ABSTRACTAzospirillumis a rhizobacterial genus containing plant growth-promoting species associated with different crops worldwide.Azospirillum brasilensestrains exhibit a growth-promoting effect by means of phytohormone production and possibly by N2fixation. However, one of the most important factors for achieving an increase in crop yield by plant growth-promoting rhizobacteria is the survival of the inoculant in the rhizosphere, which is not always achieved. The objective of this study was to develop quantitative PCR protocols for the strain-specific quantification ofA. brasilenseFP2. A novel approach was applied to identify strain-specific DNA sequences based on a comparison of the genomic sequences within the same species. The draft genome sequences ofA. brasilenseFP2 and Sp245 were aligned, and FP2-specific regions were filtered and checked for other possible matches in public databases. Strain-specific regions were then selected to design and evaluate strain-specific primer pairs. The primer pairs AzoR2.1, AzoR2.2, AzoR5.1, AzoR5.2, and AzoR5.3 were specific for theA. brasilenseFP2 strain. These primer pairs were used to monitor quantitatively the population ofA. brasilensein wheat roots under sterile and nonsterile growth conditions. In addition, coinoculations with other plant growth-promoting bacteria in wheat were performed under nonsterile conditions. The results showed thatA. brasilenseFP2 inoculated into wheat roots is highly competitive and achieves high cell numbers (∼107CFU/g [fresh weight] of root) in the rhizosphere even under nonsterile conditions and when coinoculated with other rhizobacteria, maintaining the population at rather stable levels for at least up to 13 days after inoculation. The strategy used here can be applied to other organisms whose genome sequences are available.
Draft Genome Sequences of Three Rhizospheric Plant Growth-Promoting Bacteria
