An ELISA for the detection of Bacillus subtilis L-form bacteria confirms their symbiosis in strawberry


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Abstract

Aims:To develop an ELISA for the detection of antigens derived from stable Bacillus subtilis L-form bacteria and to detect these in plants injected with L-form bacteria.Methods and Results:A sandwich ELISA was developed and its specificity was investigated using L-forms and cell-walled forms of B. subtilis, different Bacillus species and a range of bacteria isolated from glasshouse-grown strawberry plants. The detection limits of the ELISA were approximately 103 viable cells ml−1 for L-forms compared with 107 viable cells ml−1 for cell-walled forms. Results showed that L-forms survived and moved within strawberry tissues injected with L-form bacteria.Conclusion:An ELISA that selectively detects B. subtilis L-form bacteria was developed and shown to confirm the presence of L-forms in plants.Significance and Impact of the Study:This will be a valuable rapid method to further studies on L-form plant interactions.

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