While performing chloroplast transformations, I've got plants which do not carry the antibiotic resistance marker (that should have been the case with the true transformants but are growing happily in spectinomycin, even in the presence of both spectinomycin and streptomycin.
Hi Ami,
Resistance could be due to spontaneous mutation that occurred in two different regions of the chloroplast rrn 16 gene, which codes for 16S rRNA. Plants whose antibiotic resistance is due to spontaneous generation are phenotypically indistinguishable from transgenic plants. Thus it should be noted that mutant plants may arise during generation of transplastomic plants and that additional molecular analysis is necessary for their identification and elimination [Filipenko et al. (2011) Spontaneous spectinomycin resistance mutation of the chloroplast rrn 16 gene in Daucus carota Callus lines. Russian Journal of Genetics, 47(1): 35-40].
Wishing you success in your reserach.
That can also be that the concentrations of your both antibiotics are not high enough to kill the wild-type or the non-transformants. The optimum concentrations of antibiotics may be vary, depending on the plant species and even the 'genotype' of a same species. Or the antibiotics might have lost their function. These factors need to be considered.
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