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. 2023 May;13(5):220372.
doi: 10.1098/rsob.220372. Epub 2023 May 31.

Krüppel-homologue 1 regulates the development of Tuta absoluta and its cascade regulation pattern in the juvenile hormone signalling pathway

Xiaodi Wang  1 , Siyan Bi  1 , Yanhong Tang  1 , Guifen Zhang  1 , Cong Huang  1 , Fanghao Wan  1   2 , Zhichuang Lü  1 , Wanxue Liu  1
Affiliations

Krüppel-homologue 1 regulates the development of Tuta absoluta and its cascade regulation pattern in the juvenile hormone signalling pathway

Xiaodi Wang et al. Open Biol. 2023 May.

Abstract

Tomato leaf miner, Tuta absoluta (Meyrick), is one of the most destructive quarantine pests globally. It has been confirmed that Krüppel-homologue 1 (kr-h1) plays a key role in the regulation of juvenile hormone (JH). However, it is unclear how kr-h1 regulates the synthesis of JH and its cascade regulation pattern in tomato leaf miner. Here, we obtained the six JH signalling genes (kr-h1, Methoprene-tolerant, Forkhead box O, Juvenile acid methyltransferase, Juvenile hormone esterase and Fatty acid synthase 2), and applied RNA interference to explore the role of kr-h1 and the seven genes (plus Vitellogenin) regulation relationship in T. absoluta. Bioinformatics analysis revealed the structural characteristics of kr-h1 protein and JH receptor Met, which contained eight C2H2 zinc finger structures and three typical domains of the bHLH-PAS family, respectively. The expression levels of Met and Vg were upregulated after RNAi of kr-h1 gene, while the gene levels of JHAMT and FAS2 were downregulated. Furthermore, topical application of JH analogue to second instar larvae could induce the expression of kr-h1 and inhibit the expression of Met. Our study reveals the mechanism by which kr-h1 regulates JH pathway genes, which could be applied to control the growth of tomato leaf miners.

Keywords: Tuta absoluta; cascade regulation; juvenile hormones; takr-h1.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1.
Figure 1.
Relationship of juvenile hormone signalling pathway [34].
Figure 2.
Figure 2.
Expression profiles of seven JH signalling-related genes at different developmental stages of T. absoluta. The horizontal axis represents different developmental stages, A to K representing the first instar larval stage (2 d), the second instar larval stage (4 d), the third instar larval stage (6 d), the fourth instar larval stage (8 d), early pupation (14 d), late pupation (18 d), early female (21 d), early male (21 d), late female (30 d), late male (30 d), respectively. The vertical axis shows the relative expression level is expressed by mean ± s.e.m. The different letters (a, b) represent groups with significant differences according ANOVA test (Tukey's test, p < 0.05).
Figure 3.
Figure 3.
Changes in the relative expression of Takr-h1 gene and mortality of T. absoluta after feeding dskr-h1. (a) Changes in the relative expression of Takr-h1 gene. (b) Mortality of T. absoluta after feeding dskr-h1. The blue column indicates feeding dsEGFP, and the red column indicates feeding dskr-h1. Data are presented as mean ± s.e.m.; on the horizontal line represents the difference of relative gene expression under different treatment conditions (*p < 0.05).
Figure 4.
Figure 4.
Effects of feeding dskr-h1 on phenotype of T. absoluta. (a) Phenotype of T. absoluta larvae fed with dsGEFP. (b) Phenotype of T. absoluta larvae after feeding dskr-h1. (c) Feeding status of T. absoluta larvae after feeding dsEGFP. (d) The state of leaf death of T. absoluta larvae after feeding dskr-h1. (e) The phenotype of T. absoluta pupa after feeding dsEGFP. (f) The phenotype of T. absoluta pupa after feeding dskr-h1.
Figure 5.
Figure 5.
Effects of Takr-h1 silencing on expression levels of JH signalling-related genes. The relative expression levels are expressed as the mean ± s.e.m. The green column represents the control group that was fed dsEGFP, while the pink column represents the treatment group that was fed dskr-h1. Asterisks represent significance levels: *p < 0.05, **p < 0.01; n.s. indicates no significance p > 0.05.
Figure 6.
Figure 6.
Changes of relative expression of three genes after feeding dsMet and JHA. The relative expression levels are expressed as the mean ±s.e.m.; the green column represents the control group and the red column represents the treatment group.Asterisk represents significance level:*p < 0.05.
Figure 7.
Figure 7.
Regulation of genes related to juvenile hormone signalling pathway in T. absoluta.
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