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Molecular characterization of cellulolytic nitrogen-fixing bacterial species isolated from Khyber Pakhtunkhwa

By: Nausheen Abdul Malik, Bashir Ahmad, Hira Mushtaq

Key Words: Nitrogen free Carboxymethyl cellulose medium (NF-CMC), Nitrogenase gene, 16sRNA sequencing, Cellulase, Nitrogenase activities.

Int. J. Biosci. 18(3), 18-29, March 2021.

DOI: http://dx.doi.org/10.12692/ijb/18.3.18-29

Certification: ijb 2021 0174 [Generate Certificate]

Abstract

Bacteria inhabiting the soil rhizosphere had been recognized for their properties of promoting plant health. The current research was undertaken for the molecular characterization of cellulolytic nitrogen-fixing bacterial species from the rhizosphere.  Two hundred and fifty soil samples were collected from Wheat and sugarcane rhizosphere in few districts of KPK and were grown on Nitrogen free Carboxymethyl cellulose medium for cellulose and nitrogenase enzyme production. Bacterial isolates from Wheat in Congo red assay showed clearance zone ratio on CMC substrate in the range of 1.6 – 3.0mm, while bacterial isolates from Sugarcane samples showed clearance zone ratio in the range of 1.3 – 2.71mm. Quantitative analysis revealed that wheat bacterial isolates were releasing cellulase in the range of 0.751– 4.784 U/mL on NF-CMC medium (with 5% CMC+pH 7) while they produced NH4-N in the range of 0.104 – 1.115 mM in NF medium (pH 7). Bacteria from Sugarcane samples showed cellulase activities in the range of 0.418 – 4.155 U/mL and produced NH4-N in the range of 0.102 – 1.070 mM under the same conditions. The presence of nitrogenase gene only in fourteen bacterial isolates was confirmed and identified by 16sRNA sequencing as bacterial strains Achromobacter sp. MFA1, Paenibacillus pabuli HSSC 492, Streptomyces sp. strain VITBVK2, Stenotrophomonas sp. Vi65, Uncultured Actinomyces sp. clone O-71, Beijerinckia fluminensis, Alcaligenes faecalis CCM5B, Stenotrophomonas maltophilia strain CM30, Brevibacillus parabrevis strain TJ2.3, Ochrobactrum intermedium strain BE1, Brevibacillus parabrevis strain TJ2.3, Cupriavidus sp. christensen.ICD.09 and Bacillus cereus.

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Molecular characterization of cellulolytic nitrogen-fixing bacterial species isolated from Khyber Pakhtunkhwa

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Nausheen Abdul Malik, Bashir Ahmad, Hira Mushtaq.
Molecular characterization of cellulolytic nitrogen-fixing bacterial species isolated from Khyber Pakhtunkhwa.
Int. J. Biosci. 18(3), 18-29, March 2021.
https://innspub.net/ijb/molecular-characterization-of-cellulolytic-nitrogen-fixing-bacterial-species-isolated-from-khyber-pakhtunkhwa/
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