%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Couillerot, O.
%A Bouffaud, M.L.
%A Baudoin, Ezékiel
%A Muller, D.
%A Caballero-Mellado, J.
%A Moënne-Loccoz, Y.
%T Development of a real-time PCR method to quantify the PGPR strain Azospirillum lipoferum CRT1 on maize seedlings
%D 2010
%L fdi:010085705
%G ENG
%J Soil Biology and Biochemistry
%@ 0038-0717
%K ALLEMAGNE ; FRANCE
%K DUREN ; SAINT ANDRE COTE
%M ISI:000284294600030
%N 12
%P 2298-2305
%R 10.1016/j.soilbio.2010.09.003
%U https://www.documentation.ird.fr/hor/fdi:010085705
%> https://www.documentation.ird.fr/intranet/publi/2023-01/010085705.pdf
%V 42
%W Horizon (IRD)
%X Azospirillum lipoferum CRT1 is a promising phytostimulatory PGPR for maize, whose effect on the plant is cell density-dependent. A nested PCR method is available for detection of the strain but does not allow quantification. The objective was to develop a real-time PCR method for quantification of A. lipoferum CRT1 in the rhizosphere of maize seedlings. Primers were designed based on a strain-specific RFLP marker, and their specificity was verified under qualitative and quantitative PCR conditions based on successful CRT1 amplification and absence of cross-reaction with genomic DNA from various rhizosphere strains. Real-time PCR conditions were then optimized using DNA from inoculated or non-inoculated maize rhizosphere samples. The detection limit was 60 fg DNA (corresponding to 19 cells) with pure cultures and 4 x 10(4) CFU equivalents g(-1) lyophilized sample consisting of mixture of rhizosphere soil and roots. Inoculant quantification was effective down to 10(4) CFU equivalents g(-1). Assessment of CRT1 rhizosphere levels in a field trial was in accordance with estimates from semi-quantitative PCR targeting another locus. This real-time PCR method, which is now available for direct rhizosphere monitoring of A. lipoferum CRT1 in greenhouse and field experiments, could be used as a reference for developing quantification tools for other Azospirillum inoculants.
%$ 076 ; 084