International Journal of Industrial Entomology and Biomaterials

Korean Society of Sericultural Science (한국잠사학회)
권호 표지
DOI QR코드

DOI QR Code

Gene expression changes in silkworm embryogenesis for prediction of hatching time

  • Jong Woo Park (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Chang Hoon Lee (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Chan Young Jeong (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Hyeok Gyu Kwon (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Seul Ki Park (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Ji Hae Lee (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Sang Kuk Kang (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Seong-Wan Kim (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Seong-Ryul Kim (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Hyun-Bok Kim (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA) ;
  • Kee Young Kim (Department of Agricultural Biology, National Institute of Agricultural Sciences, RDA)
  • Received : 2023.01.13
  • Accepted : 2023.03.22
  • Published : 2023.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

The silkworm's dormancy and embryonic development are accomplished through the interaction of various genes. Analysis of the expression of several interacting genes can predict the embryonic stage of silkworms. In this study, we analyzed the changes in the expression level of genes at each stage during the embryonic development of dormant silkworm eggs and selected genes that can predict the hatching time. Jam123 and Jam124 silkworms were collected after egg laying, and the silkworm eggs were preserved using a double refrigeration method and expression analysis was performed for 23 genes during embryogenesis. There were 5 genes showing significant changes during embryogenesis: UDP-glucuronosyltransferases (BmUGTs), heat shock protein hsp20.8 (BmHsp20.8), Cytochromes b5-like proteins (BmCytb5), Krüppel homolog 1 (BmKr-h1), and cuticular protein RR-1 motif 41 (BmCpr41). As a result of quantitative comparison of the expression levels of these 5 genes through real-time PCR, the BmUGTs gene showed a difference between Jam123 and Jam124, making it difficult to see it as an indicator for predicting hatching time. However, the BmHsp20.8 gene had a common expression decreased at the imminent hatching stage. In addition, it was confirmed that the expression level of the BmCytb5 gene decreased to the lowest level at the time of imminent hatching, and the expression of the BmKr-h gene was made only at the time of imminent hatching. The expression of the last BmCpr41 gene can be confirmed only at the time of imminent hatching, and it was confirmed that it shows a rapid increase right before hatching. Taken together, these results suggest that expression analysis of BmHsp20.8, BmCytb5, BmKr-h1, and BmCpr41 genes can determine the stage of embryogenesis, predict hatching time, which facilitate better management of silkworm eggs.

Keywords

Acknowledgement

This study was supported by the 2023 RDA fellowship program of National Institute of Agricultural Science and was supported by a grant (No. PJ01558102) from the Rural Development Administration, Republic of Korea.

References

  1. Ahmad SA, Hopkins TL (1993) β-Glucosylation of plant phenolics by phenol β-glucosyltransferase in larval tissues of the tobacco hornworm, Manduca sexta (L.). Insect Biochem Molec Biol 23(5), 581-589. https://doi.org/10.1016/0965-1748(93)90031-M
  2. Aviles-Pagan EE, Orr-Weaver TL (2018) Activating embryonic development in Drosophila. Semin Cell Dev Biol 84, 100-110. https://doi.org/10.1016/j.semcdb.2018.02.019
  3. Brackenbury J (1997) Caterpillar kinematics. Nature 390, 453.
  4. Bukau B, Weissman J, Horwich A (2006) Molecular chaperones and protein quality control. Cell 125(3), 443-451. https://doi.org/10.1016/j.cell.2006.04.014
  5. Carter D, Locke M (1993) Why caterpillars do not grow short and fat. Int J Insect Morphol Embryol 22, 81-102. https://doi.org/10.1016/0020-7322(93)90002-I
  6. Chowdhary S, Tomer D, Dubal D, Sambre D, Rikhy R (2017) Analysis of mitochondrial organization and function in the Drosophila blastoderm embryo. Sci Rep 7(1), 5502.
  7. Denlinger DL (2002) Regulation of diapause. Annu Rev Entomol 47, 93-122. https://doi.org/10.1146/annurev.ento.47.091201.145137
  8. Gan L, Liu X, Xiang Z, He N (2011) Microarray-based gene expression profiles of silkworm brains. BMC Neurosci 19, 12-18. https://doi.org/10.1186/1471-2202-12-8
  9. Ghosal G, Lowe J (2015) Collaborative protein filaments. EMBO J 34(18), 2312-2320. https://doi.org/10.15252/embj.201591756
  10. Hwang JS, Go HJ, Goo TW, Seong SI, Yun EY, Ahn MY, et al. (2007) Molecular characterization of small heat shock protein (hsp20.8A) from the silkworm, Bombyx mori. Int J Indust Entomol 15(1), 75-78.
  11. Hong SM, Nho SK, Kim NS, Lee JS, Kang SW (2006) Gene expression profiling in the silkworm, Bombyx mori, during early embryonic development. Zoolog Sci 23(6), 517-528. https://doi.org/10.2108/zsj.23.517
  12. Hopkins TL, Kramer KJ (1992) Insect cuticle sclerotization. Annu Rev Entomol 37, 273-302.
  13. Kataoka N, Miyake S, Azuma M (2009) Aquaporin and aquaglyceroporin in silkworms, differently expressed in the hindgut and midgut of Bombyx mori. Insect Mol Biol 18(3), 303-314. https://doi.org/10.1111/j.1365-2583.2009.00871.x
  14. Kostal V (2006) Eco-physiological phases of insect diapause. J Insect Physiol 52(2), 113-127.
  15. Kraft R, Levine RB, Restifo LL (1998) The steroid hormone 20-hydroxyecdysone enhances neurite growth of Drosophila mushroom body neurons isolated during metamorphosis. J Neurosci 18(21), 8886-8899. https://doi.org/10.1523/JNEUROSCI.18-21-08886.1998
  16. Kramer KJ, Hopkins TL (1987) Tyrosine metabolism for insect cuticle tanning. Arch Insect Biochem Physiol 6, 279-301. https://doi.org/10.1002/arch.940060406
  17. Li B, Hu P, Zhang SZ, Toufeeq S, Wang J, Zhao K, et al. (2019) DNA methyltransferase BmDnmt1 and BmDnmt2 in silkworm (Bombyx mori) and the regulation of silkworm embryonic development. Arch Insect Biochem Physiol 100(3), e21529.
  18. Li T, Xia Y, Xu X, Wei G, Wang L (2020) Functional analysis of Dicer-2 gene in Bombyx mori resistance to BmNPV virus. Arch Insect Biochem Physiol 105, e21724.
  19. Lin HT, Dorfmann AL, Trimmer BA (2009) Soft-cuticle biomechanics: a constitutive model of anisotropy for caterpillar integument. J Theor Biol 256, 447-457. https://doi.org/10.1016/j.jtbi.2008.10.018
  20. Pappenheimer AM, Williams CM (1954) Cytochrome b5 and the dihydro-coenzyme I-oxidase system in the cecropia silkworm. J Biol Chem 209(2), 915-929.
  21. Park JW, Yu JH, Kim SB, Kim SW, Kim SR, Choi KH (2019) Analysis of silkworm molecular breeding potential using CRISPR/Cas9 systems for white egg 2 gene. Int J Indust Entomol 39(1), 14-21.
  22. Pecasse F, Beck Y, Ruiz Y, Richards G (2000) Kruppel-homolog, a stage-specific modulator of the prepupal ecdysone response, is essential for Drosophila metamorphosis. Dev Biol 221(1), 53-67. https://doi.org/10.1006/dbio.2000.9687
  23. Ponnuvel KM, Murthy GN, Awasthi AK, Rao G, Vijayaprakash NB (2010) Differential gene expression during early embryonic development in diapause and non-diapause eggs of multivoltine silkworm Bombyx mori. Indian J Exp Biol 48(11), 1143-1151.
  24. Qiao L, Xiong G, Wang RX, He SZ, Chen J, Tong XL, et al. (2014) Mutation of a cuticular protein, BmorCPR2, alters larval body shape and adaptability in silkworm, Bombyx mori. Genetics 196(4), 1103-1115. https://doi.org/10.1534/genetics.113.158766
  25. Sasibhushan S, Rao CGP, Ponnuvel KM (2013) Genome wide microarray based expression profiles during early embryogenesis in diapause induced and non-diapause eggs of polyvoltine silkworm Bombyx mori. Genomics 102(4), 379-387. https://doi.org/10.1016/j.ygeno.2013.07.007
  26. Teng X, Zhang Z, He G, Yang L, Li F (2012) Validation of reference genes for quantitative expression analysis by real-time rt-PCR in four lepidopteran insects. J Insect Sci 12, 60.
  27. Xuan N, Rajashekar B, Picimbon JF (2019) DNA and RNA-dependent polymerization in editing of Bombyx chemosensory protein (CSP) gene family. Agri Gene 12, 100087.
  28. Yoshiga T, Okano K, Mita K, Shimada T, Matsumoto S (2000) cDNA cloning of acyl-CoA desaturase homologs in the silkworm, Bombyx mori. Gene 246(1-2), 339-345. https://doi.org/10.1016/S0378-1119(00)00047-0
  29. Zhang R, Cao YY, Du J, Thakur K, Tang SM, Hu F, et al. (2021) Transcriptome analysis reveals the gene expression changes in the silkworm (Bombyx mori) in response to hydrogen sulfide exposure. Insects 12(12), 1110.