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Molecular diversity studies of 19 rhizobia isolates from chickpea were conducted using simple sequence repeats (SSR) and 16S rDNA-RFLP markers. Phenotypic characterization with special reference to salinity and pH tolerance was performed. These isolates were identified as different strains of Mesorhizobium, Rhizobium, Bradyrhizobium, and Agrobacterium. Twenty SSR loci of Mesorhizobium ciceri, distributed across the other rhizobial genome, clearly differentiated 19 rhizobial isolates. Analogous clustering supported the results of 16S rDNA sequence-based phylogeny. Analysis of the 16S rDNA sequences from M. ciceri strains revealed that nucleotide variables (signature sites) were located at 20 different positions; most of them were present in the first 820 bp region from 5’ terminal. Interestingly, 14 signature sites were located in two main regions, the variable region V1 (nt 527–584), and variable region V2 (nt 754–813). The secondary structure and minimal free energy were determined in these two regions. These results will be useful in characterizing the micro-evolutionary mechanisms of species formation and increase understanding of the symbiotic relationship.
Ultra-structural studies were conducted on root nodules of Samanea saman (Jacq.) Merr. collected from trees growing under natural conditions. Nodules were distributed singly as well as in clusters on the main and lateral roots. Mature nodules were elongated, branched and coralloid. Root hair curling was found but infection threads could not be observed. Rhizobia entered through the epidermis and moved intercellularly through the cortical region. Mature nodules of S. saman could be differentiated into meristem, cortex, vascular tissue and bacteroid tissue. The latter showed both infected and non-infected cells mixed together. Vascular bundles were inversely collateral and distributed around the bacteroid tissue. The bacteroids were enclosed in peribacteroid membrane in groups and showed prominent granules of polyhydroxybutyrate in their cytoplasm. Mycorrhizal hyphae were also observed along with rhizobia in the bacteroid tissue. S. saman with dual rhizobial and mycorrhizal infection is a potential tree for plantation in arid soils of Pakistan.
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