Isolation and characterization of rhizobia from the root nodule of some cultivated legume crops
Biological nitrogen fixation (BNF) as a result of mutual symbiosis between the rhizobia and the cultivated legume have a vital role to balance the nutrient paucity in the soil. Modern researches shows that the BNF can be the important factors regulating and maintaining the defendable agriculture and ensuring food security especially in the developing countries. The use of artificial nitrogen fertilizers to rise crop yield is an internment farming practice, despite its unfavorable effects and hazards to the environment and human population which can be substituted by rhizobial inoculants as a bio-fertilizers. The present study was aimed to isolate and characterize the Rhizobium from the nine different selected legumes. The Rhizobium bacterium was isolated from the nodules of the nine legume plants on YEMA medium which was found to show white translucent, circular convex colonies and characterized by the non-absorption of Congo red dye. The strains were found to be fast growing except for the rhizobial strains isolated from the Soybean and cowpeas (Bradyrhizobium) which were slow growing. The various biochemical tests of the isolated strains like catalase, bromothymol blue, Urea hydrolysis were favorable while Glucose-Peptone Agar (GPA), starch hydrolysis, Citrate utilization were found to be negative. For the ability to utilize the 2% NaCl, the strains TFR showed positive growth, the strains PSR, PhVR, VUR showed the poor tolerance while the rest of the strains showed no tolerance to the 2% NaCl.
2. Jensen ES, Peoples MB, Hauggaard-Nielsen H. Faba bean in cropping systems. Field Crops Res. 2010; 115: 203-216.
3. Köpke U, Nemecek T. Ecological services of faba bean. Field Crop Res. 2010; 115: 217-233.
4. Peoples MB, Hauggaard-Nielsen H, Jensen ES. The potential environmental benefits and risks derived from legumes in rotations. In: Emerich DW, Krishnan HB, eds. Nitrogen fixation in crop production. Madison: American Society of Agronomy, Crop Science Society of America, Soil Science Society of America. 2009: 349-385.
5. Hardarson G, Broughton WJ. Maximizing the Use of Biological Nitrogen Fixation in Agriculture. 2003: pp. 69.
6. Bala A, Ken G. Symbiotic specificity of tropical tree rhizobia for host legumes. New Phytologist. 2002; 149: 495-507.
7. Sharma PK, Kundu BS, Dogra RC. Molecular mechanism of host specificity in legume Rhizobium symbiosis. Biotechnol Adv. 1993; 11(4): 741-779.
8. Paudyal SP. Development of Bio-inoculants from the Indigenous Strains of Rhizobia Isolated from Certain Medicinal Plants. Doctoral dissertation. Dr. B.R. Ambedkar University, Agra, India. 2002.
9. Bhattarai N, Baral B, Shrestha G, Yami KD. Effects of Mycorrhiza and Rhizobium on Phaseolus vulgaris L. Sci Worlds. 2011; 9(9): 66-69.
10. Jordan DC, Allen ON. Genus II Rhizobium In Bergeys Manual of Determinative Bacteriology, 8th ed., (Buchanan RE and Gibson NE eds.). The Williams and Wilkins, Co. Baltimore. 1974; 262-264.
11. Fred EB, Baldwin IL, McCoy E. Root nodule bacteria and leguminous plants, University of Wisconsin Studies in Science No. 5. University of Wisconsin. Madison. 1932.
12. Willems A, Collins MD. Phylogenetic analysis of rhizobia and agrobacteria based on 16S rRNA gene sequences. Int J Syst Bacteriol. 1993; 43(2): 305-313.
13. Young JM, Kuykedall LD, Romero-Martinez E, Keer A, Sawada H. Classification and nomenclature of Agrobacterium and Rhizobium-a reply to Farrand et al. (2003). Int J Syst Evol. Microbiol. 2003; 53(5): 1689-1695.
14. Weir BS. The current taxonomy of rhizobia. NZ Rhizobia. 2016, https://www.rhizobia.co.nz/taxonomy/rhizobia
15. Mohammadi K, Sohrabi Y, Heidari G, Khalesro S, Mohammad M. Effective factors on biological nitrogen fixation. Afr J Agric Res. 2012; 7(12): 1782-1788.
16. Perret X, Staehelin C, Broughton WJ. Molecular basis of symbiotic promiscuity. Microbiol Mol Biol Rev. 2000; 64: 180-201.
17. Fellay R, Perret X, Viprey V, Broughton WJ, Brenner S. Organization of host inducible transcripts on the symbiotic plasmid of Rhizobium sp. NGR234. Mol Microbiol. 1995; 16: 657-667.
18. Giraud E, Moulin L, Vallenet D, Barbe V, Cytryn E, Avarre JC, et al. Legumes symbioses: Absence of Nod genes in photosynthetic Bradyrhizobia. Science. 2007; 316: 1307-1312.
19. Wadhwa Z, Srivastava V, Rani R, Bhatia T, Makkar KT, Jangra S. Isolation and characterization of Rhizobium from Chickpea (Cicer arietinum). Int J Curr Microbiol Appl Sci. 2017; 6: 2880-2893.
20. Shahzad F, Muhammad S, Abbas F, Babar S, Mohammad T, Ahmad Z. Isolation and biochemical characterization of Rhizobium meliloti from root nodules of Alfalfa (Medico sativa). J Anim Plant Sci. 2012; 22: 522-524.
21. Prajapati S, Dadke MS, Surekha S, Godika S, Krishna VP. Isolation and characterization of Rhizobium meliloti isolated from rhizosphere soil and roots of fenugreek from different locations. Int J Curr Microbiol Appl Sci. 2018; 7(6): 1460-1467.
22. Amel B, Tellah S, Faiza, Ounane S. Characterization of rhizobia from root nodule and rhizosphere of Vicia faba in Algeria. Legume Res An Int J. 2018; 41: 624-628.
23. Simon Z, Mtkei K, Gessesse A, Ndakidemi P. Isolation and characterization of Nitrogen Fixing rhizobia from cultivated and uncultivated soils of Northern Tanzania. Am J Plant Sci. 2014; 05: 4050-4067.
24. Vincent JM. A Manual for the Practical Study of Root Nodule Bacteria. Oxford, Blackwell Scientific. 1970.
25. Martínez-Romero E. Coevolution in Rhizobium-legume symbiosis? DNA Cell Biol. 2009; 28: 361-370.
26. Willems A. The taxonomy of rhizobia: an overview. Plant Soil. 2006; 287: 3-14.
27. Abrar TH, Alebejo AL. Isolation and characterization of rhizobia from rhizosphere and root nodule of Cowpea, Elephant and Lab lab plants. Int J Nov Res Interdisc Stud. 2017; 4: 1-7.
28. Datta A, Singh R, Tabassum S. Isolation, Characterization and Growth of Rhizobium strains under optimum conditions for effective Biofertilizer production. Int J Pharm Sci Rev Res. 2015; 32: 199-208.
29. Rivas R, Laranjo M, Mateos PF, Oliveria E, Martinez-Molina E, Velazquez E. Strains of Mesorhizobium amorphae and Mesorrhizobium tianshanense, carrying symbiotic genes of common chickpea endosymbiotic species, constitutue a novel biovar (Ciceri) capable of nodulating Cicer arietinum. Lett Appli Microbiol. 2007; 44: 412-418.
30. Geetha R, Maheshwari H, Patel, Sanket J. Isolation and characterization of nodule-associated Exiguobacterium sp. from the root nodules of Fenugreek (Trigonella foenum-graecum) and their possible role in plant growth promotion. Int J Microbiol. 2012: 693982.
31. Puozaa DK, Jaiswal SK, Dakora FD. Phylogeny and distribution of Bradyrhizobium symbionts nodulating cowpea (Vigna unguiculata L. Walp) and their association with the physicochemical properties of acidic African soils. Syst Appl Microbiol. 2019; 42(3): 403-414.
32. Rivas R, Garcia-Fraile P, Velazquez E. Taxonomy of bacteria nodulating legumes. Microbiol Insights. 2009; 2: 51-69.
33. Andrews M, Andrews ME. Specificity in legume-rhizobia symbioses. Int J Mol Sci. 2017; 8(4): 705.
34. Somasegaran P, Hoben HJ. Collecting nodules and isolating rhizobia. In: Handbook for Rhizobia. Methods in Legume-Rhizobium Technology. Springer. 1994; 7-23.
35. Graham PH, Parker CA. Diagnostic features in characterization of root nodule bacteria of legumes. Plant Soil. 1964; 20: 383-396.
36. Yashita, KN. Isolation, Identification and Characterization of Rhizobium from Hyacinth Bean (Lablab niger) Root Nodule and Study its Effect on Plant Growth and Nitrogen Fixation Ability. Doctoral dissertation. BRAC University. Dhaka, Bangaladesh. 2016.
37. Paudel N, Subedi S, Mandal TN, Das BD. Establishment of plant residues and inorganic fertilizer application for growth and yield of Vigna unguiculata (L.) in flood-affected cropland of Koshi Tappu Region, Eastern Nepal. Eur J Biol Res. 2021; 11(1): 75-87.
38. Saxena A, Rewari R. Differential responses of Chickpea (Cicer arietinum L.) Rhizobium combinations to saline soil conditions. Biol Fertil Soils. 1992; 13: 31-34.
39. Lindstrom K, Lehtomaki S. Metabolic properties, maximum growth temperature and phase sensitivity of Rhizobium sp compared with other fast-growing rhizobia. FEMS Microbiol Lett. 1988; 50: 277-287.
40. Kovaks N. Identification of Pseudomonas pyocyaneta by the oxidase reaction. Nature. 1956; 178: 703.
41. Paudyal SP, Gupta VNP. Bio-chemical characterization of rhizobia isolated from root nodules of Velvet bean (Mucuna pruriens L.). Our Nature. 2017; 15(1-2): 7-12.
42. Paudyal SP, Gupta VNP. Substitution of chemical fertilizer nitrogen through rhizobium inoculation technology, Our Nature. 2018; 6(1): 43-47.
43. Paudyal SP, Das BD, Paudel VR, Paudel N. Impact on productivity of preparation on rhizobial inoculant carriers. Eur J Biol Res. 2021; 11(2): 242-250.
44. Ampomah OY, Ofori-Ayeh E, Solheim B, Svenning MM. Host Range, Symbiotic Effectiveness and Nodulation Competitiveness of Some Indigenous Cowpea Bradyrhizobia Isolates from the Transitional Savanna Zone of Ghana. Afr J Biotechnol. 2008; 7: 988-996.
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