Isolation and Characterization of Chickpea (Cicer arietinum L.) Nodulating Rhizobia Collected from South Wollo Zone, Ethiopia

Abstract

This study aimed at screening rhizobial isolates of chickpea and evaluating their symbiotic nitrogen fixation efficiency and tolerance to abiotic stresses. A total of 107 (100%) isolates were collected of which 52 (48.6%) were confirmed as chickpea rhizobia using preliminary tests. Among 52 (48.6%) isolates, 46 (88.5%) have induced nodulation on chickpea under greenhouse and were screened under in vitro conditions and 6 (11.5%) of them were discarded due to fail to nodulate. The greenhouse data showed the highest nodule number (68.67 plant−1), nodule dry weight (0.17 g plant−1), and shoot dry weight (0.81 g plant−1) were scored by plants inoculated with isolates WUCR 17, WUCR 1, and WUCR 66, respectively. Among authenticated isolates, 73.9%, 21.7%, and 4.3% were highly effective, effective, and lowly effective, respectively. The physiological test showed 15% of isolates tolerated 13% salt concentration and 10.9% of isolates grew at pH 4–10 range. All isolates grew at 20–35°C and 13% grew at maximum temperature (50°C). The isolates showed better resistance to the tested antibiotics at low concentration (2.5 μg/ml) but the majorities were sensitive at high concentration (10 μg/ml). Among the isolates, 13% tolerated all tested heavy metals but 48% were sensitive to mercury. Regarding the carbohydrate utilization test, 52.2% catabolized all the tested 11 carbon sources but 6.5% of them utilized only 63.6% carbon sources. Amino acid utilization showed isolates (85%) utilized D-alanine except WUCR (14, 25, 31, 34, 39, 59, and 76) and 76% of the isolates utilized arginine and phenylalanine and 74% utilized leucine as a nitrogen source. Of the isolates, 43.5% of them utilized both D-alanine and arginine as a nitrogen source. In almost all tests conducted, isolates WUCR 1 and 5 performed top and were recommended as potential candidates for microbial inoculants.