Screening brinjal (Solanum melongena) accessions conserved in the National Genebank collected from states adjoining Bangladesh for adventitious presence of EE1 event

Kuwardadra Sahadeo Indaldas; Chithra Devi Pandey; Aparna Paliwal; Gurinderjit Randhawa

doi:10.56093/ijas.v90i8.105975

Authors

Kuwardadra Sahadeo Indaldas Ph D Scholar, Post Graduate School, ICAR-Indian Agricultural Research Institute, New Delhi
Chithra Devi Pandey Principal Scientist, Division of Germplasm Conservation, ICAR-Indian Agricultural Research Institute, New Delhi
Aparna Paliwal Senior Research Fellow, Division of Genomic Resources, ICAR-Indian Agricultural Research Institute, New Delhi
Gurinderjit Randhawa Principal Scientist and corresponding author, Division of Genomic Resources, ICAR- National Bureau of Plant Genetic Resources, New Delhi

https://doi.org/10.56093/ijas.v90i8.105975

Keywords:

Adventitious presence, GM-free Conservation, National Genebank, Polymerase Chain Reaction, Real time PCR, Transgenes

Abstract

Systematic management of Plant Genetic Resources (PGRs) is the key to sustainable agriculture for food and nutritional security and in mitigating climate change. The National Genebank (NGB) at ICAR-National Bureau of Plant Genetic Resources (NBPGR), New Delhi, acts as a repository of PGRs for future use. This study aimed at screening for the adventitious presence of transgenes in brinjal (Solanum melongena L.) accessions conserved in the NGB. The study targeted the collections made during 2007-2016 from areas adjacent to Bangladesh (Assam, Meghalaya, Mizoram, Tripura and West Bengal), where field trials of Bt brinjal event EE1 were conducted during 2005-2012 and commercial cultivation of this event was permitted in 2013.There could be an apprehension of both unintentional introgression and transboundary movement through borders. Adventitious presence of transgenes was checked in a set of 96 accessions of brinjal employing Polymerase Chain Reaction (PCR) and real-time PCR assays. As event EE1 carries cry1Ac gene for insect resistance with Cauliflower Mosaic Virus (CaMV) promoter (P-35S) and marker genes (nptII and aadA), so these genetic elements were targeted for qualitative GM testing. Based on the test results, transgenes were not detected in brinjal accessions conserved in NGB. Our study showed that brinjal and wild species collected from adjoining areas of Bangladesh, post field trials and release, do not contain the event EE1. The study presents an efficient and reliable method to ensure conservation of GM-free germplasm in the NGB.

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References

Bairwa R K, Singh M, Bhoge R K, Devi C and Randhawa G J. 2016. Monitoring adventitious presence of transgenes in ex situ okra (Abelmoschus esculentus) collections conserved in genebank: a case study. Genetic Resources and Crop Evolution 63(2): 175–184. DOI: https://doi.org/10.1007/s10722-015-0350-3

Block A, Debode F, Grohmann L, Hulin J, Taverniers I and Kluga L. 2013. The GMOseek matrix: adecision support tool for optimizing the detection of genetically modified plants. BMC Bioinformatics 14: 256. DOI: https://doi.org/10.1186/1471-2105-14-256

Bonfini L, van den Bulcke M H, Mazzara M, Ben E and Patak A. 2012. GMOMETHODS: The European Union Database of reference methods for GMO analysis. Journal of AOAC International 95(6): 1713–1719. DOI: https://doi.org/10.5740/jaoacint.12-050

Choudhary B and Gaur K. 2009. The Development and Regulation of Bt Brinjal in India (Eggplant/Aubergine). ISAAA Brief No.38. ISAAA: Ithaca, NY.

Debode F, Janssen E, Bragard C and Berben G. 2017. Detection by real-time PCR and pyrosequencing of the cry1Ab and cry1Ac genes introduced in genetically modified (GM) constructs. Food Additives & Contaminants: Part A 34(8): 1398–1409. DOI: https://doi.org/10.1080/19440049.2017.1317925

Dellaporta S L, Wood J and Hicks J B. 1983. A plant DNA minipreparation: version II. Plant Molecular Biology Reporter 1(4): 19–21. DOI: https://doi.org/10.1007/BF02712670

Gamarra L F R, Delgado J A, Villasante Y A and Ortiz R. 2011. Detecting adventitious transgenic events in a maize center of diversity. Electronic Journal of Biotechnology 14(4): 9–9. DOI: https://doi.org/10.2225/vol14-issue4-fulltext-12

Kumar P A. 2009. Bt brinjal: a pioneering push. Biotechnology News 4(6): 108–111.

Lipp M, Bluth A, Eyquem F, Kruse L, Schimmel H, Van den Eede G and Anklam E.2001. Validation of a method based on polymerase chain reaction for the detection of genetically modified organisms in various processed foodstuffs. European Food Research and Technology 212: 497–504. DOI: https://doi.org/10.1007/s002170000274

Mezzalama M, Crouch J H and Ortiz R. 2010. Monitoring the threat of unintentional transgene flow into maize gene banks and breeding materials. Electronic Journal of Biotechnology 13(2): 6–7. DOI: https://doi.org/10.2225/vol13-issue2-fulltext-5

Parimalan R, Bhoge R, Randhawa G J and Pandey C D. 2015. Assessment of transgene flow in eggplant germplasm conserved at National Genebank. Indian Journal of Biotechnology 14(3): 357–363.

Raina J and Yadav G S. 2018. Brinjal shoot and fruit borer: Bioecology and management. Journal of Pharmacognosy and Phytochemistry 7(4): 444–449.

Randhawa G J, Sharma R and Singh M. 2009. Multiplex PCR-based simultaneous amplification of selectable marker and reporter genes for the screening of genetically modified crops. Journal of Agricultural and Food Chemistry 57(12): 5167–5172. DOI: https://doi.org/10.1021/jf900604h

Randhawa G J, Sharma R and Singh M. 2010. Decaplex and real-time PCR based detection of MON531 and MON15985 Bt cotton events. Journal of Agriculture and Food Chemistry 58(18): 9875–9881. DOI: https://doi.org/10.1021/jf100466n

Randhawa G J, Sharma R and Singh M. 2012. Qualitative and event-specific real-time PCR detection methods for Bt brinjal event EE -1. Journal of AOAC International 95(6): 1733–1739. DOI: https://doi.org/10.5740/jaoacint.11-478

Randhawa G J and Singh M. 2016. Qualitative and quantitative diagnostics for EE-1 event of Bt eggplant. Genetically Modified Organisms in Food - 1st Edition, pp 87-96. Watson R R and Preedy V R (Eds). Academic Press Publication (Elsevier), USA. DOI: https://doi.org/10.1016/B978-0-12-802259-7.00009-9

Sekara A and Bieniasz M. 2008. Pollination, fertilization and fruit formation in eggplant [Solanum melongena L.]. Acta Agrobotanica 61(1): 107–113. DOI: https://doi.org/10.5586/aa.2008.014

Taberlet P, Gielly L, Pautou G and Bouvet J. 1991. Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Molecular Biology 17(5): 1105–1109. DOI: https://doi.org/10.1007/BF00037152

Tiwari S P and Randhawa G J. 2010. Strategies to monitor the adventitious presence of transgenes in ex situ collections. Indian Journal of Agricultural Sciences 80(5): 351–356.

Waiblinger H U, Ernst B, Anderson A and Pietsch K. 2008. Validation and collaborative study of a P35S and T-nos duplex real-time PCR screening method to detect genetically modified organisms in food products. European Food Research Technology 226: 1221–1228 DOI: https://doi.org/10.1007/s00217-007-0748-z

Wu Y, Wang Y, Li J, Li W, Zhang L, Li Y, Li X, Li J, Zhu L and Wu G. 2014. Development of a general method for detection and quantification of the P35S promoter based on assessment of existing methods. Science Reports 4: 7358 DOI: https://doi.org/10.1038/srep07358

http://agricoop.nic.in/sites/default/files/Horticulture%20 Statistics%20at%20a%20Glance-2018.pdf ) (last accessed on 18.11.2019)

http://www.nbpgr.ernet.in/ (last accessed on 17.11.2019)

http://www.indiaenvironmentportal.org.in/files/brinjal.pdf (last accessed on 18.11.2019)

http://www.isaaa.org/gmapprovaldatabase (last accessed on 15.11.2019)

http://www.geacindia.gov.in/decisions-of-GEAC-meetings.aspx (last accessed on 15.11.2019)

http://www.apaari.org/web/wpcontent/uploads/downloads/2018/ Bt%20Brinjal%20in%20Bangladesh%20final.pdf (last accessed on 17.11.2019)