Development of a multiplex event-specific PCR assay for detection of genetically modified rice

Lü, Juanjuan; Ji, Guo‐Zhen; Li, G.; Wu, Yuanfeng; Jing, Yang; Lin, Shu-Fang; Dian-lin, Yang; Zhao, Jie; Xiu, Weiming

doi:10.1556/0806.43.2015.043

Cited by 3 publications

(3 citation statements)

References 36 publications

(40 reference statements)

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“…Penggunaan primer spesifik dapat menjamin bahwa tanaman transforman yang diperoleh benar-benar merupakan tanaman transgenik (true transgenic plant). Deteksi transgen dengan primer spesifik ini merupakan tahapan yang selalu ada di dalam perakitan tanaman transgenik (Melloul et al 2014;Lu et al 2016;Dong et al 2017).…”

Section: Analisis Molekuler Dengan Teknik Pcrunclassified

Construction and Transformation of OsERA1 Gene into Expression Vector and Response of Nipponbare-OsERA1 Transgenic Rice to Drought Stress

Santoso¹,

Apriana²,

Sisharmini³

et al. 2018

J. AgroBiogen

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Drought stress is a major constrain which could influence rice productivity. Enhanced Response to ABA1 (ERA1) gene encoding a β-subunit farnesyltransferase enzyme plays a role to control sensitivity of the guard cells to abscisic acid (ABA), hence regulating drought stress response in plant species including rice. This study aimed to clone the OsERA1 gene into expression vector, introduce it into rice plant, and confirm the positive OsERA1-rice plants conferring drought tolerance. This study was initiated by isolation of the OsERA1 gene from rice cDNAs and cloned it to an expression vector cassette, pCAMBIA1301. The cassette harboring OsERA1 gene was introduced into rice plant cv. Nipponbare mediated by Agrobacterium tumefaciens strain LBA4404. Putative transgenic lines were detected using PCR and Southern blot analyses to confirm the inserted transgene and the positive lines were assayed their tolerance to drought. The OsERA1 gene was successfully isolated and constructed into expression vector to generate pCAMBIA1301-OsERA1. Introduction of the gene into Nipponbare has produced nine putative transgenic rice lines, of which, six lines harbored OsERA1 gene. Southern blot analysis of sixteen T2 plants from two PCR-positive transgenic lines revealed 1–3 copies of transgene were integrated into rice genome of transgenic lines. Five transgenic lines of Nipponbare-OsERA1 showed better response to drought at vegetative phase compared to control in term of recovery ability. At generative phase, the five transgenic lines yielded less unfilled grains compared to control. Overall, the transgenic lines obtained from this study could be potential candidates for developing rice varieties tolerant to drought.

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Section: Analisis Molekuler Dengan Teknik Pcrunclassified

Construction and Transformation of OsERA1 Gene into Expression Vector and Response of Nipponbare-OsERA1 Transgenic Rice to Drought Stress

Santoso¹,

Apriana²,

Sisharmini³

et al. 2018

J. AgroBiogen

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“…The KMD1 rice line is resistant to lepidopteran pest species (a synthetic cry1Ab gene) and has the potential to be approved in China. The Kefeng6 and Kefeng8 contain two insect-resistant genes, cry1Ac and cowpea trypsin inhibitor ( CpTI ) gene (Lu et al, 2016 ; Wang et al, 2012 ). The LLRice62 contains glufosinate herbicide-tolerant bar gene and phosphinothricin- N -acetyltransferase (PAT) proteins (Oberdoerfer et al, 2005 ).…”

Section: Introductionmentioning

confidence: 99%

“…In 2006, Bayer’s LLRice601 and LLRice62 have been discovered across the world (Greenpeace International, 2006 ). Bt63 was also detected in Europe and identified in imported food products according to the Rapid Alert System for Food and Feed (RASFF) (Lu et al, 2016 ; Price and Cotter, 2014 ). For these reasons, accurate and reliable detection methods are needed for the identification and analysis of unauthorized GMOs crops or products.…”

Section: Introductionmentioning

confidence: 99%

Development of a rapid detection method for genetically modified rice using the ultra-fast PCR system

Shin

Jeon

Koo

2022

Food Sci Biotechnol

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Genetically modified (GM) rice varieties containing traits such as tolerance to abiotic stress and resistance against pests and diseases continue to be developed. However, contamination incidents from unauthorized GM rice varieties have been encountered. To date, no GM rice crop has been authorized for consumption and/or commercialization in Korea. Therefore, to enhance safety management of unauthorized genetically modified organisms (GMOs), accurate and reliable detection methods are needed to identify GMOs in crops or products. In this study, we developed rapid detection methods for GM rice events (Bt63, KMD1, Kefeng6, Kefeng8, and LLRice62) using ultra-fast PCR system. Ultra-fast PCR is a state-of-the-art technology and decreases PCR run-times dramatically. However, the ultra-fast PCR is not widely used in GMO analysis. Thus, we designed a detection method for five events of GM rice and confirmed them by performing specificity, sensitivity, and applicability assays. All results demonstrate that the ultra-fast PCR system is a specific, sensitive, and reliable method to identify and monitor GM rice events. Additionally, it can be utilized as a rapid and simple method for GMO analysis in crops or processed products. This study can be used as a reference for future research on new analysis methods of unauthorized GMOs.

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Detection and occurrence of genetically modified rice and potato in the Saudi food market

Aladadh,

Ebrahim,

Elsanhoty

et al. 2024

Braz. J. Biol.

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The number of food products with genetically modified (GM) crops on the global market has increased due to advancements in genetic engineering technology. Legislation regulating the labeling and use of GM crops has increased considerably worldwide to provide consumers with health and safety assurance. It is still unclear whether genetically modified organisms (GMOs) are present in the food market of the Kingdom of Saudi Arabia due to a lack of scientific studies. This work was planned to detect GM rice and GM potatoes in the Saudi food market. One hundred non-labeled rice and rice product samples and 50 potato and potato samples were collected randomly from different market sites of Makkah, Riyadh and Jeddah during 2022-2023. The cetyl trimethyl ammonium bromide (CTAB) method was used to extract DNA. Viviants DNA extraction kit was used to extract DNA from rice starch and potato chips. To find GMOs in samples, CMOScreen 35S and NOS test kits were utilized. DNA-based qualitative and quantitative approaches were used to screen targets for PCR detection of GM rice sequences. The results indicated that 32 (32%) rice samples were positive for CaMV 35S promoter, while no positive result was detected for the NOS terminator. Besides, 30% of potato samples were positive for the CaMV 35S promoter, and the same samples were positive for the presence of the Cry V gene. It could be concluded that there were GM rice and potatoes in the Kingdom of Saudi Arabia's food markets. Establishing strong regulations and certified laboratories to monitor genetically modified foods (GMF) or crops in the Saudi market is recommended.

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Development of a multiplex event-specific PCR assay for detection of genetically modified rice

Cited by 3 publications

References 36 publications

Construction and Transformation of OsERA1 Gene into Expression Vector and Response of Nipponbare-OsERA1 Transgenic Rice to Drought Stress

Construction and Transformation of OsERA1 Gene into Expression Vector and Response of Nipponbare-OsERA1 Transgenic Rice to Drought Stress

Development of a rapid detection method for genetically modified rice using the ultra-fast PCR system

Detection and occurrence of genetically modified rice and potato in the Saudi food market

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