Frontiers in Physiology (Jan 2020)

CRISPR/Cas9-Mediated Vitellogenin Receptor Knockout Leads to Functional Deficiency in the Reproductive Development of Plutella xylostella

  • Lu Peng,
  • Lu Peng,
  • Lu Peng,
  • Lu Peng,
  • Qing Wang,
  • Qing Wang,
  • Qing Wang,
  • Qing Wang,
  • Ming-Min Zou,
  • Ming-Min Zou,
  • Ming-Min Zou,
  • Ming-Min Zou,
  • Yu-Dong Qin,
  • Yu-Dong Qin,
  • Yu-Dong Qin,
  • Yu-Dong Qin,
  • Liette Vasseur,
  • Liette Vasseur,
  • Liette Vasseur,
  • Liette Vasseur,
  • Liette Vasseur,
  • Li-Na Chu,
  • Li-Na Chu,
  • Li-Na Chu,
  • Li-Na Chu,
  • Yi-Long Zhai,
  • Yi-Long Zhai,
  • Yi-Long Zhai,
  • Yi-Long Zhai,
  • Shi-Jie Dong,
  • Shi-Jie Dong,
  • Shi-Jie Dong,
  • Shi-Jie Dong,
  • Li-Li Liu,
  • Li-Li Liu,
  • Li-Li Liu,
  • Li-Li Liu,
  • Wei-Yi He,
  • Wei-Yi He,
  • Wei-Yi He,
  • Wei-Yi He,
  • Guang Yang,
  • Guang Yang,
  • Guang Yang,
  • Guang Yang,
  • Min-Sheng You,
  • Min-Sheng You,
  • Min-Sheng You,
  • Min-Sheng You

DOI
https://doi.org/10.3389/fphys.2019.01585
Journal volume & issue
Vol. 10

Abstract

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The vitellogenin receptor (VgR) belongs to the low-density lipoprotein receptor (LDLR) gene superfamily and plays an indispensable role in Vg transport, yolk deposition, and oocyte development. For this reason, it has become a promising target for pest control. The involvement of VgR in Vg transport and reproductive functions remains unclear in diamondback moths, Plutella xylostella (L.), a destructive pest of cruciferous crops. Here, we cloned and identified the complete cDNA sequence of P. xylostella VgR, which encoded 1805 amino acid residues and contained four conserved domains of LDLR superfamily. PxVgR was mainly expressed in female adults, more specifically in the ovary. PxVgR protein also showed the similar expression profile with the PxVgR transcript. CRISPR/Cas9-mediated PxVgR knockout created a homozygous mutant of P. xylostella with 5-bp-nucleotide deletion in the PxVgR. The expression deficiency of PxVgR protein was detected in the ovaries and eggs of mutant individuals. Vg protein was still detected in the eggs of the mutant individuals, but with a decreased expression level. However, PxVg transcripts were not significantly affected by the PxVgR knockout. Knockout of PxVgR resulted in shorter ovarioles of newly emerged females. No significant difference was detected between wild and mutant individuals in terms of the number of eggs laid in the first 3 days after mating. The loss of PxVgR gene resulted in smaller and whiter eggs and lower egg hatching rate. This study represents the first report on the functions of VgR in Vg transport, ovary development, oviposition, and embryonic development of P. xylostella using CRISPR/Cas9 technology. This study lays the foundation for understanding molecular mechanisms of P. xylostella reproduction, and for making use of VgR as a potential genetic-based molecular target for better control of the P. xylostella.

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