Influence of gene flow from GM to non-GM soybeans by the size of the pollen donor

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http: //creativecommons.org/licenses/by- nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Copyright: © 2018 Korean Journal of Agrcultural Science

https://doi.org/10.7744/kjoas.20180078

PLANT & FOREST

Influence of gene flow from GM to non-GM soybeans by the size of the pollen donor

Bumkyu Lee

, Sung-Dug Oh

, Ancheol Chang

1

Department of Environment Science & Biotechnology, Jeonju University, Jeonju 55069, Korea

2

Biosafety Division, National Academy of Agricultural Sciences, Jeonju 54874, Korea

*Corresponding author: [email protected]

†

These authors equally contributed to this study as first author.

Abstract

The use of genetically modified (GM) crops has increased continuously over the world, and concerns about the potential risks of GM crops have also risen. Although, until now, GM crops have not been cultivated commercially in Korea, it is necessary to develop technology for the safe evaluation of GM crops. In this study, we investigated the influence of gene flow from GM to non-GM soybeans by the size of the pollen donor. In the experimental design, GM soybeans were placed in the center as a pollen donor and non-GM soybeans were placed in four directions as the pollen receivers. Three sizes of pollen donor were designed as 90 cm × 90 cm, 180 cm × 180 cm, and 360 cm × 360 cm. A total 22,719 seeds were collected from non- GM soybeans, and 14 hybrids were finally obtained through herbicide resistance screening and PCR analysis. The highest hybridization rate was 0.78% at a distance of 15 cm from a 360 cm × 360 cm GM pollen donor, and the farthest distance of hybridization was 180 cm from a GM pollen donor which was 360 cm × 360 cm in size. Ten hybrids were found among the 14 hybrids at the 360 cm × 360 cm pollen donor size, 3 hybrids at 180 cm × 180 cm, 1 hybrid at 90 cm × 90 cm. From these results, it could be concluded that with the larger pollen donor size, more hybridization occurred in soybeans.

Keywords: gene flow, genetically modified (GM), soybean, pollen source

Introduction

GM

genetically modified

¹⁹⁹⁴

²⁰¹⁷

²⁴

¹

⁸

⁹⁸⁰

^James

²⁰¹⁸

^GM

^{Glycine max}

²⁰¹⁷

⁹

94

¹

^GM

⁴⁹

⁶

¹²¹

⁵

⁷⁷

^James

²⁰¹⁸

^GM

²⁰¹⁷

⁹

⁶⁰¹

²⁰

¹

^GM

¹

⁰⁴³

⁴⁹⁶

³⁹⁷

¹⁵⁰

^KBCH

²⁰¹⁸

OPEN ACCESS

Accepted: September 27, 2018 Revised: September 26, 2018 Received: August 14, 2018 DOI:

Citation: Lee B, Oh SD, Chang A. 2018.

Influence of gene flow from GM to non-GM soybeans by the size of the pollen donor.

Korean Journal of Agricultural Science.

https://doi.org/10.7744/kjoas.20180078

현재 상업화 되어 재배되고 있는

^GM

⁷⁴

^{herbicide resistance}

^{insect resistance}

^GM

²⁸

^James

²⁰¹⁸

^KBCH

²⁰¹⁸

^E

^KBCH

²⁰¹⁸

GM

^Owen

²⁰⁰⁰

^{Brookes and Barfoot}

²⁰⁰⁶

^James

²⁰¹⁸

^GM

^Ellstrand

¹⁹⁹²

^{Conner et al}

²⁰⁰³

^Al

^{Ahmad and Gressel}

²⁰⁰⁶

^GM

^LMO

^{Lee and Suh}

²⁰¹¹

^LMO

^GM

^GM

^GM

콩은 대표적인 자가수분 식물로 일반적인 자연환경에서의 타식율(

outcrossing rate

⁰

⁵

^{Chiang and Kiang}

¹⁹⁸⁷

^{Nakayama and Yamaguchi}

²⁰⁰²

^{Ray et al}

²⁰⁰³

^{50 cm}

²

^{Chiang and Kiang}

¹⁹⁸⁷

^{Ahrent and Caviness}

¹⁹⁹⁴

^{Ray et al}

²⁰⁰³

^GM

^non

^GM

^{Yoshimura et al}

²⁰⁰⁶

⁴

^GM

^non

^GM

¹

^{7 m}

²

²

^{8 m}

⁴

³

^{5 m}

³

⁰

^{7 m}

⁰

¹⁹

^{Nakayama and Yamaguchi}

²⁰⁰²

^GM

^G

^soja

^{50 cm}

^GM

⁰

⁷³

^{Baek et al}

²⁰¹⁰

^{Apis mellifera L}

^{Lee et al}

²⁰¹⁵

^GM

국내에서

^GM

^{Lee et al}

^2007b

^{Lee et al}

^2009a

^{Lee et} al

²⁰¹²

^{Bae et al}

²⁰¹³

^{Oh et al}

²⁰¹⁴

^{Han et al}

²⁰¹⁵

^{Oh et al}

²⁰¹⁷

^{Kim et al}

^2009a

^{Kim et al}

^2009b

^{Baek et al}

²⁰⁰⁸

^{Kim et al}

²⁰¹⁴

^{Lee et al}

^2007a

^{Lee et al}

^2009b

^{Kim et al}

²⁰⁰⁸

^Lee et al

²⁰¹⁴

화분에 의한 유전자이동은 일반적으로 바람과 매개충 등에 의해 일어나며, 개화시기, 재배면적, 풍속, 기후 등의 환경에 의 해 영향 받는다(

^{Warwick et al}

²⁰⁰⁹

^Levin

¹⁹⁸¹

^GM

^{Chiang and Kiang}

¹⁹⁸⁷

^{Nakayama and Yamaguchi}

²⁰⁰²

^Ray et al

²⁰⁰³

^pot

GM

^{Yoshimura et al}

²⁰⁰⁶

^{22 m}

^{72 m}

⁴

^GM

^non

^GM

^{Baek et al}

²⁰¹⁰

^{4 m}

^{4 m}

^GM

^{Lee et al}

²⁰¹⁵

⁰

^{9 m}

^{5 m}

^GM

GM

^GM

기에 대한 한계치 설정과 이를 통한 안전성평가 가이드라인 개발이 요구되는 실정이다. 본 연구에서는

^GM

Materials and Methods

유전자이동평가 실험재료

실험에 사용된

^GM

^bar

^A

^β

^PAC

^{SP 7}

³

¹

^BASTA

^GM

^non

^GM

^GM

⁵⁰

^GMO

²

LMO

^RDA

^AB

²⁰¹³

⁰⁴¹

실험포장 및 실험구 배치

GM

^GM

⁴

^Fig

¹

^{15 cm}

^{90 cm}

Fig. 1. Diagrammatic representation of experimental design for gene flow. Genetically modified

화분 공급원 크기 별 유전자이동성 조사를 위해

^GM

^{90 cm}

^{90 cm}

^{180 cm}

^{180 cm}

^{360 cm}

360 cm

³

²

³

⁵

^GM

^Fig

²

^GM

^{90 cm}

^{90 cm}

¹⁴

⁷

²

^{180 cm}

^{180 cm}

³⁹

¹³

³

^{360 cm}

×

^{360 cm}

¹²⁵

²⁵

⁵

^{90 cm}

³

^GM

⁹⁰

¹⁸⁰

^{270 cm}

^{15 cm}

¹⁸

^{270 cm}

^GM

²

³

⁵

^Fig

²

GM 콩 도입유전자의 이동 조사

화분 수용체

^non

^GM

^{60 cm}

^{90 cm}

^{10 cm}

²⁰⁰

^LMO

³

⁰

⁴

Fig. 2. The experimental design of the field trial. Genetically modified (GM) soybeans were

도의 글루포시네이트 암모늄 제초제를

³

²

²

제초제로 선발된 개체에 대해

^PCR

^{100 mg}

^{DNeasy PowerPlant Pro Kit}

^QIAGEN

^Germany

^DNA

^PCR

^GM

^bar

^lectin

^PCR

^bar

^F

^{TCA ACC ACT ACA TCG AGA CAA GC}

R

^{GAT AGC GCT CCC GCA GAC}

^{105 bp}

^lectin

^F

^{TGC CGA AGC AAC} CAA ACA TGA TCC T

^R

^{TGA TGG ATC TGA TAG AAT TGA CGT T}

^{414 bp}

^PCR

⁹⁴

⁵

⁹⁴

³⁰

⁵⁵

³⁰

⁷²

¹

³⁰

⁷²

⁵

^PCR

¹

⁵

^{agarose gel}

Results and Discussion

화분 공급원인

^GM

^non

^GM

⁵

^non

^GM

^non

^GM

⁹⁰

¹⁸⁰

^{270 cm}

³

^GM

^{15 cm}

^{45 cm}

⁷

^Fig

³

^non

^GM

⁹⁶

²²

⁷¹⁹

²³⁷

Fig. 3. The example of sampling point for gene flow. Gray squares indicate the soybeans from

수확된 종자는

^{60 cm}

^{90 cm}

^{10 cm}

²⁰⁰

³

^GM

^non

^GM

^GM

^bar

^GM

²

^{Lee et al}

²⁰¹⁵

^PCR

^PCR

^GM

^bar

^lectin

^Fig

⁴

PCR

¹⁴

⁷

^GM

^{90 cm}

^{90 cm}

^{180 cm}

^{180 cm}

^{90 cm}

²

^{360 cm}

^{360 cm}

^{90 cm}

⁴

^{180 cm}

¹

^{Table 1}

⁰

⁰

³²

⁰

³²

^{360 cm}

^{360 cm}

^{90 cm}

⁰

¹

남쪽과 북쪽 방향의 경우에도 총

⁷

^GM

90 cm

^{90 cm}

^{15 cm}

¹

^{180 cm}

^{180 cm}

^{60 cm}

¹

^{360 cm}

^{360 cm}

^{15 cm}

³

^{60 cm}

²

^{60 cm}

^{180 cm}

^{360 cm}

^{360 cm}

¹⁵ cm

⁰

⁷⁸

^{90 cm}

^{90 cm}

^{15 cm}

⁰

⁴¹

교잡이 발생한 지점의 특징을 종합해보면 화분원의 크기가 가장 큰

^{360 cm}

^{360 cm}

¹⁴

¹⁰

^{180 cm}

^{180 cm}

³

^{90 cm}

^{90 cm}

¹

90 cm

^{15 cm}

^{180 cm}

^{Table 1}

²

Fig. 4. Part of hybrid detection by PCR analysis. bar gene was used to detect hybrid and lectin

Ray et al

²⁰⁰³

^GM

^non

^GM

^GM

⁰

^{9 m}

⁰

⁴¹

⁵

^{4 m}

⁰

⁰³

⁰

⁰⁵

^{Yoshimura et al}

²⁰⁰⁶

GM

^non

^GM

⁰

^{7 m}

⁰

¹

^{7 m}

⁰

⁰⁴

^{Chiang and Kiang}

¹⁹⁸⁷

^{10 cm}

⁰

⁷³⁷

^GM

^non

^GM

^{180 cm}

⁵⁰⁰

^{700 cm}

이러한 결과는 콩에서 화분을 통한 교잡은 바람, 기온 등 다양한 환경에 영향을 받으며, 특히 벌(

^{Chiang and Kiang}

¹⁹⁸⁷

^Ray et al

²⁰⁰³

^{Nakayama and Yamaguchi}

²⁰⁰²

본 연구에서

^GM

^GM

^Fig

²

Table 1. Frequency of out-crossing rates evaluated from surviving progeny in East and West sites.

Size of pollen

source (cm

) classification Distances from pollen source (cm)

East West

90 180 270 90 180 270

90 × 90 Evaluated progenies 484 785 632 753 519 423

Hybrid progenies 0 0 0 0 0 0

Out-crossing rate (%) 0 0 0 0 0 0

180 × 180 Evaluated progenies 717 961 735 991 591 846

Hybrid progenies 1 0 0 1 0 0

Out-crossing rate (%) 0.14 0 0 0.10 0 0

360 × 360 Evaluated progenies 1005 679 480 932 702 905

Hybrid progenies 1 0 0 3 1 0

Out-crossing rate (%) 0.10 0 0 0.32 0.14 0

Table 2. Frequency of out-crossing rates evaluated from surviving progeny in North and South

Size of pollen source (cm

)

classification

Distances from pollen source (cm) North and South

15 60 105 150 195 240 270

90 × 90 Evaluated progenies 241 277 422 271 434 229 480

Surviving progenies 1 0 0 0 0 0 0

Out-crossing rate (%) 0.41 0 0 0 0 0 0

180 × 180 Evaluated progenies 313 581 627 345 674 526 528

Surviving progenies 0 1 0 0 0 0 0

Out-crossing rate (%) 0 0.17 0 0 0 0 0

360 × 360 Evaluated progenies 386 755 397 651 234 615 493

Surviving progenies 3 2 0 0 0 0 0

Out-crossing rate (%) 0.78 0.26 0 0 0 0 0

단되었다(

^{Yoshimura et al}

²⁰⁰⁶

^GM

^GM

^GM

Conclusion

GM

^GM

^GM

^GM

^GM

^GM

^GM

^non

^GM

^GM

^non

^GM

⁴

^{90 cm}

^{90 cm}

^{180 cm}

¹⁸⁰ cm

^{360 cm}

^{360 cm}

³

^non

^GM

²²

⁷¹⁹

^PCR

¹⁴

⁰

⁷⁸

^{360 cm}

^{360 cm}

^{15 cm}

^{360 cm}

^{360 cm}

^{180 cm}

^{360 cm}

^{360 cm}

¹⁴

¹⁰

^{180 cm}

^{180 cm}

³

^{90 cm}

^{90 cm}

¹

^GM

Acknowledgements

본 연구는 농촌진흥청 국립농업과학원 농업과학기술 연구개발사업(과제번호:

^PJ009609

PJ011867042018

References

Ahrent DK, Caviness CE. 1994. Natural cross-pollinaion of twelve soybean cultivars in Arkansas. Crop Science 34:376-378.

Al-Ahmad H, Gressel J. 2006. Mitigation using tandem construct containing a selectively unfit gene precludes establishment of Brassica napus transgenes in hybrids and backcrosses with weedy Brassica rapa. Plant Biotech 4:23-33.

Bae HK, Oo MM, Jeon JE, Tien DN, Oh SA, Oh SD, Kweon SH, Eun MY, Park SK. 2013. Evaluation of gene flow from GM to Non-GM rice. Plant Breeding and Biotechnology 1:162-170.

Baek HJ, Sohn SI, Cho MR, Lee GS, Oh YJ, Park JS, Lee KJ, Oh SD, Suh SC, Ryu TH. 2010. Development of protocol for analyzing pollinator insect-mediated gene transfer from gm crop. Korean Journal of

International Agriculture 22:293-297. [in Korean]

Baek HJ, Won SY, Kim JK, Sohn SI, Lee K, Cho MR, Song JK, Yoon MS, Lee J, Jin YM, Ryu TH. 2008. Study on the gene introgression from GM Chinese cabbage to major crops in Cruciferae. Korean Journal of International Agriculure 20:124-129. [in Korean]

Brookes G, Barfoot P. 2006. Global impact of biotech crops: Socio-economic and environmental effects in the first ten years of commercial use. AgBioForum 9:139-151.

Chiang YC, Kiang YT. 1987. Geometric position of genotypes, honeybee foraging patterns and outcrossing in soybean. Botanical Bulletin of Academia Sinica 28:1-11.

Conner AJ, Glare TR, Nap JP. 2003. The release of genetically modified crops into the environment; Part II.

Overview of ecological risk assessment. Plant Journal 33:19-46.

Ellstrand NC. 1992. Gene flow by pollen: Implications for plant conservation genetics. Oikos 63:77-86.

Han SM, Lee B, Won OJ, Hwang KS, Suh SJ, Kim CG, Park KW. 2015. Gene flow from herbicide resistant genetically modified rice to conventional rice (Oryza sativa L.) cultivars. Journal of Ecology and Environment 38:397-403.

James C. 2018. Global status of commercialized biotech/GM crops: 2017, BRIEF 53. Accessed in http://

www.isaaa.org on 1 July 2018.

KBCH (Korea Biosafety Clearing House). 2018. Stats of GM crops in Korea. Accessed in http://

www.biosafety.or.kr on 23 July 2018.

Kim CG, Lee B, Kim DI, Park JE, Kim HJ, Park KW, Yi H, Jeong SC, Yoon WK, Harn CH, Kim HM. 2008.

Detection of gene flow from GM to non-GM watermelon in a field trial. Journal of Plant Biology 51:74- 77.

Kim CG, Kim DI, Kim HJ, Park JY, Lee B, Park KW, Jeong SC, Cho KH, An JH, Cho KH, Kim YS, Kim HM.

2009a. Assessment of gene flow from genetically modified anthracnose-resistant chili pepper (Capsicum annuum L.) to a conventional crop. Journal of Plant Biology 52:251-258.

Kim CG, Park KW, Lee B, Kim DI, Park JY, Kim HJ, Park JE, An JH, Cho KH, Jeong SC, Choi KH, Harn JH, Kim HM. 2009b. Gene flow from genetically modified to conventional chili pepper (Capsicum annuum L.).

Plant Science 176:406-412.

Kim YJ, Nam KH, Pack IS, Park JH, Jeong SC, Harn CH, Kim CG. 2014. Gene flow from GM cabbage to Non- GM control. CNU Journal of Agricultural Science 41:157-161. [in Korean]

Lee B, Kim CG, Park JY, Yi H, Park KW, Jeong SC, Yoon WK, An JH, Cho KH, Kim HM. 2007a. Survey of herbicide resistant oilseed rapes around the basin of rivers in Incheon harbor area. Korean Journal of Weed Science 27:29-35. [in Korean]

Lee SY, Kim MS, Kim HJ, Ahn JH, Baek SH, Shin WC, Kim HS. 2007b. Pollen-mediated gene flow between glufosinate ammonium-tolerant GM and Non-GM rice. Journal of Plant Biotechnology 34:47-53.

Lee B, Kim JH, Sohn SI, Kweon SJ, Chung YS, Lee SM. 2015. Influence of insect pollinators on gene transfer from GM to non-GM soybeans. Korean Journal of Agricultural Science 42:159-165. [in Korean]

Lee B, Park KW, Kim CG, Kang HG, Sun HJ, Kown YI, Song IJ, Ryu TH, Lee HY. 2014. Environmental monitoring of herbicide tolerant genetically modified zoysiagrass (Zoysia japonica) around confined field trials. Weed and Turfgrass Science 3:305-311 [in Korean].

Lee B, Suh S. 2011. A study on the trends and biosafety assessment of genetically modified crops.

Research of Environmental Law 33:1-25. [in Korean]

Lee HS, Yi GH, Kim KM. 2012. Comparison of weed characteristics and possibility of gene flow in GM rice.

Korean Journal of Weed Science 32:10-16. [in Korean]

Lee IY, Park JE, Moon BC, Suh SC, Shin KS, Woo MO, Kweon SJ. 2009a. Possibility of Gene flow and unintended escape from leaf-folder (Cnaphalocrocis medinalis) resistant rice. Korean Journal of Weed Science 29:46-55. [in Korean]

Lee YH, Kim KS, Jang YS, Cho HJ, Bang JK, Suh SJ. 2009b. Pollen-mediated gene flow from glufosinate resistant transgenic oilseed rape (Brassica napus L.) into its wild relatives. Korean Journal of International Agriculture 21:193-198.

Levin DA. 1981. Dispersal versus gene flow in plants. Annals of the Missouri Botanical Garden 68:233-253.

Nakayama Y, Yamaguchi H. 2002, Natural hybridization in wild soybean (Glycine max ssp. soja) by pollen flow from cultivated soybean (Glycine max ssp. max) in a designed population. Weed Biology and Management 2:25-30.

Oh SD, Lim MH, Ryu TH, Shon SI, Park SK, Lee K. 2014. Assessment of gene flow in disease resistant (OsCK1) genetically modified rice. Korean Journal of Breeding Science 46:44-51. [in Korean]

Oh SD, Yun DW, Shon SI, Park SK, Chang AC. 2017. Assessment of gene flow from insect-resistant genetically modified rice (Agb0101) to non-GM rice. Korean Journal of Breeding Science 49:180-189. [in Korean]

Owen MDK. 2000. Current use of transgenic herbicide-resistant soybean and corn in the USA. Crop Protection. 19:765-771.

Ray JD, Kilen TC, Abel CA, Paris RL. 2003. Soybean natural cross-pollination rates under field conditions.

Environmental Biosafety Research 2:133-138.

Warwick SI, Beckie HJ, Hall LM. 2009. Gene flow, invasiveness, and ecological impact of genetically modified crops. The Year in Evolutionary Biology 2009: Annals of the New York Academy of Sciences 1168:72-99.

Yoshimura Y, Matsuo K, Yasuda K. 2006. Gene flow from GM glyphosate-tolerant to conventional soybeans under field conditions in Japan. Environmental Biosafety Research 5:169-173.