 17~70-fold more resistance to oxyfluorfen, respectively, than the wild type. Among transgenic rice lines overexpressing Protox genes, several lines showed normal growth compared with the wild type, but several lines showed in reduction of plant height and shoot fresh weight under different light conditions. However, reduction of plant height of AP37 was much higher than other lines for the experimental period. On the other hand, the reduction of plant height and shoot fresh weight in the transgenic rice was higher in high light condition than in low light condition. Enhanced levels of Proto IX were observed in transgenic lines AP31, AP37, and H48 at 7 days after seeding (DAS) and transgenic lines PX, AP37, and H48 at 14 DAS relative to wild type. There were no differences in Mg-Proto IX of transgenic lines except for H41 and H48 and Mg-Proto IX monomethyl ester of transgenic lines except for MX, MX1, and PX. Although accumulation of tetrapyrrole intermediates was observed in transgenic lines, their tetrapyrrole accumulation levels were not enough to inhibit growth of transgenic rice. There were no differences in reactive oxygen species, MDA, ALA synthesizing capacity, and chlorophyll between transgenic lines and wild type indicating that accumulated tetrapyrrole intermediate were apparently not high enough to inhibit growth of transgenic rice. Therefore, the growth reduction in certain transgenic lines may not be caused by a single factor such as Proto IX, but by interaction of many other factors.

Relationship of Fitness and Substance of Porphyrin Biosynthesis Pathway in Resistant Transgenic Rice to Protoporphyrinogen Oxidase (Protox) Inhibitor (Protoporphyrinogen oxidase (Protox) 저해제 저항성 형질전환 벼의 적응성과 Porphyrin 생합성 경로물질과 관련성)

  • Yun, Young-Beom;Kwon, Oh-Do;Back, Kyoung-Whan;Lee, Do-Jin;Jung, Ha-Il;Kuk, Yong-In
    • Korean Journal of Weed Science
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    • v.31 no.2
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    • pp.134-145
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    • 2011
  • The objectives of this study were to investigate fitness difference in growth and rice yield in herbicide-transgenic rice overexpressing Myxococcus xanthus and Arabidopsis thaliana protoporphyrinogen oxidase (Protox) genes and non-transgenic rice. We also aimed to determine whether these fitness differences are related to ALA synthesizing capacity, accumulation of terapyrroles, reactive oxygen species, lipid peroxidation, and antioxidative enzymes at different growth stages of rice. Plant height of the transgenic rice overexpressing M. xanthus (MX) and A. thaliana (AP37) Protox genes at 43, 50, and 65 days after transplanting (DAT) was significantly lower than that of WT. Number of tiller of PX as well as MX and AP37 at 50 and 65 DAT was significantly lower than that of WT. At harvest time, culm length and yield of MX, PX and AP37 and rice straw weight of MX and AP37 were significantly low compared with WT. The reduction of yield in MX, PX, and AP37 was caused by spikelets per panicle and 1000 grain weight, ripened grain, spikelets per panicle, 1000 grain weight, and ripened grain, respectively. On the other hand, 135 the reduction of yield in MX, PX, and AP37 was also observed in another yearly variation experiment. The reduction of rice growth in MX, PX, and AP37 was observed in seedling stage as well as growth duration in field. There were no differences in tetrapyrrole intermediate Proto IX, Mg-Proto IX and Mg-Proto IX monomethyl ester, reactive oxygen species ($H_2O_2$ and ${O_2}^-$), MDA, antioxidative enzymes (SOD, CAT, POX, APX, and GR) and chlorophyll between transgenic lines and wild type, indicating that accumulated tetrapyrrole intermediate and other parameters were not related to growth reduction in transgenic rice. However, ALA synthesizing capacity in MX, PX, and AP37 at one day after exposure to light and 52 DAT was significantly lower than that of WT. Further study is required to elucidate the mechanisms underlying the growth and yield difference between transgenic and WT lines.