ISOLATION OF ROOT NODULE ENDOPHYTES FROM COWPEA (Vigna  unguiculata L.) GROWING IN JHUM FIELD IN NAGALAND (INDIA)

Authors

DOI:

https://doi.org/10.48165/abr.2024.26.01.53

Keywords:

Crop legumes, Jhum field, Root nodule endophytes, RAPD, Vigna unguiculata L.

Abstract

 Legume root nodules are inhabited by numerous symbiotic endophytes which impact the plant growth and development. Cowpea (Vigna unguiculata L.) is one of the most widely grown legume crops in the Jhum fields of Nagaland (India). However, the information on root nodule endophytes of cowpea grown in Jhum (slash and burn) ecosystem is lacking. With the aim to decipher the root nodule micro-symbionts, in the present study, 17 endophytic bacteria were isolated from the root nodules of cowpea. These isolates were clustered into two major groups, constituting eight sub-groups based on RADP-fingerprinting using nif-directed RPO1 primer. Eight unique isolates representing each sub-group were selected for 16S rRNA gene amplification and sequencing. Sequence analysis using BLASTn revealed that these isolates belonged to either Enterobacter or Bacillus species. The presence of these bacteria in root nodules suggests some vital role of non-rhizobial endophytes in the legume root nodule biology.

Downloads

Download data is not yet available.

References

Altschul, S.F., Gish, W., Miller, W., Myers, E.W. and Lipman, D.J. 1990. Basic local alignment search tool. Journal of Molecular Biology, 215: 403-410.

Anonymous (2022-2023). Statistical Handbook of Nagaland. Directorate of Economics and Statistics, Kohima, Nagaland, India.

Barbano, D.M., Clark, J.L., Dunham, C.E. and Flemin, R.J. 1990. Kjeldahl method for determination of total nitrogen content of milk: Collaborative study. Journal of the Association of Official Analytical Chemists, 73: 849-859.

Bray, R.H. and Kurtz, L.T. 1945. Determination of total organic and available forms of phosphorus in soils. Soil Science, 59: 39-45.

Etesami, H. 2022. Root nodules of legumes: A suitable ecological niche for isolating non-rhizobial bacteria with biotechnological potential in agriculture. Current Research in Biotechnology, 4: 78-86.

Ferguson, B.J., Indrasumunar, A., Hayashi S., Lin M. H., Lin Y. H., Reid D.E. and Gresshoff, P.M. 2010. Molecular analysis of legume nodule development and autoregulation. Journal of Integrative Plant Biology, 52: 61-76.

Gerding, M., Oyarzúa, P., García, L., Fischer, S., Norambuena, C., Barahona, V., del Pozo, A. and Ovalle, C. 2017. Diversity and symbiotic effectiveness of Adesmia spp. root nodule bacteria in central and southern Chile. Symbiosis, 72: 61-72. [https://doi.org/10.1007/s13199-016-0440-9].

Giller, K.E., Herridge, D.F. and Sprent, J.I. 2016. The legume-rhizobia symbiosis and assessing the need to inoculate. pp. 15-24. In: Working with Rhizobia (eds. J.G Howieson and M.J. Dilworth). Australian Centre for International Agricultural Research, Canberra, Australia.

Hanway, J.J. and Heidal, H. 1952. Soil analysis methods as used in Iowa State College soil testing laboratory. Iowa State College of Agriculture Bulletin, 57: 1-31.

Hirsch, A.M. 1992. Developmental biology of legume nodulation. New Phytologist, 122: 211- 237. [https://doi.org/10.1111/j.1469-8137.1992.tb04227.x].

Jayawardhane, J., Goyali, J.C., Zafari, S. and Igamberdiev, A.U. 2022. The response of cowpea (Vigna unguiculata) plants to three abiotic stresses applied with increasing intensity: Hypoxia, salinity, and water deficit. Metabolites, 12: 38. [doi: 10.3390/metabo12010038].

Jesus, J.G., Máguas, C., Dias, R., Nunes, M., Pascoal, P., Pereira, M. and Trindade, H. 2023. What if root nodules are a guesthouse for a microbiome? The case study of Acacia longifolia. Biology, 12: 1168. [doi: 10.3390/biology12091168]

Keen, B.A. and Raczkowski, H. 1921. The relation between the clay content and certain physical properties of a soil. Journal of Agricultural Science, 11: 441-449. [doi:10.1017/ s0021859600004469].

Maróti, G. and Kondorosi, E. 2014. Nitrogen-fixing Rhizobium-legume symbiosis: Are polyploidy and host peptide-governed symbiont differentiation general principles of endosymbiosis? Frontiers in Microbiology, 30: 5-326. [doi: 10.3389/fmicb.2014.00326].

Martínez-Hidalgo, P. and Hirsch, A.M. 2017. The nodule microbiome: N2-fixing rhizobia do not live alone. Phytobiomes, 1: 70-82.

Mukhtar, S., Hirsch, A.M., Khan, N., Malik, K.A., Humm, E.A., et al., 2020. Impact of soil salinity on the cowpea nodule-microbiome and the isolation of halotolerant PGPR strains to promote plant growth under salinity stress. Phytobiomes Journal, 4: 364-374.

Muresu, R., Polone, E., Sulas, L., Baldan, B., Tondello, A., Delogu, G., Cappuccinelli, P., Alberghini, S., Benhizia, Y., Benhizia, H. and Benguedouar, A. 2008. Coexistence of predominantly nonculturable rhizobia with diverse, endophytic bacterial taxa within nodules of wild legumes. FEMS Microbiology Ecology, 63: 383-400.

Odori, C., Ngaira, J., Kinyua, J. and Nyaboga, E.N. 2020. Morphological, genetic diversity and symbiotic functioning of rhizobia isolates nodulating cowpea (Vigna unguiculata L. Walp) in soils of Western Kenya and their tolerance to abiotic stress. Cogent Food and Agriculture, 6: 1853009. https://doi.org/10.1080/23311932.2020.1853009.

Piper, C.S. 1942. Soil and Plant Analysis: Laboratory Manual of Methods for the Examination of Soils and the Determination of the Inorganic Constituents of Plants. University of Adelaide, Adelaide, Australia.

Richardson, A.E., Viccars, L.A., Watson, J.M. and Gibson, A.H. 1995. Differentiation of Rhizobium strains using the polymerase chain reaction with random and directed primers. Soil Biology and Biochemistry, 27: 515-524.

Sadowsky, M. and Graham, P. 2006. Root and stem nodule bacteria of legumes. The Prokaryotes, 2: 818-841.

Sentimenla. 2020. Assessment of the soil chemical properties, macro and micronutrients using soil test kit and soil health card distribution in Zunheboto district of Nagaland, India. International Journal of Current Microbiology and Applied Sciences, 9: 2431-2435.

Somasegaran, P. and Hoben, H.J. 1994. Handbook for Rhizobia: Methods in Legume-Rhizobium Technology. Springer, New York, USA.

Tariq, M., Hameed, S., Yasmeen, T., Zahid, M. and Zafar, M. 2014. Molecular characterization and identification of plant growth-promoting endophytic bacteria isolated from the root nodules of pea (Pisum sativum L.). World Journal of Microbiology and Biotechnology, 30: 719-725.

van Spronsen, P.C., Gronlund, M., Pacios Bras, C., Spaink, H. and Kijne, J.W. 2001. Cell biological changes of outer cortical root cells in early determinate nodulation. Molecular Plant-Microbe Interactions, 14: 839-847.

Vincent, J.M. 1970. A Manual for the Practical Study of the Root-Nodule Bacteria. IBB Handbook No.15. Blackwell, Oxford, UK.

Walkley, A.J. and Black, I.A. 1934. Estimation of soil organic carbon by the chromic acid titration method. Soil Science, 37: 29-38.

Watts, D., Palombo, E.A., Jaimes, C.A. and Zaferanloo, B. 2023. Endophytes in agriculture: Potential to improve yields and tolerances of agricultural crops. Microorganisms, 11: 1276. [doi: 10.3390/microorganisms11051276].

Weisberg, W.G., Barns, S.M., Pelletier, D.A. and Lane, D.J. 1991. 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology, 173: 697-703. Weisz, P.R., Sinclair, T.R. 1988. A rapid non-destructive assay to quantify soybean nodule gas permeability. Plant & Soil, 105: 69-78. [https://doi.org/10.1007/BF02371144].

Downloads

Published

2024-12-12

How to Cite

ISOLATION OF ROOT NODULE ENDOPHYTES FROM COWPEA (Vigna  unguiculata L.) GROWING IN JHUM FIELD IN NAGALAND (INDIA). (2024). Applied Biological Research, 26(4), 459–465. https://doi.org/10.48165/abr.2024.26.01.53