Agrobacterium tumefaciens causes crown gall disease in plants by transferring their T-DNA which contains some oncogenic genes. At the site of the wound, these bacteria become accumulates and then transfer their T-DNA into host plant cells, which interact with host plant genomic DNA and cause tumor. With the help of knowledge, how the A.tumefaciens transfer their T-DNA to plant cells. And with the knowledge and use of the latest molecular biology techniques, A.tumefaciens is used as a tool for the transformation of our gene of interest into the host plant cells to make genetically modified plants. This review provides a working systematic review of the isolation, identification, and characterization of A.tumefaciens from the leguminous plants. This review also tells us how to transfer the T-DNA into a host plant cell and how to isolate the Agrobacterium tumefaciens from the root nodules of leguminous plants, first of all, collect the root nodules of leguminous plants then wash them properly and streak the isolates of root nodules on LB agar plates. then identify and characterize the Agrobacterium from the isolates because Agrobacterium and rhizobium stay together. For characterization, biochemical tests i.e gram staining and KOH tests performed to confirm the isolated bacteria is gram-negative. Some pathogenic tests such as carrot disk bioassay and potato disc confirm whether isolates caused a tumor or not.

Keywords: Agrobacterium tumefaciens, T-DNA, KOH tests, Antibiotic-resistant tests, Gram staining, Genetically modified plants.

[1] Ali, N., Zada, A., Ali, M., & Hussain, Z. (2016). Isolation and identification of Agrobacterium tumefaciens from the galls of peach tree. In Journal of Rural Development and Agriculture, 1(1).

[2] Finer, K. R., Fox, L., & Finer, J. J. (2016). Isolation and Characterization of Agrobacterium Strains from Soil: A Laboratory Capstone Experience. Journal of Microbiology & Biology Education, 17(3): 444–450. DOI:

[3] Hwang, H.-H., Yu, M., & Lai, E.-M. (2017). Agrobacterium-Mediated Plant Transformation: Biology and Applications. The Arabidopsis Book, 15: e0186. DOI:

[4] Koli, D. K., & Swarnalakshmi, K. (2017). Isolation and Characterization of Nodule Associated Bacteria from Chickpea and their Potential for Plant Growth Promotion. International Journal of Current Microbiology and Applied Sciences, 6(5): 1992–2004. DOI:

[5] Mir, M. I., Kumar, B. K., Gopalakrishnan, S., Vadlamudi, S., & Hameeda, B. (2021). Characterization of rhizobia isolated from leguminous plants and their impact on the growth of ICCV 2 variety of chickpea (Cicer arietinum L.). Heliyon, 7(11). DOI:

[6] Murugesan, S., Manoharan, C., Vijayakumar, R., & Panneerselvam, A. (2010). Isolation and Characterization of Agrobacterium rhizogenes from the Root Nodules of Some Leguminous Plants. International Journal of Microbiological Research, 1(3): 92–96.

[7] Pǎcurar, D. I., Thordal-Christensen, H., Pǎcurar, M. L., Pamfil, D., Botez, M. L., & Bellini, C. (2011). Agrobacterium tumefaciens: From crown gall tumors to genetic transformation. In Physiological and Molecular Plant Pathology, 76(2): 76–81, Academic Press. DOI:

[8] Rahman, S., Ahmad, K., Ali, N., Idrees, M., & Ali, W. (2020). Isolation and characterization of agrobacterium tumefaciens strains from Malakander farm, university of agriculture, Peshawar. Pakistan Journal of Zoology, 52(5): 2019–2021. DOI:

[9] Riah, N., Béna, G., Djekoun, A., Heulin, K., de Lajudie, P., & Laguerre, G. (2014). Genotypic and symbiotic diversity of rhizobium populations associated with cultivated lentil and pea in sub-humid and semi-arid regions of Eastern Algeria. Systematic and Applied Microbiology, 37(5): 368–375. DOI: 2013.12.008.

[10] Tyagi, A., Kumar, V., shottam, P., & Tomar, A. (2017). Isolation, Identification, Biochemical and Antibiotic Sensitivity Characterization of Rhizobium Strains from Vigna mungo (L) Hepper, Cicer arietinum L and Vigna radiata (L) R Wilczek in Muzaffarnagar, Uttar Pradesh, India. International Journal of Current Microbiology and Applied Sciences, 6(12): 2024–2035. DOI:

Source of Funding:

This research work did not receive any grant from funding agencies in the public or not-for-profit sectors.

Competing Interests Statement:

The authors declare no competing financial, professional, or personal interests.

Consent for publication:

The authors declare that they consented to the publication of this research work.

Authors’ Contributions:

All authors equally contributed to research and paper drafting.