Silencing PsASH2 affects embryo development in the cotton mealybug
Abstract
Here, we identified a novel histone lysine methyltransferase (KMT) gene, PsASH2, in the cotton mealybug, Phenacoccus solenopsis Tinsley. This gene was highly expressed in the ovary of female adults. Through RNA interference (RNAi) of PsASH2 by dsRNA microinjection, we found a reduction in the number of male embryos and total embryos in the ovaries of pregnant females. Continuous downregulation of PsASH2 in mated females until their death resulted in few changes in sex ratio but significant decreases in the number of both male and female offspring.
Dataset DOI: 10.5061/dryad.h70rxwdw8
Description of the data and file structure
Fig. 2: Expression profiles of PsASH2. Different developmental stages (A), tissue-specific (mated females) (B), tissue-specific (virgin females) (C), hours post-mating (D), and days of virgin female adults (E) expression profiles of PsASH2 in P. solenopsis. The 4-day-old, 5-day-old, 6-day-old, and 7-day-old virgin female adults (E) correspond to 0 h, 24 h, 48 h, and 72 h post-mating (D). Relative expression levels were normalized and visualized as mean ± SEM with three (a, b) or four (c) biological replicates. Different lower-case letters indicate statistically significant differences (P<0.05) based on ANOVA and Tukey’s multiple comparisons test.
Fig. 3: RNAi effects of dsPsASH2 on cotton mealybug embryos via dsRNA microinjection. (A) The gene expression before and after injection with dsRNA1 (left) and dsRNA2 (right). Data are presented as mean ± SEM from four biological replicates. Significance was determined by the Student’s t-test. p < 0.05. (B) Violin plots showing the effects of RNAi on the total number of embryos in the dsPsASH2* treated group. (C) Number of male embryos. (D) Proportion of male embryos. Each circle represents one female individual (n = 15–17). Significance was determined using a generalized linear model (glm) with Poisson or Binomial distributions. *p < 0.05, **p < 0.01 and ***p < 0.001.
Fig. 4: Effects of dsPsASH2 on reproductive parameters of female adults in cotton mealybug via dsRNA microinjection. (A) Effect of dsPSASH2 on the pre-oviposition period. (B) Effect on the oviposition and post-oviposition periods. (C) Effect of dsPSASH2 on adult lifespan. Each circle represents one female individual (n = 18–24). Significance was determined using a generalized linear model (glm) with Poisson or Binomial distributions. *p < 0.05, **p < 0.01 and ***p < 0.001; ns: not significant.
Fig. 5: Effects of dsPsASH2 on reproductive parameters of female adults in cotton mealybug via dsRNA microinjection. (A) Effect of dsPSASH2 on the number of total offspring. (B) Number of male offspring. (C) Number of female offspring. (D) Male ratio of offspring. Each circle represents one female individual (n = 18–24). Significance was determined using a generalized linear model (glm) with Poisson or Binomial distributions. *p < 0.05, **p < 0.01 and ***p < 0.001; ns: not significant.
Files and variables
File: ASH2.xlsx
Description:
Variables
- Expression profiles of PsASH2
- RNAi effects of dsPsASH2 on cotton mealybug embryos via dsRNA microinjection
- Effects of dsPsASH2 on reproductive parameters of female adults in cotton mealybug via dsRNA microinjection.
- Effects of dsPsASH2 on reproductive parameters of female adults in cotton mealybug via dsRNA microinjection.
Code/software
Microsoft Excel