Fig. 1: Knockdown of BPH Zfh1 leads to long-winged morphs.

a Morphology of wild-type long-winged (LW) and short-winged (SW) female adults. Arrows, forewings. Arrowheads, hindwings. b Differentially expressed transcription factors between LW-destined (Wt^LW) and SW-destined (Wt^SW) fifth-instar nymphs. Thoracic nota were dissected from fifth-instar nymphs for RNA sequencing. Genes differentially expressed between Wt^LW and Wt^SW with a false discovery rate (FDR) < 0.01 are considered to be significant. The Zfh1 protein contains a homeodomain (orange rectangle) and nine zinc-finger motifs (blue boxes). Regions targeted by dsRNAs (dsZfh1 and dsZfh1_2) are underlined. The PCR amplified region used to examine RNAi efficiency is indicated by arrows. c RNAi efficiency of dsZfh1 and dsZfh1_2. Third-instar Wt^SW nymphs were microinjected with dsZfh1 or dsZfh1_2. Individuals (n = 5) were collected for qRT-PCR assay two days after microinjection. The relative expression of Zfh1 was normalized to the expression of the ribosomal protein S11 gene (rps11). The experiments were repeated three times with similar results (circles). Data are presented as mean ± SEM. Two-tailed unpaired t-test was used for the statistical analysis (** P = 0.0028 for dsZfh1 vs dsGfp and ** P = 0.0011 for dsZfh1_2 vs dsGfp). d Morphology of dsGfp^SW, dsZfh1^LW, and dsZfh1^IMW females. dsGfp^SW, dsGfp-treated BPHs with short wings (SW). dsZfh1^LWand dsZfh1^IMW, dsZfh1-treated BPHs with long and intermediate-size wings (IMW), respectively. Scale bars in (a) and (c), 1 mm. e Number of females with different wing morphs following dsRNA treatments. The LW ratio is compared between two groups using Pearson χ² test (****P = 1.1716E-56, χ² = 251.586 and df = 1 for dsZfh1 vs dsGfp; ****P = 4.9769E-45, χ² = 198.2772 and df = 1 for dsZfh1_2 vs dsGfp). f Wing size and hind tibia length in dsZfh1^LW, Wt^LW and Wt^SW females. Each circle represents an individual female (n = 20). g Vein patterning on forewings from Wt^LW and dsZfh1^LW females. Twenty samples were repeated independently with similar results. Scale bars, 500 μm. Source data are provided as a Source Data file.