Identification and functional analysis of ApCtsk1 encoding cathepsin K in Acyrthosiphon pisum

doi:10.11686/cyxb2024147

Abstract

Abstract:

The aim of this research was to explore the function of ApCtsk1 in pea aphid （Acyrthosiphon pisum） and evaluate its potential as a target for the prevention and control of this pest. The amino acid sequence characteristics of the putative ApCtsk1 protein were analyzed using tools at various websites （NCBI， ExPASy Proteomics Server） and a phylogenetic tree was constructed using MEGA 11. The transcript levels of its encoding gene were determined in aphids at different stages of development and in different tissues by RT-qPCR analysis. The biological function of ApCtsk1 in the development and reproduction of A. pisum was investigated using an RNA interference gene knock-down strategy. In the phylogenetic analysis， ApCtsk1 was clustered with cysteine proteases from other members of the Aphididae in the Hemiptera. High transcript levels of ApCtsk1 were detected in fourth instar nymphs， and in the abdomen， ovary， and cuticle of adult aphids. Compared with A. pisum in the control group， those in the ApCtsk1-knockdown group showed an increased mortality rate （22.22% higher）， decreased body length and body width （decreased by 12.56% and 13.22%， respectively）， abnormal body color， and slower movement. In addition， the aphids with knocked-down ApCtsk1 showed the lethal phenomenon of molting failure， which affected their fecundity. These changes resulted in delayed reproduction and decreased reproduction over time. Our findings show that ApCtsk1 plays an important role in the development， survival， and reproduction of A. pisum.

Key words: Acyrthosiphon pisum, cathepsin K, fecundity, RNAi

Rui MA, Yi-ting WU, Yan LI, Zhan-zi YU, Lei LIU, Sen-shan WANG. Identification and functional analysis of ApCtsk1 encoding cathepsin K in Acyrthosiphon pisum[J]. Acta Prataculturae Sinica, 2025, 34(2): 174-183.

Figures/Tables 6

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基因 Gene	引物方向 Primer direction （5′-3′）	核苷酸序列 Nucleotide sequence （5′-3′）	产物长度 Product size （bp）
dsGFP	上游Forward	TAATACGACTCACTATAGGGCAGTTCTTGTTGAATTAGATG	436
	下游Reverse	TAATACGACTCACTATAGGGTTTGGTTTGTCTCCCATGATG	436
dsApCtsk1	上游Forward	TAATACGACTCACTATAGGGTGAATCTTTCCCTGGACCTGG	541
	下游Reverse	TAATACGACTCACTATAGGGATTGCCAAAACCCCAACTGC	541

基因 Gene	引物方向 Primer direction （5′-3′）	核苷酸序列 Nucleotide sequence （5′-3′）	产物长度 Product size （bp）
dsGFP	上游Forward	TAATACGACTCACTATAGGGCAGTTCTTGTTGAATTAGATG	436
	下游Reverse	TAATACGACTCACTATAGGGTTTGGTTTGTCTCCCATGATG	436
dsApCtsk1	上游Forward	TAATACGACTCACTATAGGGTGAATCTTTCCCTGGACCTGG	541
	下游Reverse	TAATACGACTCACTATAGGGATTGCCAAAACCCCAACTGC	541

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