Assessment of Phosphinothricin Acetyltransferase (PAT) Degradation From Transgenic Zoysiagrass Digested with Simulated Gastric Fluid (SGF)

Sun, Hui; Kang, Hong‐Gyu; Bae, Tae-Woong; Cho, Tae-Gun; Kim, Joonki; Lim, Pyung-Ok; Riu, Key Zung; Lee, Hyo-Yeon

doi:10.1007/s12374-009-9094-y

Cited by 6 publications

(6 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…As the most frequent weeds present in L. corniculatus fields, such as Sorghum halepense , Cichorium intibus and Sonchus oleraceus are damaged by spraying with 150 mg L −1 PPT (data not shown), these transgenic L. corniculatus plants with resistance to high levels of herbicide PPT provide an opportunity for more efficient weed control. The latest confirmation that the PAT protein in herbicide‐tolerant zoysiagrass plants can be safely and rapidly digested in the stomachs of humans or livestock (Sun et al , ) is an additional qualifying criterion important for the agronomical utilisation of PAT protein‐containing L. corniculatus plants. Furthermore, a report on the pleiotropic effects of the bar gene and PPT on the Arabidopsis thaliana transcriptome (Abdeen & Miki, ) and reports showing that PPT can reduce fungal diseases in PPT‐resistant bentgrasses (Wang et al , ), rice (Ahn, ) and potato (Khan et al , ), make PPT‐resistant L. corniculatus plants a suitable tool for further studies that could lead to the additional improvement of this important forage crop.…”

Section: Discussionmentioning

confidence: 99%

Fertile transgenic Lotus corniculatus resistant to the non‐selective herbicide phosphinothricin

Nikolić

Zdravković-Korać

Ninković

et al. 2013

Annals of Applied Biology

View full text Add to dashboard Cite

Resistance to the non-selective herbicide DL-phosphinothricin (PPT) was introduced into commercial Lotus corniculatus cv. Bokor by co-cultivation of cotyledons with Agrobacterium tumefaciens AGL1 harbouring the binary vector pDM805 which contains the bialaphos resistance gene (bar) from Streptomyces hygroscopicus encoding phosphinothricin acetyltransferase (PAT) and the uidA gene encoding β-glucuronidase. The half-cotyledon explants were precultured on regeneration Murashige and Skoog's (MS) medium supplemented with 6-benzyladenine (BA) and 1-naphthaleneacetic acid (NAA) at 0.5 mg L −1 each, 3 days prior to infection. Upon co-cultivation, the explants were cultured on PPT-free regeneration medium for 10 days, and then subcultured on regeneration/selection media with increasing PPT concentrations (5-7 mg L −1 ) for about 18 weeks. Out of 480 initially co-cultivated explants, 272 regenerated shoots survived the entire PPT selection procedure. Resistant shoots were grown further, multiplied by tillering that was additionally promoted by PPT and rooted on hormone-free MS medium containing 5 mg L −1 PPT. Established shoot cultures, continuously maintained on the same medium, have preserved PPT resistance up to now (more than 2 years). Transformed plants assessed in vitro and in a greenhouse were tolerant to the herbicide PPT at 300 mg L −1 equivalent to more than twofold the recommended field dosage for weed eradication. Applied PPT treatment did not affect the activities of glutamine synthetase (GS; EC 6.3.1.2) and NADH-dependent glutamate dehydrogenase (NADH-GDH; EC 1.4.1.2) in transformed plants. However, PPT did increase the mobility of glutamine synthetase isoforms GS1 and GS2 as well as the inhibition of an additional high mobility GS (hmGS) activity. In untransformed plants, PPT treatment reduced total GS activity by 4.4-fold while contrary the activity of NADH-GDH was increased by ninefold. All transformed herbicideresistant plants were phenotypically normal and exhibited genomic stability, as were the untransformed plants analysed by flow cytometry. Under greenhouse conditions, they grew to maturity, flowered and set seeds. Stable integration and expression of the bar gene in T0 and T1 plants were confirmed by Southern and Western blot analysis, while integration of the reporter uidA gene did not occur. The bar gene was inherited in a Mendelian fashion by the progeny, as detected by PPT resistance. The production of PPT-resistant plants may have significant practical applications in weed control in fields of L. corniculatus.

show abstract

Section: Discussionmentioning

confidence: 99%

Fertile transgenic Lotus corniculatus resistant to the non‐selective herbicide phosphinothricin

Nikolić

Zdravković-Korać

Ninković

et al. 2013

Annals of Applied Biology

View full text Add to dashboard Cite

show abstract

“…이 자료에 기초하여 제초제저 항성 들잔디 JG21에 도입된 바이너리벡터의 T-DNA 유 전자 구성 및 염기서열 정보를 제시하였고, 도입된 제초 제저항성 유전자 bar의 도입 및 발현 특성을 분석하여 제 시하였다. PCR에 의해 Ubiquitin promoter::bar로 재조합된 목적유전자가 도입되었음을 확인하였고, Figure 1과 같이 Southern blot분석에 의해 JG21계통 내에 1개의 삽입 복제 수가 포함되어 있음을 확인하였다 (Sun et al 2010). 삽입 복제수는 다음 세대의 제초제저항성의 분리비 (제초제저 항성:야생형=3:1)와 PCR을 이용한 도입유전자의 분리비 (동형접합형:이형접합형:야생형=1:2:1)를 통해서도 재검 증되었다 (Sun et al 2010).…”

Section: 도입유전자의 분자생물학적 특성 검정unclassified

“…PCR에 의해 Ubiquitin promoter::bar로 재조합된 목적유전자가 도입되었음을 확인하였고, Figure 1과 같이 Southern blot분석에 의해 JG21계통 내에 1개의 삽입 복제 수가 포함되어 있음을 확인하였다 (Sun et al 2010). 삽입 복제수는 다음 세대의 제초제저항성의 분리비 (제초제저 항성:야생형=3:1)와 PCR을 이용한 도입유전자의 분리비 (동형접합형:이형접합형:야생형=1:2:1)를 통해서도 재검 증되었다 (Sun et al 2010). 이러한 결과는 도입유전자가 엽록체 혹은 미토콘드리아가 아니라 핵의 염색체 내에 삽입되었음을 증명하고 있다.…”

Section: 도입유전자의 분자생물학적 특성 검정unclassified

“…이러한 결과는 도입유전자가 엽록체 혹은 미토콘드리아가 아니라 핵의 염색체 내에 삽입되었음을 증명하고 있다. 전사단계의 유전자발현은 RT-PCR 방법으로 검증되었고, 번역단계의 유전자발현은 PAT 항체를 이용한 strip kit와 Western blot 분석 방법으로 검증되었다 (Sun et al 2010). 게놈 상에서 도입 유전자의 삽입 위치를 확인하기 위한 방법으로는 Inverse PCR 방법 과 TAIL-PCR 방법이 있으나 본 실험에서는 TAIL-PCR 방 법을 이용하였다 (Liu et al 1995).…”

Section: 도입유전자의 분자생물학적 특성 검정unclassified

“…JG21 zoysiagrass genomic DNA (10 μg) was digested with HindIII, XhoI, or EcoRI and resolved on 1.0% agarose gel. DNA sizes are indicated by arrows (from Sun et al 2010 Table 2 Number of the germinated seeds tested and the hybrids identified at distances from genetically modified (GM) Zoysia grass in each plot design …”

Section: 도입유전자의 분자생물학적 특성 검정mentioning

confidence: 99%

See 2 more Smart Citations

Environmental risk assessment of genetically modified Herbicide-Tolerant zoysiagrass (Event: Jeju Green21)

Bae¹,

Kang²,

Song³

et al. 2011

Journal of Plant Biotechnology

Self Cite

View full text Add to dashboard Cite

Transgenic zoysiagrass (Zoysia japonica Steud.) expressing the bar gene inserted in the plant genome has been generated previously through Agrobacterium tumefaciensmediated transformation. The GM zoysiagrass (event: JG21) permits efficient management of weed control of widely cultivated zoysiagrass fields, reducing the frequency and cost of using various herbicides for weed control. Now we have carried out the environmental risk assessment of JG21 prior to applying to the governmental regulatory agency for the commercial release of the GM turf grass outside of test plots. The morphological phenotypes, molecular analysis, weediness and gene flow from each test plot of JG21 and wild-type zoysiagrasses have been evaluated by selectively analyzing environmental effects. There were no marked differences in morphological phenotypes between JG21 and wild-type grasses. The JG21 retained its stable integration in the host plant in T 1 generation, exhibiting a 3:1 segregation ratio according to the Mendelian genetics. We confirmed the copy number (1) of JG21 by using Southern blot analysis, as the transgenic plants were tolerant to ammonium glufosinate throughout the culture period. From cross-fertilization and gene flow studies, we found a 9% cross-pollination rate at the center of JG21 field and 0% at distances over 3 m from the field. The JG21 and wild-type zoysiagrass plants are not considered "weed" because zoysiagrasses generally are not dominant and do not spread into weedy areas easily. We assessed the horizontal gene transfer (HGT) of the transgene DNA to soil microorganisms from JG21 and wild-type plants. The bar gene was not detected from the total genomic DNA extracted from each rhizosphere soil of GM and non-GM Zoysia grass fields. Through the monitoring of JG21 transgene's unintentional release into the environment, we found no evidence for either pollen mediated gene flow of zoysiagrass or seed dispersal from the test field within a 3 km radius of the natural habitat.

show abstract

Evaluation of factors influencing Agrobacterium-mediated spinach transformation and transformant selection by EGFP fluorescence under low-selective pressure

Nguyen

Boo

Sun

et al. 2013

In Vitro Cell.Dev.Biol.-Plant

View full text Add to dashboard Cite

Assessment of Phosphinothricin Acetyltransferase (PAT) Degradation From Transgenic Zoysiagrass Digested with Simulated Gastric Fluid (SGF)

Cited by 6 publications

References 21 publications

Fertile transgenic Lotus corniculatus resistant to the non‐selective herbicide phosphinothricin

Fertile transgenic Lotus corniculatus resistant to the non‐selective herbicide phosphinothricin

Environmental risk assessment of genetically modified Herbicide-Tolerant zoysiagrass (Event: Jeju Green21)

Evaluation of factors influencing Agrobacterium-mediated spinach transformation and transformant selection by EGFP fluorescence under low-selective pressure

Contact Info

Product

Resources

About