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Open Access Research article

Symbiotic functioning and bradyrhizobial biodiversity of cowpea (Vigna unguiculata L. Walp.) in Africa

Flora Pule-Meulenberg1, Alphonsus K Belane2, Tatiana Krasova-Wade4 and Felix D Dakora3*

Author Affiliations

1 Department of Biotechnology, Tshwane University of Technology, Arcadia Campus, 175 Nelson Mandela Drive, Private Bag X680, Pretoria 0001, South Africa

2 Department of Crop Science, 1 Stateartillery Road, Tshwane University of Technology, Pretoria Campus, Private Bag X680, Pretoria 0001, South Africa

3 Department of Chemistry, Tshwane University of Technology, Arcadia Campus, 175 Nelson Mandela Drive, Private Bag X680, Pretoria 0001, South Africa

4 Laboratoire Commun de Microbiologie (LCM) IRD/ISRA/UCAD, BP 1386, CP 18524, Dakar, Senegal

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BMC Microbiology 2010, 10:89  doi:10.1186/1471-2180-10-89

Published: 23 March 2010

Abstract

Background

Cowpea is the most important food grain legume in Sub-Saharan Africa. However, no study has so far assessed rhizobial biodiversity and/or nodule functioning in relation to strain IGS types at the continent level. In this study, 9 cowpea genotypes were planted in field experiments in Botswana, South Africa and Ghana with the aim of i) trapping indigenous cowpea root-nodule bacteria (cowpea "rhizobia") in the 3 countries for isolation, molecular characterisation using PCR-RFLP analysis, and sequencing of the 16S - 23S rDNA IGS gene, ii) quantifying N-fixed in the cowpea genotypes using the 15N natural abundance technique, and iii) relating the levels of nodule functioning (i.e. N-fixed) to the IGS types found inside nodules.

Results

Field measurements of N2 fixation revealed significant differences in plant growth, δ15N values, %Ndfa and amounts of N-fixed between and among the 9 cowpea genotypes in Ghana and South Africa. Following DNA analysis of 270 nodules from the 9 genotypes, 18 strain IGS types were found. Relating nodule function to the 18 IGS types revealed significant differences in IGS type N2-fixing efficiencies. Sequencing the 16S - 23S rDNA gene also revealed 4 clusters, with cluster 2 forming a distinct group that may be a new Bradyrhizobium species. Taken together, our data indicated greater biodiversity of cowpea bradyrhizobia in South Africa relative to Botswana and Ghana.

Conclusions

We have shown that cowpea is strongly dependant on N2 fixation for its N nutrition in both South Africa and Ghana. Strain IGS type symbiotic efficiency was assessed for the first time in this study, and a positive correlation was discernible where there was sole nodule occupancy. The differences in IGS type diversity and symbiotic efficiency probably accounts for the genotype × environment interaction that makes it difficult to select superior genotypes for use across Africa. The root-nodule bacteria nodulating cowpea in this study all belonged to the genus Bradyrhizobium. Some strains from Southern Africa were phylogenetically very distinct, suggesting a new Bradyrhizobium species.