Cysteine protease ervatamin-B-like-mediated spermatophore digestion and sperm release impair fertility of Plutella xylostella females
Zheng, Junhao; Zou, Mingmin; Qian, Yurui; Li, Ruonan; Bian, Huimin; Cao, Minhui; Huang, Mengqi; Cui, Jindong; Peng, Lu
Entomologia Generalis Volume 45 Number 6 (2025), p. 1721 - 1731
publié: Dec 4, 2025
publication en ligne: Jun 17, 2025
manuscrit accepté: Apr 27, 2025
révision final du manuscrit reçu: Mar 11, 2025
révision du manuscrit demandée: Feb 9, 2025
manuscrit reçu: Jan 9, 2025
ArtNo. ESP146004506018, Prix: 29.00 €
Abstract
Mating induces post-mating responses in female insects, causing molecular, physiological, and behavioral changes. Spermatophore is a unique structure in females for seminal fluid storage in the bursa copulatrix, significant in sperm transfer and activation, and its digestion represents a crucial signal for female re-mating. Plutella xylostella, a lepidopteran pest that attacks cruciferous crops and wild plants, is a promiscuous insect that digests spermatophores rapidly. Molecular mechanisms underlying spermatophore digestion in P. xylostella are poorly understood. This study aimed to identify post-mating response proteins by using isobaric tags for relative and absolute quantification proteomics. A total of 843 proteins had significant differential expression between the reproductive glands of virgin and mated females, 720 of which were significantly upregulated after mating, including a cysteine protease family gene, cysteine protease ervatamin-B-like (PxCPEB), which is potentially related to spermatophore digestion. PxCPEB was specifically expressed in P. xylostella females, mostly in the female lower reproductive gland, but only in the bursa copulatrix and spermatheca, and PxCPEB transcription was significantly increased after mating. CRISPR/Cas9-induced PxCPEB knockout successfully produced a homozygous mutant strain with a 22-bp deletion. PxCPEB loss did not affect spermatogenesis but significantly prolonged the rate of spermatophore digestion, increased the apoptosis level of the bursa copulatrix, and inhibited sperm release and re-mating in females, significantly decreasing female fertility. PxCPEB, as a regulator of P. xylostella post-mating responses, is critical in spermatophore digestion, sperm release, re-mating, and female fertility and its potential as a promising target for genetic regulation-based pest control.
Mots-clefs
Plutellidae • bursa copulatrix • spermatheca • post-mating responses • reproductive regulation