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Penna et al. AMB Express 2011, 1:21 http://www.amb-express.com/content/1/1/21 ORIGINAL Open

Penna et al. AMB Express 2011, 1:21 http://www.amb-express.com/content/1/1/21 ORIGINAL Open Access A simple method to evaluate the number of bradyrhizobia on soybean seeds and its implication on inoculant quality control Claudio Penna1, Rosana Massa1, Florencia Olivieri1, Gabriel Gutkind2 and Fabricio Cassán3* Abstract Soybean seeds are non-sterile and their bacterial population interferes with the enumeration of beneficial bacteria, making it difficult to assess survival under different conditions. Within this context, the principal aims of this work were: (1) to improve a selective media for the enumeration of B. japonicum recovered from inoculated soybean seeds; (2) to establish the most representative mathematical function for B. japonicum mortality. | Penna et al. AMB Express 2011 1 21 http www.amb-express.eom content 1 1 21 o AMB Express a SpringerOpen Journal ORIGINAL Open Access A simple method to evaluate the number of bradyrhizobia on soybean seeds and its implication on inoculant quality control 11 12 3 Claudio Penna Rosana Massa Florencia Olivieri Gabriel Gutkind and Fabricio Cassán Abstract Soybean seeds are non-sterile and their bacterial population interferes with the enumeration of beneficial bacteria making it difficult to assess survival under different conditions. Within this context the principal aims of this work were 1 to improve a selective media for the enumeration of B. japonicum recovered from inoculated soybean seeds 2 to establish the most representative mathematical function for B. japonicum mortality on soybean seeds after inoculation 3 to evaluate if environmental or physiological conditions modify B. japonicum mortality on soybean seeds and 4 to create a new protocol for quality control of soybean inoculants. We successfully evaluated the combination of pentachloronitrobenzene and vancomycin added to the yeast-mannitol medium to inhibit most fungi and Gram-positive soybean microbiota thus producing reliable counts of B. japonicum from inoculated soybean seeds. Percentages of recovery and survival factors were obtained and used to construct a two-phase exponential decay non-linear regression function. High temperature and desiccation decreased these parameters while the optimization of temperature and the use of osmoprotective compounds with inoculants increased them. The use of this protocol minimized heterogeneity between experiments and may be considered more reliable than the simple expression of direct colony count of bacteria recovered from seeds. Keywords Bradyrhizobium japonicum inoculants soybean quality control Introduction Soybeans Glycine max L. Merr. are one of the major crops in grain-producing countries worldwide. Particularly in Argentina production for 2010 has been .