In vitro selection of tomato plants resistant to Fusarium oxysporum f. sp. lycopersici and Verticillium dahliae
Zhana Ivanova
, Katya Vasileva
Abstract: Water scarcity, low and high temperatures, pests, and diseases harm the quality and yield of tomato production worldwide. To meet the increasing demand, it is necessary to develop new high-yielding varieties that combine disease resistance and fruit quality, suitable for cultivation in diverse climatic conditions. The present study aimed to create resistance in tomato plants against soil-borne fungal pathogens, Fusarium oxysporum f. sp. lycopersici and Verticillium dahliae, through an in vitro cell line selection approach. Cotyledonary explants of two tomato variants (Ideal and L1620) were tested to obtain resistant shoots to pathogens Fusarium oxysporum f. sp. lycopersici and Verticillium dahliae. The cotyledon explants were cultured on Murashige and Skoog basal medium supplemented with 2,2 mg/L kinetin and 1.6 mg/L IAA. To the medium were added three concentrations - 30% v/v, 40% v/v, and 50% v/v of the culture filtrates from two strains of the pathogens Fusarium oxysporum and Verticillium dahliae.
A total number of 21,99 regenerated plants resistant to Fusarium oxysporum f. sp. lycopersici and 9,66 plants resistant to Verticillium dahliae were obtained from the local variety Ideal at the highest concentration of pathogens (50%). From line L1620, 17,00 regenerated plants were obtained, showing resistance to a 50% concentration of the Fusarium culture filtrate, and 22,66 plants were resistant to Verticillium dahliae.
Keywords: culture filtrate; explants; fungal pathogens; in vitro screening; Solanum lycopersicum L.; somaclonal variation
Citation: Ivanova, Zh. & Vasileva, K. (2025). In vitro selection of tomato plants resistant to Fusarium oxysporum f. sp. lycopersici and Verticillium dahliae. Bulg. J. Agric. Sci., 31(5), 983–990
| References: (click to open/close) | Acharya, B., Ingram, T. W., Oh, Y., Adhikari, T. B., Dean, R. A. & Louws, F. J. (2020). Opportunities and challenges in studies of host-pathogen interactions and management of Verticillium dahliae in tomatoes. Plants, 9(11), 1622.
Cano, E. A., Perez-Alfocea, F., Moreno, V., Caro, M. & Bolarin, M. C. (1998). Evaluation of salt tolerance in cultivated and wild tomato species through in vitro shoot apex culture. Plant Cell Tissue Organ. Cult., 53, 19 - 26.
El-Meleigy, E. A., Gabr, M. F., Mohammed, F. H. & Ismail, M. A. (2004). Responses to NaCl salinity of tomato cultivated and breeding lines differing in salt tolerance in callus cultures. Int. J. Agric. Biol., 6(1), 19 - 26.
Flores, P. S., Otoni, W. C., Dhingra, O. D., de Souza Diniz, S. P. S, Santos, T. M. D. & Bruckner, C. H. (2012). In vitro selection of yellow passion fruit genotypes for resistance to Fusarium vascular wilt. Plant Cell Tissue Organ. Cult., 108, 37 - 44.
Fradin, E. F. & Thomma, B. P. (2006). Physiology and molecular aspects of Verticillium wilt diseases caused by V. dahliae and V. alboatrum. Mol. Plant Pathol., 7(2), 71 - 86.
Frame, B., Yu, Kang-fu, Christie, B. R. & Pauls, K. P. (1991). In vitro selection for resistance to verticillium wilt in alfalfa (Medicago sativa L.) using a fungal culture filtrate. Physiological and Molecular Plant Patholog.,38(5), 325 - 340. doi:10.1016/S0885-5765(05)80134-1.
Gamborg, O. L., Miller, R. A. & Ojima, K. (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research., 50(1), 151 - 158.
García-Gonzáles, R., Quiroz, K. & Carrasco, B. (2010). Plant tissue culture: Current status, opportunities and challenges. Cienc. Investig. Agrar., 37, 5 - 30.
Hassan, N. M., Serag, M. S., El-Feky, F. M. & Nemat Alla, M. M. (2008). In vitro selection of mung bean and tomato for improving tolerance to NaCl. Ann. Appl. Biol., 152(3), 319 - 330.
He, X. H. (2007). Bio-control of root rot disease in vanilla. Ph.D. thesis, University of Wolverhampton, Wolverhampton, UK.
Hernández-Hernández, J. (2011). Vanilla diseases. In: Handbook of vanilla science and technology (Havkin-Frenkel, D., Belanger, F. C., eds). Wiley-Blackwell Publishing, Chichester, 16 - 40.
Hu, C. H., Wei, Y. R., Huang, Y. H. & Yi, G. J. (2013). An efficient protocol for the production of chit42 transgenic Furenzhi banana (Musa spp. AA group) resistant to Fusarium oxysporum. Vitro Cell Dev. Biol. Plant., 49, 584 - 592.
Ivanova, Zh., Vasileva, K. & Stoeva, V. (2023). In Vitro Regeneration from cotyledonary explants of tomatoes (Solanum Lycopersicum L.). Journal of Mountain Agriculture on the Balkans, 26(2), 491 - 505.
Ivchenko, T., Miroshnichenko, T., Mozgovska, A., Bashtan, N., Krutko, R., Kondratenko, S., Kuts, O., Semenenko, I. & Kokoiko, V. (2020). Modeling of tomato genotypes stress-tolerance by comprehensive assessment on selective media in vitro. Biol. Life Sci. Forum, 4(1), 107.
Koyyappurath, S., Conéjéro, G., Dijoux, J. B., Lapeyre-Montès, F., Jade, K., Chiroleu, F., Gatineau, F., Verdeil, J.L., Besse, B. & Grisoni, M. (2015). Differential responses of vanilla accessions to root rot and colonization by Fusarium oxysporum f. sp. radicis-vanillae. Front. Plant Sci., 6, 1125.
Kuanar, A., Nayak, P. K., Subudhi, E. & Nayak, S. (2014). In vitro selection of turmeric somaclone resistant to Fusarium oxysporum f.sp. zingiberi. Proc. Nat. Acad. Sci. India Sect. B: Biol. Sci., 84, 10.
Kumari, C., Shaunak, I., Sharma, P. & Sharma, R. (2023). Genetic improvement of tomato against fusarium wilt disease using biotechnological interventions. In: Engineering of Crop Plants for Food and Health Security (Tiwari, S., Koul, B., eds), Genetic Springer, Singapore, 407 - 420.
Lebeda, A. & Švábová, L. (2010). In vitro screening methods for assessing plant disease resistance. In: Mass screening techniques for selecting crops resistant to disease ( FAO/IAEA, eds). International Atomic Energy Agency, Vienna, 5 - 45.
Lynch, D. R., Coleman, M. C. & Lyon, G. D. (1991). Effect of Alternaria solani culture filtrate on adventitious shoot regeneration in potato. Plant Cell Rep., 9, 607 - 610.
Mahlanza, T., Rutherford, R. S., Snyman, S. J. & Watt, M. P. (2013). In vitro generation of somaclonal variant plants of sugarcane for tolerance to Fusarium sacchari. Plant Cell Rep., 32, 249 - 262.
Mazzotta, E., Muzzalupo, R., Chiappetta, A. & Muzzalupo, I. (2022). Control of the verticillium wilt on tomato plants by means of olive leaf extracts loaded on chitosan nanoparticles. Microorganisms, 10(1), 136. https://doi.org/10.3390/microorganisms10010136.
Murashige, Т. & Skoog, F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant, 15(3), 473 - 497.
Patade, V. Y., Suprasanna, P. & Bapat, V. A. (2008). Gamma irradiation of embryogenic callus cultures and in vitro selection for salt tolerance in sugarcane (Saccharum offcinarum L.). Agric. Sci. China, 7(9), 101 - 105.
Predieri, S. (2001). Mutation induction and tissue culture in improving fruits. Plant Cell Tissue Organ. Cult., 64(2), 185 - 210.
Purwati, R. D., Harran, S. & Sudarsono, S. (2007). In vitro selection of abaca for resistance to Fusarium oxysporum f. sp. cubense. Hayati J. Biosci., 14(2), 65 - 70.
Ramírez-Mosqueda, M. A., Iglesias-Andreu, L. G., Luna-Rodríguez, M. & Castro-Luna, A. A. (2015). In vitro phytotoxicity of culture filtrates of Fusarium oxysporum f. sp. vanillae in vanilla planifolia jacks. Sci. Hortic., 197, 573 - 578.
Ramírez-Mosqueda, M. A., Iglesias-Andreu, L. G., Teixeira da Silva, J. A., Luna‑Rodríguez, M., Noa‑Carrazana, J. C., Bautista‑Aguilar, J. R., Leyva‑Ovalle, O. R. & Murguía‑González, J. (2019). In vitro selection of vanilla plants resistant to Fusarium oxysporum f. sp. vanillae. Acta Physiol. Plant., 41, 40.
Rao, S. & Sandhya, H. (2016). In Vitro selection of disease-resistant plants. In: Plant Tissue Culture: Propagation, Conservation and Crop Improvement (Anis, M., Ahmad, N., eds) Springer, Singapore.
Ravikumar, R. L., Patil, B. S., Soregaon, C. D. & Hegde, S. G. (2007). Genetic evidence for gametophytic selection of wilt resistant alleles in chickpea. Theor. Appl. Genet., 114, 619 - 625.
Sharma, T., Rathour, R., Plaha, P., Katoch, V., Khalsa, G. S., Patial, V., Singh, Y. & Pathania, N. K. (2010). Induction of Fusarium wilt (Fusarium oxysporum f. sp. pisi) resistance in garden pea using induced mutagenesis and in vitro selection techniques. Euphytica, 173, 345 - 356.
Sowik, I., Michalczuk, L. & Wojcik, D. (2008). A method for in vitro testing strawberry susceptibility to Verticillium wilt. Journal of Fruit and Ornamental Plant Research., 16, 111 - 121.
Srinivas, C., Devi, D. N., Murthy, K. N., Mohan, C. D., Lakshmeesha, T. R., Singh, B., Kalagatur, N. K., Niranjana, S. R., Hashem, A., Alqarawi, A. A., Tabassum, B., Elsayed, F. A. A., Nayaka, S. Ch. & Srivastava, R. K. (2019). Fusarium oxysporum f. sp. lycopersici, causal agent of vascular wilt disease of tomato: Biology to diversity-A review. Saudi J. Biol. Sci., 26(7), 1315 - 1324.
Storti, E., Latil, C., Salti, S., Bettini, P., Bogani, P., Pellegrini, M. G., Simeti, C., Molnar, A. & Buiatti, M. (1992). The in vitro physiological phenotype of tomato resistance to Fusarium oxysporum f. sp. lycopersici. Theor. Appl. Genet., 84(1-2), 123 - 128.
Suprasanna, P., Sidha, M. & Bapat, V. A. (2009). Integrated approaches of mutagenesis and in vitro selection for crop improvement. In: Plant tissue culture, molecular markers and their role in crop productivity (Kumar, A., Shekhawat, N. S., eds). IK International Publishers, New Delhi, 73 - 92.
Švábová, A. & Lebeda, J. (2005). In vitro selection for improved plant resistance to toxin-producing pathogens. Journal of Phytopathology, 153(1), 52 - 64.
Švábová, L., Lebeda, A., Kitner, M., Sedlářová, M., Petřivalský, M., Dostálová, R., Ondřej, M., Horáček, J., Smýkalová, I. & Griga, M. (2011). Comparison of the effects of Fusarium solani filtrates in vitro and in vivo on morphological characteristics and peroxidase activity pea cultivars with different susceptibility. J. Plant Pathol., 93(1), 19 - 30.
Van Eck, J., Kirk, D. D. & Walmsley, A. M. (2006). Tomato (Lycopersicum esculentum). Methods Mol. Biol., 343, 459 - 473.
Venkatachalam, P., Geetha, N., Jayabalan, N. & Saravanababu, S. (1998). In Vitro selection of groundnut cell lines from Cercosporidium personatum culture filtrates and regeneration of resistant plants through cell culture. J. Plant Biol., 41, 318 - 323.
Yu, K., Christie, B. R. & Pauls, K. D. (1990). Effects of Verticillium albo-atrum culture filtrate on somatic embryogenesis in alfalfa. Plant Cell Rep., 8, 509 - 511. |
|
| Date published: 2025-10-28
Download full text 
