Annals of Microbiology

, Volume 68, Issue 5, pp 247–260 | Cite as

Diversity and symbiotic divergence of endophytic and non-endophytic rhizobia of Medicago sativa

  • Wenjuan Kang
  • Shangli ShiEmail author
  • Lin Xu
Original Article


Knowledge of rhizobium diversity is helping to enable the utilization of rhizobial resources. To analyze the phenotypic and genetic diversity and the symbiotic divergence of rhizobia of Medicago sativa, 30 endophytic and non-endophytic isolates were collected from different parts of five alfalfa varieties in three geographic locations in Gansu, China. Numerical analyses based on 72 phenotypic properties and restriction fragment length polymorphism (RFLP) fingerprinting indicated the abundant phenotypic and genetic diversity of the tested strains. According to the phylogenetic analysis of 16S RNA, atpD, glnII, and recA gene sequences, Rhizobium and Ensifer were further classified into four different genotypes: Rhizobium radiobacter, Rhizobium sp., Rhizobium rosettiformans, and Ensifer meliloti. The differences in architecture and functioning of the rhizobial genomes and, to a lesser extent, environment diversification helped explain the diversity of tested strains. The tested strains exhibited similar symbiotic feature when inoculated onto M. sativa cvs. Gannong Nos. 3 and 9 and Qingshui plants for the clustering feature of their parameter values. An obvious symbiotic divergence of rhizobial strains was observed in M. sativa cvs. Longzhong and WL168HQ plants because of the scattered parameter values. Their symbiotic divergence differed according to alfalfa varieties, which indicated that the sensitivity of different alfalfa varieties to rhizobial strains may differ. Most of the tested strains exhibited plant growth-promoting traits including phosphate solubilization and production of indole-3-acetic acid (IAA) when colonizing plant tissues and soil.


Medicago sativa Rhizobium Phenotypic diversity Genetic diversity Symbiotic divergence 



We thank the Key Laboratory of Grassland Ecosystem of Ministry of Education of Gansu Agricultural University for providing sampling sites and alfalfa plants. Thanks should go to Lin Xu for writing assistance.


This work is supported by the National Natural Science Foundation of China (NSFC), Fund No. 31562666.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018

Authors and Affiliations

  1. 1.College of Grassland ScienceGansu Agricultural UniversityLanzhouChina
  2. 2.Key Laboratory of Grassland Ecosystem of Ministry of EducationLanzhouChina
  3. 3.College of Agriculture and BiotechnologyHexi UniversityZhangyeChina

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