Formulation for optimizing Bacillus thuringiensis production


Affiliations

  • Indian Agricultural Research Institute (ICAR), Division of Entomology, New Delhi, Delhi, 110012, India

Abstract

Bacillus thuringiensis Berliner, a gram positive aerobic bacterium, produces parasporal crystal (Cry) toxins that are highly specific and effective against insect species. During the course of isolation of native strains, B. thuringiensis AUG-5 was found the most effective with a wide range of activity against lepidopterans. Hence, different media were evaluated for its growth and development. Increase in concentration of the Luria Bertani [(LB), composed of casein, yeast extract and sodium chloride in 2:1:2 w/w)] medium in the fermentation broth from 1 to 2% increased colony forming unit (CFU), spore and also Cry1Ac and Cry2Ab toxin content. However, further increase of LB concentration to 3% adversely affected bacterial growth and development. Addition of 1% Wesson salt in 1% LB broth significantly increased spore, CFU counts, and also that of Cry1Ac but not of Cry2Ab. Spore and CFU counts in media were positively correlated and cell mass negatively correlated with Cry1Ac and Cry2Ab contents. Of all media substituting LB with agro products, medium consisting of 2% wheat flour, 2% soybean meal and 1% Wesson salt could be considered as an alternative to LB medium to achieve economy of largescale production costs. Spore-crystal complexes of Medium II and III were most toxic to the neonates of cotton bollworm, Helicoverpa armigera and tobacco caterpillar, Spodoptera litura at 10 μg/g, and differed significantly from those of Medium LB-2X and LB-3X and Cry2Ab2. Cry1Ac was most toxic to H. armigera at 1 μg/g and less toxic to S. litura than Cry2Ab.

Keywords

Agro byproducts, Bacillus thuringiensis, endotoxin production, culture media, insecticidal activity, Helicoverpa armigera, Spodoptera litura

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References

Abdel- Hameed A, Carlberg G, El-Tayeb OM. 1991. Studies on the Bacillus thuringiensis H-14 strains isolated in Egypt-IV Characterization of fermentation conditions for δ endotoxin production. World J Micro Biot. 7: 231–236.

Alves LFA, Alves SB, Capalbo DMF. 1997. Selection of Agroindustrial by-products as components of media used for production of Bacillus thuringiensis var. kurstaki Berliner. An Soc Entomol Bras. 26(2): 379–382.

Bravo A, Likitvivatanavong S, Gill SS, Soberon M. 2011. Bacillus thuringiensis: A story of a successful bioinsecticide. Insect Biochem Mol Biol. 41(7): 423–431.

Crickmore N, Zeigler DR, Feitelson J, Schnepf E, Van Rie J, Lereclus D, Baum J, Dean DH. 1998. Revision of nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol Mol Biol Rev. 62(3): 807–813.

Crickmore N, Baum J, Bravo A, Lereclus D, Narva K, Sampson K, Schnepf E, Sun M, Zeigler, DR. 2016. Bacillus thuringiensis toxin nomenclature. (http://www.btnomenclature.info/); www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/holo2.html).

Dhingra H, Chaudhary K. 2011. Production of Bacillus thuringiensis S6 biomass using different cheap nitrogen sources. Acta Agric Serb. 16(31): 3–8.

Dregval OA, Cherevach NV, Andrienko OE, Vinnikov AI. 1999. The effect of a soil extract on the development of Bacillus thuringiensis on its synthesis of an insecticidal endotoxin. Mikrobiol Z. 61(4): 40–44.

Dregval OA, Cherevach NV, Vinnikov AI. 2002. Influence of composition of the nutrient medium on growth and development of entomopathogenic bacteria Bacillus thuringiensis. Mikrobiol Z. 64(2): 44–48.

Dulmage HT. 1971. Production of delta-endotoxin by eighteen isolates of Bacillus thuringiensis serotype 3 in fermentation media. J Invertebr Pathol. 18(3): 353–358.

Dulmage HT, Correa JA, Martinez AJ. 1970. Co-precipitation with Lactose as a means of recovering the spore-crystal complex of Bacillus thuringiensis. J Invertebr Pathol. 15(1): 15–20.

Entwistle PF, Cory JS, Bailey MJ, Higgs S. 1993. Bacillus thuringiensis: an environment biopesticide, theory and practice. Chichester: John Wiley & Sons, USA. 193–220 pp.

Gujar GT, Kumari A, Kalia V, Chandrashekar K. 2000. Spatial and temporal variation in susceptibility of the American bollworm, Helicoverpa armigera (Hübner) to Bacillus thuringiensis var. kurstaki in India. Curr Sci India. 78(8): 995–1001.

Harish J. 2006. Development and evaluation of different formulations of Bacillus thuringiensis for management of Helicoverpa armigera. Ph.D. Dissertation, CCS Haryana Agriculture University, Hisar, India.

Hofte H, Whiteley HR. 1989. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev. 53(2): 242–255.

Hwang DL, Yang WK, Foard DE, Lin KTD. 1978. Rapid release of protease inhibitors from soybeans. Plant Physiol. 61: 30–34.

Johnson V, Shah RN, Shah DN, Patel KA, Mehta MH. 1994. Production of Bacillus thuringiensis based bioinsecticide: Influence of various nitrogen sources on process economics, pp. 161–167, In: Proceedings, Microbes for better living: proceedings of MICON-International-94 and 35th Annual Conference of Association of Microbiologists of India, 9-12 November 1994, DFRL, Mysore, India.

Kalia V, Sethi T, Gujar GT. 2013. Susceptibility of Brinjal shoot and fruit borer, Leucinodes orbonalis (Guenee) to Bacillus thuringiensis and its Cry toxins. Biopestic Int. 9(1): 88–92.

Kumar A, Sra K, Sangodkar UMX, Sharma VP. 2000. Advances in the bio-control of mosquito vectors utilizing Bacillus sphaericus and Bacillus thuringiensis var. israelensis. Proceedings of National Academy of Science, India. 70(1): 1–20.

Khodair TA, Abdelhafez AAM, Sakr HM, Ibrahim MMM. 2008. Improvement of Bacillus thuringiensis bioinsecticide production by fed-batch culture on low cost effective medium. Res J Agric Biol Sci. 4(6): 923–935.

Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacterioohage T4. Nature (Lond.) 227: 680–685.

Lu Q, Cao G-C, Zhang L-l, Liang G-M, Gao X-W, Zhang Y-J, Guo Y-Y. 2013. The binding characterization of Cry insecticidal proteins to the brush border membrane vesicles of Helicoverpa armigera, Spodoptera exigua, Spodoptera litura and Agrotis ipsilon. J Integr Agr. 12(9): 1598–1605.

Meena RK, Krishna kumara GG, Alpana, Gujar GT, Kaur S. 2012. Screening of Bacillus thuringiensis isolates recovered from diverse habitats in India for the presence of cry1A-type Genes and cloning of a cry1Ac33 gene toxic to Helicoverpa armigera (American Bollworm). Asian J Biotechnol. 4(2): 53–69.

Moraes IO, Santana MHA, Hokka CO. 1981. The influence of oxygen concentration on microbial insecticide production. In: Proceedings of 8th International Forum Symposium: Advances in Biotechnology. Vol 1, London, U.K. pp. 75–79.

Morris ON, Kanagaratnam P, Converse V. 1997. Suitability of 30 agricultural products and by–products as nutrient sources for laboratory production of Bacillus thuringiensis subsp. aizawai. J Invertebr Pathol. 70(2): 113–120.

Mummigatti SG, Raghunathan AN. 1990. Influence of media composition on the production of delta-endotoxin by Bacillus thuringiensis var. thuringiensis. J Invertebr Pathol. 55(2): 147–151.

Navon A. 2000. Bacillus thuringiensis insecticides in crop protection- reality and prospects. Crop Prot. 19(8-10): 669–676.

Nester EW, Thomashow LS, Metz M, Gordon M. 2002. 100 years of Bacillus thuringiensis: a critical scientific assessment. American Society for Microbiology, Washington, DC.(http://academy.asm.org/images/stories/documents/100yearsofbtcolor.pdf).

Oliveira HD, Sousa DOB, Oliveira JTA, Carlini CR, Oliveira HP, Pereira ML, Rocha RO, Morais JKS, Gomes-Filho E, Vasconcelos IM. 2010. Gm-Tx, a new toxic protein from soybean (Glycine max) seeds with potential for controlling insect pests. Process Biochem. 45: 634–640.

Poopathi S, Kumar KA. 2003. Novel fermentation media for production of Bacillus thuringiensis israelensis. J Econ Entomol. 96(4): 1039–1044.

Prabakaran G, Balaraman K. 2006. Development of a cost-effective medium for the large scale production of Bacillus thuringiensis var. israelensis. Biol Control. 36: 288–292.

Ramanujam B, Rangeshwaran R, Sivakmar G, Mohan M, Yandigeri MS. 2014. Management of insect pests by microorganisms. Proc Indian Natn Sci Acad. 80(2):455–471

Salama HS, Foda MS, Dulmage HT, Sharaby EL. 1983. Novel fermentation medium for production of δ-endotoxin from Bacillus thuringiensis. J Invertebr Pathol. 41(1): 8–19.

Sanchis V, Bourguet D. 2008. Bacillus thuringiensis: applications in agriculture and insect resistance management. A review. Agron Sustain Dev. 28(1): 11–20.

SAS Institute. 1998. SAS system for elementary statistical analysis surveys. For this research site, using the two technolo- SAS Institute Inc., Cary, NC.

Saravanan L, Gujar GT. 2006. Distribution of Bacillus thuringiensis Berliner in the samples of warehouse and cadavers. J Entomol Res (New Delhi). 30(1): 1–4.

Sarrafzadeh MH, Guiraud JP, Lagneau C, Gaven B, Carron A, Navarro JM. 2005. Growth, sporulation, delta–endotoxins synthesis, and toxicity during culture of B. thuringiensis H 14. Curr Microbiol. 51(2): 75–81.

Scherrer P, Lüthy P, Brüno T. 1973. Production of δ-endo-toxin by Bacillus thuringiensis as a function of glucose concentration. Appl Microbiol. 25(4): 644–646.

Schnepf E, Crickmore N, Van Rie J, Lereclus D, Baum J, Feitelson J, Zeigler DR, Dean DH. 1998. Bacillus thuringiensis and its pesticidal crystals proteins. Microbiol Mol Biol Rev. 62(3): 775–806.

Shojaaddini M, Moharramipour S, Khodabandeh M, Talebi AA. 2010. Development of a cost effective medium for production of Bacillus thuringiensis bio-insecticide using food barley. J Plant Prot Res. 50(1): 9–14.

Travers RS, Martin PAW, Reichelderfer CF. 1987. Selective process for efficient isolation of soil Bacillus spp. Appl Environ Microbiol. 53(6): 1263–1266.

Valicente FH, Tuelher EDS, Leite MIS, Freire FL, Vieira CM. 2010. Production of Bacillus thuringiensis biopesticide using commercial lab medium and agricultural byproducts as nutrient sources. Revista Brasileira De Milho E Sorgo 9(1): 1–11.

Whalon ME, Mota-Sanchez D, Hollingworth RM. 2008. Global pesticide resistance in arthropods. CABI International, Wallingford, U.K.

Whalon ME, Mota-Sanchez D, Hollingworth RM. 2013. Arthropod Pesticide Resistance Database. (http://www.pesticideresistance.com/index.php).

WHO [World Health Organization]. 1999. Microbial Pest Control Agent: Bacillus thuringiensis. Environment Health Criterion 217, World Health Organization, Geneva.

Yadav K, Dhiman S, Baruah I, Singh L. 2011. Development of cost effective medium for production of Bacillus sphaericus strain isolated from Assam, India. Microbiol J. 1(2): 65–70.

Zouari N, Bensikali S, Jaoua S. 2002. Production of delta-endotoxins by Bacillus thuringiensis strains exhibiting various insecticidal activities towards Lepidoptera and Diptera in gruel and fish meal media. Enzyme Microbiol Technol. 31(4): 411–418.


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