• Title/Summary/Keyword: herbicide resistant gene

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Herbicide Resistant Turfgrass(Zoysia japonica cv. 'Zenith') Plants by Particle bombardment-mediated Transformation

  • Lim Sun-Hyung;Kang Byung-Chorl;Shin Hong-Kyun
    • Asian Journal of Turfgrass Science
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    • v.18 no.4
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    • pp.211-219
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    • 2004
  • Transgenic zoysiagrass (Zoysia japonica cv. Zenith) plants have been obtained by particle bombardment of embryogenic callus with the plasmid pSMABuba, which contains hygromycin resistance (hpt) and bialaphos resistance (bar) genes. Parameters on DNA delivery efficiency of the particle bombardment were partially optimized using transient expression assay of a chimeric $\beta-glucuronidase$(gusA) gene driven by the CaMV 35S promoter. Stably transfarmed zoysiagrass plants were recovered with a selection scheme using hygromycin. Transgenic zoysiagrass plants were confirmed by PCR analysis with specific primer for bar gene. Expression of the transgene in transformed zoysiagrass plants was demonstrated by Reverse transcriptase (RT)-PCR analysis. All the tested transgenic plants showed herbicide BastaR resistance at the field application rate of $0.1\%-0.3\%$.

Development of Seashore Paspalum Turfgrass with Herbicide Resistance

  • Kim, Kyung-Moon;Song, In-Ja;Lee, Hyo-Yeon;Raymer, Paul;Kim, Beom-Seok;Kim, Wook
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.4
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    • pp.427-432
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    • 2009
  • Seashore Paspalum (Paspalum vaginatum Swartz) is a warm season grass and indigenous to tropical and subtropical regions of coastal areas worldwide. The species is used as feed for cattle and horses and has been very successful for golf courses worldwide. One of the most outstanding characteristics of seashore paspalum is its tolerance to saline soils compared to other warm season turfgrasses. The development of new seashore paspalum cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to product for herbicide resistant seashore paspalum using Arobacterium-mediated transformation and this study is the first report on transformation and herbicideresistant transgenic plants in seashore paspalum. Embryogenic calli were induced from the seeded variety of pseashore paspalum. Embryogenic calli were transformed with Agrobacterium tumefaciens strain EHA105 carrying the binary vector pCAMBIA3301 with two genes encoding gusA and bar. Transformed calli and plants were selected on medium containing 3 mg/l PPT. PCR detected the presence of the gusA and bar gene, indicating both genes are integrated into the genome of seashore paspalum. A chlorophenol red assay was used to confirm that the bar gene was expressed. By application of herbicide BASTA, the herbicide resistance in the transgenic seashore paspalum plants was confirmed.

Production of transgenic potato exhibiting enhanced resistance to fungal infections and herbicide applications

  • Khan, Raham Sher;Sjahril, Rinaldi;Nakamura, Ikuo;Mii, Masahiro
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.13-20
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    • 2008
  • Potato (Solanum tuberosum L.), one of the most important food crops, is susceptible to a number of devastating fungal pathogens in addition to bacterial and other pathogens. Producing disease-resistant cultivars has been an effective and useful strategy to combat the attack of pathogens. Potato was transformed with Agrobacterium tumefaciens strain EHA101 harboring chitinase, (ChiC) isolated from Streptomyces griseus strain HUT 6037 and bialaphos resistance (bar) genes in a binary plasmid vector, pEKH1. Polymerase chain reaction (PCR) analysis revealed that the ChiC and bar genes are integrated into the genome of transgenic plants. Different insertion sites of the transgenes (one to six sites for ChiC and three to seven for bar) were indicated by Southern blot analysis of genomic DNA from the transgenic plants. Expression of the ChiC gene at the messenger RNA (mRNA) level was confirmed by Northern blot analysis and that of the bar gene by herbicide resistance assay. The results obviously confirmed that the ChiC and bar genes are successfully integrated and expressed into the genome, resulting in the production of bialaphos-resistant transgenic plants. Disease-resistance assay of the in vitro and greenhouse-grown transgenic plants demonstrated enhanced resistance against the fungal pathogen Alternaria solani (causal agent of early blight).

Expression of Phosphinothricin Acetyltransferase Gene in Transgenic Rice Plants (형질전환체 벼에서 phosphinothricin acetyltransferase 유전자 발현)

  • Lee, Soo-In;Lee, Sung-Ho
    • Journal of Life Science
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    • v.14 no.2
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    • pp.368-373
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    • 2004
  • We have obtained fertile transgenic rice plants resistant to the broad spectrum herbicide Bast $a^{(R)}$ (active ingredient phosphinothricin, PPT) by PEG-mediated transformation procedure. The plasmid pCaMV35S::Bar was used to deliver the bar gene into embryogenic suspension culture-derived protoplasts of rice (Oryza sativa L.). Transformed plants were regenerated and selected on medium containing 15 mg/l of phosphinothricin. Stable integration and expression of the bar gene in transgenic rice plants was confirmed by Southern and Northern blot analysis. Transgenic $R_1$ plants were also confirmed by assays for phosphinothricin acetyltransferase (PAT) activity. The bar gene was expressed in the primary transgenic rice plants and in the next generation progeny, in which it showed a 3 : 1 Mendelian inheritance pattern. Transgenic $R_1$ and $R_2$ plants were resistant to the herbicide Bast $a^{(R)}$ when sprayed at rates used in field practice.ice.

An Effective Selection of PAT Gene Transformed Populus alba $\{times}$ Populus glandulosa No.3 using Herbicide Basta Treatment (제초제 Basta를 이용한 Phosphinothricin Acetyltransferase 유전자로 형질전환된 현사시 3호의 효율적인 선발)

  • 오경은;문흥규;박재인;양덕춘
    • Korean Journal of Plant Resources
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    • v.17 no.1
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    • pp.28-33
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    • 2004
  • This study was conducted to simple transformants selection by herbicide Basta treatment after transformation with Agrobacteium tumefaciens MP90/PAT in hybrid poplar(Populus alba ${\times}$ P. glandulosa No. 3). In preliminary study, we determined that the lethal concentration of herbicide Basta was 1.0mg/L in callus culture, adventitious bud formation and axillary bud elongation experiment. By the treatment of 1.0mg/L Basta, we could be selected the transformed shoots effectively from the various cultures. In addition, the treatment was useful on selection of transformants which are growing in soil pot. Finally, we also confirmed the transformation by PAT assay, Above results show that the herbicide Basta treatment and PAT assay can be a very simple and effective method for the identification of PAT gene transformed hybrid poplar.

Gene flow from herbicide resistant genetically modified rice to conventional rice (Oryza sativa L.) cultivars

  • Han, Sung Min;Lee, Bumkyu;Won, Ok Jae;Hwang, Ki Seon;Suh, Su Jeoung;Kim, Chang-Gi;Park, Kee Woong
    • Journal of Ecology and Environment
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    • v.38 no.4
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    • pp.397-403
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    • 2015
  • Rice (Oryza sativa L.) is an important feeding crop in Asia, and utilization of genetically modified (GM) rice is highly demanding. For co-existence of GM rice and non-GM rice, the proper confinement measures should be provided. Thus, we surveyed gene flow from herbicide resistant GM rice to the conventional rice cultivars in the field tests. Gene flow frequency decreased with increasing distance between the pollen donor and recipients and did not exceed more than 1% even at the nearest distance. In single recipient model plot, a maximum gene flow frequency was observed at the shortest distance and hybrid was detected up to 12 m from the pollen donor. The direction of gene was coincided with the dominant wind direction. Gene flow assessment to multiple recipient plots was conducted under the high raining season by chance, and abrupt decline of gene flow frequency and maximum distance were resulted. According to the survey results, current regulation for isolation distance is reasonable for environmental safety or for general crop production. However, we suggest an alternative measure for GM rice cultivation that should be supplemented to overcome the out of estimation and in the environment asking higher security levels.

High-efficiency development of herbicide-resistant transgenic lilies via an Agrobacterium-mediated transformation system (고효율의 아그로박테리움 형질전환법을 이용한 제초제저항성 나리 식물체 개발)

  • Jong Bo Kim
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.56-62
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    • 2023
  • Transgenic lilies have been obtained using Agrobacterium tumefaciens (AGL1) with the plant scale explants, followed by DL-phosphinothricin (PPT) selection. In this study, scales of lily plants cv. "red flame" were transformed with the pCAMBIA3301 vector containing the gus gene as a reporter and the blpR gene as a selectable marker, as well as a gene of interest showing herbicide tolerance, both driven by the CaMV 35S promoter. Using a 20-minute infection time and a 5-day cultivation period, factors that optimized and demonstrated a high transformation efficiency were achieved. With these conditions, approximately 22-27% efficiency was observed for Agrobacterium-mediated transformation in lilies. After transformation with Agrobacterium, scales of lilies were transferred to MS medium without selective agents for 2 weeks. They were then placed on selection MS medium containing 5 mg/L PPT for a month of further selection and then cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets after transferring into hormone-free MS medium. Also, most survived putatively transformed plantlets indicated the presence of the blpR gene by PCR analysis and showed a blue color indicating expression of the gus gene. In conclusion, when 100 scales of lily cv. "red flame" are transformed with Agrobacterium, approximately 22-27 transgenic plantlets can be produced following an optimized protocol. Therefore, this protocol can contribute to the lily breeding program in the future.

Selection of Herbicide Resistant Potatoes Transformed with Phosphinothricin Acetyltransferase Gene (Phosphinothricin Acetyltransferase 유전자 도입에 의한 제초제 저항성 감자의 선발)

  • Han, S.S.;Jeong, J.H.;Bang, K.S.;Yang, D.C.
    • Korean Journal of Weed Science
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    • v.17 no.4
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    • pp.390-399
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    • 1997
  • This experiment was conducted to introduce PAT (phosphinothricin acetyltransferase, non-selective herbicide bialaphos resistant gene) gene into potato (Solanum tuberosum. cv. Desiree). Optimal shoot regeneration from leaf discs and stem segments was obtained in MS medium supplemented with 0.1 mg/L IBA and 0.5 mg/L BA, and the frequency of shoot regeneration was 54% in left discs and 46% in stem segments. In this condition, leaf discs and stem segments of potato were co-cultivated with A. tumefaciens MP90 which contained binary vector with GUS: :NPTII gene and PAT gene. Transgenic shoots were regenerated from leaf and stem-derived calli on selection medium with 100mg/L kanamycin. The 100${\mu}M$ acetosyringone treatment during the co-cultivation highly enhanced(4 times than the control) the shoot regeneration on selection medium. When the putative transgenic plants were transferred to medium with 10mg/L basta, all of them were survived. After PCR. GUS test, and Southern blot analysis of the survived plant, we confirmed that the gene was stably integrated into the potato genome and expressed. After the transgenic plants were transplanted in soil, and the transgenic plants were sprayed with the herbicide basta (300ml/10a), the transgenic plants remained green but control plants were died.

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Report on the 54th annual meeting of the weed science society of Japan (일본잡초학회에서 본 일본의 잡초연구 동향)

  • Hwang, Jae-Bok;Kim, Kyung-Hyun
    • Korean Journal of Agricultural Science
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    • v.42 no.4
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    • pp.335-339
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    • 2015
  • This paper reviews current status of weed science in Japanese regional agricultural systems based on the 54th Annual Meeting of the Weed Science Society of Japan. About 300 researchers from 5 countries including Korea participated in the Conference and presented 100 papers in research areas. This congress has an purpose to discuss new troubles, findings and results of weed science. Weed science faces big challenges such as increase in herbicide-resistant weeds, gene-flow from transgenic crops, and invasive weeds. Major research topics were invasive weeds and their ecology, allelopathy, weed management in paddy field, weed management in field crops, and herbicide resistance. Weed control and herbicide resistance management in paddy field were a main object of research. To prevent the increase of problematic weeds and to overcome food crisis, the importance of weed-related researches has been raised. Therefore it is expected that various weed management systems and control of herbicide resistant weeds should be studied continuously in the weed science.

Evaluation of Crossability, Seed Dormancy and Overwintering Ability in Glufosinate Ammonium-Resistant GM Rice and Their Hybrids with Non-GM and Weedy Rice

  • Lee, Seung-Yeob;Kim, Min-Soo;Kim, Hyo-Jin;Han, Seong-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.1
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    • pp.53-58
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    • 2006
  • This study was conducted to investigate the crossability, seed dormancy and overwintering ability of rice plant in GM (glufosinate ammonium-resistant lines. Iksan 483 and Milyang 204) and non-GM (their parents) or red rice (Andongaengmi). Seed-setting rate was not significantly different between GM and non-GM rice varieties. Iksan 483 and Milyang 204 showed the similar level of seed germination rate from 30 to 50 days after heading as compared to non-GM rice varieties. After overwintering in paddy field, seed germination rate of GM and non-GM rice varieties ranged from 14.3 % to 57.6 % in dry soil condition, but there was no germination in wet-soil except red rice. The result in wet-soil condition may help to set up a strategy for reducing the risk of gene flow of transgene via dispersal of seeds of GM plants. The crossability, seed dormancy and seed overwintering of Iksan 483 and Milyang 204, herbicide resistant GM rice varieties, were not significantly different compared to non-GM rice varieties. The results might be helpful to reduce the risk of transgene dispersal from GM crop via seeds and pollens.