RNAi-Mediated Silencing of Putative Halloween Gene Phantom Affects the Performance of Rice Striped Stem Borer,

Overview

Journal Insects

Specialty Biology

Date 2022 Aug 25

PMID 36005356

Authors

Muhammad Faisal Shahzad

Atif Idrees

Ayesha Afzal

Jamshaid Iqbal

Ziyad Abdul Qadir

Azhar Abbas Khan

Ayat Ullah

Jun Li

Affiliations

Soon will be listed here.

Abstract

The physiological and biochemical characterization of the "Halloween" genes has fundamental importance in the biosynthesis pathway of ecdysteroids. These genes were found to catalyze the final phases of ecdysteroid biosynthesis from dietary cholesterol to the molting hormone 20-hydroxyecdysone. We report the characterization of the in a major insect pest in agriculture, the rice striped stem borer, (). A full-length transcript of was amplified with an open reading frame (ORF) of 478 amino acids through 5' and 3' RACE. shows five insect-conserved P450 motifs: Helix-C, Helix-I, Helix-K, PERF, and heme-binding motifs. Phylogenetic analysis clearly shows high similarity to Lepidoptera and evolutionary conservation in insects. The relative spatial and temporal transcript profile shows that is highly expressed in the prothoracic glands and appears throughout the larval development, but with low expression at the start of the larval instar. It seems to peak in 3-4 days and decreases again before the larvae molt. Double-stranded RNA (dsRNA) injection of Cs-Phm at the larval stage efficiently knocked down the target gene and decreased its expression level. The dsRNA-treated group showed significantly decreased ecdysteroid titers, which leads to delayed larval development and higher larval mortality. Negative effects of larval development were rescued by treating 20E in the dsRNA-treated group. Thus, in conclusion, our results suggest that is functionally conserved in and encodes functional CYP that contributes to the biogenesis of 20E.

Citing Articles

Lethal, Sub-Lethal and Trans-Generational Effects of Chlorantraniliprole on Biological Parameters, Demographic Traits, and Fitness Costs of (Lepidoptera: Noctuidae).

Akhtar Z, Afzal A, Idrees A, Zia K, Qadir Z, Ali S Insects. 2022; 13(10).

PMID: 36292828 PMC: 9603994. DOI: 10.3390/insects13100881.

References

Peng L, Wang L, Zou M, Vasseur L, Chu L, Qin Y . Identification of Halloween Genes and RNA Interference-Mediated Functional Characterization of a Halloween Gene in . Front Physiol. 2019; 10:1120. PMC: 6724230. DOI: 10.3389/fphys.2019.01120. View

Moulos P, Samiotaki M, Panayotou G, Dedos S . Combinatory annotation of cell membrane receptors and signalling pathways of Bombyx mori prothoracic glands. Sci Data. 2016; 3:160073. PMC: 5004587. DOI: 10.1038/sdata.2016.73. View

Yoshiyama-Yanagawa T, Enya S, Shimada-Niwa Y, Yaguchi S, Haramoto Y, Matsuya T . The conserved Rieske oxygenase DAF-36/Neverland is a novel cholesterol-metabolizing enzyme. J Biol Chem. 2011; 286(29):25756-62. PMC: 3138242. DOI: 10.1074/jbc.M111.244384. View

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S . MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011; 28(10):2731-9. PMC: 3203626. DOI: 10.1093/molbev/msr121. View

Niwa R, Sakudoh T, Namiki T, Saida K, Fujimoto Y, Kataoka H . The ecdysteroidogenic P450 Cyp302a1/disembodied from the silkworm, Bombyx mori, is transcriptionally regulated by prothoracicotropic hormone. Insect Mol Biol. 2005; 14(5):563-71. DOI: 10.1111/j.1365-2583.2005.00587.x. View

Zhang X, He Q, Zhang T, Wu H, Zhang J, Ma E . Characteristics of Halloween genes and RNA interference-mediated functional analysis of LmCYP307a2 in Locusta migratoria. Insect Sci. 2021; 29(1):51-64. DOI: 10.1111/1744-7917.12907. View

Pfaffl M . A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001; 29(9):e45. PMC: 55695. DOI: 10.1093/nar/29.9.e45. View

Niwa R, Matsuda T, Yoshiyama T, Namiki T, Mita K, Fujimoto Y . CYP306A1, a cytochrome P450 enzyme, is essential for ecdysteroid biosynthesis in the prothoracic glands of Bombyx and Drosophila. J Biol Chem. 2004; 279(34):35942-9. DOI: 10.1074/jbc.M404514200. View

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

10.

Yin C, Liu Y, Liu J, Xiao H, Huang S, Lin Y . ChiloDB: a genomic and transcriptome database for an important rice insect pest Chilo suppressalis. Database (Oxford). 2014; 2014. PMC: 4082153. DOI: 10.1093/database/bau065. View

11.

Rewitz K, Rybczynski R, Warren J, Gilbert L . The Halloween genes code for cytochrome P450 enzymes mediating synthesis of the insect moulting hormone. Biochem Soc Trans. 2006; 34(Pt 6):1256-60. DOI: 10.1042/BST0341256. View

12.

Ruang-Rit K, Park Y . Endocrine system in supernumerary molting of the flour beetle, Tribolium freemani, under crowded conditions. Insect Biochem Mol Biol. 2018; 101:76-84. DOI: 10.1016/j.ibmb.2018.08.002. View

13.

Rewitz K, Gilbert L . Daphnia Halloween genes that encode cytochrome P450s mediating the synthesis of the arthropod molting hormone: evolutionary implications. BMC Evol Biol. 2008; 8:60. PMC: 2276477. DOI: 10.1186/1471-2148-8-60. View

14.

Wan P, Jia S, Li N, Fan J, Li G . RNA interference depletion of the Halloween gene disembodied implies its potential application for management of planthopper Sogatella furcifera and Laodelphax striatellus. PLoS One. 2014; 9(1):e86675. PMC: 3904942. DOI: 10.1371/journal.pone.0086675. View

15.

Rewitz K, Rybczynski R, Warren J, Gilbert L . Developmental expression of Manduca shade, the P450 mediating the final step in molting hormone synthesis. Mol Cell Endocrinol. 2006; 247(1-2):166-74. DOI: 10.1016/j.mce.2005.12.053. View

16.

Christiaens O, Iga M, Velarde R, Rouge P, Smagghe G . Halloween genes and nuclear receptors in ecdysteroid biosynthesis and signalling in the pea aphid. Insect Mol Biol. 2010; 19 Suppl 2:187-200. DOI: 10.1111/j.1365-2583.2009.00957.x. View

17.

Petryk A, Warren J, Marques G, Jarcho M, Gilbert L, Kahler J . Shade is the Drosophila P450 enzyme that mediates the hydroxylation of ecdysone to the steroid insect molting hormone 20-hydroxyecdysone. Proc Natl Acad Sci U S A. 2003; 100(24):13773-8. PMC: 283497. DOI: 10.1073/pnas.2336088100. View

18.

Gilbert L, Warren J . A molecular genetic approach to the biosynthesis of the insect steroid molting hormone. Vitam Horm. 2006; 73:31-57. DOI: 10.1016/S0083-6729(05)73002-8. View

19.

Warren J, Petryk A, Marques G, Jarcho M, Parvy J, Dauphin-Villemant C . Molecular and biochemical characterization of two P450 enzymes in the ecdysteroidogenic pathway of Drosophila melanogaster. Proc Natl Acad Sci U S A. 2002; 99(17):11043-8. PMC: 123207. DOI: 10.1073/pnas.162375799. View

20.

Miller W, Auchus R . The molecular biology, biochemistry, and physiology of human steroidogenesis and its disorders. Endocr Rev. 2010; 32(1):81-151. PMC: 3365799. DOI: 10.1210/er.2010-0013. View