Vitelline membrane protein gene ZcVMP26Ab and its role in preventing water loss in Zeugodacus cucurbitae (Coquillett) embryos

Li, Weijun; Song, Yujia; Xu, Hui-Qian; Wei, Dong; Wang, Jinjun

doi:10.1127/entomologia/2021/1037

Cited by 14 publications

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“…Total RNA and cDNA preparation was performed as described above. Ribosomal protein subunit 3 ( Rps3 ) and ribosomal protein L13 ( Rpl13 ) were selected as the internal reference genes to explore the expression patterns of ZcVgs at different developmental stages [ 31 ]. The tissue samples from the midgut, fat body, Malpighian tubules, and ovary were dissected from 5 d old virgin female adults to determine the expression pattern of ZcVgs .…”

Section: Methodsmentioning

confidence: 99%

“…Four biological replicates were prepared, and each sample was dissected from 20 individuals. α-Tubulin ( α-Tub ) and β-tubulin 1 ( β-Tub1 ) were selected as the internal reference genes to analyze the expression in various tissues of female adults [ 31 ]. The primers for the RT-qPCR were designed by Primer Premier 5.0 ( Table 1 ), followed by the amplification efficiency determination.…”

Section: Methodsmentioning

confidence: 99%

“…Total RNA and cDNA preparation was performed as described above. Rps3 and α-Tub were selected as reference genes to normalize the expression of ZcVgs on day 5 after RNAi [ 31 ]. Ovaries were dissected to determine the size on day 5.…”

Section: Methodsmentioning

confidence: 99%

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Expression and Role of Vitellogenin Genes in Ovarian Development of Zeugodacus cucurbitae

Chen

Han

et al. 2022

Insects

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Vitellogenin (Vg) genes encode the major egg yolk protein precursor in arthropods. In this study, four Vgs were identified in Zeugodacus cucurbitae (Coquillett). Sequence analysis showed that four ZcVgs had the conserved Vg domain. Phylogenetic analysis indicated that four ZcVgs were homologous to the Vgs of Tephritidae insects. The temporal and spatial expression patterns of ZcVgs were analyzed by quantitative real-time polymerase chain reaction (RT-qPCR), and the four ZcVgs showed high expression levels in female adults, especially in the fat body. The expression of ZcVg1 and ZcVg3 was down-regulated by a low dosage (0.5 μg) of 20-hydroxyecdysone (20E), and ZcVg2, ZcVg3, and ZcVg4 were up-regulated by a high dosage (1.0 and 2.0 μg) of 20E. The expression of ZcVg1 and ZcVg2 was up-regulated by 5 μg of juvenile hormone (JH), while all of the ZcVgs were down-regulated by a low and high dosage of JH. Expression of ZcVgs was down-regulated after 24 h of starvation and recovered to normal after nutritional supplementation. After micro-injection of the gene-specific double-stranded RNA, the ZcVgs’ expression was significantly suppressed, and ovarian development was delayed in Z. cucurbitae females. The results indicate that RNA interference of reproduction-related genes is a potential pest control method that works by manipulating female fertility.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Expression and Role of Vitellogenin Genes in Ovarian Development of Zeugodacus cucurbitae

Chen

Han

et al. 2022

Insects

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“…The PCR amplification procedure was as follows: an initial denaturation at 95 °C for 3 min, followed by 40 cycles of 95 °C for 5 s and 60 °C for 15 s. Finally, the specificity of each pair of primers was ensured by recording a melting curve analysis from 60 to 95 °C. Ribosomal protein subunit 3 and Ribosomal protein L13 were used as the reference genes for lncRNA expression analysis at the different developmental stages, alpha‐tubulin and beta‐tubulin 1 were used as the reference genes for lncRNA expression analysis among different tissues (Li et al ., 2021). Data were analysed by qBase plus software (Hellemans et al ., 2007).…”

Section: Methodsmentioning

confidence: 99%

lnc94638 is a testis‐specific long non‐coding RNA involved in spermatozoa formation in Zeugodacus cucurbitae (Coquillett)

Wei

Han

et al. 2021

Insect Molecular Biology

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Long non‐coding RNAs (lncRNAs) generally display tissue‐specific distributions, and testis‐specific lncRNAs form the highest proportion of lncRNAs in many species. Here, we presented a detailed analysis of testis‐specific lncRNAs in the melon fly, Zeugodacus cucurbitae, a highly destructive insect pest of cucurbitaceous and other related crops. Most testis‐specific lncRNAs were found to be long intergenic non‐coding RNAs (lincRNA). The size distribution of these lncRNAs ranged between 600 and 1000 nucleotides. Testis‐specific lncRNAs that harboured one isoform number and two exons were the most abundant. Compared to other male tissues, the testis had more highly expressed lncRNAs. The quantitative real‐time polymerase chain reaction results of 10 randomly selected testis‐specific lncRNAs showed expression patterns consistent with RNA‐seq data. Further analysis of the most highly expressed testis‐specific lncRNA, lnc94638, was undertaken. Fluorescent in situ hybridization assays localized lnc94638 to the apical region of the testis that contains mature spermatozoa. RNA interference‐mediated knockdown of lnc94638 expression reduced spermatozoa numbers and impaired the fertility of Z. cucurbitae male. This study provides a catalogue of testis‐specific lncRNAs, shows that the testis‐specific lnc94638 is involved in spermatogenesis and has the potential to be used for treating male sterility.

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“…Two VMPs are found in Anopheles gambiae [ 15 ]. In Bombyx mori , six putative VMP genes ( VMP15a-1/2/3 , VMP30 , VMP90 , and EP80 ) have been identified, and three VMP genes in Anastrepha obliqua [ 16 ], Bactrocera dorsalis [ 17 , 18 ], and Zeugodacus cucurbitae [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Vitelline Membrane Protein 26 Mutagenesis, Using CRISPR/Cas9, Results in Egg Collapse in Plutella xylostella

Zhai

Dong

Zou

et al. 2022

IJMS

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Vitelline membrane proteins (VMPs) are the main proteins that form the inner shell (vitelline membrane layer) of insect eggs and are an integral part of egg formation and embryo development. Here, we characterized the molecular structure and expression patterns of the VMP26 gene and analyzed its reproductive functions in diamondback moth, Plutella xylostella (L.), a worldwide migratory pest of cruciferous plants. The PxVMP26 gene was shown to be a single exon gene that contained an open reading frame of 852 base pairs (bp) encoding 283 amino acids. Both qPCR and western blot analyses showed that PxVMP26 was specifically expressed in female adults and was significantly highly expressed in the ovary. Further anatomical analysis indicated that the expression level of PxVMP26 in the ovarian tube with an incomplete yolk was significantly higher than that in the ovarian tube with a complete yolk. CRISPR/Cas9-induced PxVMP26 knockout successfully created two homozygous strains with 8- and 46-bp frameshift mutations. The expression deficiency of the PxVMP26 protein was detected in the mutant strains using immunofluorescence and western blot. No significant difference was found in the number of eggs laid within three days between wild and mutant individuals, but there was a lower egg hatchability. The loss of the PxVMP26 gene changed the mean egg size, damaged the structure of the vitelline membrane, and increased the proportion of abnormal eggs due to water loss, resulting in egg collapse. This first analysis of the roles of the VMP gene in the oocyte formation and embryonic development of P. xylostella, using CRISPR/Cas9 technology, provides a basis for screening new genetic control targets of P. xylostella.

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Vitelline membrane protein gene ZcVMP26Ab and its role in preventing water loss in Zeugodacus cucurbitae (Coquillett) embryos

Cited by 14 publications

References 0 publications

Expression and Role of Vitellogenin Genes in Ovarian Development of Zeugodacus cucurbitae

Expression and Role of Vitellogenin Genes in Ovarian Development of Zeugodacus cucurbitae

lnc94638 is a testis‐specific long non‐coding RNA involved in spermatozoa formation in Zeugodacus cucurbitae (Coquillett)

Vitelline Membrane Protein 26 Mutagenesis, Using CRISPR/Cas9, Results in Egg Collapse in Plutella xylostella

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