Four multiplex PCR Sets of 11 LM Maize for LMO environmental monitoring in Korea

Shin, Su Young; Lim, Hae-Song; Seol, Min-A; Jung, Young Jun; Kim, Il Ryong; Song, Hae Ryoung; Lee, Jung Ro; Choi, Wonkyun

doi:10.5010/jpb.2016.43.4.473

Cited by 4 publications

(7 citation statements)

References 10 publications

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“…For high-sensitive detection, large amount of DNA as template is used; however, environmental sample of LMO cannot ensure DNA quality and quantity. Several testing methods released by developers or regulators and several previously reported GMO detection methods recommend that the minimum amount of purified DNA required is 40-100 ng in total reaction volume [6,10,11]. In this study, we detected LMOs using the newly developed multiplex PCR method, which required only 12.5 ng of purified DNA.…”

Section: Discussionmentioning

confidence: 96%

“…The multiplex PCR results with randomly mixed RM mixtures suggested that this method can identify all LMO stacks, including the four LM cotton varieties that will be approved in the future. The multiplex PCR method was applied to identify LM samples, which were collected from environmental monitoring [9][10][11]. To verify the application of multiplex PCR method in LMO monitoring, we used LM cotton samples collected from an LMO monitoring project in the Republic of Korea in 2018.…”

Section: Verification Of the Efficiency Of Multiplex Pcrmentioning

confidence: 99%

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Event-specific multiplex PCR method for four genetically modified cotton varieties, and its application

et al. 2019

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Multiplex polymerase chain reaction (PCR) methods have been developed and validated for screening, tracing, and regulating genetically modified (GM) crops in quarantine and environmental monitoring. In this study, we aimed to develop a method to simultaneously detect four GM cotton varieties in order to establish a screening system for cotton volunteers. Based on the sequence of DNA in the junction between introduced gene and flanking genomic DNA of four GM cotton events, herbicide-tolerant MON88701 and DAS-81910-7 and insect-resistant COT102 and T304-40, event-specific primers were designed and a multiplex detection method was developed. The simplex PCR results supported the multiplex PCR results; the amplification efficiency of the novel multiplex PCR method was increased compared with that of the Joint Research Centre (JRC) method. Based on the accuracy and efficiency, the method can be applied to detect and identify randomly mixed reference materials and suspected cotton volunteers. To apply this multiplex PCR method to living modified (LM) environmental monitoring samples, we performed additional PCR analysis to identify whether the volunteers were the four LM cotton varieties. As a result, 66 cotton volunteers were identified with stack event, comprising one or two of the four LM cotton events, and all stacks have been approved in South Korea for food, feed, and processing. These results indicated that our novel multiplex method is suitable for LMO identification.

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Section: Discussionmentioning

confidence: 96%

Section: Verification Of the Efficiency Of Multiplex Pcrmentioning

confidence: 99%

Event-specific multiplex PCR method for four genetically modified cotton varieties, and its application

et al. 2019

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“…Based on the simplex PCR results, we optimized a novel multiplex PCR analysis for four LM soybeans (CV127, MON87705, FG72 and MONM87701). Previous studies revealed that optimization of multiplex PCR was very delicate and difficult because of insertion gene similarity, difference of primer and PCR condition [8][9][10]. In this study, we tried to fix primer concentration and PCR reaction condition to optimize all PCR reaction products.…”

Section: Establishment Of Simultaneous Multiplex Pcrmentioning

confidence: 99%

“…[7,8]. Moreover, the detection methods were further elaborated for the multiplex PCR which was applicable to the analysis of LMO monitoring samples [9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Development and application of a novel multiplex PCR method for four living modified soybeans

Choi

Seol

et al. 2018

Appl Biol Chem

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Since the early 1990s when the first commercialization of living modified organism (LMO), LMO has been developed to improve nutrient quality and productivity of crops. As the self-sufficiency rate of soybean has gradually decreased in South Korea, most of soybeans have been imported. The cultivation and trade of LM crops are regulated in many countries and authorizations for the use are mandatory in most. In South Korea, the cultivation of LM crop is not allowed and unintentional release of LMO into the natural environment is prohibited. In this study, we developed a novel multiplex PCR method for four LM soybean events (CV127, MON87705, FG72 and MON87701) which were approved recently in South Korea. Multiplex PCR primers were designed for PCR amplification of four LMO event-specific fragments, and we analyzed 41 environmental monitoring samples to confirm the efficiency of this method. These results indicated that the multiplex PCR detection method is sufficient for four LM soybeans found in the natural environment. Based on our finding, we suggest that the new technique may be useful as a lead tool for the development of a detection method for various LMO/GMOs.

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“…Since 2009, the MOE and National Institute of Ecology (NIE) have been conducting The Environmental LMO Monitoring and Post-Management Project [9,10]. In the project, the natural environmental leakage of LM crops such as cotton, maize, soy bean, and canola was monitored and the risk assessment and management of released LMOs was performed [11][12][13]. A specific LMO detection method is crucial to determine whether the samples collected in this project contain LMOs [14,15].…”

Section: Introductionmentioning

confidence: 99%

Development of a Multiplex PCR Assay to Monitor Living Modified Cottons in South Korea

Kim

Lim

et al. 2019

Applied Sciences

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Cotton has been cultivated worldwide and is a useful crop for humans. However, all living modified organisms (LMOs), including living modified (LM) cotton, are not cultivated in South Korea and are imported from overseas. LM cotton imports are on the rise and most of the imported cotton is used as livestock feed. In particular, it is commonly used to feed Holstein breeds that produce milk, because cotton improves the quality of milk. However, as the cotton imports increase, the possibility of unintentional outflows in the distribution process also increases. Consequently, there is an increasing concern about unintentional release of LM cotton into the natural environment. Therefore, environmental monitoring and post-management of LMOs are very important steps. Recently, a total of 30 LM crop events were approved for LM cotton import in South Korea. A single detection method has been used to monitor individual events. However, a single method of detection for collected samples requires a large number of PCRs, with obvious disadvantages. Therefore, a simultaneous detection method was developed for 8 representative events (GHB119, GHB614, MON88913, MON15985, LLCOTTON25, MON1445, 281-3006, and MON531) in an effort to monitor 26 of them and facilitate the identification of LM cotton. The results suggest that our new multiplex PCR method may be useful for monitoring and post-management of LM cotton.

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Four multiplex PCR Sets of 11 LM Maize for LMO environmental monitoring in Korea

Cited by 4 publications

References 10 publications

Event-specific multiplex PCR method for four genetically modified cotton varieties, and its application

Event-specific multiplex PCR method for four genetically modified cotton varieties, and its application

Development and application of a novel multiplex PCR method for four living modified soybeans

Development of a Multiplex PCR Assay to Monitor Living Modified Cottons in South Korea

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