Scholarly record
ALLELIC POLYMORPHISM OF SYMBIOTICALLY ESSENTIAL GENES OF SINORHIZOBIUM MELILOTI NATIVE TO LEGUME PLANTS ORIGIN AT THE CAUCASUS
Abstract
Legumes are a unique source of protein for food and feed industry. Legumes synthesize protein valuable for living organisms using atmospheric nitrogen which they can fix due to their symbiosis with root nodule bacteria. The formation of plant-microbial symbiotic systems depends on complementary interaction of signaling molecules of both symbionts. Rhizobia signaling molecules are lipochitooligosaccharides, so called Nod factors. The synthesis of core part of Nod factor molecule is determined by nodA, nodB and nodC genes, that was shown for a vast majority of rhizobia species. There is the evidence that structural changes in one as well in three nod genes could lead to significant changes in symbiotic activity of rhizobia like nodule formation or specificity toward plant host. The aim of this work was to assess allelic polymorphism of nod genes in relation to symbiotic activity of Sinorhizobium meliloti strains native to the origin of cultivated plants at the Caucasus. The analysis was done on the 70 S. meliloti strains which were originally isolated from pink nodules (N-isolates) of wild-growing alfalfa or sweet clover plants and on the 63 strains trapped by alfalfa plants in sterile plant tests (T-isolates). From 3 to 5 alleles were identified for each above mentioned genes when DNA from isolates were tested by PCR-RFLP approach. The alleles of nodA, nodB and nodC genes similar to reference were revealed among 44% of N-isolates. While, most of N-isolates and almost 70% of T-isolates had alleles different from the reference (divergent alleles). The certain combinations of nodA/nodC alleles were identified in N- and T-isolates, while the specific combination of nodA/nodB alleles was revealed only for T-isolates (according to linkage disequilibrium test; LD). The level of diversity of genes of nod operon was higher for N-isolates than for T-isolates (N=0.78, 0.80, 0.84, and N=0.60, 0.65, and 0.77, respectively). In total the 8 distinct allelic combinations (PCR-nod types from I till VIII) were revealed for nodABC operon and the occurrence of those types between N- and T-group of isolates was significantly different (P<0.05). Thus, the nod type-IV was in 1.5 times more frequently determined among T-isolates, while the nod type-II was 2 times more frequently detected among N-isolates (P<0.05). The four particular nodABC genotypes were determined in majority of N- and T-isolates and coming from that the "trap" approach should be considered as valid method for genotypic diversity analysis in rhizobia native populations. We have shown for the first time that a rare nodABC/nodH genotypes were revealed almost six times more among N-isolates, while unique nod genotypes were identified only among T-isolates. This data is supporting the evidence that host plant contributes to genetic diversity of the microsymbiont. Summarizing, the entire pool of alleles of symbiotically essential genes was obtained as a result of the study of both nodule and soil subpopulations of rhizobia.
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