Abstract
Aim:
The present study aimed to select a powerful tool to evaluate the antibiotic
susceptibility of endosymbionts of papaya mealybug.
Methodology: Antibiotic Ofloxacin (0.25µg) was used to disrupt
the endosymbionts of mealybug. The antibiotic susceptibility of endosymbionts
was studied by conventional method; isolation of symbionts in Luria Bertani
Agar and Nutrient Agar and also metagenomic analysis was conducted using
nanopore sequencing of 16S rRNA gene.
Results:
Molecular identification of isolates showed Gram-positive bacteria i.e., Bacillus
cluasii, B. altitudinis, B. siamensis and Gram-negative bacteria, i.e., Serratia
marcescens and Stenotrophomonas maltophilia. Metagenomic analysis
using 16S amplicon sequencing resulted in identification of Candidatus
Tremblaya Princeps (55%) was more abundant followed by Candidatus
Mikella endobia (23%) in untreated mealybug population. Their population
was reduced to 7% in Candidatus Tremblaya Princeps and below 1% in Candidatus
Mikella endobia.
Interpretation: The current study confirms that the
metagenomic analysis acts as a power lens to evaluate the antibiotic
elimination of endosymbionts, hence with this method future works on
developing aposymbiotic mealybugs can be done.
Key words: Endosymbiont, Metagenomic analysis, Ofloxacin,
Papaya mealybug
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