Scientists

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I. Technologies Developed

1. Loop mediated Isothermal amplification (LAMP) assays for GM detection have been developed for the first time in India, under my supervision in GM detection laboratory at NBPGR. This easy to use developed assay enables visual detection, when combined with efficient DNA extraction method can be used at the port of the entry by the custom officials or on-site in farmer’s fields, as it does not require many laboratory equipments and specialized expertise.
2. Technology developed for initial screening of GM crops

• Hexaplex PCR assay for simultaneous detection of six marker genes, i.e., aadA, bar, hpt, nptII, pat and uidA to check the GM status of a sample irrespective of the crop and GM trait:
• Heptaplex PCR assay simultaneously amplifying marker genes; nptII, aadA, pat, uidA and regulatory elements, viz., CaMV 35S, nos promoters and nos terminator.
• Visual and real-time LAMP assays for commonly employed promoters: P-35S and P-FMV, marker genes: nptII, aadA and uidA, transgenes: cry1Ac, cry2Ab2 and cp4-epsps have been developed for on-site screening for the GM status of planting material at the ports of entry or on farmers’ fields when combined with efficient DNA extraction method.
• A GMO matrix based on the information on 141 GM events of 21 crops with 106 transgenic elements has been developed as a decision support system. Ten most frequently present targets have been identified to screen these events using a GMOseek algorithm.
• Multi-target TaqMan® real-time PCR-based screening system, simultaneously targeting 47 targets, including 21 GM events, 5 constructs, 11 transgenes, 4 control elements and 6 endogenous genes to test an unknown sample for these targets in duplicate. Limit of detection (LOD) is 0.1-0.01% depending upon the target.

3. Technology developed for detection of more than 40 GM events of 12 crops

• Technology for detection of commercialized Bt Cotton events: Decaplex PCR technology with limit of detection (LOD) upto 0.01%, has been developed to differentiate and to check the adulterants in two major commercialized Bt cotton events in India, viz., MON531 (Bollgard^® I) and MON15985 (Bollgard^® II). Recently, another decaplex PCR method with LOD upto 0.1%, differentiating five commercialized events of Bt cotton, viz., MON531, MON15985, Event1, GFM-cry1A and MLS-9124 has been developed.

Event-specific visual and real-time LAMP assays for detection of MON531 and MON15985 events have been developed for the first time.

• Technology developed for detection of Bt Brinjal: Event-specific conventional and real-time PCR assays with LOD upto 0.01% have been developed for detection of Bt brinjal event EE1.
• Event-specific LAMP assays for six GM maize events, Bt11, GA21, MON810, MON89035, NK603 and TC1507 have been developed.
• Multiplex PCR assays to detect and identification of specific transgenic elements in 12 GM crops, viz. brinjal, cabbage, cauliflower, cotton, maize, mustard, okra, potato, rice, soybean, tomato and wheat have been developed.
• Real-time PCR assays for quantification of 24 GM events in eight crops, viz., cotton, maize, rice, brinjal, tomato, potato, wheat and cauliflower, targeting specific events/constructs/transgenes have been developed using TaqMan and SYBR®-Green I chemistry, in simplex and multiplex formats.

II. Transfer and Commercialization of Technologies

DNA based GM detection technologies have been transferred to :

1. M/s DSS Imagetech Private Limited, Delhi, facilitated by Agrinnovate India Limited (AgIn) on 19th August, 2015 

 Five DNA-based GMO screening technologies:
(i) Hexaplex PCR targeting six marker genes (aadA, bar, hpt, nptII, pat and uidA)
(ii) Duplex TaqMan Real-time PCR targeting P-35S and T-nos
(iii) Visual Loop-mediated Isothermal Amplification (LAMP)-based technology targeting eight transgenic elements (P-35S, T-nos, aadA, nptII, uidA, cry1Ac, cry2Ab, cp4-epsps)
(iv) Real-time LAMP-based technology targeting eight transgenic elements (P-35S, T-nos, aadA, nptII, uidA, cry1Ac, cry2Ab, cp4-epsps)
(v) TaqMan Real-time PCR-based multi-target system covering 47 targets for screening

Hexaplex PCR technology simultaneously targeting six marker genes, and duplex TaqMan Real-time PCR simultaneously targeting P-35S and T-nos, would be able to detect more than 90 per cent of globally commercialized GM events, including all commercialized GM cotton events in India
Visual and real-time LAMP technologies targeting eight transgenic elements can collectively screen more than 80 per cent of globally commercialized GM events, including all commercialized GM cotton events in India, facilitating rapid/cost-efficient on-site GMO screening.
TaqMan Real-time PCR based multi-target technology detecting 47 targets would assist in rapid GMO screening for a wide range of GM crops/events including cotton, maize, rice, brinjal, potato and soybean
These technologies provide efficient GMO screening tools to check the GM status of a sample irrespective of GM crop/trait, in a rapid/cost-efficient way. Visual and real-time LAMP technologies, when combined with fast DNA extraction.

2. Basmati Export Development Foundation (BEDA), APEDA, Meerut on non-exclusive basis.

3. Punjab Biotechnology Incubator (Agri and Food Testing Laboratory) Mohali, on non-exclusive basis.

3. Amar Immunodiagnostics Pvt Ltd. Hyderabad, in May 2010, on non-exclusive basis.                                                                                                                                                   

III. Consultancy Services for developing expertise in the area of  GMO Testing and Designing of GM Testing laboratory as per international Standards ISO/IEC 17025:2005  are being provided to:

1. Punjab Biotechnology Incubator (PBTI), (Agri and Food Testing Laboratory), Mohali.
               MoA, was signed on 6^th February, 2013
2. Basmati Export Development Foundation (BDEF), Modipuram, Meerut.
              MoA, was signed on 15^thJuly, 2013,

IV. International validations for quality assurance and global harmonization since 2010

• Participation in 26 Proficiency testings as per ISO/IEC 17043: 2010 for testing the unknown levels of GM events of maize, soybean, oilseed rape, rice using Real-time PCR assays and Ring trial for validation, since 2010:

• Thirteen proficiency testings organized by the European Union Reference Laboratory for GM Food and Feed, European Commission, Joint Research Centre, Italy

• Twelve proficiency testings of Grain Inspection, Packers and Stockyards Administration, United States, Department of Agriculture (GIPSA-USDA).
• One proficiency testing of Programas Interlaboratoriais e Materiais de Referência – PRIMAR Laboratório Nacional Agropecuário-LANAGRO/MG Ministério da Agricultura, Pecuária e Abastecimento, Brazil.

• Participated in ring trial organized by Federal Office of Consumer Protection and Food Safety, Germany in 2012 for the validation of two real-time PCR methods, viz., cry1Ab/Ac and pubi-cry frequently present in insect resistant GM rice lines and can be utilized to check GM contamination in rice.
• Participated in ring trial organized by EC-JRC, Italy in 2013 for detection and quantification of Golden Rice 2 (GR2) event.
• Validated multi-target Real-time PCR plate developed by the Institute for Health and Consumer Protection, EC-JRC, Italy in 2010-11 for individual detection of 39 GM events of 7 plants species (maize, cotton, rice, oilseed rape, soybean, sugar beet and potato) with 13 imported GM events of cotton and maize along with their species specific genes.

• Awarded “Panjabrao Deshmukh Outstanding Women Scientist Award, 2020”, National Award for Excellence in Agricultural Research by Indian Council of Agricultural Research, New Delhi
• Elected as “NAAS Fellow 2020” by National Academy of Agricultural Sciences, New Delhi.
• Awarded Honoray Fellow of the Academy (F.P.A.S.) under Agricultural Sciences during 23rd Punjab Science Congress in recognition of outstanding research contributions and achievements in the field of Agricultural Biotechnology.
• Agriculture Leadership Award 2015 by Agriculture Today for leading role in Biotechnology Research and Development on 18 September, 2015, Hotel Taj Palace, New Delhi.
• Best Scientist Award 2015 by Science and Technology, EET CRS, Research Wing for excellence in Professional Education and Industry, New Delhi on 14 June, 2015 at Bangalore.
• Recognition by OMICS International, USA for outstanding contributions in 7th Indo Global Summit and Expo on Food and Beverages, from 8-10 October, 2015.

International Recognitions

• Expert member of Secretariat, Convention on Biological Diversity, Montreal, United Nations Environment Programme, Joint Research Centre, Ispra, Italy.
  On invited sponsored visit to participate in Workshop of the Network of Laboratories for the Detection and Identification of LMOs at Ispra, Italy from 9-11 June, 2015.
  (i) Key note address delivered “Current Status of GMO Detection in India and Asian Region”
  (ii) While designing the outline for capacity-building workshops on the detection and identification of LMOs, a session was moderated on Reporting of analytical results in compliance with national and international regulations and practices.
•  Invited keynote speaker by Korean Society of Food Science and Technology as make a keynote address on “Cost-effective Novel Technologies to Check Unauthorized GM Events in Food Chain” in 81st Annual Meeting and International Conference in the session “New technologies in GM food detection for the future of food safety” at Republic of Korea from 25th- 27th August, 2014.
• Expert Member nominated by Ministry of Environment and Forests, Govt. of India and invited by Secretariat of the Convention on Biological Diversity (SCBD), United Nations Environment Programme, Montreal for the Workshop of the Network of Laboratories for the Detection and Identification of Living Modified Organisms at European Commission’s Joint Research Centre, Institute for Health and Consumer Protection, Ispra, Italy, from 25 to 27 November 2013.
• Expert Member of FAO (Regional Office for Asia and the Pacific), for GM testing and Biosafety: As a Resource person made three presentations during the Regional workshop on strengthening ‘Regional Cooperation and National Capacity Building on Biosafety in Asia’ at Bangkok, from 17-20 June 2013.
• Expert Member in ‘International workshop of GMO-analysis and networking’ and in brainstorming on ‘The present and future of DNA-based diagnostics within the broader context of bio-based economy’, sponsored by European Commission at  Joint Research Centre, Ispra, Italy, 8-10 April, 2013.
• Expert Member of FAO, Rome for GM testing and Biosafety: As Resource person for  GMO

Testing and Biosafety for Third Regional Workshop to exchange of information on GMO detection practices and techniques and their progressive harmonization at regional level at Beirut, Lebanon  from 24^th-27^th January, 2011.

Chaired two sessions
(i) Technical session on Biosafety
(ii) Plenary session

Delivered two key note addresses on:
(i) Harmonization of GMO Detection Strategies at Regional and Global Level;
(ii) Biosafety Clearing House in India

• Expert Member of European Networking on GM Detection Laboratories (ENGL) Ispra, Italy of  European Commission, Brussels, Belgium from  December, 2009-till date

Participated in four plenary meetings of European Networking on GM detection Laboratories (ENGL) at Ispra, Italy:
(i) 15^th ENGL meeting in May, 2011;
(ii) 14^th ENGL meeting in November, 2010; 
(iii) 13^th ENGL meeting in May, 2010;  
(iv) 12^th ENGL meeting in December, 2009

Key note address was made on Harmonisation of GMO Analysis in 13^th ENGL meeting held at Ispra, Italy, on 18^th May, 2010.

• Expert Member of European Commission-Asia Regional Network Meeting on GMO Analysis of European Commission, Brussels, Belgium from June, 2009-till date

Participated in Four European Commission-Asia Regional Network Meeting on GMO Analysis:

(i) 4^th European Commission-Asia Regional Network Meeting on GMO Analysis in June 2012 at Philippines;

(ii) 3^rd European Commission-Asia Regional Network Meeting on GMO Analysis in June 2011 at Singapore;

(iii) 2^nd European Commission-Asia Regional Network Meeting on GMO Analysis in June 2010 at Singapore;

(iv) 1^st European Commission-Asia Regional Network Meeting on GMO Analysis in June, 2009 at Malaysia.

Key note address on GMO Status and GMO Analysis in India was made in 2^nd European Commission-Asia Regional Network Meeting on GMO Analysis at Singapore on 22^nd June, 2010.

• Expert Member of FAO, Rome for GMO Testing: FAO's consultancy TCDC Expert, GCP/RAS/185/JPN, for ten days from 24^th November-5^th December, 2009.
• Resource person for GM testing in FAO sponsored Regional workshop on Biosafety at Bangkok from 30^thNovember- 4^th December, 2009. Delivered two key note addresses on: (i) GMO Detection: Emerging Scenario; (ii) Global and Regional initiatives in GM detection

• Group leader of Indian team for implementation of bilateral project on “Novel cost-effective methods for GMO detection” under Indo-Slovenian Inter Governmental Programme of Cooperation in Science and Technology by the Department of Science and technology (DST), Govt. of India and National Institute of Biology- Department of Biotechnology and Systems Biology, Slovenia. First and second exchange visits to Department of Biotechnology and System Biology at National Institute of Biology (NIB), Ljubljana, Slovenia were undertaken by me from 1^st-30^th September, 2011 and from 7^th June -6^th July, 2012. Two scientists from NIB, Slovenia visited twice in my GM detection laboratory in November, 2011 and December 2012, for sharing knowledge and know how in the area of GM detection.
• Expert Member of Indian Delegation to participate in International meeting Conference of Parties (COP)-Members of Parties (MOP)-6 of Convention on Biological Diversity (CBD), Montreal, held at Hyderabad, nominated by Ministry of Environment and Forests, (MoEF) for the theme areas of (i) Handling, Transport, Packing and Identification of LMOs  (ii) Capacity building and use of roster of experts. 1-5 October, 2012.

National Recognitions

1. Member as technical expert of five member Indian delegation Nominated by Director General, Bureau of Indian Standards, for participation in:

• 3^rd Plenary Meeting of “Horizontal methods for the detection of biomarkers in: foods; seeds and food crops; fruit; vegetables and derived foods” ISO/TC 34/ SC 16 at Beltsville, MD, USA from 25th-27th October, 2011.
• 2^nd Meeting of ISO/TC 34/ SC 16 “Horizontal methods for the detection of biomarkers Analysis at Tokyo, Japan from 9th-11th February, 2010.
• 1^st Meeting of “Horizontal methods for the detection of biomarkers in: foods; seeds and food crops; fruit; vegetables and derived foods” ISO/TC 34/ SC 16 at Chicago, IL, US from 11^th-13^th November, 2008.

2. Technical Expert Member of four member Indian delegation nominated by Ministry of Environment and Forests, Govt. of India under Global Environment Facility-(GEF) & World Bank Project for the Implementation of Cartagena Protocol on Biosafety & Ministry of Environment and Forests, Govt. of India. To discuss for further collaborations and linkages in the area of different biosafety and regulatory aspects of GM crops, and in United States visited: (i) Michigan State University, Michigan (ii) US Department of Agriculture (USDA) (iii) World Bank, GEF Secretariat Washington DC (iv) Cornell University, Cornell.
   In Canada, visited: (i) Canadian Food Inspection Agency (ii) Environment Canada, Ottawa (iii) Canadian Seed Growers’ Association, Ottawa (iv) Secretariat, Convention on Biological Diversity, Montreal.

Member of National Committees

1. Member of Scientific Panel on Genetically Modified Organisms and Foods of Food Safety and Standards Authority of India (FSSAI) 2020.
2. Member of Research Advisory Committee of Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow 2021.
3. Technical Expert member of Accreditation Committee of National Accreditation Board for testing and Calibration of Laboratories (NABL), Ministry of Science and Technology, New Delhi, from July, 2012 till date.
4. Technical Expert member in the area of GM testing of National Accreditation Board for testing and Calibration of Laboratories (NABL), Ministry of Science and Technology, Govt. of India, to evaluate the technical competence of national public and private sector laboratories involved in GMO testing from 2007 till date. As Technical Expert for GMO testing of NABL conducted assessment of seven National Laboratories for GM testing for issuance of NABL certificate for GM testing.
5. Technical Member of National Committee of National Accreditation Board for Testing and Calibration (NABL) for revising 102 NABL Guidelines for Biological Testing Laboratories specifically for GM testing.Technical inputs were extended in six meetings held at NABL Secretariat between April 2011-December, 2011.
6. Technical Expert member of Biotechnology for Food & Agriculture Sectional Committee, FAD 23, of Bureau of Indian Standards (BIS) Manak Bhavan, New Delhi, to evaluate the Indian Standards especially for testing of GM  crops and  GM  food products from 2010.

Patents Granted by Indian Patent Office New Delhi

1. Patent No. 258165 Randhawa GJ and Firke PK (2013) “Diagnostic kit based on polymerase chain reaction for detection of cry1Ac gene in Bt cotton Bollgard-I”
   2069/DEL/2006; Priority date: 20^th September, 2006
2. Patent No. 254341 Randhawa GJ et al, (2012) “Process enabling simultaneous detection of two transgenes namely human serum albumin (HAS) and bar genes using a Multiplex Polymerase Chain Reaction utilizing a combination of novel primers and Polymerase Chain Reaction in transgenic wheat”
   3530/DEL/2005; Priority date 30^th December, 2005
3. Patent No. 245749 Randhawa GJ et al, (2011) “Process enabling simultaneous detection of two transgenes namely 5 enolpyruvylshikimate-3-phosphate synthase gene and CaMV 35S promoter using a multiplex polymerase chain reaction utilizing combination of novel primers and polymerase chain reaction in transgenic maize”
   3451/DEL/2005; Priority date: 23^rd December, 2005

Research Publications (During the last five years)

1. M Singh, KK Sodhi, A Paliwal, S Sharma, GJ Randhawa (2021) Efficient DNA Extraction Procedures for Processed Food Derivatives – A Critical Step to Ensure Quality for GMO Analysis. Food Analytical Methods 14: 2249–2261 https://doi.org/10.1007/s12161-021-02051-y (NAAS rating: 8.67; Impact Factor: 3.366)*
2. SI Kuwardadra, KC Bhatt, A Paliwal, P Sood, PK Malav, KK Sodhi & GJ Randhawa (2020) Monitoring adventitious presence of transgenes in brinjal (Solanum melongena L.) collections from the regions in India bordering Bangladesh: a case report. Genetic Resources and Crop Evolution, 67: 1181–1192. https://doi.org/10.1007/s10722-020-00903-w
3. SI Kuwardadra, CD Pandey, A Paliwal & GJ Randhawa (2020) Screening brinjal accessions conserved in the National Genebank collected from North Eastern states of India for adventitious presence of EE1 event. Indian Journal of Agricultural Sciences. (Accepted).
4. M Singh, D Pal, P Sood & GJ Randhawa (2020) Construct-specific loop-mediated isothermal amplification: Rapid detection of genetically modified crops with insect resistance or herbicide tolerance. Journal of AOAC International https://doi.org/10.1093/jaoacint/qsaa043.
5. M Singh, GJ Randhawa, RK Bhoge, Sushmita, A Kak & O Sangwan (2020) Absence of adventitious presence of transgenes in cotton collections from genebank and experimental plots: Ensuring GM-free conservation and cultivation of genetic resources. Agricultural Research, https://doi.org/10.1007/s40003-019-00449-z.
6. M Singh, D Pal, P Sood & GJ Randhawa (2019) Loop-mediated isothermal amplification assays: Rapid and efficient diagnostics for genetically modified crops. Food Control, 106: 106759.
7. M Singh, RK Bhoge, S Nain & GJ Randhawa (2019) Loop-mediated isothermal amplification: A rapid detection method for rice actin and nopaline synthase promoters in genetically modified crops. Journal of Plant Biochemistry & Biotechnology. 28 (3): 353-356. 
8. M Singh, RK Bhoge & GJ Randhawa (2018) Loop-mediated isothermal amplification for detection of endogenous Sad1 gene in cotton: An internal control for rapid on-site GMO testing. Journal of AOAC International, 101(5):1657-1660.
9. M Singh, RK Bhoge & GJ Randhawa (2017) Real-time and visual loop-mediated isothermal amplification: Efficient GMO screening targeting pat and pmi genes. Food Control, 71: 248-254.
10. Randhawa GJ, Singh M & Sood P (2016) DNA-based methods for detection of genetically modified events in food and supply chain. Curr. Sci. 110 (6): 1000-1009.
11. Singh M & Randhawa GJ (2016) Transboundary movement of genetically modified organisms in India: Current scenario and a decision support system. Food Cont. 68: 20-24.
12. Singh M, RK Bhoge & GJ Randhawa (2016) Crop-specific GMO matrix-multiplex PCR: A cost-efficient screening strategy for GM maize and cotton events approved globally. Food Cont. 70: 271-280.
13. Bairwa RK, Singh M, Bhoge RK, Devi C & Randhawa GJ (2016) Monitoring adventitious presence of transgenes in ex situ okra (Abelmoschus esculentus) collections conserved in genebank: a case study. Genetic Resou. Crop Evol. 63 (2): 175-184.
14. Bhoge RK, Singh M, Chhabra R, Sathiyabama  M & Randhawa GJ (2016) Multiplex real-time PCR-based detection and quantification of GM maize events employing SYBR® Green I and TaqMan® chemistries. Curr. Sci. 110(8) 1446-1451.