Infection of Bacterial Endosymbionts in Insects: A Comparative Study of Two Techniques viz PCR and FISH for Detection and Localization of Symbionts in Whitefly, Bemisia tabaci

• Published in: PloS one Vol. 10; no. 8; p. e0136159
• Main Authors: Raina, Harpreet Singh, Singh, Ambika, Popli, Sonam, Pandey, Neeti, Rajagopal, Raman
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 19.08.2015; Public Library of Science (PLoS)
• Subjects: Aleyrodidae; Animals; Aphidoidea; Arsenophonus; Arthropods; Bacteria; Bacteria - genetics; Bacteria - isolation & purification; Bemisia tabaci; Biology; Comparative analysis; Comparative studies; Cotton; DNA Probes - genetics; DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; DNA, Ribosomal - genetics; DNA, Ribosomal - isolation & purification; Electrophoresis; Endosymbionts; Health aspects; Hemiptera; Hemiptera - microbiology; Hemiptera - pathogenicity; Hybridization; Identification; In Situ Hybridization, Fluorescence; India; Insects; Laboratories; Localization; Medical research; Methods; Microscopy; Morphology; Pests; Polymerase Chain Reaction; Rickettsia; Rickettsiales; Scholarly publishing; Symbionts; Symbiosis; Viruses; Whiteflies; Wolbachia; Zoology; India
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0136159

Bacterial endosymbionts have been associated with arthropods and large number of the insect species show interaction with such bacteria. Different approaches have been used to understand such symbiont- host interactions. The whitefly, Bemisia tabaci, a highly invasive agricultural pest, harbors as many as seven different bacterial endosymbionts. These bacterial endosymbionts are known to provide various nutritional, physiological, environmental and evolutionary benefits to its insect host. In this study, we have tried to compare two techniques, Polymerase chain reaction (PCR) and Flourescence in situ Hybridisation (FISH) commonly used for identification and localization of bacterial endosymbionts in B. tabaci as it harbors one of the highest numbers of endosymbionts which have helped it in becoming a successful global invasive agricultural pest. The amplified PCR products were observed as bands on agarose gel by electrophoresis while the FISH samples were mounted on slides and observed under confocal microscope. Analysis of results obtained by these two techniques revealed the advantages of FISH over PCR. On a short note, performing FISH, using LNA probes proved to be more sensitive and informative for identification as well as localization of bacterial endosymbionts in B. tabaci than relying on PCR. This study would help in designing more efficient experiments based on much reliable detection procedure and studying the role of endosymbionts in insects.