engleski

Genotypic and symbiotic characterisation of indigenous field pea rhizobia in cultivated soils of the north-west Croatia

The natural population of Rhizobium leguminosarum bv. viciae was sampled from cultivated soils in north-west Croatia. The isolates were characterised with the following genotypic methods: (i)repetitive extragenomic palindromic – PCR, (ii) randomly amplified polymorphic DNA (RAPD) method, (iii)nodC gene amplification and (iiii)PCR-restriction fragment length polymorphism (RFPL) analysis of nodulation nodD gene. After genetic identification, isolates and reference strains were tested in greenhouse for evaluation of their symbiotic efficiency. All rhizobial isolates were identified as R. leguminosarum bv. viciae by specific amplification of nodC gene. Analysis of intraspecies relationship among strains by using the RAPD and rep-PCR showed significant differences between reference strains and all field isolates but also among R. leguminosarum bv. viciae isolates. Two indigenous strains, which are different from all other strains, in greenhouse experiment revealed high symbiotic efficiency. Based on results of restriction analysis of nodD gene regions, two indigenous strains, as well as two reference strains determined significantly different nodD gene types from all tested strains. So, classification of strains by the PCR- amplified symbiotic gene region methods were partly in correlation with classification of strains based on RAPD and rep-PCR. In greenhouse, indigenous strains with significantly different nodD gene type determined the highest nodule dry weight from all tested strains so it could be characterised as highly infective strains.

Rhizobium leguminosarum bv. viciae; indigenous strains; rep-PCR; RAPD; nodC gene; PCR-RFLP of nodD gene; symbiotic characterisation

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