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.
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.
Gut Biology Laboratory, Department of Zoology, University of Delhi, Delhi, India.
Harpreet Singh
HS
Raina
Gut Biology Laboratory, Department of Zoology, University of Delhi, Delhi, India.
Ambika
A
Singh
Gut Biology Laboratory, Department of Zoology, University of Delhi, Delhi, India.
Sonam
S
Popli
Gut Biology Laboratory, Department of Zoology, University of Delhi, Delhi, India.
Neeti
N
Pandey
Gut Biology Laboratory, Department of Zoology, University of Delhi, Delhi, India.
Raman
R
Rajagopal
D000818:Animals; D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification*; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D006430:Hemiptera / Q000382:microbiology* / Q000472:pathogenicity; D017404:In Situ Hybridization, Fluorescence; D007194:India; D016133:Polymerase Chain Reaction; D013559:Symbiosis
D015342:DNA Probes; D004269:DNA, Bacterial; D004275:DNA, Ribosomal