RNAi-Mediated Knockdown of Imaginal Disc Growth Factors (IDGFs) Genes Causes Developmental Malformation and Mortality in Melon Fly, Zeugodacus cucurbitae

Ahmad, Shakil; Jamil, Momana; Fahim, Muhammad; Zhang, Shujing; Ullah, Farman; Lyu, Baoqian; Luo, Yunbo

doi:10.3389/fgene.2021.691382

Cited by 6 publications

(11 citation statements)

References 79 publications

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“…Significant reduction of Ago1 at an early stage indicates its effectiveness at this stage to cause loss of pigmentation and ovarian defects in Z. cucurbitae . A similar response was observed to Ago2 and Dicer2 upon dsRNA exposure in Anastrepha fraterculus and ds IDGF4_1 feeding in Z. cucurbitae ( Dias et al, 2019 ; Ahmad et al, 2021 ). In addition to the core RNAi machinery genes, such phenomena have been extensively noticed in several other mechanisms of target gene silencing including dsRNA uptake and degradation ( Zhu and Palli 2020 ).…”

Section: Discussionsupporting

confidence: 73%

“…Therefore, focusing on the functions of key genes involved in development and reproduction is necessary for future pest control. In previous research, the knockdown of imaginal disc growth factor (IDGF) genes caused developmental defects in Z. cucurbitae (Ahmad et al, 2021). However, to control this destructive pest, the identification of lethal genes is encouraging.…”

Section: Introductionmentioning

confidence: 99%

“…In previous research, the knockdown of imaginal disc growth factor ( IDGF ) genes caused developmental defects in Z . cucurbitae ( Ahmad et al, 2021 ). However, to control this destructive pest, the identification of lethal genes is encouraging.…”

Section: Introductionmentioning

confidence: 99%

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Argonaute1 and Gawky Are Required for the Development and Reproduction of Melon fly, Zeugodacus cucurbitae

et al. 2022

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Argonaute family genes encode a highly conserved group of proteins that have been associated with RNA silencing in both animals and plants. This study investigates the importance of microRNA biogenesis key regulators Argonaute1 (Ago1) and Gawky genes in the post-embryonic and ovarian development of the melon fly, Zeugodacus cucurbitae. The expression levels of these genes were mapped in all developmental stages and different adult tissues. Their roles in development were investigated using RNA interference (RNAi) via two different dsRNA delivery techniques. Embryo microinjection and oral feeding of third instar larvae successfully knocked down and greatly reduced the expression level of the target genes. Additionally, ex vivo essays revealed the stability of dsRNA in food was sufficient for gene silencing, although its integrity was affected in midgut. A wide range of phenotypes were observed on pupation, segmentation, pigmentation, and ovarian development. RNAi-mediated silencing of Gawky caused high mortality and loss of body segmentation, while Ago1 knockdown affected ovarian development and pigmentation. Developmental abnormalities and ovarian malformations caused by silencing these genes suggest that these genes are crucial for viability and reproductive capacity of Z. cucurbitae, and may be used as potential target genes in pest management.

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Section: Discussionsupporting

confidence: 73%

Section: Introductionmentioning

confidence: 99%

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Argonaute1 and Gawky Are Required for the Development and Reproduction of Melon fly, Zeugodacus cucurbitae

et al. 2022

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“…Among these, we selected eight DEGs based on their putative function described in the literature (Vlachou et al, 2006;Ragland et al, 2011;Mizoguchi and Okamoto, 2013;Krüger et al, 2015;Lekha et al, 2015;Chang et al, 2019;Pérez-Lluch et al, 2020;Saraswathi et al, 2020), and we knocked down these genes individually to assess their role in the development of Z. cucurbitae. Successful knockdown of orally delivered dsRNA for target gene in artificial diet was reported previously in Z. cucurbitae (Ahmad et al, 2021b;Jamil et al, 2022). The knockdown of two genes, ImpE2 and Tpk-tok, caused major developmental defects.…”

Section: Genes Silencing Effects On Insect Development and Survivalmentioning

confidence: 71%

Comparative transcriptomics of the irradiated melon fly (Zeugodacus cucurbitae) reveal key developmental genes

et al. 2023

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Irradiation can be used as an insect pest management technique to reduce post-harvest yield losses. It causes major physiological changes, impairing insect development and leading to mortality. This technique is used to control the melon fly Zeugodacus cucurbitae, a major pest of Cucurbitaceae in Asia. Here, we applied irradiation to melon fly eggs, and the larvae emerged from irradiated eggs were used to conduct comparative transcriptomics and thereby identify key genes involved in the development and survival. We found 561 upregulated and 532 downregulated genes in irradiated flies compared to non-irradiated flies. We also observed abnormal small-body phenotypes in irradiated flies. By screening the 532 downregulated genes, we selected eight candidate genes putatively involved in development based in described functions in public databases and in the literature. We first established the expression profile of each candidate gene. Using RNA interference (RNAi), we individually knocked down each gene in third instar larvae and measured the effects on development. The knockdown of ImpE2 ecdysone-inducible gene controlling life stage transitions–led to major body size reductions in both pupae and adults. The knockdown of the tyrosine-protein kinase-like tok (Tpk-tok) caused severe body damage to larvae, characterized by swollen and black body parts. Adults subject to knockdown of the eclosion hormone (Eh_1) failed to shed their old cuticle which remained attached to their bodies. However, no obvious developmental defects were observed following the knockdown of the heat shock protein 67B1-like (Hsp67), the insulin receptor (Insr), the serine/threonine-protein kinase Nek4 (Nek4), the tyrosine-protein kinase transmembrane receptor Ror (Ror_1) and the probable insulin-like peptide 1 (Insp_1). We argue that irradiation can be successfully used not only as a pest management technique but also for the screening of essential developmental genes in insects via comparative transcriptomics. Our results demonstrate that ImpE2 and Eh_1 are essential for the development of melon fly and could therefore be promising candidates for the development of RNAi-based pest control strategies.

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“…Adult flies were fed with a mixture of sugar and yeast in a ratio of 3:1. Pupae in a Petri dish were placed in 45 × 45 × 50 cm cages for adult emergence ( Ahmad et al 2021 ). After hatching, the first instar from the control and irradiated treatments was given an artificial diet consisting of wheat bran (80 g), inactive dry yeast (16 g), sugar (32 g), agar powder (16 g), water (1000 ml), and 3.7 % formalin (HCHO) (10 ml) ( Somogyi et al 2007 ).…”

Section: Methodsmentioning

confidence: 99%

60Co-γ Radiation Alters Developmental Stages of Zeugodacus cucurbitae (Diptera: Tephritidae) Through Apoptosis Pathways Gene Expression

Ahmad

Hussain

Ullah

et al. 2021

Journal of Insect Science

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Radiation is considered as a promising insect pest control strategy for minimizing postharvest yield losses. Among various techniques, irradiation is a method of choice as it induces lethal biochemical or molecular changes that cause a downstream cascade of abrupt physiological abnormalities at the cellular level. In this study, we evaluated the effect of 60 Co-γ radiation on various developmental stages of Zeugodacus cucurbitae Coquillett and subsequent carry-over effects on the progeny. For this purpose, we treated eggs with 30- and 50-Gy radiation doses of 60 Co-γ. We found that radiation significantly affected cellular antioxidants, insect morphology, and gene expression profiles. Our results indicate that in response to various doses of irradiation reactive oxygen species, catalase, peroxidase, and superoxide dismutase activities were increased along with a significant increase in the malondialdehyde (MDA) content. We observed higher mortality rates during the pupal stage of the insects that hatched from irradiated eggs (50 Gy). Furthermore, the life span of the adults was reduced in response to 50 Gy radiation. The negative effects carried over to the next generation were marked by significantly lower fecundity in the F 1 generation of the irradiation groups as compared to control. The radiation induced morphological abnormalities at the pupal, as well as the adult, stages. Furthermore, variations in the gene expression following irradiation are discussed. Taken together, our results signify the utility of 60 Co-γ radiation for fruit fly postharvest management.

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RNAi-Mediated Knockdown of Imaginal Disc Growth Factors (IDGFs) Genes Causes Developmental Malformation and Mortality in Melon Fly, Zeugodacus cucurbitae

Cited by 6 publications

References 79 publications

Argonaute1 and Gawky Are Required for the Development and Reproduction of Melon fly, Zeugodacus cucurbitae

Argonaute1 and Gawky Are Required for the Development and Reproduction of Melon fly, Zeugodacus cucurbitae

Comparative transcriptomics of the irradiated melon fly (Zeugodacus cucurbitae) reveal key developmental genes

60Co-γ Radiation Alters Developmental Stages of Zeugodacus cucurbitae (Diptera: Tephritidae) Through Apoptosis Pathways Gene Expression

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