DOI: 10.5061/dryad.h70rxwdw8
title: Silencing PsASH2 affects embryo development in the cotton mealybug

description: 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.
description: # Silencing PsASH2 affects embryo development in the cotton mealybug
 Dataset DOI: [10.5061/dryad.h70rxwdw8](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*&lt;0.05) based on ANOVA and Tukey’s multiple comparisons test. Fig.
 3: RNAi effects of ds*PsASH2* 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* &lt; 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* &lt; 0.05, ***p* &lt; 0.01 and ****p* &lt; 0.001. Fig. 4: Effects of
 ds*PsASH2* on reproductive parameters of female adults in cotton mealybug
 via dsRNA microinjection. (A) Effect of ds*PSASH2* on the
 pre-oviposition period. (B) Effect on the oviposition and
 post-oviposition periods. (C) Effect of ds*PSASH2* 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* &lt; 0.05, ***p* &lt; 0.01 and ****p* &lt;
 0.001; ns: not significant. Fig. 5: Effects of ds*PsASH2* on reproductive
 parameters of female adults in cotton mealybug via dsRNA microinjection.
 (A) Effect of ds*PSASH2* 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* &lt; 0.05, ***p* &lt; 0.01 and
 ****p* &lt; 0.001; ns: not significant. ### Files and variables #### File:
 ASH2.xlsx **Description:**  ##### Variables * Expression profiles of
 *PsASH2* * RNAi effects of ds*PsASH2* on cotton mealybug embryos via dsRNA
 microinjection * Effects of ds*PsASH2* on reproductive parameters of
 female adults in cotton mealybug via dsRNA microinjection. * Effects of
 ds*PsASH2* on reproductive parameters of female adults in cotton mealybug
 via dsRNA microinjection. ## Code/software Microsoft Excel