Abstract
Induction of L-form bacteria from Bacillus thuringiensis was investigated with the long-term view of their use as targeted biological control agents within plants. L-forms were successfully induced from B. thuringiensis subspecies israelensis 36-3 by preparing pour plates of exponential cultures in L-phase medium (LPM) containing μg ml-1 cephalosporin C. The induced L-forms were then subcultured and maintained in L-phase medium supplemented with inactivated mycoplasma screened 5% (v/v) horse serum (5HS) with a combination of 225 μg ml-1 cephalosporin C and 0.6 mg ml-1 penicillin G, using the push block method. Induced L-forms showed typical features with pleiomorphic cells that contained granular particles and small vacuoles but no parasporal bodies. These L-forms were capable of growing on solidified media but failed to grow in liquid media. Ceylon Journal of Science (Bio. Sci.) 41 (2): 137-144, 2012 DOI: http://dx.doi.org/10.4038/cjsbs.v41i2.5383
Highlights
Bacterial L-forms may be defined as a special type of growth derived or induced from a bacterium following suppression of the rigid cell wall (Madoff, 1986)
Screening for inducing agents Inhibition zones were observed in all surface seeded L-phase medium (LPM) + 5% (v/v) horse serum (5HS) plates with the four B. thuringiensis isolates after overnight incubation
The initial research undertaken concerned the selection of inducing agents for the initiation of B. thuringiensis L-forms with the choice of agents being made from those used routinely in the Aberdeen laboratory, for Bacillus spp
Summary
Bacterial L-forms may be defined as a special type of growth derived or induced from a bacterium following suppression of the rigid cell wall (Madoff, 1986). They were originally isolated from natural sources but nowadays are mainly induced artificially by treatment of parental cell-walled bacteria with different cell wall inhibiting agents such as penicillin and lysozyme (Madoff, 1986). Experience by the Aberdeen Group (i.e. Paton and Allan) have indicated that the concentration of inhibitors often has to be altered during the induction process and that in some scenarios, the use of two inhibitors is more effective at maintaining the L-form state
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