Isolation and characterization of endophytic bacteria from Pigeon pea (Cajanus cajan L.)
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Date
2019
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Indira Gandhi Krishi Vishwavidyalaya
Abstract
The present study entitled “Isolation and characterization of endophytic bacteria from pigeon pea” was carried out at ICAR -National Institute of Biotic Stress Management (NIBSM), Baronda, Raipur.A total of 45 bacterial endophytes were isolated from seven pigeonpea lines and five plant tissues namely root, stem, leaf, flower and pods. The number of isolates isolated tissue wise (19) 42.2%, (11) 24.4%, (11) 24.4%, (2) 4.4% and (2) 4.4% from root, stem, leaf, flower and pod/seed tissues, respectively. Morphological characteristics of bacterial endophytes showed small to medium sized irregular shape colony and the size of most of the isolates varied from pin to medium. Most of the bacterial endophytes (36) have single bacilli and (8) cocco-bacilli, while (1) bacteria had capsulated form. Gram’s reaction of bacterial isolates showed that the (28) of bacterial isolates were Gram positive and (17) were gram negative. Biochemical characterization showed that the number of isolates was positive for Oxidase (24), Catalase (30), Citrate utilization (21), Methyl Red (9) and Vogus Proskauer (3), Urease (10), Nitrate reduction (18). The motility and gases producing ability of isolates showed that 22were motile and 6 isolates produced gas.
Molecular characterization of bacterial endophytes was performed by amplification and sequencing of 16S rDNA gene. On the basis of nucleotide sequences generated by sequencing of 16S rDNA amplicon bacterial isolates were identified and grouped in different genera’s namely Bacillus, Fictibacillus, Pseudomonas, Enterobactor, Klebsiella, Beijerinckia, Pantoea,and Serretia. The gene sequences of all the 45 bacterial endophytes have been deposited to Gene bank, NCBI and their accession numbers were obtained.
The bacterial isolates were screened for their plant growth-promoting (PGP) activities viz., production of indole acetic acid (4), siderophore (19), phosphate solubilization (11), DNase (13), HCN production (1) were found positive. Bacterial isolates were screened for antibiotics sensitivity pattern using 14 numbers of multispectral antibiotics showed highest resistance to Ceftriaxone followed by Cifixime, Methicillin, Polymyxin and Ampicilin, while highest sensitivity to Gatifloxacin followed by Gentamicin.
Antagonistic activity test identified bacterial isolates having antagonism against soil borne fungal pathogens viz, Sclerotium rolfsii (19) Fusarium sps (14) and Rhizoctonia solani (17). Highest antagonistic activities against soil borne fungal pathogens were exhibited by bacterial endophytes isolated from root tissues. The potential isolates exhibiting antifungal activities may be used for the development of bio control formulations for controlling multiple biotic stresses.
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