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Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis
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Tudor knockdown disrupts ovary development in Bactrocera dorsalis
Y-F Xie1,2, F Shang1,3, B-Y Ding1,3
1Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing, China.
The tudor gene is crucial for reproduction in the oriental fruit fly, Bactrocera dorsalis. Its knockdown results in underdeveloped ovaries and reduced reproductive success, highlighting its essential role.
Area of Science:
- Molecular Biology
- Genetics
- Entomology
Background:
- The piwi-interacting RNA pathway is vital for germline transposon silencing.
- Tudor superfamily proteins are key regulators in this silencing mechanism in insects.
Purpose of the Study:
- To identify and characterize the tudor gene in Bactrocera dorsalis.
- To investigate its expression patterns and functional role in reproduction.
Main Methods:
- Transcriptome analysis to identify the tudor homologue.
- Phylogenetic analysis for confirmation.
- Quantitative expression profiling across developmental stages and tissues.
- RNA interference (RNAi) for functional analysis.
Main Results:
- A tudor homologue with 10 Tudor domains and an ATP synthase delta subunit signature was identified.
- Tudor expression significantly increased in adult and pupal stages, predominantly in ovaries, Malpighian tubules, and fat body.
- Gene knockdown led to underdeveloped ovaries, reduced copulation rates, and decreased oviposition.
Conclusions:
- The tudor gene plays a critical role in Bactrocera dorsalis ovary development and reproductive functions.
- This study elucidates the function of tudor in insect reproduction.
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