Diversity And Symbiotic Characteristics Of Cowpea Brad Yrhizobium Strains In Ghanaian Soils

ABSTRACT This study reports investigation of the biodiversity of bradyrhizobia isolates that nodulate cowpea in Ghanaian soils. As a prelude, some components of nitrogen fixation of cowpea in the various soils were examined through: (1) assessment of the natural nodulation of 45 cowpea cultivars in 20 soils sampled from 5 ecozones (coastal savanna, tain forest, semi deciduous forest, forest savanna transition and guinea savanna), (2) determination of the numbers of bradyrhizobial isolates in the soils and (3) determination of the response of cowpea to nitrogen fertilization. The results of the ability of 45 cowpea cultivars to nodulate naturally in different soil types showed large variability among the cultivars. Counts of the indigenous bradyrhizobia population in the soils showed that most of the soils in Ghana harbour large populations of bradyrhizobia (in the range of 0.6 x 10 to 31 x 103) capable of nodulating cowpea. Response of cowpea to nitrogen fertilizer differed in the different soils. In general all the cultivars showed significant responses to increasing levels of nitrogen, an indication that nitrogen fixation was not supplying the plants with all the external nitrogen required for maximum yield. A combination of morpho-physiological and molecular analysis was used to assess the diversity of the bradyrhizobia isolates. A total of 100 isolates were assessed. The results of the morphophysiological analysis indicated that cowpea is nodulated by both fast and slow growing rhizobia. The results also showed that the isolates were versatile and could survive under different soil conditions particularly acidity and salt stress. A cross inoculation study of the isolates with nine legume species produced seven major groupings with 28 subgroups based on distinct nodulation patterns. Results of the serology (ELISA) assay indicated that only a small fraction of the isolates reacted strongly with antisera of each other. The greater proportion showed no cross reactivity. Analysis of the 16S rRNA gene of the isolates by PCR-RFLP identified 20 different composite genotypes. Diversity among the genomic species identified was very high, reaching 80% diversity. The various methods used indicated large diversity among the isolates, but the groupings of the isolates by the various methods were inconsistent, due to the different levels of resolution by the various methods. Diversity of the isolates in symbiotic effectiveness showed that some of the isolates had high nitrogen fixing capabilities that were comparable to plants fertilized with inorganic fertilizer nitrogen. Some of the isolates even showed superiority in symbiotic effectiveness relative to the standard strain TAL 169, suggesting that the native isolates may be useful strains for cowpea inoculation. The Gus A marker gene technique was used to assess the competitive abilities of the effective and ineffective isolates. Competition between the isolates was examined at different population ratios. The results obtained indicated that competitive ability was not directly related to effectiveness of strains. Inoculation of cowpea with indigenous bradyrhizobia isolates increased the number of nodules, shoot dry weight and total nitrogen of plants. The method of inoculation was observed to influence these parameters The results indicated that response of cowpea to inoculation in the presence of native rhizobia in some soils is possible.