Heterologous expression of nifA or nodD genes improves chickpea-Mesorhizobium symbiotic performance

  • José Rodrigo da-Silva
  • Esther Menéndez
  • Fernando Eliziário
  • Pedro F. Mateos
  • Ana AlexandreEmail author
  • Solange Oliveira
Regular Article



The aim of this study was to investigate whether the overexpression of NifA and NodD regulators contribute to the symbiotic improvement of chickpea mesorhizobia.


The native strains V-15b, ST-2, and PMI-6 were transformed with extra copies of nifA or nodD genes and several plants trial were performed.


Plant growth assays showed that nifA overexpression was able to improve the symbiotic effectiveness of V-15b, while nodD overexpression lead to the improvement of ST-2 and PMI-6. Hydroponic assays showed that plants inoculated with V15bnifA+ and PMI6nodD+ started developing nodules earlier than those inoculated with the corresponding control strains. In addition, the number of nodules was always higher in plants inoculated with the strains overexpressing the symbiotic genes. Analysis of histological sections of nodules formed by V15bnifA+ showed a more developed fixation zone when compared with control. On the other hand, nodules induced by PMI6nodD+ did not show a senescent zone, which was observed in nodules from plants inoculated with the control strain. Plants inoculated with PMI6nodD+ and ST2nodD+ showed a higher number of infection threads than the corresponding control inoculations.


These results indicate that overexpressing nifA and nodD may be an important tool to achieve the improvement of the symbiotic performance of mesorhizobia.


Overexpression Nodulation Nitrogen fixation Legume Rhizobia Symbiotic effectiveness 



The authors thank Dr. Alvaro Peix (IRNASA-CSIC) for providing pMP4661 plasmid and Dr. Doroteia Campos for her help with the real-time PCR experiments performed in the Molecular Biology Laboratory-ICAAM. The authors also thank G. Mariano for technical assistance.

Funding information

This work was financed by FEDER Funds through the Operational Program for Competitiveness Factors—COMPETE and National Funds through FCT (Fundação para a Ciência e a Tecnologia), under the Strategic Project UID/AGR/00115/2013, Project n° FCOMP-01-0124-FEDER-028316 (PTDC/BIA-EVF/4158/2012), Project POCI-01-0145-FEDER-016810 (PTDC/AGR-PRO/2978/2014) and InAlentejo ALENT-07-0262-FEDER-001871. J. Rodrigo da-Silva acknowledges a PhD fellowship (1254-13-8) from CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior).

Supplementary material

11104_2019_3950_MOESM1_ESM.docx (1.6 mb)
ESM 1 (DOCX 1642 kb)


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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Laboratório de Microbiologia do Solo, Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Instituto de Investigação e Formação Avançada (IIFA)Universidade de ÉvoraÉvoraPortugal
  2. 2.Departamento de Microbiología y Genética, Centro Hispano Luso de Investigaciones Agrarias, Unidad Asociada CSIC-USALUniversidad de SalamancaSalamancaSpain

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