Stacking of cry1Ab and Chitinase Genes in Commercial Cotton Varieties through Crossing

Document Type : Original Research

Authors
S. Mirzaei 1
S. Dezhsetan 1
M. Tohidfar 2
1 Department of Agronomy and Plant Breeding, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Islamic Republic of Iran.
2 Department of Plant Biotechnology, Faculty of Life Science and Biotechnology, Shahid Beheshti University, GC, Tehran, Islamic Republic of Iran.
Abstract
Cotton (Gossypium hirsutum) is one of the most important fiber crops with a raw material value of $5.5 billion per year worldwide. Cotton breeders with the help of conventional breeding programs have improved the cotton germplasm for resistance to pests and pathogens. However, there has been little success in producing cotton lines resistant to biotic stresses, due to limited cotton germplasm with adequate resistance to biotic stresses, especially pests and diseases. In this study, cry1Ab (resistance to cotton bollworm) and chitinase (resistance to cotton Verticillium wilt) genes were transferred to Iranian commercial cotton varieties (Varamin, Khordad, Sahel, and Bakhtegan), by crossing transgenic lines containing cry1Ab and chi transgenes with Iranian commercial varieties. To recover genetic background of commercial varieties, the obtained progenies were backcrossed with own commercial parent. The progenies of backcrosses were assessed by western blot and PCR analysis. Insect and fungus in vitro bioassay showed resistance against cotton bollworm and Verticillium dahliae in plants containing the stacked transgenes. According to the results, cry1Ab and chi genes were successfully transferred and stacked in Iranian commercial cotton varieties. Also, plants with two transgenes and plants with one transgene were protruded to BC1 and BC2 generations, respectively.

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Subjects

1. Agapito-Tenfen, S. Z., Vilperte, V., Benevenuto, R. F., Rover, C. M., Traavik, T. I. and Nodari R. O. 2014. Effect of Stacking Insecticidal Cry and Herbicide Tolerance Epsps Transgenes on Transgenic Maize Proteome. BMC Plant Biol., 14: 346.
2. Agrawal, D., Banerjee, A., Kolala, R., Dhage, A., Kulkarni, A., Nalawade, S., Hazra, S. and Krishnamurthy K. 1997. In Vitro Induction of Multiple Shoots and Plant Regeneration in Cotton (Gossypium hirsutum L.). Plant Cell Rep., 16: 647-652.
3. Awan, M. F. Abass, M. A., Muzaffar, A., Ali, A., Tabassum, B., Rao, A. Q., Ahmad Nasir, I. and Husnain, T. 2015. Transformation of Insect and Herbicide Resistance Genes in Cotton (Gossypium hirsutum L.). J. Agr. Sci. Tech., 17: 287-298.
4. Bakhsh, A., Siddique, S. and Husnain, T. 2012. A Molecular Approach to Combat Spatio-Temporal Variation in Insecticidal Gene (Cry1Ac) Expression in Cotton. Euphytica, 183: 65-74.
5. Bakhsh, A., Anayol, E., Ozcan, S.F., Hussain, T., Aasim, M., Khawar, K.M. and Ozcan, S. 2015. An Insight into Cotton Genetic Engineering (Gossypium hirsutum L.): Current Endeavors and Prospects. Acta Physiol. Plant, 37: 1-17.
6. Cheng, H., Jian, G., Ni, W., Yang, H., Wang, Z., Sun, W., Zhang, B., Wang, X., Ma, C. and Jia, S. 2005. Increase of Fusarium-and Verticillium-Resistance by Transferring Chitinase and Glucanase Gene into Cotton. Sci. Agric. Sinica, 38: 1160-1166.
7. Datta, K., Baisakh, N., Thet, K.M., Tu, J. and Datta, S. K. 2002. Pyramiding Transgenes for Multiple Resistance in Rice Against Bacterial Blight, Yellow Stem Borer and Sheath Blight. Theor. Appl. Genet., 106: 1-8.
8. Doyle, J. 1991. DNA Protocols for Plants Molecular Techniques in Taxonomy. Springer, PP. 283-293.
9. Gallic, D. R., Walbot, V. and Hershey, J. W. 1988. The Ribosomal Fraction Mediates the Translational Enhancement Associated with the 5′-Leader of Tobacco Mosaic Virus. Nucl. Acids Res., 16: 8675-8694.
10. Halpin, C. 2005. Gene Stacking in Transgenic Plants: The Challenge for 21st Century Plant Biotechnology. Plant Biotechnol. J., 3: 141-155.
11. James, C. 2015. Global Status of Commercialized Biotech/GM Crops: 2015 ISAAA Brief No. 51. ISAAA, Ithaca, NY.
12. Katageri, I., Vamadevaih, H., Udikeri, S., Khadi, B. and Kumar, P.A. 2007. Genetic Transformation of an Elite Indian Genotype of Cotton (Gossypium hirsutum L.) for Insect Resistance. World Cotton Research Conference-4, International Cotton Advisory Committee (ICAC), 10-14 September 2007, Lubbock, Texas, USA.
13. Khan, G.A., Bakhsh, A., Ghazanfar, M., Riazuddin, S. and Husnain, T. 2013. Development of Transgenic Cotton Lines Harboring a Pesticidal Gene (cry1Ab). EJFA, 25: 434.
14. Lukonge, E. P., Labuschagne, M. T. and Herselman, L. 2008. Combining Ability for Yield and Fibre Characteristics in Tanzanian Cotton Germplasm. Euphytica, 161: 383-389.
15. Ma, Z., Liu, J. and Wang X. 2013. Agrobacterium-Mediated Transformation of Cotton (Gossypium hirsutum) Shoot Apex with a Fungal Phytase Gene Improves Phosphorus Acquisition. Methods Mol. Biol., 958: 211-22.
16. Maqbool, S., Riazuddin, S., Loc, N., Gatehouse, A. R., Gatehouse, J. and Christou, P. 2001. Expression of Multiple Insecticidal Genes Confers Broad Resistance Against a Range of Different Rice Pests. Mol. Breed., 7: 85-93.
17. McCarty, J. C., Jenkins, J. N. and Wu, J. 2004. Primitive Accession Derived Germplasm by Cultivar Crosses as Sources for Cotton Improvement. Crop Sci., 44: 1226-1230.
18. Rao, A. Q., Irfan, M., Saleem, Z., Nasir, I.A., Riazuddin, S. and Husnain, T. 2011. Overexpression of the phytochrome B Gene from Arabidopsis thaliana Increases Plant Growth and Yield of Cotton (Gossypium hirsutum). J. Zhejiang Univ. Sci. B, 12: 326-334.
19. Rios, R. O. 2015. Plant Breeding in the Omics Era. Springer, PAGES???
20. Satyavathi, V., Prasad, V., Lakshmi, B. G. and Sita, G. L. 2002. High Efficiency Transformation Protocol for Three Indian Cotton Varieties via Agrobacterium tumefaciens. Plant Sci., 162: 215-223.
21. Senthilkumar, R., Cheng, C. -P. and Yeh, K. -W. 2010. Genetically Pyramiding Protease-Inhibitor Genes for Dual Broad-Spectrum Resistance Against Insect and Phytopathogens in Transgenic Tobacco. Plant Biotechnol. J., 8: 65-75.
22. Sharma, K. K. and Ortiz, R. 2000. Program for the Application of Genetic Transformation for Crop Improvement in the Semi-Arid Tropics. In Vitro Cell Dev. Biol. Plant, 36: 83-92.
23. Smith, R. H., Ulian, E. and Gould, J. H. 1990. Transformation of Plants via the Shoot Apex Plant Cell and Tissue Culture. Springer, PP. 335-340.
24. Taverniers, I., Papazova, N., Bertheau, Y., De Loose, M. and Holst-Jensen, A. 2008: Gene Stacking in Transgenic Plants: Towards Compliance Between Definitions, Terminology, and Detection Within the EU Regulatory Framework. Environ. Biosafety Res., 7: 197-218.
25. Tohidfar, M., Mohammadi, M. and Ghareyazie, B. 2005. Agrobacterium-Mediated Transformation of Cotton (Gossypium hirsutum) using a Heterologous Bean Chitinase Gene. Plant Cell Tiss. Organ Cult., 83: 83-96.
26. Tohidfar, M., Hossaini, R., Sokhandan Bashir, N. and Tabatabaei, M. 2012. Enhanced Resistance to Verticillium dahliae in Transgenic Cotton Expressing an Endochitinase Gene from Phaseolus vulgaris. Czech J. Genet Plant Breed., 48: 33-41.
27. Tohidfar, M., Ghareyazie, B., Mosavi, M., Yazdani, S. and Golabchian, R. 2008. Agrobacterium-Mediated Transformation of Cotton (Gossypium hirsutum) using a Synthetic cry1Ab Gene for Enhanced Resistance Against Heliothis armigera. IJB, 6: 164-173.
28. VafaieTabar, M. and Tajik Khave, Z. 2013. Evaluation of Quantitative Traits Correlation and Diversity of Early Cotton Varieties (Gossypium hirsutumL.) in Varamin Region. Iran. J. Agric. Sci., 1: 97-114. (in Persian)
29. Vinodhana, N. K., Ganesan, N. M. and Rajeswari, S. 2014. Development of Insect Resistant Transgenic Lines Mediated by Agrobacterium in Cotton (Gossypium hirsutum L.). Mol. Plant Breed., 5. 24-28.
30. Wang, Z., Yu, C. and Jiang, L. 2012. Segregation and Expression of Transgenes in the Progenies of Bt Transgenic Rice Crossed to Conventional Rice Varieties. Afr. J. Biotechnol., 11. 7812-7818.
31. Zeng, L., Meredith, W. R. and Boykin, D. L. 2011. Germplasm Potential for Continuing Improvement of Fiber Quality in Upland Cotton: Combining Ability for Lint Yield and Fiber Quality. Crop Sci., 51: 60-68.
32. Zhao, Q. C., Liu, M. H., Zhang, X. W. Lin, C. Y., Zhang, Q. and Shen, Z. C. 2015. Generation of Insect-Resistant and Glyphosate-Tolerant Rice by Introduction of a T-DNA Containing Two Bt Insecticidal Genes and an EPSPS Gene. J. Zhejiang Univ. Sci. B, 16: 824-31.
33. Zhu, Z., Xu, X., Cao, B., Chen, C., Chen, Q., Xiang, C., Chen, G. and Lei, J. 2015. Pyramiding of AtEDT1/HDG11 and Cry2Aa2 into Pepper (Capsicum annuum L.) Enhances Drought Tolerance and Insect Resistance Without Yield Decrease. Plant Cell Tiss. Organ Cult., 120: 919-932.
Journal of Agricultural Science and Technology
Volume 20, Issue 6
November and December 2018
Pages 1259-1268