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:: Volume 14, Issue 1 (9-2025) ::
gebsj 2025, 14(1): 89-101 Back to browse issues page
Transformation of human IFN-α (HU-IFN-α) gene to Urmia cultivar of tomato using Agrobacterium tumefaciens
Vahid Mehrizadeh , Ebrahim Dorani *
Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Iran , dorani@tabrizu.ac.ir
Abstract:   (863 Views)
Interferons, especially human interferon Alpha, have high therapeutic and economic value and there is a strong interest in producing them from stable, safe and inexpensive sources such as plants. The aim of this study was investigated the possibility of transformation of tomato with human interferon Alpha2b. For this purpose, some of the effective factors on the transfer of the human IFN-α gene to tomato, such as the type of explant, Agrobacterium concentration (OD600nm = 0.2, 0.4, 0.6, 0.8 and 1), inoculation time (5, 10, 15, 20 and 25 minutes), co-cultivation time (1, 2, 3 and 4 days) and different concentration of the hygromycin (0, 7.5, 10, 15 and 20 mg/l) were investigated. The results showed that the cotyledon explants were the most suitable explant and 15 mg/L of hygromycin was the effective concentration of the antibiotic in transformation of tomato. Also, the highest percentage of transformation of tomato explants was observed in the OD600nm= 0.6 of Agrobacterium, 20 minutes of inoculation and 3 days of co-cultivation. Finally, the presence of human IFN-α gene in the probable transgenic plants was confirmed by the PCR technique and specific primers.
Keywords: Agrobacterium, Transformation, Interferon Alpha, Recombinant Protein, Tomato
Full-Text [PDF 701 kb]   (372 Downloads)    
Type of Study: Research | Subject: Plant
Received: 2025/03/11 | Accepted: 2025/08/27 | Published: 2025/10/28
References
1. Afroz, A., Chaudhry, Z., Rashid, U., Rashid, M. and Muhammad, C. (2010). Enhanced regeneration in explants of tomato (Lycopersicon Esculentum L.) with the treatment of coconut water. African Journal of Biotechnology, 9(24), 3634-3644.
2. Ahmed, S., Wan-Azizan, W.A.S., Akhond, M.A.Y., Juraimi, A.S., Ismail, S.I., Ahmed, R. and Md-Hatta, M.A. (2023). Optimization of in vitro regeneration protocol of tomato cv. MT1 for genetic transformation. Horticulturae, 9 (800), 1-21. doi.org/10.3390/horticulturae9070800 [DOI:10.3390/horticulturae9070800]
3. Bamishaiye, E.I., Balakrishnan, N., Udayasuriyan, V., Varanavasiappan, S. and Sudhakar, D. (2017). A rapid Agrobacterium-mediated transformation protocol for tomato (Solanum lycopersicum L.) cv. PKM-1. International Journal of Chemical Studies, 5(6), 1024-1030. [DOI:10.20546/ijcmas.2017.612.495]
4. Bharathi, J.K., Suresh, P., Prakash, M.A.S. and Muneer, S. (2024). Exploring recent progress of molecular farming for therapeutic and recombinant molecules in plant systems. Heliyon, 10(18), 1-30. doi.org/10.1016/j.heliyon. 2024.e37634 [DOI:10.1016/j.heliyon.2024.e37634] [PMID] [PMCID]
5. Doranie-uliaie, E. and Mehrizadeh, V. (2017). Optimization of Agrobacterium tumefaciens-mediated transformation of barley and production of fertile transgenic plants. Genetic Engineering and Biosafety Journal, 5(2), 131-142. dor: 20.1001.1.25885073.1395.5.2.7.4
6. Ferdous, M.E.M., Datta, A. and Islam, A. (2022). Evaluation of factors influencing in vitro regeneration and transformation protocols to produce salinity tolerant tomato (Solanum lycopersicum L.). Journal of Applied Biotechnology Reports, 9(3), 726-739. doi 10.30491/JABR.2022.336979.1519
7. Fischer, R. and Buyel, J.F. (2020). Molecular farming - The slope of enlightenment. Biotechnology Advances, 40(1), 1-16. doi.org/10.1016/j.biotechadv.2020.107519 [DOI:10.1016/j.biotechadv.2020.107519] [PMID] [PMCID]
8. Gadir, I.K.A., Wahid, H.H.A., El-Hussein, A.A. and El-Siddig, M.A. (2018). Effect of Agrobacterium-mediated transformation on regeneration efficiency of tomato explants. International Journal of Biotechnology and Bioengineering, 4 (5), 106-112.
9. Gerszberg, A., Hnatuszko-Konka, K., Kowalczyk, T. and Kononowicz, A.K. (2015). Tomato (Solanum lycopersicum L.) in the service of biotechnology. Plant Cell, Tissue and Organ Culture, 120(3), 881-902. doi:10.1007/ s11240-014-0664-4 [DOI:10.1007/s11240-014-0664-4]
10. Guo, M., Zhang, Y.L., Meng, Z.J. and Jiang, J. (2012). Optimization of factors affecting Agrobacterium-mediated transient expression of Micro-Tom tomatoes. Genetics and Molecular Research, 11(1), 661-671. doi: 10. 4238/2012.March.16.4 [DOI:10.4238/2012.March.16.4] [PMID]
11. Hassan, N., El-Shafey, N.M., Khodary, S.E.D.A., El- Shabrawi, H. and Badr, A. (2021). Expression of OsDREB2A in transgenic tomato improves drought tolerance. Romanian Biotechnological Letters, 26(6), 3145-3154. doi: 10.25083/rbl/26.6/3145-3154 [DOI:10.25083/rbl/26.6/3145-3154]
12. Honda, C., Ohkawa, K., Kusano, H., Teramura, H. and Shimada, H. (2021). A simple method for in planta tomato transformation by inoculating floral buds with a sticky Agrobacterium tumefaciens suspension. Plant Biotechnology, 38(1), 153-156. doi: 10.5511/plantbiotechnology.20.0707a [DOI:10.5511/plantbiotechnology.20.0707a] [PMID] [PMCID]
13. Jan, S.A., Ali, G.M., Ali, S., Shah, S.H. and Ahmad, N. (2018). Genetic improvement in tomato (Solanum lycopersicum) against salt stress. Indian Journal Biotechnology, 17(3), 459-465.
14. Jiang, M.C., Hu, C.C., Lin, N.S., and Hsu, Y.H. (2019). Production of human IFN-γ protein in Nicotiana benthamiana plant through an enhanced expression system based on bamboo mosaic virus. Viruses, 11(509), 1-18. doi:10. 3390/v11060509 [DOI:10.3390/v11060509] [PMID] [PMCID]
15. Kaul, R., Thangaraj, A., Sharda, S. and Kaul, T. (2025). Optimization of tissue culture and Cas9 transgene expression in tomato: A step towards CRISPR/Cas9-based genetic improvement. Plant Science, 352, 1-14. doi.org/10.1016/ j. plantsci.2024.112324 [DOI:10.1016/j.plantsci.2024.112324] [PMID]
16. Kim, M.J., Baek, K. and Park, C.M. (2009). Optimization of conditions for transient Agrobacterium-mediated gene expression assays in Arabidopsis. Plant Cell Reports, 28(8), 1159-1167. doi: 10.1007/s00299-009-0717-z [DOI:10.1007/s00299-009-0717-z] [PMID]
17. Kumar, S., Jindal, S.K., Sarao, N.K. and Dhaliwal, M.S. (2017). Development of an efficient in vitro regeneration protocol in tomato (Solanum lycopersicum L.). Agriculture Research Journal, 54 (4), 475-479. doi:10.5958/2395-146X.2017.00091.6 [DOI:10.5958/2395-146X.2017.00091.6]
18. Lee, H., Auh, C.K., Kim, D., Lee, T.K. and Lee, S. (2006). Exogenous cytokinin treatment maintains cyclin homeostasis in rice seedlings that show changes of cyclin expression when the photoperiod is rapidly changed. Plant Physiology and Biochemistry, 44(4), 248-52. doi.org/10.1016/j.plaphy.2006.03.006 [DOI:10.1016/j.plaphy.2006.03.006] [PMID]
19. Lopez, E., Proano, K., Jadan, M. and Mihai, R. (2015). Callus tissue induction and analysis of GUS reporter gene expression in tomato (Solanum lycopersicum L.) transformed with Agrobacterium tumefaciens. Romanian Biotechnological Letters, 20(2), 10205-10211.
20. Manamohan, M., Prakash, M.N., Sharath Chandra, G., Asokan, R. and Nagesh, S.N. (2011). An improved protocol for rapid and efficient Agrobacterium mediated transformation of tomato (Solanum lycopersicum L.). Journal of Applied Horticulture, 13(1), 3-7. doi:10.37855/jah. 2011.v13i01.01 [DOI:10.37855/jah] [PMCID]
21. Mathews, H., Clendennen, S.K., Caldwell, C.G., Liu, X.L., Connors, K., Matheis, N., Schuster, D.K., Menasco, D.J., Wagoner, W., Lightner, J., and Wagner, D.R. (2003). Activation tagging in tomato identifies a transcriptional regulator of anthocyanin biosynthesis, modification, and transport. The Plant Cell, 15(8), 1689-1703. doi:10. 1105/tpc.012963 [DOI:10.1105/tpc.012963] [PMID] [PMCID]
22. Mehrizadeh, V., Dorani, E., Mohammadi, A. and Ghareyazi, B. (2021). Study on the factors affecting Agrobacterium-mediated transformation of soybean. Genetic Engineering and Biosafety Journal, 9(2), 225-236. dor:20.1001.1.25885073.1399.9.2.13.8
23. Mikulecky, P., Zahradnik, J., Kolenko, P., Cerny, J., Charnavets, T., Kolarova, L., Necasova, I., Pham, P.N., Schneider, B. (2016). Crystal structure of human interferon-gamma receptor 2 reveals the structural basis for receptor specificity. Acta Crystallographica Section D: Structural Biology, 72(9), 1017-1025. doi: 10.1107/ S2059798316012237 [DOI:10.1107/S2059798316012237] [PMID] [PMCID]
24. Mohajel-Kazemi, E., Pazhouhandeh, M., Jonoubi, P. and Kazemian, M. (2020). The optimization of gene transfers to tomato and the study of expression possibility of salt-tolerance gene (SOS3). Nova Biologica Reperta, 7(1), 76-84. doi: 10.29252/nbr.7.1.76 [DOI:10.29252/nbr.7.1.76]
25. Paramesh, H., Fakrudin, B. and Kuruvinashetti, M.S. (2010). Genetic transformation of a local variety of tomato
26. using gus gene: an efficient genetic transformation protocol for tomato. Journal of Agricultural Technology, 6(1), 87-97.
27. Prihatna, C., Chen, R., Barbetti, M.J. and Barker, S.J. (2019). Optimisation of regeneration parameters improves transformation efficiency of recalcitrant tomato. Plant Cell, Tissue and Organ Culture, 137, 473-483. doi:10.1007/s11240-019-01583-w [DOI:10.1007/s11240-019-01583-w]
28. Qiu, D., Diretto, G., Tavarza, R. and Giuliano, G. (2007). Improved protocol for Agrobacterium mediated transformation of tomato and production of transgenic plants containing carotenoid biosynthetic gene CsZCD. Scientia Horticulturae, 112 (2), 172-175. doi.org/10.1016/j.scienta.2006.12.015 [DOI:10.1016/j.scienta.2006.12.015]
29. Rajesh, N., Siva-Kumar, J., Prashanth, J.E.P. and Osman-Basha, P. (2016). An establishment of efficient Agrobacterium-mediated transformation in tomato (Solanum lycopersicum). International Journal of Recent Scientific Research, 7(1), 8583-8591.
30. Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A. and Allard, R.W. (1984). Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proceedings of the National Academy of Sciences of the United States of America, 81, 8014-8018. [DOI:10.1073/pnas.81.24.8014] [PMID] [PMCID]
31. Sambrook, J. and Russell, D.W. (2001). Molecular cloning. A laboratory manual. 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 16.1-16.62
32. Sandhya, D., Jogam, P., Venkatapuram, A.K., Savitikadi, P., Peddaboina, V., Allini, V.R. and Abbagani, S. (2022). Highly efficient Agrobacterium-mediated transformation and plant regeneration system for genome engineering in tomato. Saudi Journal of Biological Sciences, 29(6), 1-13. doi: 10.1016/j.sjbs.2022.103292 [DOI:10.1016/j.sjbs.2022.103292] [PMID] [PMCID]
33. Schillberg, S. and Finnern, R. (2021). Plant molecular farming for the production of valuable proteins - Critical evaluation of achievements and future challenges. Journal of Plant Physiology, 258, 1-7. doi.org/10.1016/j. jplph.2020.153359 [DOI:10.1016/j.jplph.2020.153359] [PMID]
34. Sharma, M.K., Solanke, A.U., Jani, D., Singh, Y. and Sharma, A.K. (2009). A simple and efficient Agrobacterium-mediated procedure for transformation of tomato. Journal of Biosciences, 34(3), 423-433. doi: 10.1007/s12038-009-0049-8 [DOI:10.1007/s12038-009-0049-8] [PMID]
35. Soltanmohammadi, B., Jalali-Javaran, M., Rajabi-Memari, H. and Mohebodini, M. (2014). Cloning, transformation and expression of proinsulin gene in tomato (Lycopersicum esculentum Mill.). Jundishapur Journal of Natural Pharmaceutical Products, 9(1), 9-15. doi: 10.17795/jjnpp-7779 [DOI:10.17795/jjnpp-7779] [PMID] [PMCID]
36. Somleva, M.N., Tomaszewski, Z. and Conger, B.V. (2002). Agrobacterium mediated transformation of switchgrass. Crop Science, 42(6), 2080-2087. doi. 10.2135/cropsci2002.2080 [DOI:10.2135/cropsci2002.2080]
37. Stavridou, E., Τzioutziou, N.A., Madesis, P., Labrou, N.E. and Nianiou-Obeidat, I. (2019). Effect of different factors on regeneration and transformation efficiency of tomato (Lycopersicum esculentum) hybrids. Czech Journal of Genetics and Plant Breeding, 55(3), 120-127. doi.org/10.17221/61/2018-CJGPB [DOI:10.17221/61/2018-CJGPB]
38. Su, X., Wang, B., Geng, X., Du, Y., Yang, Q., Liang, B., Meng, G., Gao, Q., Yang, W., Zhu, Y. and Lin, T. (2021). A high-continuity and annotated tomato reference genome. BMC Genomics, 22(1), 1-12. doi: 10.1186/s12864-021-08212-x [DOI:10.1186/s12864-021-08212-x] [PMID] [PMCID]
39. Sujatha, M., Vijay, S., Vasavi, S., Veera-Reddy, P. and Chander-Rao, S. (2012). Agrobacterium-mediated transformation of cotyledons of mature seeds of multiple genotypes of sunflower (Helianthus annuus L.). Plant Cell Tissue and Organ Culture, 110(2), 275-287. doi:10.1007/s11240-012-0149-2 [DOI:10.1007/s11240-012-0149-2]
40. Sun, S., Kang, X.P., Xing, X.J., Xu, X.Y., Cheng, J., Zheng, S.W. and Xing, G.M., (2015). Agrobacterium-mediated transformation of tomato (Lycopersicon esculentum L. cv. Hezuo 908) with improved efficiency. Biotechnology and Biotechnological Equipment, 29(5), 861-868. doi.org/10.1080/13102818.2015.1056753 [DOI:10.1080/13102818.2015.1056753]
41. Van, D.T., Ferro, N. and Jacobsen, H.J. (2010). Development of a simple and effective protocol for Agrobacterium tumefaciens mediated leaf disc transformation of commercial tomato cultivars. GM crops and food, 1(5), 312-321. doi:10.4161/gmcr.1.5.14703 [DOI:10.4161/gmcr.1.5.14703] [PMID]
42. Vats, S., Shivaraj, S.M., Sonah, H. Patil, G. Roy, J., Sharma, T.R. and Deshmukh, R. (2023). Efficient regeneration and Agrobacterium mediated transformation method for cultivated and wild tomato. Plant Molecular Biology Reporter, 41, 405-416. doi.org/10.1007/s11105-023-01374-w [DOI:10.1007/s11105-023-01374-w]
43. Wang, Y. and Demirer, G. S. (2023). Synthetic biology for plant genetic engineering and molecular farming. Trends in Biotechnology, 41(9), 1182-1198. doi: 10.1016/j.tibtech.2023.03.007 [DOI:10.1016/j.tibtech.2023.03.007] [PMID]
44. Yasmin, A. and Debener, T. (2010). Transient gene expression in rose petals via Agrobacterium infiltration. Plant Cell Tissue and Organ Culture, 102(2), 245-250. doi:10.1007/s11240-010-9728-2 [DOI:10.1007/s11240-010-9728-2]
45. Zhu, Q., Tan, J. and Liu, Y.G. (2022). Molecular farming using transgenic rice endosperm. Trends in Biotechnology, 40(10), 1248-1260. doi: 10.1016/j.tibtech.2022.04.002 [DOI:10.1016/j.tibtech.2022.04.002] [PMID]
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Mehrizadeh V, Dorani E. Transformation of human IFN-α (HU-IFN-α) gene to Urmia cultivar of tomato using Agrobacterium tumefaciens. gebsj 2025; 14 (1) :89-101
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دوفصل نامه علمی-پژوهشی مهندسی ژنتیک و ایمنی زیستی Genetic Engineering and Biosafety Journal
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