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

Wonkyun Choi,Jung Ro Lee,Min-A Seol,Beom-Ho Jo,Il Ryong Kim

doi:10.1007/s13765-018-0399-8

Abstract

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.

Highlights

According to the recent report from the International Service for the Acquisition of Agribiotech Application, the global cultivation areas of LM soybeans (94.1 million hectares) occupy approximately 50% of the total GM crop cultivation area [1]
We developed the multiplex PCR detection method of four LM soybean events (CV127, MON87705, FG72 and MON87701) (Fig. 1)
All primers used for multiplex PCR analysis were designed to be located in the soybean genomic flanking region and transgenic region (Fig. 1)

Summary

Introduction

According to the recent report from the International Service for the Acquisition of Agribiotech Application, the global cultivation areas of LM soybeans (94.1 million hectares) occupy approximately 50% of the total GM crop cultivation area [1]. Production of soybeans in Republic of Korea has been diminished continuously because of the decrease in cultivation areas and poor harvests. In 2017, the amount of soybeans which cultivated domestically occupied 75,000 tones and it comprises approximately 25% of the entire domestic consumption (300,000 tones) a year [2]. In the recent report of KBCH, 17 single events and 10 stack traits of LM soybean were approved in Republic of Korea [3]. CV127 is an event developed by BASF and resistant to imidazolinone herbicides due to insertion of CSR1 gene from Arabidopsis. It was approved as only single event in Republic of Korea, and currently has been used in 21 countries. MON87701 developed by Monsanto was approved for a single event and two stack events

Methods

Results

Conclusion