Genomic Diversity, Plant Growth Promoting Properties and Symbiotic Effectiveness of Mesorhizobum Spp. Nodulating Chickpea (Cicer Arietinum L.) on Acidic Soils of Ethiopia: Implication for Inoculant Development to Enhance Production Under Low Inputs Agriculture
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Date
2020-07-09
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Addis Ababa University
Abstract
Nitrogen and phosphorus deficiencies are the two most limiting factors in crop
production. Although chickpea fixes nitrogen in association with diverse Mesorhizobium
species, effectiveness in the amount of nitrogen fixed depends upon the inherent character
and other factors of the endosymbionts. The nitrogen-fixing process requires more
phosphrous and phosphorus deficiency aggravated under acidic soils because of sorption.
Thus, selection of acid tolerant rhizobia with effective nitrogen fixation and phosphate
solubilizing activity is essential in order to improve phosphorus and nitrogen fertility to
enhance the growth and production of the crop in acidic soil. This study was initiated to
collect chickpea rhizobia from acidic soils of central, western, southern and northern
parts of Ethiopia and determine their genomic diversity, and further screen for low pH
tolerance, solubilization of insoluble phosphate on tri-calcium phosphate medium and
their potential to ecological adaptations and symbiotic effectiveness with host varieties on
sand culture at low pH and acidic soil culture under greenhouse conditions. The genomic
diversity was studied using illumina sequencing a whole-genome sequence (WGS)
approach. The results showed that 81 Mesorhizobium strains were obtained and estimated
with average genome sizes 7Mbp, average depth of 143 X coverage. Annotation of the
assembled genome predicted an average of 6725 protein-coding genes (CDSs). Genome
completeness showed that 81 strains are near complete (>99% complete). Concatenation
of 400 universal phylophlan marker genes, present in all genomes (strains) allowed
detailed phylogenetic analysis, from which eight well-supported phylogenetic groups were
identified. Phylogenetic reconstruction based on the symbiosis-related (nod C and nif H)
genes were generally different to those shown by the phylophlan marker genes. The
pattern of genomic diversity indicates the existence of multiple, broadly distributed
phylotypes, with no relationship between geographic and genetic distance (r=0.10,
p<0.01). The result indicated that 62 strains grew well at low pH 5; and 47 (76%) of them
were phosphate solubilizers. The strains showed wide diversity in their substrate
utilization and tolerance to salinity, high temperatures, Mn 2+ and Al3+ toxicity, heavy
metals and antibiotics. Symbiotic characterization also showed a wide diversity among
these mesorhizobial strains. Moreover, most of the phosphate solublizing strains had
multiple growth promoting characteristics (production of indole acetic acid, hydrogen
cyanide, siderophore, ACC diaminase and inhibition of Fusarium oxysporum f.sp.ciceris
growth) under in vitro conditions. Under field conditions, the mesorhizobial inoculants
(a.117L2, a.71 and a.15star) showed significant (P<0.01) improvement in biological
nitrogen fixation, growth, yield, nitrogen and phosphorus uptakes of chickpea compared
to the commercially available local reference strain Cp41. The indigenous mesorhizobial
strain a.117L2 was superior inoculant for almost all the tested parameters. Generally, the
present study indicated that Ethiopian acidic soils contain symbiotically effective,
phosphate solublizing, chickpea nodulating Mesorhizobium spp. endowed with different
plant growth promoting attributes which are diverse in their genomic and taxonomic
identities. Therefore, there is a potential advantage using these mesorhizobial inoculants
to enhance chickpea production in acidic soils by improving phosphorus and nitrogen
fertility; and provide that further field trials recommended over several seasons and sites
in Ethiopia.
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Keywords
Inoculants, Whole-Genome Sequence, Yields