2.8.3.

Crop rotation assay peanut-maize-peanut in test pots

Growth promotion of peanut and maize plants by selected PSB were tested in a
simulated crop rotation assay (peanut-maize-peanut). Plastic pots (30 cm-diameter,
35 cm height) that contained sieved unsterilized soil with low P content from the
peanut cultivation area of Córdoba (organic matter: 1.40% (Walkley-Black method),
pH: 6.82 (Potenciometry 1:2.5), N: 11.5 μg/g (phenolsulfonic acid), P: 8.5 μg/g
(Kurtz and Bray I method)) were used. The soil was supplemented with 40 mg/kg
Ca3(PO4)2. Firstly, peanut seeds (var. Granoleico) were surface disinfected as
described previously, germinated and transferred to the pots. The treatments were:
plants inoculated with (1) Pantoea sp. J49; (2) Bacillus sp. L55; (3) Serratia sp.
S119; (4) Enterococcus sp. L191; (5) Serratia sp. J260; (6) Pantoea sp. J49 +
Serratia sp. S119; (7) Bacillus sp. L55 + Enterococcus sp. L191; (8) Uninoculated
plants (negative control plants, C−); (9) uninoculated plants supplemented
regularly with 20 mM KH2PO4 (plants fertilized with P; C+) and (10) plants
inoculated with commercial strain P. fluorescens PMT1.

Bacterial inocula were obtained as described previously in the microcosm maize

assay. Bacterial inoculation was performed one time at the beginning of the trial.
Peanut plants were grown under controlled environmental conditions, watered
regularly with sterilized tap water and, once a week, with Hoagland nutrient
solution which was devoid of soluble P. Peanut plants were extracted at 45 days
post inoculation (first harvest of peanut). Subsequently, disinfected maize seeds
were sown in the same pots and at 21 days (66 days post inoculation), maize plants
were extracted. Subsequently peanut disinfected seeds were sown and at 45 days (111
days post inoculation) peanuts plants were extracted (second harvest of peanut). In
each plant harvest the following parameters were determined: length (cm), dry
weight (g/plant) and P content (mg/g) of aerial and root tissues, as described
above, as well as P content (μg/ml) and pH of soil.

2.9. Data analysis

The data were subjected to analysis of variance (ANOVA) and differences among
treatments were detected by Tukey test (P < 0.05) for biochemical parameters and
LSD test (P < 0.05) for plant assays. Pearson correlation coefficient between
soluble P released by bacteria and pH was calculated. In all cases Infostat
software was used.