Abstract

The study of bacteria associated with plants of agricultural importance has increased recently;
considering the potential of these microorganisms for use as inoculants. The objective of this
research was to study the diversity of bacteria associated with the Agave americana L. and select
strains with potential for use as fertilizers in this species of agave. The bacterial strains were
isolated from rhizospheric soil as well as the leaves of the plants, and were subsequently
characterized phenotypically, through a microscopic analysis and differential staining of Gram. The
microscopic study showed that the isolates were made up mainly by bacteria that have the form of
short, Gram negative bacilli, rapid growth, and that in the YEM formed various forms of colonies.
The potential of the strains for use as biofertilizers, was evaluated through evidence of inoculation
in the greenhouse using seedlings of A. americana L. as a model plant. A total of 235 strains were
isolated, which were analyzed by use of genomic fingerprints ERIC, ARDRA and by the
amplification of 16s rDNA chromosomal gene to have a taxonomic approach. The phylogenetic
analysis based on 16S rDNA gene allowed grouping strains within the genera Achromobacter,
Acinetobacter, Agrobacterium, Comamonas, Enterobacter, Klebsiella, Novoshingobium,
Pseudomonas, Rhizobium y Stenotrophomonas. Several of these genera include species that are
characterized by their high potential of biological fixation of N2 and by the ability to solubilize
phosphate and synthesize auxin (IAA). The strains of Rhizobium sp. ACO-34A and Acinetobacter
calcoaceticus ACO-40, influenced significantly (p<0.05) on the growth of plants of A. americana
and in the content of sugars (fructose, glucose, sucrose and principally fructans). The greater
amount of sugars type fructan was registered on the leaves of plants inoculated with the strain of
Rhizobium sp. ACO-34A; which was sequenced using the platform of massive sequencing of
Pacific Biosciences (PacBio RSII). The assembly of the genome revealed the presence of 5
replicones that correspond to 1 chromosome of 4.75 Mbp and 4 plasmids (516, 494, 305 y 213 Kb),
these results are consistent with the pattern of plasmids observed through the methodology of
Ekcardt. The analysis of the genome of this strain showed that contains genes related to the
biological fixation of N2 as are the nifHDK, and although there were some genes related to the
process of nodulation, not essential genes were found to carry this biological process. In this work it
was demonstrated that the bacterial strains play an important role in the biosynthesis of sugars in
plants of A. americana L. inoculated, which represents an important alternative for the end of
cultivation, agro-industrial production and utilization of this agave.

Key words: Fructans, diazotrophic bacteria, nitrogen fixation.