Mutation of Bdpaired induces embryo lethality in the oriental fruit fly, Bactrocera dorsalis

Abstract

BACKGROUND Pair‐rule genes were identified and named for their role in segmentation in animal embryos. Paired, a homolog of mammalian PAX3, acts as one of several pair‐rule genes and is key in defining the boundaries of future parasegments and segments during insect embryogenesis. RESULTS We cloned the paired gene from the oriental fruit fly, Bactrocera dorsalis, and then applied CRISPR/Cas9‐mediated genome editing to investigate its physiological function in the embryonic stage of this pest. We identified one transcript for a paired homolog in B. dorsalis, which encodes a protein containing a Paired Box domain and a Homeobox domain. Phylogenetic analysis confirmed that the paired gene is highly conserved and the gene was highly expressed at the 12–14 h‐old embryonic stage. Knock‐out of Bdpaired led to lack of segment boundaries, cuticular deficiency, and embryonic lethality. Sequence analysis of the CRISPR/Cas9 mutants exhibited different insertion and deletions in the Bdpaired locus. In addition, the relative expression of Wingless (Wg) and Abdominal A (Abd‐A) genes were significantly down‐regulated in the Bdpaired mutant embryos. CONCLUSION These results indicate that Bdpaired gene is critical for the embryonic development of B. dorsalis, and could be a novel molecular target for genetic‐based pest management practices to combat this serious invasive pest. © 2019 Society of Chemical Industry

CRISPR‐mediated mutagenesis of the odorant receptor co‐receptor (Orco) gene disrupts olfaction‐mediated behaviors in Bactrocera dorsalis

“…In the present study, we used the CRISPR/Cas9 system to knock out Orco in B. dorsalis to generate the BdorOrco −/− mutants. However, the hatching rate of injected eggs was higher than previous studies (Bai et al ., 2019; Zheng et al ., 2019; Wang et al ., 2020). During embryo injection, previous studies used double‐sided adhesive tape to fix the eggs on slides.…”

“…The CRISPR/Cas9 system has been successfully used in Spodoptera littoralis , Locusta migratoria , Bombyx mori , and Harpegnathos saltator (Koutroumpa et al ., 2016; Li et al ., 2016; Liu et al ., 2017; Yan et al ., 2017) to study their olfactory systems. In B. dorsalis , CRISPR/Cas9‐based genome editing was initially established to study genes with visible or measurable phenotypes, including lack of segment boundaries, cuticular deficiency, embryonic lethality (Wang et al ., 2020), eyes and head spot color alterations (Bai et al ., 2019), muscle weakness, and flightless phenotypes (Zheng et al ., 2019).…”

miR-309a is a regulator of ovarian development in the oriental fruit fly Bactrocera dorsalis

“…Similarly, GATAβ4 promotes Vg transcription and egg formation in B. mori [16], while GATA acts as a transcriptional activator of Vg, leading to the regulation of vitellogenesis and reproduction in the Asian longhorned tick Haemaphysalis longicornis [23]. Regrettably, we could not effectively knockdown the expression of pnr through RNAi to better evaluate its effect on the expression of Vg-related genes (S2D Fig) . The availability of other tools such as CRISPR/Cas9 for genome editing in B. dorsalis [24,25] could be exploited to achieve this in future experiments. Nevertheless, our results indicate that miR-309a targets pnr to regulate Vg-related genes, which in turn directly affects ovarian development in B. dorsalis.…”

“…The availability of other tools such as CRISPR/Cas9 for genome editing in B . dorsalis [ 24 , 25 ] could be exploited to achieve this in future experiments. Nevertheless, our results indicate that miR-309a targets pnr to regulate Vg -related genes, which in turn directly affects ovarian development in B .…”

First Report on CRISPR/Cas9-Based Genome Editing in the Destructive Invasive Pest Tuta Absoluta (Meyrick) (Lepidoptera: Gelechiidae)

“…For example, cinnabar -targeting small guide RNA (sgRNA) molecules have been used to produce red eyes in Nasonia vitripennis ( Li et al, 2017 ) and Nilaparvata lugens ( Xue et al, 2018 ). However, even though the CRISPR/Cas9 system and cinnabar mutagenesis have been successfully used to generate and verify heritable mutations in other insects, respectively ( Adrianos et al, 2018 ; Xue et al, 2018 ; Wang et al, 2020 ; Zhu et al, 2020 ; Zhan et al, 2021 ), their application to T. absoluta have yet not to be reported.…”