• Title/Summary/Keyword: Transgenic rice plant.

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Comparison of Insect Fauna in Transgenic and Common Rice Paddy Fields (형질전환 벼와 일반 벼간 곤충상 비교)

  • Park, Taechul;Choe, Hojeong;Jeong, Hyoujin;Jang, Hojung;Kim, Jukon;Park, Jung-Joon
    • Korean Journal of Environmental Biology
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    • v.36 no.4
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    • pp.488-497
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    • 2018
  • Quantitative classification of insect fauna in transgenic rice in 2 LMO(Living genetically Modified Organism) isolated paddy fields (Gyeongsang National University, Gyeongbuk National University) and rice in 4 paddy fields(Duryang 1, Duryang 2, Hwagye 1, Hwagye 2), were evaluated for consecutive 5 years (2013 to 2017) in Sacheon, Gyeongnam province, and for 2 consecutive years (2015 to 2016) in Gunwi Gyeongbuk province. Sampling insect fauna were evaluated by black light trapping, sticky trapping, visual surveying, and sweeping methods in each target paddy field, respectively. A total of 37,941 individuals, of 464 species from 15 orders, were collected in Sacheon for 5 years. A total of 10,030 individuals, of 366 species from 13 orders, were collected in Gunwi for 2 years. Based on results of comparison between transgenic and non-transgenic paddy fields for 5 consecutive years, the similarity index between LMO paddy field and common paddy field is not different. Thus, this difference is not due to the environment, not the LMO.

Methods for environmental risk assessment of rice transgenic plants expressing small non-coding RNA (Small non-coding RNA를 발현하는 형질전환 벼의 환경위해성 평가 방법)

  • Jin, Byung Jun;Chun, Hyun Jin;Cho, Hyun Min;Lee, Su Hyeon;Choi, Cheol Woo;Jung, Wook-Hun;Baek, Dongwon;Han, Chang-deok;Kim, Min Chul
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.205-216
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    • 2019
  • Since the RNA interference (RNAi) had been discovered in many organisms, small non-coding RNA-mediated gene silencing technology, including RNAi have been widely applied to analysis of gene function, as well as crop improvement. Despite the usefulness of RNAi technology, RNAi transgenic crops have various potential environmental risks, including off-target and non-target effects. In this study, we developed methods that can be effectively applied to environmental risk assessment of RNAi transgenic crops and verified these methods in 35S::dsRNAi_eGFP rice transgenic plant we generated. Off-target genes, which can be non-specifically suppressed by the expression of dsRNAi_eGFP, were predicted by using the published web tool, pssRNAit, and verified by comparing their expressions between wild-type (WT) and 35S::dsRNAi_eGFP transgenic rice. Also, we verified the non-target effects of the 35S:: dsRNAi_eGFP plant by evaluating horizontal and vertical transfer of small interfering RNAs (siRNAs) produced in the 35S::dsRNAi_eGFP plant into neighboring WT rice and rhizosphere microorganisms, respectively. Our results suggested that the methods we developed, could be widely applied to various RNAi transgenic crops for their environmental risk assessment.

A Routine System for Generation of Fertile Transgenic Rice Plants Using Biolistic Method

  • Lee Soo-In;Kim Cha-Young;Lim Chae-Oh;Choi Young-Ju;Kim Ho-Il;Lee Sang-Yeol;Lee Sung-Ho
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.163-168
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    • 2003
  • A routine system based on particle bombardment of embryogenic callus for recovery of fertile transgenic rice (Oryza sativa L.) plants was developed. Embryogenic callus was established within 2-3 months from calli derived from mature seeds of Korean rice cultivar, Nagdongbyeo. The callus was bombarded with the plasmid pRQ6 containing the $\beta$-glucuronidase gene (gusA) and hygromycin phosphotransferase gene (hph, conferring resistance to hygromycin B), both driven by CaMV 35S promoter. Placement of cells on an osmoticum-containing medium (0.2 M sorbitol and 0.2 M mannitol) 4 hrs prior to and 16 hrs after bombardment resulted in a statistically significant increase with 3.2-fold in transient expression frequency gusA. In five independent experiments, the average frequency of transformation showing GUS activities was $8.86\%$. A large number of morphologically normal, fertile transgenic rice plants were obtained. Integration of foreign gene into the genome of $R_0$ transgenic plants was confirmed by Southern blot analysis. GUS and HPT were detected in $R_1$ progeny and Mendelian segregation of these genes was observed in $R_1$ progeny.

Disease Assessment in Transgenic Rice (CPPO06) Resistant to the Protox-inhibiting Herbicide (Protox 저해 제초제 내성 형질전환 벼(CPPO06)에서의 병 발생 조사)

  • Kim, Ga-Hye;Lee, Ho-Taek;Park, Se-Jung;Kim, A-Hyeong;Gwon, Hyeon-Wook;Kim, Joo-Hyung;Kim, Heung-Tae
    • Research in Plant Disease
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    • v.18 no.1
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    • pp.24-28
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    • 2012
  • The rice diseases occurring on the transgenic rice plant carrying protox gene (CPPO06) was assessed and compared with other varieties of rice such as Dongjin, Chucheong, Ilpeum, and Onnuri in the fields located in Cheongwon of Chungbuk, Suwon of Gyeunggi, and Gwangju of Chonnam. In the field of Cheongwon, the diseases such as blast, leaf spot, sheath blight and Fusarium blight were observed. False smut were observed only in the field of Suwon, where the ratio of diseased plants was 0.28% in CPPO06 and 0.37% in Onnuri, respectively. In the field of Gwangju, leaf spot caused by Cochlioborus miyabeanus was the most severely occurring disease among rice diseases mentioned above. Fusarium blight occurred in all the 3 fields, which were more severe in CPPO06 plants treated with or without oxadiazon as the herbicide. Except for Fusarium blight, there was no significant difference in the rice diseases as blast, leaf spot, sheath blight and false smut between CPPO06 and other rice varieties.

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.

Expression of hpa1 Gene Encoding a Bacterial Harpin Protein in Xanthomonas oryzae pv. oryzae Enhances Disease Resistance to Both Fungal and Bacterial Pathogens in Rice and Arabidopsis

  • Choi, Min-Seon;Heu, Sunggi;Paek, Nam-Chon;Koh, Hee-Jong;Lee, Jung-Sook;Oh, Chang-Sik
    • The Plant Pathology Journal
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    • v.28 no.4
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    • pp.364-372
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    • 2012
  • Xanthomonas oryzae pv. oryzae causing bacterial leaf blight disease in rice produces and secretes Hpa1 protein that belongs to harpin protein family. Previously it was reported that Hpa1 induced defense responses when it was produced in tobacco. In this study, we expressed hpa1 gene in rice and Arabidopsis to examine the effects of Hpa1 expression on disease resistance to both fungal and bacterial pathogens. Expression of hpa1 gene in rice enhanced disease resistance to both X. oryzae pv. oryzae and Magnaporthe grisea. Interestingly, individual transgenic rice plants could be divided into four groups, depending on responses to both pathogens. hpa1 expression in Arabidopsis also enhanced disease resistance to both Botrytis cineria and Xanthomonas campestris pv. campestris. To examine genes that are up-regulated in the transgenic rice plants after inoculation with X. oryzae pv. oryzae, known defense-related genes were assessed, and also microarray analysis with the Rice 5 K DNA chip was performed. Interestingly, expression of OsACS1 gene, which was found as the gene that showed the highest induction, was induced earlier and stronger than that in the wild type plant. These results indicate that hpa1 expression in the diverse plant species, including monocot and dicot, can enhance disease resistance to both fungal and bacterial plant pathogens.

CaPUB1, a Hot Pepper U-box E3 Ubiquitin Ligase, Confers Enhanced Cold Stress Tolerance and Decreased Drought Stress Tolerance in Transgenic Rice (Oryza sativa L.)

  • Min, Hye Jo;Jung, Ye Jin;Kang, Bin Goo;Kim, Woo Taek
    • Molecules and Cells
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    • v.39 no.3
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    • pp.250-257
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    • 2016
  • Abiotic stresses such as drought and low temperature critically restrict plant growth, reproduction, and productivity. Higher plants have developed various defense strategies against these unfavorable conditions. CaPUB1 (Capsicum annuum Putative U-box protein 1) is a hot pepper U-box E3 Ub ligase. Transgenic Arabidopsis plants that constitutively expressed CaPUB1 exhibited drought-sensitive phenotypes, suggesting that it functions as a negative regulator of the drought stress response. In this study, CaPUB1 was over-expressed in rice (Oryza sativa L.), and the phenotypic properties of transgenic rice plants were examined in terms of their drought and cold stress tolerance. Ubi:CaPUB1 T3 transgenic rice plants displayed phenotypes hypersensitive to dehydration, suggesting that its role in the negative regulation of drought stress response is conserved in dicot Arabidopsis and monocot rice plants. In contrast, Ubi:CaPUB1 progeny exhibited phenotypes markedly tolerant to prolonged low temperature ($4^{\circ}C$) treatment, compared to those of wild-type plants, as determined by survival rates, electrolyte leakage, and total chlorophyll content. Cold stress-induced marker genes, including DREB1A, DREB1B, DREB1C, and Cytochrome P450, were more up-regulated by cold treatment in Ubi:CaPUB1 plants than in wild-type plants. These results suggest that CaPUB1 serves as both a negative regulator of the drought stress response and a positive regulator of the cold stress response in transgenic rice plants. This raises the possibility that CaPUB1 participates in the cross-talk between drought and low-temperature signaling pathways.

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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Overexpression of a Rice Diacylglycerol Kinase Gene OsBIDK1 Enhances Disease Resistance in Transgenic Tobacco

  • Zhang, Weidong;Chen, Jie;Zhang, Huijuan;Song, Fengming
    • Molecules and Cells
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    • v.26 no.3
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    • pp.258-264
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    • 2008
  • A rice diacylglycerol kinase (DGK) gene, OsBIDK1, which encodes a 499-amino acid protein, was cloned and characterized. OsBIDK1 contains a conserved DGK domain, consisting of a diacylglycerol kinase catalytic subdomain and a diacylglycerol kinase accessory subdomain. Expression of OsBIDK1 in rice seedlings was induced by treatment with benzothiadiazole (BTH), a chemical activator of the plant defense response, and by infection with Magnaporthe grisea, causal agent of blast disease. In BTH-treated rice seedlings, expression of OsBIDK1 was induced earlier and at a higher level than in water-treated control seedlings after inoculation with M. grisea. Transgenic tobacco plants that constitutively express the OsBIDK1 gene were generated and disease resistance assays showed that overexpression of OsBIDK1 in transgenic tobacco plants resulted in enhanced resistance against infection by tobacco mosaic virus and Phytophthora parasitica var. nicotianae. These results suggest that OsBIDK1 may play a role in disease resistance responses.