The survival of plant growth promoting microorganisms in peat inoculant as measured by selective plate counting and enzyme-linked immunoassay

Achieving specific counting of plant growth promoting (PGP) microorganisms maintained at high numbers in inert carriers such as peat is an important objective for the inoculation of field crops such as cereals. In this paper, methods based on selective media together with strain-specific counting using enzyme-linked immunoassay (ELISA) were examined. Selective plate counting was developed by screening four commercial PGP biofertiliser strains for resistance to antibiotics. ELISAs for each strain were developed and calibrated by purifying polyclonal antibodies, testing sample pre-treatment strategies, and investigating the effect of culture age on standard curves. Selective plate counting proved to be more accurate than the ELISA methodology, confirming that all microbial strains survived for at least 1 month in sterile peat without loss in viable numbers, and further demonstrated growth inhibition of the strain Candida tropicalis HY when co-inoculated with the other strains Pseudomonas fluorescens 1 N, Bacillus amyloliquefaciens E19 and Bacillus subtilis B9. This is the first known study to have investigated the dynamics of PGP microorganisms in multi-strain inoculants and demonstrates the utility and hitherto unmentioned drawbacks of two different low-cost counting methods for biofertiliser quality control. Such information is vital for the adoption and success of non-rhizobial PGP biofertilisers for sustainable agriculture.