Induction of hairy roots in tomato (Solanum lycopersicum L.) and assessment of tolerance to Fusarium oxysporum culture filtrates

Badia Abdul Razzaq Malla Obaida, Nadeem Ahmed Ramadan

Abstract


Tomato plants (Lycopersicon esculentum Mill) were shown to be genetically compatible with Agrobacterium rhizogenes R1601 to form transgenic hairy roots of sterile seedlings without any natural root system. A bacterial suspension (96×108 cells/mL) was used to inoculate stem segments directly with a transformation efficiency of 94.28 percent and an average of 3.0 hairy roots per segment in 47 days. Several hairy root cultures were prepared with successive subculture in MS medium with 400mg/L cefotaxime which proved to be effective in removing the bacterial contamination. These were found to have a high growth rate, negative geotropism, high level of branching with color being white to pale-yellow. On solid medium with the Fusarium oxysporum filtrates being used at 5, 10, 15 and 20 percent concentration, the hairy roots were observed to be highly resistant, especially at the concentration of 15 and 20 percent indicating successful genetic transformation and tolerance to fungal stress.


Keywords


Agrobacterium rhizogenes, Hairy root transformation, Fusarium oxysporum, Tomato (Solanum lycopersicum L.), Fungal culture filtrate tolerance

Full Text:

PDF

References


Abdullah, A., 2000. Formation of some plants resistant to fusarium wilt, Fusarium oxysporum and Fusarium solani, from calluses of different parts of the genetic tomato plant, Lycopersicon esculentum Mill. University of Al Mosul.

Abdullah, R.M., Salih, S.M., Al-Nema Qutaiba, S., 2021. Plant regeneration from transformed tissues of broccoli (Brassica oleracea var. italica) by Agrobacterium rhizogenes ATCC13332 and their retention of rol-genes. European Academic Research 9(9), 6006-6017.

Aggarwal, P.R., Nag, P., Choudhary, P., Chakraborty, N., Chakraborty, S., 2018. Genotype-independent Agrobacterium rhizogenes-mediated root transformation of chickpea: a rapid and efficient method for reverse genetics studies. Plant Methods 14(1), 55.

Al-Sahuki, M., Abdel Basset, A., 2020. Plant genomics and breeding. University of Baghdad.

Al-Sanjari, A.M.H.S., 2022. Recruitment of copper oxide nanoparticles by tissues of sunflower plants transgenic with the vector Agrobacterium rhizogenes ATCC 15834. University of Mosul, Mosul, Iraq.

Al-Taie, A.A.R., 2022. Agrobacterium rhizogenes R1601 is a natural vector for genetic transformation of Solanum nigrum L. University of Al Mosul.

Bahari, Z., Sazegari, S., Niazi, A., Afsharifar, A., 2020. The application of an Agrobacterium-mediated in planta transformation system in Catharanthus roseus, a medicinal plant. Czech Journal of Genetics and Plant Breeding 56(1), 34-41.

Bahramnejad, B., Naji, M., Bose, R., Jha, S., 2019. A critical review on use of Agrobacterium rhizogenes and their associated binary vectors for plant transformation. Biotechnology 37(7), 107405.

Chakrabarty, S., Jin, M., Wu, C., Chakraborty, P., Xiao, Y., 2020. Bacillus thuringiensis vegetative insecticidal protein family Vip3A and mode of action against pest Lepidoptera. Pest Management Science 76(5), 1612-1617.

Ebrahim, H.S., El Ashry, A.A., Rabie, S.A.A., Ismail, I.A., El-Bahr, M.K., Gabr, A.M.M., 2025. Establishment of hairy root cultures by Agrobacterium rhizogenes-mediated transformation of Gardenia jasminoides variegata for enhancing phenolic and antioxidant accumulation capacity. Egyptian Pharmaceutical Journal 24(1), 131-140.

Fan, Y., Zhang, X., Zhong, L., Wang, X., Jin, L., Lyu, S., 2020. One-step generation of composite soybean plants with transgenic roots by Agrobacterium rhizogenes-mediated transformation. BMC Plant Biology 20(1), 208.

Garagounis, C., Georgopoulou, M.-E., Beritza, K., Papadopoulou, K.K., 2020. An Agrobacterium rhizogenes-mediated hairy root transformation protocol for fenugreek. MethodsX 7, 101098.

Georgiev, M.I., Agostini, E., Ludwig-Müller, J., Xu, J., 2012. Genetically transformed roots: from plant disease to biotechnological resource. Trends in Biotechnology 30(10), 528-537.

Guillon, S., Tremouillaux-Guiller, J., Pati, P.K., Rideau, M., Gantet, P., 2006. Hairy root research: recent scenario and exciting prospects. Current Opinion in Plant Biology 9(3), 341-346.

Kidd, S., Halliday, C., Alexiou, H., Ellis, D., 2016. Descriptions of medical fungi. CABI.

Lacroix, B., Citovsky, V., 2019. Pathways of DNA transfer to plants from Agrobacterium tumefaciens and related bacterial species. Annual Review of Phytopathology 57(1), 231-251.

Meyer, A.D., Tempé, J., Costantino, P., 2000. Hairy root: a molecular overview. Plant-Microbe Interactions 5, 1-39.

Mohamed, R.Y., Al-Mallah, M.K., Ramadan, N.A., 2015. A protocol for the production of chickpea plants resistant to Fusarium wilt from genetically transformed hairy roots. European Academic Research 3(3), 2833-2842.

Mohammed, A.A.-H., 2020. Uptake of copper oxide nanoparticles by different tissues of fenugreek (Trigonella foenum-graecum L.) plant. Plant Archives 20(2), 8121-8126.

Mohammed, A.A.-H., Masyab, H.M., 2020. Genetic transformation of Nigella sativa L. plants with Agrobacterium rhizogenes 35S GUS R1000 and estimation of thymoquinone level in transformed hairy root cultures. Plant Archives 20 (Supplement 1), 3649-3652.

Morgan, A.J., Cox, P.N., Turner, D.A., Peel, E., Davey, M.R., Gartland, K.M.A., Mulligan, B.J., 1987. Transformation of tomato using an Ri plasmid vector. Plant Science 49(1), 37-49.

Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15(3), 473-497.

Peterson, A., Kishchenko, O., Zhou, Y., Vasylenko, M., Giritch, A., Sun, J., Borisjuk, N., Kuchuk, M., 2021. Robust Agrobacterium-mediated transient expression in two duckweed species (Lemnaceae) directed by non-replicating, replicating, and cell-to-cell spreading vectors. Frontiers in Bioengineering and Biotechnology 9, 5.

Saeed, W., Naseem, S., Gohar, D., Ali, Z., 2019. Efficient and reproducible somatic embryogenesis and micropropagation in tomato via novel structures-rhizoid tubers. PLoS ONE 14(5), e0215929.

Shanks, J.V., Morgan, J., 1999. Plant ‘hairy root’ culture. Current Opinion in Biotechnology 10(2), 151-155.

Sultan, S.J., Mohammed, A.A., 2020. Genetic transformation of broccoli (Brassica oleracea var. italica) by plasmids of Agrobacterium rhizogenes R1601 and their molecular detection. Biochemical and Cellular Archives 20(2), 5491-5496.

Tzfira, T., Citovsky, V., 2007. Agrobacterium: from biology to biotechnology. Springer Science and Business Media.

Venkatachalam, L., Lokesh, V., Bhagyalakshmi, N., 2011. A rare event of Agrobacterium rhizogenes-assisted genetic transformation of ‘Silk’ banana (genotype AAB). Journal of Microbial and Biochemical Technology 3(1), 013-017.

Wang, D., Liu, Y., Zhang, Y., Huang, X., Wang, J., Wang, Y., Liu, Y., Yan, C., Lv, B., Jia, Y., 2025. Establishment of transient transformation systems in Welsh onion (Allium fistulosum L.): hairy root induction and protoplast transformation. Plants 14(17), 2664.

Yassin, A., Fredrick, K., Mankin, A.S., 2005. Deleterious mutations in small subunit ribosomal RNA identify functional sites and potential targets for antibiotics. Proceedings of the National Academy of Sciences 102(46), 16620-16625.




DOI: https://doi.org/10.33804/pp.010.02.6029

Refbacks

  • There are currently no refbacks.