Isolation and characterization of bacteria from tissue culture of Cenchrus purpureus CT-608
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Abstract
The objective of the present work consisted of reporting advances in the isolation and characterization of bacteria present in the tissue culture of Cenchrus purpureus CT-608. The study was developed in the Biotechnology Laboratory of the Agrarian University of Havana. Fragments of bacterial colonies from contaminated microplants were used and isolated in test tubes with Nutrient Agar medium. The colonies were described according to their macroscopic and microscopic characteristics through Gram staining and optical microscopy at 1000× magnification. The following descriptors were evaluated: cellular shape, size, grouping arrangement, reaction to Gram staining, cyst production, capsule production, endospore production. Bacterial sensitivity to the set of antibiotics was determined: Penicillin, Amoxicillin, Ampicillin, Cephalexin, Cefixime, Ceftriaxone, Nalidixic Acid, Gentamicin, and Chloramphenicol. Antibiotic response was assessed using the Kirby-Bauer technique. Two bacterial colonies were isolated, where one Gram-positive morphotype (MF1) and two Gram-negative morphotypes (MF2 and MF3) were obtained from one colony. The bacillary cellular form predominated, with a size of 3-4 mm, and the grouping arrangement varied from bacillus to irregular rosette-shaped. The morphotypes showed sensitivity to Nalidixic Acid and Cephalexin, and resistance to Penicillin, Amoxicillin, and Ampicillin. Three bacterial morphotypes with diverse morphological and physiological characteristics were isolated from C. purpureus CT-608 microplants. Their late appearance in the culture medium and their confinement around the explant indicate they may be endophytic microorganisms, an aspect that should be verified through chemotaxis.
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