Introduction: For many years, chickpeas were considered a restricted host species for nodulation, although recent studies describe greater bacterial diversity associated with the crop, allowing for increased knowledge of this symbiosis to obtain isolates that are efficient in their contribution. Objective: To characterize and identify an isolate obtained from chickpea plant nodules and evaluate its effectiveness under field conditions. Materials and methods: The research was conducted in the laboratories of two centers: the Instituto de Investigaciones Fundamentales en Agricultura Tropical “Alejandro de Humboldt”, Cuba, and the Centro Nacional de Recursos Genéticos, Mexico. The research was also conducted under field conditions in two campaigns during 2018 and 2020 to demonstrate its effectiveness in crop interaction at the first center. Isolate R3 was identified at the taxonomic level by sequencing four genes. A completely randomized design was used for laboratory experiments, while plant trials were conducted using a randomized block design. The results were subjected to an analysis of variance and the means were compared according to the Tukey test (p < 0.05). Results: Isolate R3 showed variability in its morphophysiological, physiological, and biochemical characteristics. Furthermore, it responded significantly differently than the other treatments in the growth and yield indicators evaluated in chickpeas. Conclusions: Rhizobium pusense was identified for the first time in Cuba associated with chickpea nodules. The strain showed positive growth at pH levels between 5.5 and 9.0 and temperatures ranging from 29 °C to 38 °C, demonstrating its tolerance to these factors. Its inoculation in chickpeas stimulated nodule formation and increased yield variables.

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