National Academy Science Letters

, Volume 41, Issue 4, pp 249–253 | Cite as

Optimization and Standardization of Conditions for Production of Commercially Viable Formulation of Native Agrobacterium sp. UHFBA-218

  • Aditi Sharma
  • A. K. GuptaEmail author
  • Rishi Mahajan
Short Communication


Studies were carried out for the mass multiplication of native Agrobacterium sp. UHFBA-218 (MCC 2101, NCBI: KC488176; deposited as UHFBA-218[Cherry 2E-2-2]) and Rhizobium rhizogenes strain K84 at different pH, temperature, incubation period, broth media and at different air supply. The most effective pH, temperature, incubation period, broth medium and air supply for both the isolates was 7.0, 25 °C, 4 days, yeast extract mannitol broth medium and 20 L/min, respectively. These parameters were further used for mass multiplication of these strains and mixing the multiplied bacterial inoculum for studying the viability of Agrobacterium sp. UHFBA-218 and R. rhizogenes strain K84 in different carrier media—white stone powder, carboxymethyl cellulose and activated charcoal during 6 months of storage at room temperature at three different locations, mixed in one part of broth culture in exponential phase having 270 ± 10 × 1012 cfu/ml to two part of carrier and one part of broth culture mixed to five part of carrier and were compared for viable counts observed in different formulations kept at 4 °C for reference. White stone powder was superior to other carrier media tested under these studies. There were appreciable counts ranging from 34.67 to 103.3 × 108 cfu/g after 6 months of room temperature storage at different locations. However, irrespective of strains, storage at 4 °C temperature provided maximum viability of 108.50 and 155.7 × 108 cfu/g in white stone powder mixed in 1:5 and 1:2 ratios, respectively.


Agrobacterium sp. UHFBA-218 Bioformulation Carrier media Viability 



The authors are thankful to the Dean, College of Horticulture, Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan-173 230, Himachal Pradesh for providing lab facilities.


  1. 1.
    Gelvin SB (2003) Agrobacterium-mediated plant transformation: the biology behind the “gene-jockeying” tool. Microbiol Mol Biol Rev 67:16–37CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Mousavi SA, Osterman J, Wahlberg N, Nesme X, Lavire C, Vial L, Paulin L, de Lajudie P, Lindstrom K (2014) Phylogeny of the Rhizobium-Allorhizobium-Agrobacterium clade supports the delineation of Neorhizobium gen. nov. Syst Appl Microbiol 37:208–215CrossRefPubMedGoogle Scholar
  3. 3.
    Mousavi SA, Willems A, Nesme X, de Lajudie P, Lindström K (2015) Revised phylogeny of Rhizobiaceae: proposal of the delineation of Pararhizobium gen. nov., and 13 new species combinations. Syst Appl Microbiol 38:84–90CrossRefPubMedGoogle Scholar
  4. 4.
    Petersen SG, Stummann BM, Olesen P, Henningsen KW (1989) Structure and function of root-inducing (Ri) plasmids and their relation to tumor-inducing (Ti) plasmids. Plant Physiol 77:427–435CrossRefGoogle Scholar
  5. 5.
    Kerr A (1972) Biological control of crown gall: seed inoculation. J Appl Bacteriol 35:493–497CrossRefGoogle Scholar
  6. 6.
    Moore LW, Warren G (1979) Agrobacterium radiobacter strain K84 and biological control of crown gall. Ann Rev Phytopathol 17:163–179CrossRefGoogle Scholar
  7. 7.
    Anonymous (2012) Completion report of NFBSRA/PCN/AP-18/2007-08 project ‘‘Molecular analysis of agrocin producing Agrobacterium radiobacter for biological control of crown gall in stone fruits”. Department of Mycology and Plant Pathology, Dr. Y.S.Parmar University of Horticulture and Forestry, Nauni, Solan, p 38Google Scholar
  8. 8.
    Dua A, Sangwan N, Kaur J, Saxena A, Kohli P, Gupta AK, Lal R (2013) Draft genome sequence of Agrobacterium sp. strain UHFBA-218, isolated from rhizosphere soil of crown gall-infected cherry rootstock Colt. Genome Announc 1(3):e00302-13. CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Okereke GU, Okeh O (2007) Survival of cowpea bradyrhizobia in carrier materials and inoculation response in soil. Afr Crop Sci Soc Conf Proc 8:1183–1186Google Scholar
  10. 10.
    Hoben HJ, Somasegaran P (1982) Comparison of the pour, spread and drop plate method for enumeration of Rhizobium spp. In inoculants made from pre sterilized peat. Appl Environ Microbiol 44:1246–1247PubMedPubMedCentralGoogle Scholar
  11. 11.
    Gomez KA, Gomez AA (1983) Statistical procedures for agricultural research, 2nd edn. Wiley, New York, pp 357–427Google Scholar
  12. 12.
    Smith EF, Townsend CO (1907) A plant tumor of bacterial origin. Science 25:671–673ADSCrossRefPubMedGoogle Scholar
  13. 13.
    Moore LW, Canfield M (1996) Biology of Agrobacterium and management of crown gall disease. In: Hall R (ed) Principles and practice of managing soil borne plant pathogens. APS Press, St. Paul, pp 151–191Google Scholar
  14. 14.
    Pesenti-Barili B, Ferdani E, Mosti M, Degli- Innocenti F (1991) Survival of Agrobacterium radiobacter K84 on various carriers for crown gall control. Appl Environ Microbiol 57:2047–2051PubMedPubMedCentralGoogle Scholar
  15. 15.
    Kloepper JW, Schroth MN (1981) Development of a powder formulation of rhizobacteria for inoculation of potato seed pieces. Phytopathology 71:590–592CrossRefGoogle Scholar
  16. 16.
    Kaljeet S, Keyeo F, Amir HG (2011) Influence of carrier materials temperature on survivability of rhizobial inoculants. Asian J Plant Sci 10(6):331–337CrossRefGoogle Scholar
  17. 17.
    Thomson J A (1984) Production and quality control of carrier based inoculants. International crop research institute for semi-arid tropics.

Copyright information

© The National Academy of Sciences, India 2018

Authors and Affiliations

  1. 1.Department of Plant PathologyDr. Y. S. Parmar University of Horticulture and ForestryNauni, SolanIndia

Personalised recommendations