Trackable CEMB-Klean Cotton Transgenic Technology: Affordable Climate Neutral Agri-biotech Industrialization for Developing Countries

Zahida Qamar, Muhammad Tariq, Tahir Rehman, Muhammad Shahzad Iqbal, Muhammad Bilal Sarwar, Muhammad Nauman Sharif, Zohaib Hassan, Ayesha Ahmad, Aiman Zahra, Ayesha Latif, Bushra Rashid, Bushra Tabassum, Sameera Hassan, Allah Baksh, Saira Azam, Shafique Ahmed, Kamran Shahzad Bajwa, Mudassar Fareed Awan, Naila Shahid, Arfan Ali, Abdul Munim Farooq, Abdul Qayyum Rao, Muhammad Saleem Haider, Tassawar Hussain Malik, Idrees Ahmad Nasir

Abstract


Background: Transgenic technology reflects the incorporation of novel useful traits in crop plants like cotton for economic benefits by overcoming the problems including insects’ pests and weeds in special. The present study is the success story of the continuous effort of CEMB team started back in the 1990s.

Methods: This study includes characterization of a large number of Bacillus thuringiensis (Bt) strains taken from local soil and subjected to direct transformation of isolated BT genes into local cotton cultivars. Protocols for transformation into cotton plants were optimized and validated by the development of double gene codon optimized (Cry1Ac and Cry2A) transgenic cotton varieties.

Results: The resulting GMOs in the form of CEMB-33, CA-12, CEMB-66 have been approved by Punjab Seed Council in 2013 and 2016 respectively. Double Bt and weedicide resistant cotton harboring CEMB-Modified and codon optimized cp4EPSPS (GTGene). These varieties can tolerate glyphosate spray @ 1900ml per acre without the appearance of necrotic spots/shedding and complete removal of all surrounding weeds in the cotton field is a significant advance to boost cotton production without spending much on insecticides and herbicides.

Conclusion: In the current report, two unique sets of primers which amplify 1.1 Kb for CEMB-double Bt genes and 660 bp product for CEMB-Modified cp4EPSPS (GTGene) were tested. CEMB cotton variety CKC-01 is specially designed as low cost and easy to use by local farmer’s technology has the potential to revolutionize the cotton growing culture of the country.


Keywords: Event detection; Bt Cotton; CEMB transgenic technology; GTGene


Full Text:

PDF

References


Shahid AA, Shaukat MS, Bajwa KS, Rao AQ, Husnain T. Detection of Transgenes in Cotton (Gossypium hirsutum L.) by Using Biotechnology/Molecular Biological Techniques. International Journal of Biological, Biomolecular, Agricultural, Food and Biotechnological Engineering, (2015); 9(1): 87-92.

Puspito AN, Rao AQ, Hafeez MN, Iqbal MS, Bajwa KS, et al. Transformation and Evaluation of Cry1Ac+Cry2A and GTGene in Gossypium hirsutum L. Front Plant Sci, (2015); 6(943): 1-13.

Abdullah A. An analysis of Bt cotton cultivation in Punjab, Pakistan using the agriculture Decision Support System (ADSS). AgBioForum, (2010); 13(3): 274-287.

Spielman DJ, Zaidi F, Zambrano P, Khan AA, Ali S, et al. What are farmers really planting? Measuring the presence and effectiveness of Bt cotton in Pakistan. PloS one, (2017); 12(5): e0176592.

Asi MR, Afzal M, Anwar SA, Bashir MH. Comparative Efficacy Of Insecticides Against Sucking Insect Pests of Cotton. Pakistan J Life Soc Sci, (2008); 6(2): 140-142.

Farooq A, Farooq J, Mahmood A, Shakeel A, Rehman KA, et al. An overview of cotton leaf curl virus disease (CLCuD) a serious threat to cotton productivity. Australian Journal of Crop Science, (2011); 5(13): 1823-1831.

Kamle M, Kumar P, Patra JK, Bajpai VK. Current perspectives on genetically modified crops and detection methods. 3 Biotech, (2017); 7(3): 1-15.

Latif A, Rao AQ, Khan MA, Shahid N, Bajwa KS, et al. Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal. BMC Res Notes, (2015); 8(1): 1-8.

Naqvi RZ, Asif M, Saeed M, Asad S, Khatoon A, et al. Development of a Triple Gene Cry1Ac-Cry2Ab-EPSPS Construct and Its Expression in Nicotiana benthamiana for Insect Resistance and Herbicide Tolerance in Plants. Front Plant Sci, (2017); 8(55): 1-9.

Muzaffar A, Kiani S, Khan MA, Rao AQ, Ali A, et al. Chloroplast localization of Cry1Ac and Cry2A protein--an alternative way of insect control in cotton. Biol Res, (2015); 48(1): 14.

Sarwar MB, Batool F, Rashid B, Aftab B, Hassan S, et al. Integration and expression of heat shock protein gene in segregating population of transgenic cotton for drought tolerance. Pakistan Journal of Agricultural Sciences, (2014); 51(4): 1-11.

Cressey D. Transgenics: A new breed. Nature, (2013); 497(1): 27-29.

Dong H, Li W. Variability of endotoxin expression in Bt transgenic cotton. Journal of Agronomy and Crop Science, (2007); 193(1): 21-29.

Rashid B, Saleem Z, Husnain T, Riazuddin S. Transformation and inheritance of Bt genes in Gossypium hirsutum. Journal of Plant Biology, (2008); 51(4): 248-254.

Pray CE, Naseem A. Supplying crop biotechnology to the poor: opportunities and constraints. The Journal of Development Studies, (2007); 43(1): 192-217.

Dill GM. Glyphosate-resistant crops: history, status and future. Pest Manag Sci, (2005); 61(3): 219-224.

Riazuddin S, Husnain T, Khan E, Khanum F. Insect resistant transgenic basmati rice. Rice Biotechnology Quarterly, (1995); 23(1): 7-8.

Husnain T. Transformation of indica rice with synthetic cry1Ac gene. Biologica, (1998); 44(1): 180-192.

Maqbool SB, Husnain T, Riazuddin S, Masson L, Christou P. Effective control of yellow stem borer and rice leaf folder in transgenic rice indica varieties Basmati 370 and M 7 using the novel δ-endotoxin cry2A Bacillus thuringiensis gene. Molecular Breeding, (1998); 4(6): 501-507.

Husnain T, Asad J, Maqbool SB, Datta SK, Riazuddin S. Variability in expression of insecticidal Cry1Ab gene in Indica Basmati rice. Euphytica, (2002); 128(1): 121-128.

Azam S, Samiullah TR, Yasmeen A, ud Din S, Iqbal A, et al. Dissemination of Bt cotton in cotton growing belt of Pakistan. Adv life sci, (2013); 1(1): 18-26.

Nazli H, Sarker R, Meilke KD, Orden D (2010) Economic performance of Bt cotton varieties in Pakistan. Denver, Colorado Agricultural and Applied Economics Association’s 2010 AAEA, CAES & WAEA Joint Annual Meeting.

Höfte H, Whiteley HJM, Reviews MB. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev, (1989); 53(2): 242-255.

Islam R, Malik T, Husnain T, Riazuddin S. Strain and cultivar specificity in the Agrobacterium-chickpea interaction. Plant Cell Rep, (1994); 13(10): 561-563.

Husnain T, Malik T, Riazuddin S, Gordon MPJPc, tissue, culture o. Studies on the expression of marker genes in chickpea. Plant Cell, Tissue and Organ Culture, (1997); 49(1): 7-16.

Husnain TJB. Transformation of indica rice with synthetic cry1Ac gene. (1998); 44180-192.

Chaudhry B, Yasmin A, Husnain T, Riazuddin SJPMBR. Mini-scale genomic DNA extraction from cotton. Plant Molecular Biology Reporter (1999); 17(3): 280-280.

MAJEED A, HUSNAIN T, Riazuddin S. Transformation of virus-resistant genotype of Gossypium hirsutum L. with pesticidal gene. Plant Biotechnology, (2000); 17(2): 105-110.

Allah B, Rao A, Khan G, Bushra R, Shahid A, et al. Insect resistance studies of transgenic cotton cultivar harboring cry1Ac and cry2A. CAB Direct, (2012); 5(2): 167-171.

Pandey A, Kamle M, Yadava L, Muthukumar M, Kumar P, et al. Genetically modified food: its uses, future prospects and safety assessments. Biotechnology, (2010); 9(4): 444-458.

Delaney B. Safety assessment of foods from genetically modified crops in countries with developing economies. Food Chem Toxicol, (2015); 86(2015): 132-143.

Fraiture M-A, Herman P, Taverniers I, De Loose M, Van Nieuwerburgh F, et al. Validation of a sensitive DNA walking strategy to characterise unauthorised GMOs using model food matrices mimicking common rice products. Food Chemistry, (2015); 173(15): 1259-1265.

Fraiture MA, Herman P, Lefevre L, Taverniers I, De Loose M, et al. Integrated DNA walking system to characterize a broad spectrum of GMOs in food/feed matrices. BMC Biotechnol, (2015); 15(76): 1-11.

Zhang X, Tang Q, Wang X, Wang Z. Structure of exogenous gene integration and event-specific detection in the glyphosate-tolerant transgenic cotton line BG2-7. PLoS One, (2016); 11(7): e0158384.

James D, Schmidt AM, Wall E, Green M, Masri S. Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis. J Agric Food Chem, (2003); 51(20): 5829-5834.

Mäde D, Degner C, Grohmann LJEFR, Technology. Detection of genetically modified rice: a construct-specific real-time PCR method based on DNA sequences from transgenic Bt rice. European Food Research and Technology, (2006); 224(2): 271-278.

Mannerlöf M, Tenning P. Screening of transgenic plants by multiplex PCR. J Plant Molecular Biology Reporter, (1997); 15(1): 38-45.


Refbacks

  • There are currently no refbacks.